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Emergency at the Zoo: An Orangutan Bite Resulting in Thumb Amputation and Forearm Laceration by Shaza Aouthmany MD

Abstract

Zoos and aquariums receive over 180 million visitors per year. Although this amount of traffic is equivalent to over half the US population, traumatic injuries at modern zoo enclosures are rare. Despite adequate safety standards, proper training, and experience with animals, incidents may occur that require extensive medical management. We report the case of a 57-year-old woman who presented to the emergency department after being bitten by an orangutan resulting in right disarticulation of her right thumb and a laceration to her right forearm.

Introduction

Primates are an order within the class of Mammalia and include humans, gorillas, orangutans, chimpanzees, among many others. Simians are an infraorder within the Primate order but encompass the species stated above [1,2]. Such exotic animals are kept in zoos all over the world and require close human interaction in order to be fed and to be treated medically. These interactions pose a risk for serious injury. While there are countless studies on bites from dogs, cats, and other animals, there are no case reports on orangutan bites. We present a case of thumb amputation and forearm laceration from an orangutan bite to an animal caretaker.

Case Summary

A 57-year-old, right-handed, female patient with no significant past medical history arrived via EMS from a local zoo for right thumb amputation and forearm laceration after being bitten by an orangutan. The patient, an animal caretaker, was feeding the primate when it grabbed the patient’s right upper extremity through its cage. Orangutan initially bit into right hand disarticulating patient thumb and then into into right medial forearm . EMS reported an arterial bleed and applied a tourniquet on the scene.

On presentation patient was alert and oriented x4, had blood pressure 149/71 mmHg, pulse rate 95 beats/min, respirations 12 breaths/min, and pulse oximetry 100% on room air. Bleeding was well controlled with the tourniquet in place.. There was obvious amputation of the right thumb which was hanging by approximately 15 cm of avulsed extensor pollicis longus and flexor pollicis longus tendons. The right forearm had significant soft tissue damage from laceration on the medial aspect with musculature and tendon exposed. There was no evidence of compartment syndrome and the distal circulation was intact. The patient was able to move her right second through fifth digits. No sensory deficits were elicited. The remainder of the physical exam was unremarkable.

The patient was given a 0.5 mL Tdap injection IM, 3000 mg IV ampicillin-sulbactam was given along with 50 mcg IV fentanyl and the tourniquet was removed. Radiograph of the right hand revealed complete amputation of the first digit at the metacarpophalangeal joint (Figure 1). Radiographs of the right forearm demonstrated minimally displaced distal ulnar styloid fracture and large soft tissue laceration with associated soft tissue hematoma (Figure 2). Orthopedic surgery and vascular surgery were consulted and patient was taken emergently to the operating room for wound exploration, replantation, and wound closure.

Discussion and Conclusion

Patients with bite injuries comprise about 1% of visits to the emergency department annually [3,4]. In 2009, animal-related injuries accounted for 1.3 million emergency department visits, 60,800 of which led to in-patient stays. Of these in-patient stays, 9,700 were related to animals other than dogs [5]. These ‘other animal’ injuries may not be as well studied as dog bites, but information available allows for physicians to make clinically sound decisions. As far as we know, there are no published studies on orangutan bites specifically, and as such we review information regarding the complications, treatment of, and risk of infection from nonhuman primate bites.

Bite injuries from mammals have the potential for a wide range of damage to the recipient, from a simple abrasion to the more concerning laceration, avulsion, fracture, and amputation [4]. To put the potential for injury from an orangutan bite into perspective, consider that an African lion has a bite force of over 4000 N, an orangutan almost 2000 N, and a human about a third of an orangutan [6].

The most common complication of mammal bite injuries is infection, which in turn can progress to osteomyelitis, tenosynovitis, and cellulitis.4 Current data suggests a similarity between the bacteria isolated from simian bites and that which is isolated from human bites which includes alpha-hemolytic streptococci, S. aureus, Neisseria species, Eikenella corrodens, anaerobes, and Enterobacteriaceae [7,8]. Left untreated, human bites have an infection rate of 48%, so it is imperative to adequately treat orangutan bites [4].

Treatment principles outlined for animal bites include some combination of irrigation, prophylactic antibiotics, targeted antibiotics, Tetanus immunization, debridement, wound closure, and reporting if legally necessary [3,8-10]. Prophylactic antibiotics should be given based on the propensity for risk of the bite itself, the animal and its immunization status, and the time elapsed from injury to presentation. When choosing antibiotics, selection should be based on coverage for the aforementioned organisms and further guided by results from aerobic and anaerobic cultures. Medication with proper coverage includes oral amoxicillin and clavulanate, IV ampicillin-sulbactam, or doxycycline if allergic to penicillin [3,8,10]. An orangutan bite on the magnitude that our patient suffered was indeed high risk, and as such, 3000 mg IV ampicillin-sulbactam was administered upon arrival to the emergency department.

The situation regarding zoo animal bites and getting immunizations or post-exposure prophylaxis is interesting. While they are exotic animals, primates in captivity undergo similar vaccination schedules as do humans. Orangutans share 97% of human DNA, so it’s no surprise that they can also transmit and can become infected with similar respiratory and gastrointestinal diseases. In fact, the Orangutan Care Manual published by the Association of Zoos and Aquariums recommends following the American Academy of Pediatrics immunization schedule. In the case of an orangutan not being immunized early, the AZA strongly recommends all orangutans get yearly influenza vaccines, tetanus toxoid every 5-10 years, a one-time pneumococcus vaccine, and Haemophilus influenzae type b vaccine. Lastly, since rabies vaccines are not routine in humans, the decision to vaccinate an orangutan against rabies should be based on the exhibits exposure risk [11]. The CDC recommends a rabies preexposure vaccine in high risk groups such as veterinarians and animal handlers. Although this does not completely eliminate the need for therapy after rabies exposure, it eliminates the need for rabies immunoglobulin and reduces the number of vaccine doses [12]. It is reasonable to conclude that the decision to immunize an individual bitten by an orangutan should be based on a combination of vaccination history of the patient as well as the animal. In the emergency setting this information may not be immediately available and so it is prudent for the physician to act on their clinical acumen. In our patient’s case, we chose only to administer the Tdap vaccine.

The patient experienced a tragic encounter while feeding an orangutan at a local zoo and suffered tremendous damage to her arm and thumb. Following her evaluation in the emergency department she was taken to the operating room where orthopedic surgeons extensively irrigated and debrided the wound. Surgeons then closed the forearm laceration and replanted the right thumb. Since 1990, there have been 16 primate escape or attacks at AZA accredited facilities resulting in human injury and zero reported cases of primate attacks resulting in death [13]. This indicates the safeguards currently in place are generally effective. It is difficult to ascertain what precautions should be instituted in the future to prevent such incidents from recurring but most emergency medicine departments are equipped to acutely treat them.

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Bioresorbable Polymers Market Opportunities on the Horizon by 2029

Research Nester published a report titled “Bioresorbable Polymers Market: Global Demand Analysis & Opportunity Outlook 2029” which delivers detailed overview of the global bioresorbable polymers market in terms of market segmentation by type, application, and by region.

Further, for the in-depth analysis, the report encompasses the industry growth indicators, restraints, supply and demand risk, along with detailed discussion on current and future market trends that are associated with the growth of the market.

The global bioresorbable polymers market is estimated to garner a considerable CAGR over the forecast period, i.e., 2021–2029 owing to the increasing use of these polymers in the healthcare sector. Bioresorbable polymers are biodegradable, and they eventually dissolve in the body, which eliminates the need for a second surgery to remove the implants from the body. Furthermore, the growing patient pool of people suffering from orthopedic or musculoskeletal problems, is estimated to boost the market growth.

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The global bioresorbable polymers market is segmented on the basis of application into sutures, orthopedic fixation devices, dental implants, tissue staples, drug delivery devices, and others, out of which, the orthopedic fixation devices segment is foreseen to experience notable growth over the forecast period, on the back of rising incidences of orthopedic ailments, such as, fractures. Moreover, the use of these polymers, reduces the post-surgery complications, especially among the elders, which is estimated to boost the growth of the segment. On the other hand, the dental implants segment is projected to garner a significant market share during the forecast period owing to the increasing dental problems among people, as they are consuming foods high in sugar.

Regionally, the bioresorbable polymers market is segmented into five major regions including North America, Europe, Asia Pacific, Latin America and Middle East & Africa region. The market in the North America region is anticipated to witness a noteworthy growth over the forecast period on the back of the advanced healthcare facilities, and high R&D investment in the region. Moreover, presence of medical reimbursement and refund policies, is also estimated to impact the market growth positively. The market in the Europe region, is estimated to grow significantly during the forecast period, on the back of high geriatric population in the region, combined with improving healthcare sector and rising disposable income. According to the report of the World Bank, over one fifth of the total population, precisely 20.463% population is aged 65 years or above in the European Union.

Growing Application of Bioresorbable Polymers in Healthcare Likely to boost the Market Growth

The use of bioresorbable polymers as tissue staples, implants and drug delivery devices, is rapidly gaining prominence, owing to its biodegradability, which reduces the risk of complications after a surgery. This is estimated to boost the market growth. Moreover, the increasing cases of such ailments, owing to the increasing global geriatric population, and changing lifestyle, is estimated to promote the market growth.

However, the lack of proper medical facilities in the developing nations is estimated to hamper the growth of the market over the forecast period.

This report also provides the existing competitive scenario of some of the key players of the global bioresorbable polymers market which includes company profiling of Evonik Industries AG (ETR: EVK), Corbion NV (AMS: CRBN), Ashland Global Holdings Inc. (NYSE: ASH), Foster Corporation, Zeus Industrial Products, Inc., Merck Group (ETR: MRK), Gebrüder Martin GmbH & Co. KG, Polymedicure Ltd. (NSE: POLYMED), Biogeneral, Inc., and Koninklijke DSM N.V. (AMS: DSM).

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The profiling enfolds key information of the companies which encompasses business overview, products and services, key financials and recent news and developments. On the whole, the report depicts detailed overview of the global bioresorbable polymers market that will help industry consultants, equipment manufacturers, existing players searching for expansion opportunities, new players searching possibilities and other stakeholders to align their market centric strategies according to the ongoing and expected trends in the future. 

[athiranair23]

MEA Technetium-99m Market Analysis (2020-2027)

Technetium-99m is a short-lived form of Technetium-99, an isotope of technetium. It is found as a component of nuclear waste and is used as a medical diagnostic tool. It is used as a radioactive tracer and can be detected in the body by medical equipment such as gamma cameras and Single Photon Emission Computed Tomography (SPECT).

MEA technetium-99m market is estimated to account for US$ 394.6 Mn in terms of value and is expected to reach US$ 688.5 Mn by the end of 2027.

MEA Technetium-99m Market: Drivers

High prevalence of congenital heart disease is expected to propel growth of MEA technetium-99m market over the forecast period. For instance, according to the study, ‘Patterns, prevalence, risk factors, and survival of newborns with congenital heart defects in a Saudi population: a three-year, cohort case-control study’, published in March 2019, the prevalence of congenital heart defects was 14.8/1000 births in the study population.

Moreover, increasing investment in the production of molybdenum-99 is also expected to aid in growth. For instance, in July 2019, NorthStar Medical Radioisotopes, LLC announced completion of construction on its 20,000 square foot molybdenum-99 processing facility.

Gamma Camera held dominant position in MEA technetium-99m market in 2019, accounting for 57.3% share in terms of volume, followed by SPECT, respectively.

Figure 1. MEA Technetium-99m Market Share (%), by Value, by Isotopic Application, 2019

MEA Technetium-99m Market: Restraints

Availability of 99mTc depends on the production of Mo-99 from the small number of nuclear reactors worldwide. The major producing reactors have already aged and on the verge of shutting down. This creates an issue related to production disruption assuming the need for extended shutdowns for planned or unplanned maintenance work, which is expected to hinder growth of MEA technetium-99m market.

Adoption of alternative imaging modalities is also expected to limit growth of the market. The expected shortage of Tc-99m in the near future has led to increase in adoption of PET or other imaging modalities.

MEA Technetium-99m Market: Opportunities 

Increasing adoption of dual modality imaging technique or hybrid imaging is expected to offer lucrative growth opportunities for players in MEA technetium-99m market. The adoption of hybrid imaging consisting of single-photon emission computed tomography (SPECT) and X-ray transmission computed tomography (CT) scan is increasing globally as it offers short acquisition time and provides accurate attenuation correction and fusion imaging leading to increase in diagnostic sensitivity and specificity.

Moreover, non-reactor production of Mo-99 without using highly enriched uranium is also expected to aid in growth of the market.

MEA technetium-99m market was valued at US$ 394.6 Mn in 2019 and is forecast to reach a value of US$ 688.5 Mn by 2027 at a CAGR of 8.3% between 2020 and 2027.

Figure 2. MEA Technetium-99m Market Value (US$ Mn), and Y-o-Y Growth (%), 2019-2027

Market Trends/Key Takeaways

Players in the market are focused on extending patent protection for their products. For instance, in March 2020, Navidea Biopharmaceuticals, Inc. received the U.S. Patent extension for Lymphoseek (technetium [Tc 99m] tilmanocept) until 2025.

Tc-99m can find application in myocardial metabolism imaging. For instance, in November 2019, researchers from Institute of Nuclear and Radiological Sciences and Technology, Greece, reported the synthesis and characterization of two novel derivatives of undecanonoic and hexadecanonoic acid with the aim of developing 99mTc-labeled fatty acids intended for myocardial metabolism imaging.

Regulations                                                         

·  It is essential to attain the approval of the national atomic energy agencies before each step. The supplier has to apply separately to regulatory body for No Objection Certificate / Type Approval for the medical cyclotron.

·  The various stages of clearances needed from national atomic energy agencies which mainly includes:

o    Site Assessment and Approval: The first stage of process is to assess the suitability of the site which includes compliance with the geological and soil characteristics such as seismic condition, geo-hydrology and existing infrastructure etc.

o ��  Design and Construction Approval: The medical cyclotron should be installed in a room with adequate shielding with adequate isolation and access control. The licensee needs to submit Quality Assurance reports during construction and installation to the concerned regulatory body

o    Commissioning: For granting license for commissioning, the regulatory body evaluates the system performance and the shielding adequacy of the installation

o    Decommissioning: When the medical cyclotron operation is ceased, the permission for decommissioning is required

MEA Technetium-99m Market: Competitive Landscape

Major players operating in MEA technetium-99m market include, GE Healthcare, IBA, Sumitomo Corporation, Advanced Cyclotron Systems, Inc., NorthStar Medical Radioisotopes, and Siemens Healthineers.

MEA Technetium-99m Market: Key Developments

Major players in the market are focused on adopting partnership strategies to enhance their market share. For instance, in March 2020, NorthStar Medical Technologies, LLC partnered with Capella Imaging, LLC, a BioGenerator company, under which NorthStar has licensed exclusive rights to FibroScint (Tc-99m F4A), a fibrin-targeted diagnostic imaging agent for cardiac imaging.

Major players in the market are also focused on labeling its products. For instance, in February 2020, BWX Technologies, Inc. successfully labeled technetium-99m derived from its proprietary molybdenum-99 production process and its proprietary Tc-99m generators with cold kits.

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Open access Journal of Biogeneric Science and Research_BGSR

Biogeneric Publishers

Our main dictum of establishing Our Open Access Journal of BGSR is it’s an unique platform which cover all scientific subjects that provides imperative information for all research communities. We at Biogeneric sciences are an open access platform including peer-reviewed Journal articles like Research, Reviews, Conceptual Papers, Case Reports, PPT in seminars, and more effective video articles for easy sympathetic and cramming for all educated & untutored.

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The Provision of the Modern and Special Features of Arealing of the Mountains and Black Lands in the Gedebey District by Aliyev Zakir Huseyn Oglu

Open Access Journal of Biogeneric Science and Research

It is well known that the classification of brown soils was first made in 1949 by IP Gerasimov. Although 69 years have passed since the first classification to date, brown soil has not yet been able to occupy its place in the classification. Thus, to substantiate the aforementioned idea, we have many years of theoretically and practically analyzed data. It is known that brown soils are widely formed in the Balkan Peninsula and around the Mediterranean (European and African coasts).  This information was published in the works of the 6th International Congress of Soil Science, held in Paris in 1956. The last classification of Azerbaijani lands was given by ME Salayev [1], where the author was especially interested in the search for correlation between the nomenclature system of the land and the system of nomenclature of the world with a more detailed study of the diagnostic parameters of soils.  

Thus, when analyzing the results of SE Salayev's observations, attention is drawn to the specificity of mountain and brown soils from other types of soil and the relief and climatic complexity of the mountainous terrain. As noted by Salayev ME, brown soils were formed on both flat and mountainous background. However, it should be borne in mind that the brown soils in Azerbaijan have largely formed their genetic characteristics in the complex exposition of the mountainous relief and retained their classic morphological features. You don't have to go very far to justify this idea. Thus, it is possible to encounter mountainous brown and mountainous lands in the northeastern part of the Lesser Caucasus, with a large range of Gadabay. Brown soils are associated with the gray-brown soils formed in the foothills and forests of the Gadabay region from the south. In the upper border, brown mountain-forest lands create a transition to dark brown soils. The brown area of ​​the forest zone has developed from 400-500m up to 1100-1200m, mainly under oak and partly forest.

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Membership in Open Access Journal of Biogeneric Science and Research

Membership in Open Access Journal of Biogeneric Science and Research

Open Access Journal of BGSR refers to free and unrestricted online access to scientific and scholarly information. Peer-reviewed article is freely available without subscription or price barriers, article is immediately released in open access format like PDF, HTML, e-book and free of charge. The objective of  Open Access Journal of Biogeneric Journal Articles is to provide a scientific communication medium to discuss the highest advancements in the domain of medical, Business, Social, Life sciences & Technological. In fact, our Open Access Journal mainly aims to assemble and reserve precise, specific, detailed data on this immensely significant subject. Open Access Journal of BGSR is a self-supporting, with no dependency on any other external sources like Research centers, universities for funds and strives for the best and enhanced quality publications compete for the worldwide open access publishing. All the publication articles, which will be submitted by young researchers or experts, will undergo a peer review process under the guidance of our expert editors. We always rely on the support from the members of our team that is relevantly our Authors, Editorial Committee members, advisory board, Reviewers Board and all the technical support teams all over the globe. We accept all the submissions of significant articles like Research, reviews, Case Studies, Ppts in Seminars, Mini reviews, Short Communications, Conceptual Papers, Editorials, Perspectives etc. which covers advanced research output aiding in forwarding the Science are utmost welcome. To Read more about our Journal click on Open access Journal Publishers: Biogeneric Publishers Visit us: https://biogenericpublishers.com/

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Editor & Author Guidelines in Open access of Biogeneric Science and Research

Editor & Author Guidelines in Open access of Biogeneric Science and Research

EDITORIAL GUIDELINESEditors play an important role in any publishers. Our Open Access Journal of BGSR welcomes prominent personalities in various research fields to improve the quality of the Journal. Roles And ResponsibilitiesAble to write an editorials frequently based on their feasibility on current trends of research field.Based on the field of research (or) subject, editor we will assign submitted manuscript to accurate editor for the peer review process.Editorial board members are most welcome to give their valuable suggestions for assigned manuscript and also for the publishers.Ensures feedback provided to authors is constructive, fair, and timely.The decision to accept or reject a manuscript for publication is only based on the editor’s decision taking parameters such as importance, relevance, validity of the research, and clarity into consideration.The originality of article submissions and to be alert to redundant publication and plagiarism.Should be able to resolve any conflicts. AUTHOR GUIDELINESWho wish to submit a manuscript to Our Open Access Journal of BGSR has some guidelines: Authors’ can submit any topic relevant to science which are not published and peer reviewed too.Before Submission of manuscript authors are requested to review and ensure the accuracy and validity of all the results >=70 and Submitted Manuscript should be in the form of Doc, File PDF, Videos, PPts.No restrictions on word count, Figures, Tables and References and is based on their research work. (i.e. article exceeding >30pages considered as e-Book).Along with Research work, cover letter is compulsory.All authors are requested to submit the copyright transfer form once they receive the acceptance of article for the publication.Author’s must accept the comments from the Editorial Board members and ready to modify the submitted manuscript.

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Short Course Digoxin in Acute Heart Failure by Nouira Semir in Open Access Journal of Biogeneric Science and Research

ABSTRACT

Background Despite many critical voices regarding its efficacy and safety, digoxin may still have a role in the management of heart failure. The objective of this study was to evaluate the efficacy and safety of a short course digoxin therapy started in the emergency department based on clinical outcome after 30 days post hospital discharge.

Methods From Great Tunisian registry, acute decompensated heart failure (ADHF) patients from January 2016 to January 2018 were identified. Patients with incomplete data were excluded. Digoxin treated and non-treated patients were compared in a matched control study with respect to primary outcomes of all-cause mortality and HF readmission. Secondary outcomes included changes of cardiac output (CO) and left ventricular ejection fraction (LVEF) after 72 hours of hospital admission.

Results The study population comprised 104 digoxin treated and 229 matched non-treated with a median age of 67.4±12.8. After 72 hours of ED admission, there was a larger increase of CO (17.8 % vs 14%; p=0.015) and LVEF (14.4% vs 3.5%; p=0.003) in digoxin group compared to control group. At 30-day post-hospital discharge 34 (10.2%) patients died and 72 (21.6%) patients were readmitted. Use of digoxin was associated with decreased risk of death and hospital readmission [odds ratio, 0.79 (95% CI, 0.71-0.89)].

Conclusion In ADHF patients, treatment with digoxin was associated with a significant decrease risk of 30-day mortality and hospital readmission with an improvement of cardiac output and left ventricular ejection fraction.

Key words: Acute heart failure; digoxin; mortality; rehospitalization; emergency department.

INTRODUCTION

Heart failure (HF) is a major worldwide health problem and one of the most important causes of hospital admissions [1,2]. These hospitalizations are responsible for an important economic burden and are associated with high mortality rates, up to 20% following hospital discharge [3,4]. Acute decompensated HF (ADHF) management is difficult given the heterogeneity of the patient population, incomplete understanding of its pathophysiology and lack of evidence-based guidelines. Although the majority of patients with ADHF appear to respond well to initial therapies consisting of loop diuretics and vasoactive agents, these first line treatments failed to decrease post-discharge mortality and readmission rate [5,6]. Investigations of novel therapies such as serelaxin did not show a significant clinical benefit. In a recent multicenter, double-blind, placebo-controlled trial including patients who were hospitalized for acute heart failure, it was shown that the risks of death at 180 days were not lower in patients who had received intravenous serelaxin for 48 hours than in patients who had received placebo [7]. Numerous other clinical trials have been published on ADHF treatment and their results were disappointing in term of efficacy and/or safety [8-11]. Digoxin is one of the oldest compounds in cardiovascular medicine but its beneficial effect is very controversial [12]. Yet, digoxin has many potential beneficial properties for heart failure as it is the only oral inotrope available that did not alter blood pressure neither renal function. Despite its useful hemodynamic, neurohormonal, and electrophysiological effects in patients with chronic congestive HF, concerns about digoxin safety were constantly highlighted [13]. Consequently, the use of digoxin has decreased considerably, in the last 15 years [12]. Digoxin under prescribing is problematic for several reasons. First, it underestimated the substantial beneficial effect of digoxin on the reduction of hospital admissions in HF patients. Second, for its low cost, the favorable cost-effectiveness ratio of digoxin is highly desirable in low-income countries. Moreover, the question whether a short course of digoxin is useful in ADHF was not previously investigated in the era of new heart failure therapies including β-blockers, angiotensin converting enzyme inhibitors and angiotensin-receptor blockers [12]. The objective of this study is to assess the efficacy and safety of a short course digoxin in patients admitted to the ED with ADHF (Figure1 and Figure 2).

PATIENTS AND METHODS

Data Source

We conducted a retrospective matched case-control study to assess the association between digoxin treatment and 30-day outcome in patients with ADHF. The ADHF patients were identified from the Great Tunisian database between January 2016 and January 2018. The patients included are residents of a community of 500,000 inhabitants in the east of Tunisia, served by 2 university hospitals (Fattouma Bourguiba Monastir, and Sahloul Sousse). ADHF was defined as an acute onset of symptoms within 48 hours preceding presentation, dyspnea at rest or with minimal exertion, evidence of pulmonary congestion at chest radiograph or lung ultrasound, NT-proBNP ≥1400 pg/ml. This electronic medical recording system provided detail of each patient admitted to emergency department (ED) for acute undifferentiated non traumatic dyspnea.

Study Population

Patients were included if the following data are available: demographic characteristics, comorbidities, current drug use, baseline NYHA functional class, physical exam findings, standard laboratory tests, brain natriuretic peptide levels at ED admission; echocardiographic results, bioimpedance measured cardiac output at ED admission and at hospital discharge, digoxin daily dose, 30-day follow-up information including ED readmission and survival status. A patient who received at least 0.25 mg of oral digoxin (1 tablet) for three days during hospital stay was defined as case; those who did not receive digoxin treatment were selected as control. The protocol used in this study was approved by the ethics committee of our institution, and all subjects gave their written informed consent to be included in the data base. All the listed criteria have to be fulfilled for patient’s inclusion. Exclusion criteria included ongoing treatment with digoxin, pregnant or breast-feeding women, patients with known severe or terminal renal failure (eGFR<30 ml/min/1.73m2), alteration of consciousness (Glasgow coma score <15) and patients needing immediate hemodynamic or ventilatory support. Cases were matched first for sex, then for age (±2 years) and NYHA functional class. We performed an individual matching; we matched each patient under digoxin (case) with 2 patients who did not receive digoxin (control) for age, gender and New York Heart Association (NYHA) classification. Reviewers were limited to matching criteria data only (e.g., blinded to 30-day outcomes) to eliminate potential sources of bias. Patients who were treated with digoxin and those who did not receive digoxin were clinically managed the same way.

Outcome Measures

The main end points included death or rehospitalization within 30 days after hospital discharge, and 30-day combined death-rehospitalization outcomes. Secondary end-points included CO change from baseline and length of stay in the hospital during the index episode.

Statistical Analysis

Baseline characteristics were compared between groups to detect any differences between cases and controls; independent t-tests were performed for normally distributed variables; Mann Whitney U tests were performed for continuous non-normally distributed variables; and chi square analyses were performed for categorical variables. Logistic regression analysis was performed to identify the odds ratios (ORs) and 95% confidence intervals (95% CIs) for hospital readmission and/or death risk with respect to digoxin treatment. Data are reported as means ± standard deviations, unless otherwise noted, and a p-value less than 0.05 via two-sided testing was considered statistically significant. Data were analyzed using the statistical software package SPSS version 18.

RESULTS

The initial study population comprised 1727 participants who were registered in the database. From this initial population, we excluded 956 with non-cardiac cause of dyspnea, and 211 with incomplete data. From the remaining patients, 104 were included in the digoxin group and 229 in the control group. Digoxin was orally administered once a day and almost all of our patients received the same dose (0.25 mg, one tablet) each day during at least three days. Only few patients received a lower (0.125 mg) or a higher (0.5mg) dose. Baseline characteristics of both groups are shown in table 1. Demographic characteristics were comparable among both study’s groups. There were no relevant differences in age, sex, or NYHA classification. The NYHA class collected was related to base line medical status (within three months before the ongoing exacerbation). Cardiovascular medical history was comparable for both groups. There were no significant differences between cases and controls regarding underlying other comorbidities. Fifty-two percent of the patients had ischaemic cardiomyopathy as the primary aetiology of their heart failure (47-57%) (Table1). Principal baseline medication consisted of diuretics, angiotensin converting enzyme-inhibitors, beta-blockers, and nitrates. Mean vital signs values at baseline were comparable among the 2 groups with respect to heart rate, respiratory rate, and blood pressure. NT-proBNP levels ranged from 1412 to 8615 pg/ml between; 61% in digoxin group and 59% in control group had reduced LVEF (<45%) (p=0.77). After 72 hours of ED admission, there was a larger increase of CO (17.8% vs 14%; p=0.015) and LVEF (14.4% vs 3.5%; p=0.003) in digoxin group compared to control group (Figure 1); NTpro BNP levels decreased and in digoxin group (2%) and in control group (1.2%) but the difference was not significant (p=0.06). Digoxin treatment was associated with a reduced length of hospital stay (10.1±7.2 days versus 6.6± days; p<0.01). At 30-day follow-up, digoxin group showed a significantly lower all-cause (p=0.04) and heart failure (p=0.02) hospital readmission rate compared to control group, and lower mortality (11.8% versus 6.7%; p=0.03) (Table 2). Digoxin treatment was found to significantly decrease the odds for the combined events of mortality and hospital readmission [odds ratio, 0.79 (95% CI, 0.71-0.89)]. No major side effects were observed in relation to digoxin therapy.

DISCUSSION

Our results demonstrated that digoxin is associated with a lower risk of 30-day hospital readmission among ED patients with decompensated HF. Compared with control group, LVEF and cardiac output increased and length of hospital stay decreased significantly in digoxin-treated group. Most available studies analyzed long-term effect of digoxin in patients with chronic heart failure, but data on the effect of short course digoxin on early clinical outcome and physiological related parameters in patients with acute heart failure are scarce. The concordance between physiological and clinical outcomes was in favor of the validity of our results. Digoxin is one of the oldest drugs used in cardiology practice, and few decades ago, it was prescribed in more than 60% of heart failure patients in the United States [14]. Digoxin is the only inotropic drug known to increase cardiac output and to reduce pulmonary capillary pressure without increasing heart rate or lowering blood pressure in contrast to other oral inotropes. However, despite the evidence of its beneficial effects on hemodynamic, neuro-hormonal and electrophysiological parameters, a great concern regarding its safety profile has been raised and the use of digoxin has declined significantly over the past two decades [15]. Indeed, in the ESC guidelines (2016), digoxin indication was limited only to patients with AF and rapid ventricular rate [16]. This could be understandable given the scarcity of randomized trials specifically aimed at testing digoxin safety in heart failure patients. The Digitalis Investigation Group (DIG) trial, the only large randomized trial of digoxin in heart failure, reported a significant reduction in heart failure hospitalizations [17]. Most of the identified studies against the use of digoxin had many potential sources of bias requiring careful assessment. In fact, digoxin safety concern comes from very heterogeneous studies and non-experimental observational studies carrying a high risk of misinterpretation [18-20]. A recent study concluded that prescription of digoxin is an indicator of disease severity and not the cause of worse prognosis which means that a significant prescription bias might be caused by the fact that sicker patients, having a higher mortality risk receive additional treatment with digoxin [21]. Currently, in DIG trial, there is no evidence of an increased risk with digoxin treatment. Importantly, DIG trial demonstrated that beneficial digoxin effects were mainly observed in patients with HFrEF and those with serum digoxin concentration ≤0.9ng/ml. Digoxin efficacy may be attributed in part to the neurohormonal‐inhibiting properties of digoxin, especially in lower doses; it may also be related to its synergistic effects with beta-blockers as pro‐arrhythmic effects of digoxin would be expected to be attenuated by β‐blockers [22].

Our study has several limitations. First, as this is a retrospective analysis, we should clearly highlight that our results only describe associations and not causality. Second, our study is limited by its small sample size. Third, as in all case control studies; bias due to unmeasured confounders remains possible. We should have used the propensity-score matching to better match our two groups but we should point out that most of confounding variables influencing outcome were well balanced between the 2 groups of our study. Third, we had no data regarding post-discharge adherence to prescribed treatment nor we had informations on neither serum digoxin concentration nor the incidence of digoxin toxicity. We acknowledge that this important information would be a valuable support to our findings in demonstrating a correlation between serum digoxin levels and their clinical outcome in our patients. In addition, in our study only 30% of our patients were receiving aldosterone antagonists, and none were receiving cardiac resynchronization therapy, which may limit generalizability of our results.

CONCLUSIONS

Our findings provided an additional data to support the association between use of digoxin and clinical benefit in HF patients with reduced LVEF. Digoxin may potentially serve as an inexpensive tool for the reduction of short-term mortality and hospital readmissions which is an important objective especially in low-income countries in the health system.

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Noise Pollution is One of the Main Health Impacts in Big Cities Today by Tamaz Patarkalashvili* in Open Access Journal of Biogeneric Science and Research

ABSTRACT

Noise pollution today is one of the biggest health risks in big cities along with air pollution. It must be admitted that noise pollution was overlooked by scientists and city authorities lately. Noise pollution has adverse effect on all living organisms. Scientists confirm that noise incentives central nervous system that stimulus to release some hormones which increases risk of hypertension. Hypertension is related with many other cardiovascular and cerebrovascular diseases like infarction and strokes. Nowadays this tendency is being changed at last and noise pollution is often considered not only as harmful as air pollution but sometimes even more. European and North American countries have taken a number of measures to reduce noise level in big cities. Examples of popular measures include replacement of older paved roads with smoother asphalt, better management of traffic flows and reducing speed limits to 30 km. per hour, using less-noisy models of transport, like electric vehicles, cycling and walking.

KEYWORDS: Noise; Pollution; Health; Traffic; Aviation; Vehicle; Electric Car; Cycling; Walking

INTRODUCTION

Noise pollution is a constantly growing problem in all big cities of the world. Many people may not be aware of its adverse impacts on their health. Noise pollution is a major problem both for human health and the environment [1,2]. Long-term exposure to noise pollution can induce variety of adverse health effects like increasing annoyance, sleep disturbance, negative effects on cardiovascular and metabolic system, as well as cognitive impairment in children. Millions of people in big cities suffer from chronic high annoyance and sleep disturbance. It is estimated that school children suffer reading impairment as a result of aircraft noise. Despite the fact that noise pollution is one of the major public health problems in most big cities of the world there was a tendency of underestimating it making accent mostly on-air pollution [3].

World Health Organization (WHO) guidelines for community noise recommends less than 30 A-weighted decibels dB(A) in bedroom during night for good quality sleep and less than 35 dB dB(A) in classrooms to allow good teaching and learning conditions. The WHO guidelines for night noise recommend less than 40 dB (A) 0f annual average (L night) outside of bedrooms to prevent adverse health effects from night noise.

According to European Union (EU) publication:

about 40% of the population in EU countries is exposed to road traffic noise at levels exceeding 55 dB (A)

20% is exposed to levels exceeding 65 dB (A) during daytime and

more than 30 % is exposed to levels exceeding 55 dB (A) at night

Some groups of people are more vulnerable to noise. For example, children spending more time in bed than adults are more exposed to night noise. Chronically ill and elderly people are more sensitive to disturbance. Shift workers are at increased risk because their sleep structure is under stress. Nuisance at night can lead to increased visits in medical clinics and extra spending on sleeping pills that effects family’s budgets and countries’ health expenditure [4,5].

FACTS AND ANALYSIS

Adverse effect of noise is defined as a change in the morphology and physiology of an organism that results in impairment of functional capacity. This definition includes any temporary or long-term lowering of the physical, psychological or social functioning of humans or human organs. The health significance of noise pollution is given according to the specific effects: Noise-induced hearing impairment, Cardiovascular and physiological effects, Mental health effects, Sleep disturbance and Vulnerable groups.

Noise-Induced Hearing Impairment

The International Organization for Standardization (ISO 1999) standard 1999 gives a method for calculation noise-induced hearing impairment in populations exposed to all types of noise (continu-ous, intermittent, impulsive) during working hours. Noise exposure is characterized by LAeq over 8 hours (LAeq, 8h). In the standard, the relationships between Laeq, 8h and noise-induced hearing impairment are given for frequencies of 500-6000Hz and for exposure time of up to 40 years. These relations show that noise-induced hearing impairment occurs predominantly in the high-frequency range of 3000-6000Hz, the effect being largest at 4000Hz [6,7].

Hearing impairment in young adults and children were assessed by Laeq on 24h time basis [7-9]. It includes pop music in discotheques and rock-music concerts [8]. Pop music through headphones [10,11], music played by brass bands and symphony orchestras [11,12]. There is literature showing hearing impairment in people exposed to specific types of non-occupational noise. These noises originate from shooting, motorcycling, using noisy toys by children, fireworks’ noise [13,14].

In Europe environmental noise causes burden that is second in magnitude to that from air pollution. At least 113million people are suffered from traffic-related noise above 55dB Lden that costs the EU about E57.1 billion a year. Additionally, 22 million Europeans are exposed to railway noise, 4 million to aircraft and about 1 million to industrial noise. All these exposures to noise pollution cause about 1.6 million of life lost annually, about 12000 premature deaths and 48000 cases of ischemic heart diseases. About22 million people suffer from chronic high annoyance and 6.5 million from sleep disturbance [15-17].

Cardiovascular and Physiological Effects

Laboratory studies of workers exposed to occupational noise and noisy streets, indicate that noise can have temporary as well as, permanent impacts on physiological functions in people. Acute noise exposures activate autonomic and hormonal systems, leading to temporary changes such as hyper- tension and ischemic heart diseases associated with long-term exposure to high sound pressure levels [7,11,18]. The magnitude and duration of the effects are determined in part by individual characteristics, lifestyle behaviors and environmental conditions. Sounds also evoke reflex responses, particularly when they are unfamiliar and have a sudden onset. The most occupational and community noise studies have been focused on the possibility that noise may be a risk factor for cardiovascular disease. Studies in occupational settings showed that workers exposed to high levels of industrial noise for many years at their working places have increased blood pressure and risk for hypertension, compared to workers of control areas [19,20]. Cardiovascular adverse effects are associated to long-term exposure of LAeq, 24h. values in the range of 65-70 dB or more, for both air and road-traffic noise.

Mental Health Effects

Environment noise accelerates and intensifies development of adverse effects on mental health by variety of symptoms, including anxiety, emotional stress, nervous complains, nausea, headaches, changes in mood, increase in social conflicts, psychiatric disorders as neurosis, psychosis and hysteria [21-32]. Noise adversely effects cognitive performance. In children environmental noise impairs a number of cognitive and motivational parameters [20,22]. Two types of memory deficits were identified under experimental noise exposure: incidental memory and memory for materials that observer was not explicitly instructed to focus on during learning period. Schoolchildren in vicinity of Los Angeles airport were found to be deficient in proofreading and persistence with challenging puzzles [20]. It has been documented following exposure to aircraft noise that in workers exposed to occupational noise it adversely effects cognitive task performance. In children too environmental noise impairs a number of cognitive and motivational parameters in children too [21-24].

Sleep Disturbance

Annoyance in populations exposed to environmental noise varies not only with the acoustical characteristics of the noise, but also with many non-acoustical factors of social, psychological, or economic nature [17,7]. These factors include fear associated with the noise source, conviction that the noise could be reduced by third parties, individual noise sensitivity, the degree to which an individual feels able to control the noise.

At nights environmental noise starting at Lnight levels below 40 dB, can cause negative effects on sleep such as body movements, awakenings, sleep disturbance, as well as effects on the cardiovascular system that becomes apparent above 55dB [24-27]. It especially concerns vulnerable groups such as children, chronically ill and elderly people. All these impacts contribute to a range of health effects, including mortality. During the COVID-19 pandemic European cities experienced sufficient reduction in noise pollution due to reduced road traffic movement.

The WHO recommends reduction of road traffic noise levels to 53dB during daytime (Lden) and 45dB during the night (Lnight). Though, the Environment Noise Directive (END) sets mandatory reporting for noise exposure at 55dB Lden and 50 dB Lnight [26-28]. It means that we don’t yet have accurate understanding of exact number of people exposed to harmful noise levels as defined by the WHO [5,6].

Vulnerable Groups

Vulnerable groups of people include people with decreased abilities like: people with particular diseases and medical problems; blind people or having hearing impairment; babies and small children; elderly and old-aged people. These people are less able to cope with impairments of noise pollution and are at greater risk to harmful effects. People with impaired hearing are most effected to speech intelligibility. From 40 years aged people demonstrate difficulties to understand spoken messages. Therefore, majority of this population can be belonged to vulnerable group of people. Children are also included in vulnerable group of noise exposure [29]. So, monitoring is necessary to organize at schools and kindergartens to protect children from noise effects. Specific regulations and recommendations should be taken into account according to types of effects for children like, communication, recreation, listening to loud music through headphones, music festivals, motorcycling, etc.

CONCLUSIONS

Our cities have already witnessed welcome period of unusual quiet during confinement periods due to Covid-19 pandemic, but noise pollution is rising again and, in some cases, even more than precrisis levels. It is clear that we cannot live without sound or noise and reducing noise pollution to zero level is unrealistic. However, we must work to make sure that noise be reduced to less harmful levels to environment and human health. Examples of measures include: installing road and rail noise barriers; optimizing aircraft movements around airports and urban planning measures. But the most effective actions to reduce exposure can be reduction of noise at source, namely by reducing number of vehicles, introducing quieter tires for road vehicles and laying quieter road surfaces. Anyway, it is unlikely that noise pollution will decrease significantly in near future and that transport demand is expected to increase. Air traffic noise is also predicted to increase along with city inhabitants. Effective measures against this situation can be raising awareness and changing people’s behavior by using less-noise models of transport, such as electric vehicles, cycling and walking. Zero emission buses must be welcomed in big cities as well, as refuse collection trucks and municipal vans. Required infrastructure of safe cycling must be constructed in cities for safe cycling and available public bike fleet. Such types of transport as motorcycles and scooters must be banned in big cities because they produce the most terrible and loud noise that adversely impacts on citizens. Municipalities and mayors of big cities must organize so-called quiet city areas, like commodity parks and other green spaces, where people can go to escape city noise.

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Effect of Qishan Formula Granules on Interventing Obesity Intestinal Microflora and Immune-Inflammatory by Wei Yan in Open Access Journal of Biogeneric Science and Research

ABSTRACT

Objective: To investigate the effects of Qishan Formula Granule on Simple Obesity and Intestinal Microflora-Inflammatory Immune Pathway. Methods: Eighty patients with simple obesity in our hospital were randomly divided into two groups: traditional Chinese medicine group and placebo group. The Chinese medicine group was treated with lifestyle intervention + Qishan formula granule, while the placebo group was treated with lifestyle intervention + placebo. The therapeutic effect, biochemical indexes, clinical symptoms, the number and composition of bacteria, the proportion of Th17/Treg cells in serum and inflammatory factors were measured before and after treatment. Results: After treatment, the total effective rate of simple obesity patients in Chinese medicine group was significantly higher than that in placebo group. (P ＜05). Compared with the placebo group after treatment, the biochemical indexes and clinical symptoms of the Chinese medicine group improved significantly after treatment. Further tests showed that Qishan Formula Granule could significantly improve the intestinal bacterial abundance, species and quantity of simple obesity patients. The levels of IL-17, TNF-alpha, Th17/Treg and LPS in patients with simple obesity in traditional Chinese medicine group were significantly lower than those in placebo group before and after treatment (P ＜ 0.05). Conclusion: Qishan Formula Granule can alleviate clinical symptoms of simple obesity and improve treatment efficiency through intestinal flora-inflammatory immune pathway.

KEYWORDS: Qishan formula granule; Obesity; Intestinal flora; Immune inflammatory; Th17/Treg

In recent years, with the change of people's lifestyle, the incidence of obesity has increased rapidly. For the chronic metabolic disorders celected to obesity and overweight, the prevalence of diseases such as diabetes and cardiovascular and cerebrovascular diseases has increased year by year [1]. Obesity cannot only lead to diabetes or high incidence of cardiovascular and cerebrovascular events, but also closely relate to cancer, depression, asthma, apnea syndrome, infertility, osteoarthropathy, fatty liver and many other diseases [2-5]. Therefore, it has become a serious impact on people's health, it’s urgent to find reasonable and effective intervention measures. At present, the main drugs of weight loss treatment include non-central drugs, central drugs and hypoglycemic drugs, which have many problems such as low effective response rate, large side effects and weight rebound after stopping [6]. The combination of diet and exercise is often difficult to adhere to for a long time, and compliance is poor [7]. Seeking effective drugs or methods to treat simple obesity has become an important research hotspot in recent years.

It is believed that intestinal flora plays an important role in the regulation of immune inflammation and glycolipid metabolism [8,9]. Studies have shown that there is chronic low-level inflammation in obese people, and chronic low-level inflammation caused by obesity may promote the occurrence development of metabolic disorders [10]. At the same time, it has been found that the disorder of intestinal flora and its metabolites in obesity patients and the obvious imbalance of proportion can affect the formation and differentiation of immune cells such as Th17 cells and Treg cells, thus leading to chronic low-level immune inflammation and obesity [11-13]. At present, a number of studies have shown that traditional Chinese medicine has an important effect on intestinal flora, berberine, Gegenqinlian decoction, tonifying traditional Chinese medicine and so on have a certain degree of adjustment of bacterial dysbiosis [14-15]. Qishan formula granules from Gegenqinlian modification, is a national famous old Chinese medicine experience prescription. The results of the previous study showed that Qishan formula has the effect of reducing blood sugar, improving insulin resistance and reducing body weight. However, the mechanism is not clear, and the simple obesity population has not been studied. Therefore, this study explores the efficacy of Qishan Formula Granule in the treatment of simple obesity and its effect on intestinal flora immune inflammatory pathway, in order to provide reference for traditional Chinese medicine in the treatment of obesity.

INFORMATION AND METHODOLOGY

General Information

In this study, A randomized (randomized digital approach), double-blind（The subjects, researchers, surveyors or data analysts did not know the treatment allocation, placebo-controlled, prospective study approach was selected for simple obese patients through health check-ups, community population screening, and outpatient visits. All patients were treated with lifestyle intervention（In a low sugar diet, 200-350g of main food should be eaten every day, and the ratio of carbohydrate to total calories should be 50% - 65%. Low fat diet, fat intake within 50g, about 30% of the total calories. Protein balance, about 15% of the total heat. Encourage the intake of foods rich in dietary fiber and vitamins. The daily total heat is controlled within 100kj / kg. Adhere to moderate intensity aerobic exercise, i.e., heart rate + 170 age after exercise, at least 3-5 days a week. It lasts for half a year; those with diabetes were treated with glizat sustained-release tablets; and those with hypertension were treated with amlodipine. The study included 80 patients who still met the following criteria after the 1-month elution period. a random number table was established using excel software. the standard 80 patients were randomly averaged into two groups: the traditional medicine group (40) and the placebo group (40). In the group of traditional Chinese medicine,14 cases were male and 26 cases were female; the age was 25-50 years, the average age was 38.74±10.23 years; and the average course of disease was 6.63±3.55 years. in the placebo group,16 men and 24 women; age 25-50 years, mean age 39.61±9.83 years; and mean course of disease 6.17±3.82 years.

Inclusion and Exclusion Criteria

Inclusion Criteria

(1)Patients with simple obesity (male waist ≥90 cm, female ≥85 cm, and BMI≥25 kg/m2) in accordance with the 2011 edition of the Expert Consensus on the Prevention and Control of Adult Obesity in China;(2) age greater than 25 years of age less than 70 years of age;(3) classification of TCM syndrome differentiation as obesity-wet-heat accumulation of spleen syndrome ：The body is fat（25kg/m2≤BMI≤28kg/m2：+1score, 28kg/m2≤BMI≤30kg/m2：+2scores, 30kg/m2≤BMI：+3scores,）; the abdomen is full(+1score); the food is little and tired(+1score); the head is heavy as wrap(+1score),The loose stool is not good(+1score); the urine color is yellow(+1score); and the whole body is hot and humid jaundice(+1score);The tongue is fat(+2scores); with yellow and greasy fur(+2scores);and smooth veins(+2scores);(4) discontinuation of drugs affecting weight for 4 weeks; (5) signing of informed consent.

Exclusion Criteria

(1)Weight gain due to drugs, endocrine diseases or other diseases;(2) severe liver and kidney dysfunction or other severe primary diseases;(3) history of acute cardiovascular and cerebrovascular events or myocardial infarction within 6 months;(4) stress state or secondary blood glucose elevation or secondary hypertension;(5) weight-loss surgery within one year;(6) severe dyslipidemia;(7) unwillingness of cooperators (who cannot cooperate with dietary control or do not use drugs as prescribed);(8) mental illness, tumor patients;(9) women with or breast-feeding, and women with planned or unplanned contraception;(10) possible allergy to gestational drugs;(11) patients with diabetes who have received medication.

Treatment

(1) The Chinese medicine group was given lifestyle intervention + qishan formula granules orally, one pack at a time, twice a day. (The formulas are: Pueraria root 15g, Scutellaria baicalensis 10g, Coptis chinensis 10g, Rhubarb 3g, gynostemma pentaphyllum 10g, Shengqi 20g, Huai yam 20g, Atractylodes chinensis 15g, Poria cocos 15g, fried Fructus Aurantii 10g, Raw Hawthorn 10g, Chuanxiong 10g, produced by the preparation room of traditional Chinese medicine in our hospital). (2) The placebo group was given a lifestyle intervention plus a placebo oral dose, twice daily. (The formulas are: starch, pigment and adhesive, which are produced by the traditional Chinese medicine preparation room of our hospital).

Indicator Measurements

Fasting blood glucose (FPG), blood lipids, blood pressure, waist-to-hip ratio, body mass index (BMI), body fat content, tcm symptom score (and other biochemical indicators, clinical symptoms were measured every 4 weeks. HbA1c, fasting insulin (FINS), fecal intestinal flora, proportion of serum th17/treg cells and serum IL-17, TNF-α, liver and kidney function, blood routine, urine routine, electrocardiogram were measured at week 0 and 12.

Determination of Flora Size and Composition

Quantitative intestinal excreta were diluted and inoculated into bs medium (bifidobacterium isolate) anaerobic culture for 48 h, bbe medium (bacillus isolates) anaerobic culture for 24h, lactic acid bacteria selective medium for 24h, enterococcal agar for 24h, fs medium (clostridium isolates) for 72h, kf streptococcus agar (streptococcus isolates) anaerobic culture for 24 h, and iridium agar for 24h. After the growth of the colony, the desired target bacteria were identified by colony morphology, Gram staining and biochemical reaction. On a variety of different media, the colonies were identified, and the number of each bacteria was compared with the reference value, and the B/E value was calculated to evaluate the number and composition of intestinal flora in patients with simple obesity after oral intervention of Qishan without sugar.

PCR-DGGE Analysis Intestinal Flora Composition

Fecal specimen collection and DNA extraction: The collection of feces of simple obese patients by aseptic method was about 1 g in 2 mL EP tube, and the stool genomic DNA was extracted according to the instructions of the DNA extraction kit.pcr: the v3 section of bacterial 16srdna was amplified by universal primers. the amplification conditions were 94°c for 3 min predenaturation,94° c for 1 min denaturation,55°c for 1 min annealing,72°c for 1 min extension, a total of 36 cycles,72°c for 10 min extension, and 4°c preservation.PCR products were detected by 2% agarose gel electrophoresis and stored at -20°C.dgge: the pcr product was separated on 8% polyacrylamide glue, the gel was stained by gelred after the end, the gs-800 grayscale scanner was imaged, and the correlation analysis of dgge molecular fingerprint was performed by biomerics software.

Quantitative Quantitative PCR Analysis of Intestinal Microflora

Fecal specimen collection and DNA extraction: The collection of feces of simple obese patients by aseptic method was about 1 g in 2 mL EP tube, and the stool genomic DNA was extracted according to the instructions of the DNA extraction kit. PCR primer design: according to Bifidobacterium, Lactobacillus, Escherichia coli, Bacillus, Clostridium, Streptococcus 16SrDNA gene sequence, the corresponding bacterial PCR primer was designed, and the specificity of the corresponding bacterial sequence was compared in the BLAST gene bank. Preparation of the standard curve: the PCR products of each bacteria in the control group were purified according to the instructions of the DNA purification kit, and the absorbance (A value) and concentration of the purified product were determined, and the copy number of each standard product 1μl was converted to be used to make the standard curve.

Detection of Biochemical Indexes

Blood sugar, blood lipids and other biochemical indicators were determined by the Olimpas 2000 large automatic biochemical instrument. serum insulin, hba1c was determined by our advia centur®xp fully automated chemiluminescence immunoanalyzer. Serum LPS was detected by ELISA. Methods for the determination of intestinal flora and SCFAs:2g of fresh feces of patients were frozen at -20°C refrigerator with a toilet provided by the Institute of Microbiology of Zhejiang Province (containing stabilizer), and the Institute was commissioned to test it. Body fat content was determined by the department's own body fat tester.HOMA-IR is calculated by formula HOMA-IR = FPG × FINS /22.5, HOMA-IS is calculated by formula HOMA-IS =1/HOMA-IR=22.5/ FPG × FINS.

Anges Of Th17/Treg Cells Before and After Intervention

The peripheral blood of the two groups of patients was collected, standing for 1 h,2000 rpm,4°c, centrifuged for 10 min, collected and packed into supernatant, and stored at -20°c when not detected in time. the content of serum il-17, tnf- α was determined by double antibody sandwich enzyme-linked immunosorbent assay (elisa), and the specific operation was carried out strictly according to the instructions of elisa kit. cd3-pecy7 and cd4-pe 0.5μg each, after oscillating and mixing evenly, incubated at room temperature for 30 min;300 g centrifuged for 5 min. after washing with cold pbs,1 ml of diluted fixed, membrane-penetrating agent was added. after reaction for 50 min, the concentration of th17 and treg cells was determined by flow cytometry.

Safety Evaluation and Adverse Reaction Management

If there is an alt increase during medication, the principle of adjusting the drug dose or interrupting treatment is:1 If the alt increase is within 2 times the normal value, continue to observe. If ALT rises at 2-3 times the normal level and is taken in half, continue to observe if ALT continues to rise or remains between 80-120 U.L-1 and interrupt treatment. 3 If ALT rises above 3 times the normal value, stop the drug. After the withdrawal of drugs return to normal can continue to use, and strengthen the treatment of liver protection and follow-up. If leukopenia occurs during medication, the principles for adjusting the drug dose or interrupting treatment are as follows:1 If leukopenia is not lower than 3.0 x 109·L-1, continue to take medication to observe. If the white blood cell drops between (2.0 and 3.0) ×109·L-1, observe in half. Most patients can return to normal during continued medication. If the review of leukocytes is still below 3.0 x 109·L-1, the treatment is interrupted. 3 If leukopenia falls below 2.0 x 109·L-1, interrupt treatment.

CRITERIA FOR EVALUATION OF SYNDROME EFFICACY

Clinical recovery: TCM clinical symptoms, signs disappear or basically disappear, syndrome score reduction ≥90% and weight lost by ≥ 15%. Remarkable effect: the clinical symptoms and signs of TCM were obviously improved, and the score of syndromes was reduced by more than 70% and weight lost by ≥ 10%. Effective: TCM clinical symptoms, signs are improved, syndrome score reduction≥30% and weight lost by ≥ 5%. Invalid: TCM clinical symptoms, signs are not significantly improved, or even aggravated, syndrome score reduction <30% or weight lost by<5%. Total effective = (clinical recovery + significant + effective)/ total number *100%.

Statistical Analysis

Statistical software SPSS 17.0 is used to compare and analyze the indexes in this paper. The measurement data are expressed in form, and the measurement data are normally distributed, which meet the t-test standard.T-test to compare the counting data with xs test comparison; the count data are compared by Χ2 test; when p < 0.05, the statistics have significant differences.

Estimation of Sample Size

Estimation of sample size: according to the estimation method of sample size in clinical experimental research, the sample size of two sample mean comparison is estimated. Check the "sample size table required for two sample mean comparison". According to the bilateral α = 0.05, the test efficiency (1- β) = 0.9, μ 0.05 = 1.96, μ 0.1 = 1.28, according to the previous research experience, σ is the estimated value of the overall standard deviation of two samples = 16, δ is both samples. The difference of number = 3.0, and the result is n = 34. Considering the loss rate of sample 15%, 40 cases in the experimental group and 40 cases in the control group were preliminarily determined.

RESULTS

The rate of abscission and baseline were compared between the two groups. In the placebo group, 40 cases were enrolled, 2 cases were dropped, 38 cases were observed, and the drop rate was 5%. In the treatment group of traditional Chinese medicine, 40 cases were enrolled, 3 cases fell off, 37 cases were observed, the rate of falling off was 7.5%. There were 12 diabetic patients in placebo group, 15 hypertensive patients, 13 diabetic patients and 14 hypertensive patients in traditional Chinese medicine treatment group. There was no significant difference between the two groups in the proportion of diabetic and hypertensive patients. The age gender, baseline FPG, total cholesterol, triglyceride, LDL, BMI, body fat content, HbA1c, fins, waist to hip ratio, TCM syndrome score and the number of specific intestinal flora were comparable.

Groups Comparison of Clinical Effect of Simple Obesity Patients After Treatment

Compared with the placebo group after treatment, the total effective rate of simple obese patients in the traditional chinese medicine group was significantly increased after treatment (81.1% vs 50.0 %), and there was a significant difference in comparison (p <0.05). the clinical efficacy of the two groups was compared in table 1. Further observation found that the two groups of patients did not have liver function, renal function, abnormal white blood cell level and other adverse reactions Table 1.

Two Groups Comparison of Biochemical Indexes of Simple Obesity Patients Before and After Treatment

Compared with the previous treatment, the indexes of the patients with simple obesity in placebo group did not change significantly after treatment, and there was no significant statistical difference (P >0.05). The scores of FPG, total cholesterol, triglyceride, low-density lipoprotein, BMI, body fat, HbA1c, FINS, waist-to-hip ratio and TCM syndromes were significantly lower in patients with simple obesity than before and after treatment group of placebos, whereas HDL was significantly higher than that before and after treatment placebo group, there was a significant difference in comparison (P <0.05); the comparison of biochemical indexes before and after treatment in the two groups was shown in Table 2.

Groups Patients with simple obesity before and after treatment Comparison of intestinal flora

The colony numbers of Bifidobacterium, Bacillus fragilis, Lactobacillus, Enterococcus and Escherichia coli in the placebo group were not significantly different after treatment compared with that before and after treatment (P >0.05). The colony numbers of Bifidobacterium and Bifidobacterium in the traditional Chinese medicine group were significantly higher than those in the comfort group before and after treatment (P <0.05). A similar result was found for the detection of bacterial copy number of each stool by real-time fluorescence quantitative PCR, and the real-time fluorescence quantitative PCR is shown in Table 4.

Two Groups Comparison of Inflammatory Indexes in Simple Obesity Patients Before and After Treatment

Compared with pre-treatment, the inflammatory indexes of patients with simple obesity in placebo group did not change significantly after treatment (P >0.05)The levels of IL-17, TNF-α and Th17/Treg were significantly lower in patients with simple obesity than those before and after treatment placebo group, there was a significant difference in comparison (P <0.05); the comparison of biochemical indexes before and after treatment in the two groups was shown in Table 5.according to the relationship between th17/treg level and normal range after treatment of qishan formula granules, it was divided into normal group (th17/treg level was within normal range) and high level group (th17/treg level was higher than normal range). Further statistics found that the total amount of intestinal flora after treatment in the normal group was significantly higher than that in the high-level group (P <0.05, Fig.1) Table 3.

DISCUSSION

In recent years, with the rapid development of today's society, people's life style and diet, nutrition structure has undergone great changes, so the incidence of obesity remains high, and the incidence of diabetes, cardio-cerebrovascular diseases and other diseases caused by obesity is increasing year by year. Several studies have shown that Qishan formula granules play an important role in reducing body weight. Therefore, this study discussed the effect of Qishan formula granule on simple obesity and further discussed its mechanism.

This study found that the total effective rate of simple obesity patients was significantly increased after the intervention of qishan formula granules. at the same time, the intervention of qishan formula granules could improve the clinical symptoms and biochemical indexes such as blood sugar and blood lipids in simple obesity patients. The application of Qishan formula granules is beneficial to the treatment of simple obese patients. The theory of traditional Chinese medicine believes that obesity is due to dietary fat, inactivity,dysfunction of spleen in transportation, accumulation of phlegm and dampness cream. Body diseases characterized by obesity and fatigue. Its most common symptom is damp-heat accumulation spleen syndrome [16,17]. Qishan formula granules from Gegenqinlian decoction and six gentleman decoctions reduced. Raw astragalus, huai yam, poria qi invigorating spleen, Scutellaria baicalensis, Coptis chinensis Qingzhongjiao dampness and heat, rhubarb, gynostemma pentaphyllum to remove dampness, Atractylodes aromatization and dampness, Fructus Aurantiii, Hawthorn Qi digestion, phlegm elimination, Pueraria, Chuanxiong heat Qingjin Qi Huoxue. Thus, it has the effect of invigorating qi and invigorating spleen, clearing away heat and removing dampness and removing turbidity. It can make temper health transport and water Tianjin four cloth, phlegm turbidity inside the heat clear and fat in the full elimination. The purpose of removing Glycyrrhiza in the original prescription of Gegen Qinlian decoction is to prevent the rise of blood sugar and the storage of water and sodium.Therefore, the application of Qishan formula granules can significantly improve the total effective rate of patients and significantly improve clinical symptoms, blood sugar, blood lipids and other biochemical indicators. However, the mechanism of cell biology and molecular biology has not been elucidated Table 5.

Several studies have pointed out that differences in the composition of the intestinal flora are one of the most important causes of obesity, and their mechanisms mainly involve activating inflammatory responses, promoting energy absorption and regulating intestinal permeability [18,19] Studies have shown that obese patients are often accompanied by inflammatory signaling pathway activation, immune cell infiltration and other pathological changes [20]. therefore, stopping from the intestinal flora-inflammatory immune pathway will be helpful to elucidate the mechanism of qishan formula granules to improve simple obesity. this study examined the number and composition of intestinal flora and found that the colony numbers lactic acid bacteria, bifidobacterium and bacteroides were significantly higher in patients with simple obesity than in the placebo group before and after treatment after treatment after treatment with qishan formula granules. The results showed that Qishan formula could significantly affect the intestinal flora of simple obese patients. He Xuyun and other studies have found that Astragalus polysaccharide, the main ingredient of Astragalus membranaceus, can significantly inhibit the formation of obesity in mice and significantly restore intestinal flora disorders [21]. At the same time, several studies have pointed out that Radix Puerariae, Scutellariae, Coptis, Huai yam, Ligusticum chuanxiong, Poria cocos and Astragalus membranaceus It can affect the composition and richness of intestinal flora [22-25]. thus further confirming the effect of Qishan formula granules on intestinal flora.Intestinal flora plays an important regulatory role in the balance of immune cells. Fang Qian et al found a linear positive correlation between Bifidobacterium/ Escherichia coli ratio and Treg/Th17 in children with asthmatic bronchitis [26]. At the same time, numerous studies have found that berberine regulates the intestinal flora and the balance of Th17/Treg cells in rats, while the disruption of the balance between pro-inflammatory Th17 cells and inhibitory Treg cells is a key factor in many immune and metabolic diseases [27-29]. Therefore, the effect of Qishan formula granules on inflammatory cells and inflammatory factors was further examined. the results showed that il-17, tnf-α, th17/treg and levels were significantly reduced in patients with simple obesity after intervention of qishan formula granules, indicating that qishan formula granules could affect the inflammatory response in patients with simple obesity. further exploring the relationship between th17/treg levels and intestinal flora found that the total amount of intestinal flora in patients with th17/treg levels in the normal range was significantly higher than that of patients with th17/treg levels than normal, indicating a association between intestinal flora and treg/th17 values in patients with simple obesity. In summary, Qishan formula granules can improve the symptoms of obesity by increasing the richness and diversity of intestinal flora in simple obese patients and inhibiting the inflammatory response.

To sum up, this study found that Qishan formula granules can alleviate the clinical symptoms of simple obesity and improve the treatment efficiency through the intestinal flora-inflammatory immune pathway. Qishan formula granules can regulate the proportion of Th17, Treg cells and the secretion level of inflammatory factors by influencing the composition and richness of intestinal flora, so as to reshape the body shape and improve the biochemical index, and then achieve the purpose of treating simple obesity. However, there are still some shortcomings in this study, and it is necessary to further explore the main components of its efficacy and how the intestinal flora affects the inflammatory response.

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Complex Kinesiological Conundrum Could Microzyman Machinations/Inhibitions Explain the Relative Tardiness of Initial Infantile Human Locomotion? Seun Ayoade* in  Open Access Journal of Biogeneric Science and Research

ABSTRACT

“The onset of walking is a fundamental milestone in motor development of humans and other mammals, yet little is known about what factors determine its timing. Hoofed animals start walking within hours after birth, rodents and small carnivores require days or weeks, and nonhuman primates take months and humans approximately a year to achieve this locomotor skill”.

Introduction

We, mankind, are the tardiest living thing in terms of the age we start walking. This is highly embarrassing. It is embarrassing to evolutionists who declare man to be the most biologically advanced and evolved species. It is equally embarrassing to creationists who insist that man was made in the Image of God. If man is the most advanced and evolved animal why do our babies take so long to learn to walk? Why are we, the “peak of God’s creation” carried around by our mothers for a year while the zebras and goats and horses are proudly walking and cavorting just hours after delivery? Creationists have a ready excuse-the fall of man and his expulsion from the Garden of Eden caused man to become genetically degraded [1]. After all, they argue, the first humans Adam and Eve walked and talked the very day they were created. Evolutionists on the other hand put forth other arguments for the very embarrassing ambulatory limitations of Homo sapiens. I hereby refute these arguments viz-Refuting the gestation argument-This argument states that humans are pregnant for 9 months unlike those other animals that are pregnant for shorter periods. However, baby elephants walk hours after birth and the gestation period in elephants is 18 to 22 months! Refuting the Life Span Relativity Argument This argument states that because horses and dogs have shorter life spans than we humans their apparent early walking is not really that early [2]. I refute this argument in the table below by showing at what age human babies would walk if we had the life span of cats and dogs etc table 1.

Refuting the Brain Development Argument

This argument states that all animals start walking when their brains reach a particular stage of development [3]. Then why do humans reach the stage so late if we are the most evolved animal?

Refuting the Bipedal Argument

This argument states that walking on two legs involves much more balance and coordination than walking on all fours and so should take longer. If this argument was true human babies would start crawling hours after birth! Human babies don’t crawl till 4 -7 months! Also studies by Francesco Lacquaniti at the University of Rome Tor Vergata, Italy have shown that despite homo sapiens’ unique gait, the motor patterns controlling walking in other animals are nearly identical to that in man!

Intelligence Argument

This argument claims that since humans are more intelligent than other animals we have to start walking later because we have so many other things to do with our minds apart from walking [4-7]. However, ravens are very intelligent birds yet raven chicks walk and fly at one month old. Monkeys are intelligent and yet start waking at 6 weeks!

MY HYPOTHESIS AND PROPOSAL

The key to cracking this mystery will be to do a comparative study of the cellular dust [8-10] of various animals. This is not likely to happen any time soon however as the mainstream scientific community continues to deny the existence of the microzymas [11].

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Learning Difficulties and Reading Comprehension in the First Grades of Primary School by Theofilidis Antonis* in  Open Access Journal of Biogeneric Science and Research

ABSTRACT

This paper is a study on the concept of learning disabilities and reading comprehension. Specifically, it studies the learning difficulties and the reading ability in terms of the school performance of the students of the first grades of primary school.

Aim: In our work we try to analyze and present the following topics:

The definition of the term "Learning Disabilities", their etiology and the correlation of learning difficulties and reading.

What learning difficulties do,what we face in reading and writing and how do they affect the school context and students' performance.

How to diagnose learning disabilities in reading ability and interventions that need to be implemented in the classroom to reduce them.

Method: We followed the most up-to-date literature on the subject

Conclusions: Learning difficulties can also cause emotional problems in children as they feel that they are lagging behind compared to the rest of the class. Our goal is to include children with learning disabilities in the classroom by adapting the lesson to the children and not the children in it. Our main concern should be the valid and timely diagnosis of difficulties and effective intervention to address them.

Keywords: Learning difficulties, reading comprehension, primary school

LEARNING DIFFICULTIES - DEFINITION AND ETIOLOGY

Learning disabilities are a generalized expression of some of the individual difficulties encountered by students. More specifically, the concept of learning disabilities refers to a variety of heterogeneous disorders, which result in difficulty in learning, speaking, writing, reading, information processing, mathematical computation, attention retention and in the coordination of movements. Every disorder that is part of the learning disability is differentiated in terms of the intensity of its manifestation, the nature and the symptoms of the difficulties, as well as the consequences that they have. These disorders can cause problems throughout the life of the individual [1]. An important point in the study of learning difficulties concerns the impossibility of having common symptoms of expression of these difficulties, a fact that leads to their delayed recognition, which usually occurs during school age [2].

The goal of any educational system is the success of students in their academic performance, as well as their acceptance by the school environment. Many children, however, who have learning difficulties do not have the expected school performance required. The learning difficulties that can occur to each child individually vary, which makes the role of the teacher difficult, since he is called to deal with each case separately [3].

The origin of learning difficulties seems to be mainly due to dysfunction in the central nervous system of the individual [5]. Thus, learning disabilities come through the pathogenesis of the individual himself. However, apart from the neurological factors that seem to play a major role in the development of learning disabilities, the great heterogeneity of the symptoms of learning disabilities leads, however, many researchers to conclude that the etiology of these difficulties is very likely to be multifactorial and epigenetic factors also play a role. More specifically, researchers should not neglect the study of environmental, cultural and emotional factors, processes that seem to play a significant role in the occurrence of learning disabilities. Such factors may be an inappropriate school environment, a difficult family environment, depression, anxiety, the child's personality, psychological neglect, etc. Still, we should not overlook the cognitive factors that emerge through low performance in almost all learning activities. In addition, there seems to be a differentiation of learning difficulties in relation to the sex of the child. In contrast to girls, boys have a higher rate of learning difficulties especially in the behavioral field and language learning [4]. Therefore, as learning is a complex, multifactorial process, all the complex factors that may affect it should be considered. However, all of the above have not yet been clarified in the literature whether they are factors that cause learning disabilities or simply predisposing or risk factors [6].

Papadakou, Palili, Gini, (2014) studied some epidemiological factors that seem to be directly related to the occurrence or not of learning difficulties. The research process showed that children diagnosed with learning disabilities had at least one first-degree relative with learning disabilities. The researchers also found that learning disabilities were directly linked to sleep disorders, attention deficit hyperactivity disorder (ADHD), and a variety of other emotional and social problems. Such problems may be related to anxiety, depression, adaptive disorders and social interaction, etc. The researchers emphasize that the above factors could be considered as prognostic and gain important function in the design of targeted early interventions, with the aim of better academic and social development [7].

Finally, the correlation of learning difficulties with various emotional problems that children may manifest is considered important. Although no precise explanation has been given, it seems that children with learning disabilities tend to develop less positive and more negative emotions, which reduce their willpower and prevent them from making the necessary effort in the school context. (Gugumi, 2015). Behavioral problems can be characterized as internal and external. Problems such as stress, melancholy, depression, obsessive-compulsive disorder, dysthymia and social phobia, as well as disorders such as bulimia, nightmares, anorexia, shyness and isolation are considered internal, while problems such as aggression are considered. Disasters, negativity, rudeness, attention deficit hyperactivity disorder, adjustment and conduction disorders, hostility and theft, are considered external problems [8]. Bornstein, Hahn & Suwalsky, 2013). Children's intrapersonal or interpersonal adjustment is directly affected by these emotional and behavioral problems, which may be due to students' low self-esteem, which stems from the learning difficulties they face (Koliadis, 2010).

LEARNING DISABILITIES AND READING

Most of the children who have Learning Disabilities have problems in the cognitive process of reading and understanding the written text. The process of reading is a complex task of cognitive function, which refers to the processing and analysis of graphs, phonemes and semantic information of the written language. It is closely related to a variety of other cognitive functions of the child, which must be activated for the full understanding of reading, such as the degree of ability of his phonological awareness and the capacity of his short-term memory, perception, concentration, attention, language and thinking, as well as sensory skills such as vision, motor and reading ability [9] Natália Jordão, Adriana de Souza Batista Kida, Danielle Dutenhefner de Aquino, Mariana de Oliveira Costa, & Clara Regina Brandão de Avila. 2019). More specifically, reading refers to the process in which the student decodes the written symbols of our language and converts them into speech. The graphic processing of these written symbols can contain phonemic, phonological and semantic information about the receiver (Porpodas, 2002).

In order for the reading process to take place correctly, it is necessary not only to decode the imprinted symbols, but also to understand their conceptual content. It is thus an important process of processing and extracting information, deeply connected and dependent on decoding and understanding (Porpodas, 2002. Tsesmeli, S. 2012).

We understand, then, that decoding and comprehension are the two most important cognitive functions that play the most important role in the reading process. More specifically, decoding is the ability to recognize written symbols and automatically convert them to a phonological representation. An important role in the correct decoding is played by the state of long-term memory, access to it and retrieval of any information necessary for the correct letter-phonemic matching [19] (Tzivinikou, S. 2015. Kim, MK, Bryant, DP, Bryant, BR, & Park, Y. 2017) [11]. Comprehension, which is the second equally important cognitive function of reading, presupposes the recognition of the semantic content of words, which can come from the knowledge of the meaning of words, the understanding of their grammatical pronunciation as well as their syntactic structure (Kokkinaki 2014. Westwood, 2016). For the correct understanding of the text, the cognitive strategies should be used correctly, the words of the text should be recognized and combined with the previous knowledge [12] (Westwood, 2016. Tzivinikou, 2015) [13]. According to Kaldenberg, Watt, & Therrien, (2015) reading comprehension is directly related to the reader's prior knowledge, as the information obtained from a text in order to be generalized and understood, must be processed and to connect based on the knowledge already possessed by the individual. Lack of this knowledge leads to inability to use metacognitive strategies needed for reading (Kaldenberg et al., 2015) [14].

Reading, then, can be said to be the product of these two factors presented above, decoding and comprehension. As a result, any malfunction even in one of these two factors can lead to the so-called Reading Difficulties. Difficulty in reading comprehension can manifest itself throughout the levels of the reading process, from the simple learning of individual graphs to the reading, comprehension and retention of the acquired textual information. The reading difficulties that children present are mainly based on neurological abnormalities, they can be combined with delayed speech and generally with language problems (Kokkinaki 2014).

LEARNING DIFFICULTIES AND READING IN PRIMARY SCHOOL

In terms of reading comprehension in the school performance of primary school students we must keep in mind the basic cognitive development of the reading process. A student is expected to have successfully completed the decoding ability by the second grade of elementary school. By the third grade, a child should not only be able to easily decode the written text, but also understand the meaning of what they are reading. With this in mind, we can talk about learning difficulties related to reading comprehension, only if the child has received the appropriate education for the stage of the class in which he is. The usual deviation of students with learning disabilities from the rest of the class is about one to two years.

Learning disabilities seem to slow down children's performance at school. The motivation and enthusiasm that each student has for learning, does not seem to exist in students with learning difficulties, resulting in a low academic level [15] (Lama, 2019). We understand, then, that the early diagnosis and treatment of learning difficulties in a child in the first grades of primary school is vital. The main problem lies in the field of decoding since the difficulty of the reader to decode the words hinders his entry into the process of comprehension (Kokkinaki 2014) [16].

LEARNING DIFFICULTIES AND WRITING SPEECH PROBLEMS IN PRIMARY SCHOOL

The role of writing in school is very important since apart from being a means of communication it is also one of the basic skills that will accompany the child throughout his student years and later in life. A necessary criterion for the production of written speech is the linguistic, metalanguage but also the cognitive, metacognitive skills of the individual. An important role in the production of written speech is played by the already existing knowledge and experiences of the individual, his motivations, feelings and goals (Panteliadou 2000. Vasarmidou, D., & Spantidakis, I. I. 2015). Students with learning disabilities have difficulty using metacognitive strategies that should be employed to produce written communication. These strategies would give the student the opportunity in the writing process, would help him in the production and in the end would give him the opportunity to check his result and evaluate it, allowing him to make the necessary corrections (Panteliadou, 2000. Vasarmidou , 2015).However, in addition to cognitive and metacognitive skills, difficulties may also arise in the student's mechanistic skills. These skills include handwriting, spelling, vocabulary development and the use of punctuation, accentuation, writing and the use of uppercase and lowercase letters. Difficulties in some of these skills or in all, create problems in writing [17] (Vasarmidou, 2015. Panteliadou, 2000). Also, features of written speech difficulty can be the reversal or confusion of letters, omissions or additions of letters, illegible letters and permutations. Identifying more serious issues related to speech and reading in combination with writing is much more complicated than the first grade of elementary school, since the difficulty in organizing speech is something common in preschool children (Tzouriadou, 2008) [18].

DIAGNOSIS AND TEACHING INTERVENTIONS IN CHILDREN WITH LEARNING DISABILITIES

There are many cases of students with learning difficulties who have the ability to keep up with the class schedule without particular problems. However, in most cases, the difficulties are quite intense, as a result of which the content needs to be adjusted in order to be able to follow it [20] (Tzouriadou, 2011).

Learning Disabilities are a common problem for many students, but due to their special nature, they enable us to carry out effective intervention programs. This intervention must be timely, in order to deal with the current difficulty in giving birth, as well as not to create negative feelings regarding the self-esteem, self-image and self-confidence of children for their abilities and school performance. Proper intervention comes from a valid assessment of students' dysfunctions and weaknesses and is a multifactorial process, which requires adequate knowledge of the child's weaknesses, strengths and personality, cooperation with parents, study of social, family and cultural environment of the child, as well as many other important information (Porpodas, 2002). Assessment in terms of cognitive objects should be based on phonological awareness, short-term memory, decoding and finally, comprehension of the reported text (Kokkinaki 2014).

Therefore, a necessary condition for an effective didactic intervention is the correct diagnosis. Specifically, for reading ability, starting with a well-targeted assessment based on the difficulties we detect in the child, we give a specialized approach to the teaching of reading. In order to make this assessment, we must take as a guide the level of reading ability that the student already possesses. The correct evaluation in the kindergarten and in the first grades of elementary school leads to a timely intervention and prevention of the student's difficulties [19] (Tzivinikou, S., 2015).

Today, we are given the opportunity, with the use of appropriate tools, to understand the learning difficulties that a child faces from an early age. Some of the most common screening tests that are mainly related to reading ability are:

The Predictive Assessment of Reading (PAR). It is considered one of the most reliable and valid tests for kindergarten children, through which we have the ability to predict the reading ability of children up to high school.

The Texas Primary Reading Inventory (K-2). It focuses on children from pre-school to the third grade and has the ability to recognize and evaluate the developmental stages of their reading [21].

The Dynamic Indicators of Basic Early Literacy Skills (DIBELS). It is a test that deals with the processes of phonemic awareness, the alphabetic principle and phonological awareness, the ease and accuracy of reading a text, the development of vocabulary and the process of comprehension.

Finally, the AIMS-Web test, which enables the use of RtI programs and multilevel teaching in schools. It refers to children from kindergarten to high school and gives a basis for the detailed curriculum in mathematics and reading [21] (Tzivinikou, S., 2015)

So, after completing our diagnosis and evaluating the capabilities and needs of the student, we turn to the right teaching intervention. In order to eliminate the differences between the students, we focus on adapting the curriculum to the needs of the student and not the other way around. Thus, the goals we have set in a classroom do not change according to the abilities of each student, but the way they are approached and their degree of difficulty differ (Skidmore 2004). The ultimate goal is to develop learning strategies and use them within the classroom. Strategies such as group research or peer-to-peer teaching can be particularly useful, not only for students with difficulties but also for all students in the class (Tzouriadou, 2011). By learning to use cognitive and metacognitive strategies, students have the opportunity to process and use the information they will receive, to think and perform a task, and to evaluate their performance in it [22] (Luke 2006).

Students with reading difficulties can improve their reading skills through various educational approaches applied during teaching. Indicatively, we can refer to direct teaching, the formation of small groups that enhance discussion and support, vocal thinking, etc. (Tzivinikou, 2015). Also, strategies that have to do with reading are:

The Reading Analysis, Merge and Decoding strategy. It refers to students with learning difficulties aged 7 to 12 years and has to do mainly with the connection of sounds with voices.

Auditory Discrimination in Depth. It emphasizes the posture of the mouth and they learn the feeling that each sound gives at the time of its pronunciation. Thus, students analyze the words and recognize the sounds according to the placement of the tongue and mouth.

Analysis for Decoding Only. A strategy that is implemented in order to teach students to analyze letter patterns in small words that they often come across. For example, the word "beyond" combined with the words "day, wedding ring, good morning".

The Read-By-Ratio Approach. This strategy is based on phonemes aimed at word recognition. Through the same spelling patterns of words, students are taught how to analyze and decode unknown words (Tzivinikou, 2015).

CONCLUSIONS

In recent years, there has been a rapid increase in learning disabilities. The percentage of children diagnosed with learning disabilities, which can cause serious developmental problems within and outside the school context, is increasing. Children who have been diagnosed with learning disabilities appear to have serious difficulty adapting to the classroom and thus lagging behind in their academic performance. In the past there was a perception that these children were "lazy", "bad students", "stupid", but now we know that this perception is wrong, since we are not talking about bad or lazy students, but about students who due to a nervous disorder their system, do not have the same capabilities as the rest. Learning difficulties can also cause emotional problems in children as they feel that they are lagging behind compared to the rest of the class. Our goal is to include children with learning disabilities in the classroom by adapting the lesson to the children and not the children in it. Our main concern should be the valid and timely diagnosis of difficulties and effective intervention to address them.

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Forest Biodiversity Degradation: Assessment of Deforestation in Ohaji Egbema Forest Reserve, Imo State, Nigeria Using GIS Approach by Egbuche Christian Toochi in Open Access Journal of Biogeneric Science and Research

ABSTRACT

This research is focused on a spatial analysis of a reserved forest deforestation over a period of time using a GIS approach in Ohaji Egbema Local Government Area Imo state, Nigeria. It aimed at assessing and analyze deforestation in Ohaji Egbema forest reserve and examined the possible effects of deforestation on the forest environment. The assessment concentrated on when and where have forestlands changed in the reserved forest programmed within the period of 1984 - 2040 forecast. The key objectives were to assess the impact of land use and land cover changes on forest cover for the past 36 years, while sub objectives were dedicated to achieve in mapping out different land cover in Ohaji Egbema forest reserve, to assess land cover changes in the forest reserve susceptible to long term degradation from 1984 to 2020 of about 20 years. To evaluate forest loss in the area for the past 36years, and to predict the state of the land cover (forest) for the next 20 years (2040). Primary and secondary data employed using (200 ground truth points) were systematically collected from four different LULC classes in the study area using geographical positioning system (GPS), the secondary data (Satellite Landsat Imageries of 1984, 2002 and 2020) of the study area was acquired. The imageries were processed, enhanced and classified into four LULC classes using supervised classification in Idrisi and ArcGis software Ground truth points were utilized to assess the accuracy of the classifications. The data collected was analyzed in tables and figures and represented with a bar chart and pie chart graphs. Results showed that forest land, built up, grassland and water body were the four LULC classified in the study area. Kappa coefficient values of 91%, 85% and 92% for 1984, 2002 and 2020 respectively shows the accuracy of the classifications. Classifying the land uses into built-up and forest lands revealed that the built-up lands constantly rose while the forest lands kept dropping. The built-up lands increased by 49.30% between 1984 and 2000, 50.00% between 2002 and 2020 and 28.40% between 2020 and 2040 at the expense of the forest portion of the area which fell by 33.88% between 1984 and 2000,46.45% between 2002 and 2020, and 49.22% between 2020 and 2040. Increase in population, per capita income, and land use activities and by extension urban expansion were found to be the major factors causing deforestation in the forest reserve, it is likely that in the nearest future the remaining forest lands would be gradually wiped out and consequently the environmental crisis would be aggravated. Based on the findings of the study, there is need to urgently limit and control the high rate of deforestation going on in Ohaji Egbema forest reserve and embark on tree replanting campaigns without delay. There is need and recommended that a higher quality satellite imagery that offers up to 4m resolution should be used and a forest relic analysis should be conducted.

KEYWORDS: Biodiversity; Forest degradation; GIS; Forest Reserve; LULC; Deforestation and Satellite Imagery

Introduction

Deforestation constitutes one of the serious threats to forest biodiversity and pose a global development challenges of long-term environmental problem at both regional level and the world at large. According to [1] and [2] the degradation of the forest ecosystem has obvious ecological effects on the immediate environment and forested areas. Deforestation can result in erosion which in turn may lead to desertification. The economic and human consequences of deforestation include loss of potential wood used as fuel wood for cooking and heating among others. The transformation of forested lands by human actions represents one of the great forces in global environmental change and considered as one of the great drivers of biodiversity loss. Forests are cleared, degraded and fragmented by timber harvest, conversion to agriculture, road-construction, human-caused fire, and in myriad of other ways of degradation. According to [5], deforestation refers to the removal of trees from afforested site and the conversion of land to another use, most often agriculture. There is growing concern over shrinking areas of forests in the recent time [7]. The livelihoods of over two hundred million forest dwellers and poor settlers depend directly on food, fibre, fodder, fuel and other resources taken from the forest or produced on recently cleared forest soils. Furthermore, deforestation has become an issue of global environmental concern, in particular because of the value of forests in biodiversity conservation and in limiting the greenhouse effect [8]. Globally, deforestation by this trend has been described as the major problem facing the forest ecosystem. The extent of deforestation in any particular location or region can be viewed from economic, ecological and human consequences as well as scramble for land. Forest degradation may in many ways be irreversible, because of the extensive nature of forest degradation which the impact of activities altering their condition may not be immediately apparent and as a result they are largely ignored by those who cause them. Forest is often perceived as a stock resource and always and freely available for conversion to other uses without considering the consequences for the production services and environmental roles of the forest. As environmental degradation and its consequences becomes a global issue, the world is faced with the danger that the renewable forest resources may be exhausted and that man stands the risk of destroying his environment if all the impacts of deforestation are allowed to go unchecked. It becomes therefore important to evaluate the level of deforestation and degradation in Ohaji Egbema forest reserve using a GIS application. The effect of deforestation and degradation of the only forest reserve in South east Nigeria has recently become a serious problem. It has been identified that in the area is mostly the quest for fuel wood, grazing and for agricultural use. One of the effects of deforestation is global warming which occurs as a result of deforestation as trees uses carbon dioxide during photosynthesis. Deforestation leads to the increase of carbon dioxide in the environment which traps heat in the atmosphere leading to global warming. I become very objective to assess the impact of land use and land cover changes on forest cover for the past 36 years with further interest to map out different land cover in the Ohaji Egbema forest reserve, assess land cover changes from 1984 to 2020 at 20 years, evaluate forest loss in the area for the past 36 years and make a prediction of the state of the land cover (forest) for the next 20 years (2040). It is known that deforestation and degradation of the forest has posed a serious problem especially at this era of global climatic change.

United Nations Food and Agricultural Organization [12], deforestation can be defined as the permanent destruction of forests in other to make the land available for other uses. Deforestation is said to be taking place when forest is being cut down on a massive scale without making proportionate effort at replanting. Also, deforestation is the conversion of forest to an alternative permanent non-forested land use Such as agriculture, grazing or urban development [5]. Deforestation is primarily a concern for the developing countries of the tropics [6] as it is shrinking areas of the tropical forests [3] causing loss of Biodiversity and enhancing the greenhouse effect [8]. Forest degradation occurs when the ecosystem functions of the forest are degraded but where the area remains forested rather cleared [9]. Available literatures shows that the causes of forest deforestation and degradation are caused by expansion of farming land, logging and fuel wood, overgrazing, fire/fire outbreak, release of greenhouse gases and urbanization/industrialization as well as infrastructural provisions. More so agents of deforestation in agricultural terms include those of slash and burn farmers, commercial farmers, ranchers, loggers, firewood collectors etc. Generally, the center of biodiversity and conservation (CBC 1998) established the remote sensing and geographic information system (RS/GIS) facilities as technologies that will help to identify potential survey sites, analyze deforestation rates in focal study areas, incorporate spatial and non-spatial databases and create persuasive visual aids to enhance reports and proposals. Change detection is the process of identifying differences in the state of an object or phenomenon by observing it at different times [11]. Change detection is an important process in monitoring and managing natural resources and urban development because it provides quantitative analysis of the spatial distribution of the population of interest.

Study Area

Ohaji Egbema lies in the southwestern part of Imo state and shares common boundaries with Owerri to the east, Oguta to the north Andogba/Egbema/Ndoni in Rivers state in the southwest. The 2006 census estimated the study area to over 182,500 inhabitants but recently due to industrialization and urbanization, Ohaji/Egbema has witnessed a great deal of population influx. The study area lies within latitudes 50 11’N and 50 35’N and longitudes 6037’ ad 6057’. It covers an area of about 890km2.

The study area is largely drained by the Otammiri River and other Imo river tributaries. The study area belongs to a major physiographic region- the undulating lowland plain which bears a relationship with its geology. The low land areas are largely underlain by the younger and loosely consolidated Benin formation [12]. The vegetation and climate of the study area has been delineated to have 2 distinct seasons both of which are warm, these are the dry and rainy season.

Climate and Vegetation

The dry season occurs between November and March, while the rainy season occurs between April and October. The high temperatures, humidity and precipitation of the area favour quick plant growth and hence vegetation cover of the area that is characterized by trees and shrubs of the rainforest belt of Nigeria. Geology and Soil

The study area is located in the Eastern Niger delta sedimentary basin, characterized by the three lithostratigrapgic units in the Niger delta. These units are – Akata, Agbada and Benin formation in order of decrease in age [13]. The overall thickness of the tertiary sediments is about 10,000 meters.

Method of Data Collection

Data are based on field observation and from monitoring the real situation, they are collected as fact or evidence that may be processed to give them meaning and turn them into information in line with [14]. Heywood (1988). Geographical Positioning System (GPS) was used to collect fifty (50) coordinate points at each land use land cover, totaling 200 points for the four major land use and land cover identified in the study area. Landsat Imageries of one season (path 188, 189 and row 56) were acquired from United State Geological Survey (USGS) in time series; 1984 Thematic Mapper (Tm), 2002 Enhanced Thematic Mapper (ETM) and 2020 Operational Land Imager (OLI) as shown in the table one below table 3.

Data Analysis and Data Processing

The acquired landsat imageries were pre-processed for geometrical corrections, stripes and cloud removal. Image enhancement was carried out on the acquired imageries employing bands 4, 3, 2 for LANDSAT TM and ETM while bands 5, 4, 3 for LANDSAT OLI/TIRS to get false colour composite using Idrisi and Arcgis softwares. In the resultant false colour composite, built up areas appear in cyan blue, vegetation in shades of red differentiating dense forest and grass or farm lands, water bodies from dark blue to black, bare lands from white to brown [15]. This was necessary to enhance visualization and interpretability of the scenes for classification. The study area was clipped out using administrative map of Nigeria containing Imo State and Local Government shape files in Arc map (Table1 and table 2).

Land Use Land Cover Classification

The false Colour composite images were subjected to supervised classification which was based on ground-based information. Maximum likelihood was adopted to define areas of Landsat images that represented thematic classes as determined by maximal spectral heterogeneity according to [16]. Maximum likelihood algorithm considers the average characteristics of the spectral signature of each category and the covariance among all categories, thus allowing for precise discrimination of categories. Hence the land covers were classified into four major land use land cover classes: Built up, forest cover, grass cover and water body. Forest vegetation are the areas dominated by trees and shrubs; grass land are the areas dominated by grasses, including farm lands and gardens; water body are the areas occupied by streams, rivers, inland waters; while built-up areas are the areas occupied by built structures including residential, commercial, schools, churches, tarred roads and those land surface features devoid of any type of vegetation cover or structures including rocks. Four applications (ArcGis 10.5, Idris software, Excel and Microsoft word) were also applied in this study.

Accuracy Assessment of The Classification

The aim of accuracy assessment is to quantitatively assess how effectively the pixels were sampled into the correct land cover classes. Confusion matrix was used for accuracy assessment of the classification procedure in accordance with the training samples and the ground truth points as a reference point. This approach has also been adopted effectively in similar studies by [17]; [18]. The accuracy assessments of the classified maps for 1984, 2002 and 2020 were evaluated using the base error matrix. The base error matrix evaluates accuracy using parameters such as agreement/accuracy, overall accuracy, commission error, omission error and the Kappa coefficient. The agreement/accuracy is the probability (%) that the classifier has labeled an image pixel into the ground truth class. It is the probability of a reference pixel being correctly classified. The overall accuracy specifies the total correctly classified pixels and is determined by dividing the total number of correctly classified pixels by the total number of pixels in the error matrix. Commission error represent pixels that belong to another class but are labeled as belonging to the class; while the Omission error represent pixels that belong to the truth class but fail to be classified into the proper class. Finally, the Kappa coefficient (Khat) measures the agreement between classification map and reference data, as expressed below:

kappa coefficient t= (Observed Accuracy-Chance agreement)/ (1-Chance agreement)

It is stated that Kappa values of more than 0.80 (80%) indicate good classification performance. Kappa values between 0.40 and 0.80 indicate moderate classification performance and Kappa values of less than 0.40 (40%) indicate poor classification performance [19].

Change Detection Analysis

Spatio-temporal changes in the four classified land use classes for the past 36 years were analyzed through comparison of area coverage of the classified maps. Change detection was carried out in each of the classes to ascertain the changes over time in terms of area and percentage coverage according to [18]. This was done by computing the area coverage for each of the feature class in each epoch from the classified images in idrisi and Arcmap softwares following the expression below:

Area (m2) = (Cell Size x Count)/10,000

Percentage cover (%) = Area/ (Total) x100

Where cell size and count were gotten from the properties of the raster attributes.

The extent of land use land cover over change, land use encroachment as well as gains and losses experienced within the study period were analyzed and presented in maps and charts.

Prediction Analysis

The classified land use imageries were subjected for Land Change Modeling in idrisi software using Cellular Automata and Markov Chain algorithm for prediction. Then land cover scenario under prevailing conditions for the year 2040 was modeled table 6.

RESULTS AND DISCUSSION LAND COVER LAND USE CHANGES LASSIFICATION FROM 1984-2040

The result of the work is presented starting with the land use and land cover classification in the years 1984,2002,2020 and 2040 presented in Figures 4.1,4.2,4.3 and 4.4 below. Dark green colour represents forest vegetation, light green represents grass lands, blue represent water body, while orange colour represent built-up area. In figure 4.1 which is the LULC classification of 1984 shows that the study area is largely covered with deeply dark green which is forest vegetation, patches of scattered light green and orange colour which is grass land and built-up while the blue colour which is the water bodies covers a little part of the study area. This depict that the study area was more of forest vegetation in 1984 table 7.

In figure 4.2, it is observed that dark green colour is reduced, there was a slight increase in blue colour, and there was a slight reduction inlight green colour, while the orange colour was at an increase rate mainly at Obofia, Awarra, Amafor, Ohoba, Umukani, Ohaji Egbema forest reserve and Adapalm axis. This indicated that by the year 2002, reasonable forest lands were deforested and converted to residential, commercial, Agricultural and other land uses, and this could be attributed to infrastructural development, urbanization, industrialization and human population increase in the area which is the causes of deforestation table 4 and table 5.

In fig 4.3, deforestation continued. There were more of orange colours and light green colors were observed more at Umukani and Umuakpu axis in the map, while the dark green colour is gradually decreasing and it’s been observed that the blue colour was rarely seen in the map. These indicated that as the years passes by, there are more built ups, and grass lands while the forest land is gradually degraded and used for built-up and agricultural purposes.

In figure 4.4 below, the whole map is mostly covered with orange colour, with scattered patches of light green, and dark green colour being deforested, while the blue colour is hardly seen in the map. This shows that the forest land cover has been on the regular decrease, while built ups, grass lands have been on the regular increase.

Area coverage, percentage cover and change detection land use and land cover 1984 - 2040

The area coverage and percentage cover of different land use classes are represented below. It is observed that the forest land covers about 723.26km2 with a percentage cover of 81.31% which was the major land cover of the study area in 1984. This implies that more than half of the study area was under forest cover in 1984, In the areas of built-up, it covers about 128.40km2 in 1984 with a percentage cover of 14.43%. Areas covered by grass land (either by sparse vegetation, farmland or grasses) was at minimal in 1984 with an area cover of 32.57km2 and percentage cover of 3.66%, while the water bodies cover an area of 5.34km2 and a percentage cover of 0.6%.

Figure: 5. The land use land cover classification of 2040

Figure 4.7 and 4.8 below show the area coverage and percentage coverage of the year 2002. As time goes on, the forest land decrease from area coverage of 723.26km2 in 1984 to 699.68km2 in 2002, with a percentage cover of 81.31% in 1984 to 78.68% in 2002 which depict that the forest cover was at a loss while the built–up was at increase from 128.40km2 to162.46km2 and a percentage cover of 18.26%. Areas covered by grass land gradually decrease to 21.41km2 with a percentage cover of 2.41% in 2002, and a slight increase of the water bodies from an area cover of 5.34km2 to 5.80km2 with a percentage cover of 0.65%.

In the year 2020, it is shown that the forest land covers about 589.73km2 with a percentage cover of 66.30% which shows that there was a decrease within 2002 to 2020 while in the area of built up it increased to an area cover of 280.98km2 with a percentage cover 31.59%. An area covered by grass land covers about 15.53km2 with a percentage cover of 1.75% and the water bodies cover about 3.27km2 and a percentage cover of 0.37%. This shows that the built-up area which is at increase were initially forest lands and water bodies in the past years.

In 2040 it was predicted that the forest cover about 497.67km2 with a percentage cover of 55.95%, the built up was predicted to be on the increase with an area cover of 334.11km2 and a percentage cover of 37.56%. Area covered by grassland was at predicted to be on increase with an area cover of 55.92km2 and a percentage cover 6.29% and this grass land was formally forest land in the past years and this change occurred mainly at Ohoba, Awarra, Umuakpu, Umukani and Ohaji Egbema forest reserve the only forest reserve in the south east of Nigeria which has been deforested and use for agricultural purposes. The water bodies cover about 1.82km2 with a percentage cover of 0.20%. This implies that the forest land has been deforested and degraded to other land uses in the study area within the study periods. All this are shown in figure 4.11 and figure 4.12.

Change Detection Observed Between (1984-2040)

Approximately the change detected in the forest land from 1984 to 2002 in the area coverage is 23.41km2 with a percentage change of 33.88% which shows it was at decrease, the built up from 1984 to 2002 the change detected in area is -34.06km2 with a percentage change of 49.30% which is at increase while in the area of grassland, the change observed is 11.16km2 with a percentage change of 16.15%. The change detected in the area coverage of water bodies from 1984 to 2002 is -0.46km2 with a percentage coverage of 0.67% which was at increase.

The change detection observed in table 2 below shows that between 2002 to 2020 the forest cover was at a high decrease with about 110.12km2 of change observed and a percentage change of 46.45% and the built up was about -118.53km2 and a percentage change of 50.00% which shows there was an increase in the area. The change observed in the grass was at 5.88km2 with a percentage cover of 2.48% which is at a decrease while for the water bodies the change observed is 2.53km2 with a percentage change of 1.07% which implies that the area of built up has been on the high increase over other land classes.

In the 2020 to 2040 change detection table shows that there was more of built up in the study area which is observed to cover about -53.12km2 with a percentage change of 28.40% which shows that the built up was at increase, and the forest cover was at decrease with the change observed at 92.06km2 and a percentage change of 49.22%.Also the grassland was detected to be on a increased with about -40.39km2 with a percentage change of 21.60% and the water bodies was at a decrease with the change detected at the study period to be about 1.45km2 and a percentage change of 0.78% which implies that the water body has been lost to other land uses.

Land Use Land Cover Classification Accuracy

The result of the accuracy assessment for 1984, 2002 and 2020 were presented in table 4 to table 6below. The overall accuracy for 1984 classification was 0.94%, with overall kappa accuracy of 91%, in 2002 classification, overall accuracy 0.89%, kappa accuracy was 85%, in 2020 classification, overall accuracy was 0.94%, kappa accuracy was 92%.

Discussion

Findings from the study showed that land cover of the study area has been heavily deforested and degraded within the study period 1984, 2002 and 2020, which will continue if control measures are not taken into consideration. Forest land was on the decrease while built ups and grass lands were on the increase. This is in line with other findings of [16] and [23] and [30]. These outrageous changes in the origin all and cover in the study area could be linked to human population, unsustainable human activities in the study area as well as unsustainable environmental management practices and weak environmental policies. As the human population increases, more lands were needed for settlements and many other commercial activities, which gradually led to rapid industrialization, infrastructural development and urbanization. Increase in human population could also increase the levels of anthropogenic activities such as deforestation, intensive farming and sand mining. In other words, the large spread of forest land in 1984 could be linked to low population and productivity, less socio-economic activity. The forest lands have been drastically reduced to build-ups and other land uses in the study area, without consideration to the many environmental needs that forest provides. Hence loss of biodiversity, land degradation, noise pollution, air pollution and climate change could be rooted to changes in the land cover. It can observe that in the past two centuries the impact of human activities on the land has grown enormously, altering entire landscapes, and ultimately impacting the earth's nutrient and hydrological cycles as well as climate. The classification accuracy for the 3 years represents strong agreement. According to [31] values between 0.4 and 0.8 represent moderate agreement, values below 0.4 represent poor agreement and values above 0.81 represent strong agreement.

CONCLUSION AND RECOMMENDATION

In this study, four land use land cover classes were identified as they change through time. However, the result shows a rapid change in the vegetation cover of the study area between 1984 to 2040. Within this period, 225.59km2 of forest land areas and 3.52km2 of water body were lost and converted to other land uses in the study area. Whereas built up and grassland was at increase covering part of the forest and water body. However, if these patterns of degradation continue in the study area, it is likely that in the nearest future the remaining forest land would be wiped out and environmental crisis would be aggravated. Therefore, the assessment of the level of deforestation in Ohaji Egbema using GIS is thus a vital tool for sustainability of the forest management and environmental planning of the area especially at the only forest reserve in the South east, Nigeria.

Based on the findings, there is need to urgently limit and control the high rate of deforestation going on in Ohaji Egbema and embark on tree planting campaigns without delay. It is also recommended that an Environmental Impact statement (EIS) should be carried out. Furthermore, policy makers should ensure that the existing/future polices with regard to environmental and forest degradation is utmost implemented. There is need to create an awareness programme for all stakeholders on the issues at hand and the need to adopt sustainable use of natural resources, sustainable living habits and minimizing impact on the environmental. Finally, [2] having conducted species relics in this forest reserve further research should be conducted on higher quality satellite imagery that offers up to 4m resolution within as well as forest relic analysis.

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Wilms’s Tumor Gene Mutations: Loss of Tumor Suppresser Function: A Bioinformatics Study by Uzma Jabbar in Open Access Journal of Biogeneric Science and Research

ABSTRACT

Introduction: Mutation in the Wilms’s Tumor (WT1) gene product has been detected in both sporadic and familial cases suggesting that alteration in WT1 may disrupt its normal function. The study aims to find the protagonist amino acid in WT1 proteins by mutating these residues with other amino acids.

Material and Methods: The 3D modeling approach by MODELLER 4 was utilized to build a homology of WT1 proteins. Quality of the WT1 model was verified by predicting 10 models of WT1 and hence selecting the best one. Stereochemistry of model was evaluated by PROCHECK. Mutational studies were done by WHAT IF. Five human WT1 mutations were modeled which were Lys371→Ala371, Ser415→Ala415, Cys416→Ala416, His434→Asp434 and His434→Arg434.

Result: Based on active side of WT1 protein and its role in DNA binding mutation. No significant change was observed when Lys371 was mutated to Ala371, Ser415 was mutated to Ala415. Significant change was observed in Cys416 mutated to Ala416. In mutant Ala416, loss of coordination with the metal ion Zn was also predicted. In case of Mutants His434→Asp434, there was a loss of coordination of metal ion (Zn203) with mutant Asp434. In case of mutant His434→Arg434, there was a loss of Zn203 coordination with Arg434. His434 does not interact directly with any DNA base, whereas mutated Arg434 is predicted to interact directly with DNA base.

Conclusion: It is concluded that mutation of amino acid residue Cys416→Ala416, His434→Asp434 and His434→Arg434 may lose the proto-oncogenic function of WT1.

Keywords: WT1 protein, MODELLER9.0, Mutation, Active side residues

Introduction

WT1, is a protein, which in humans is encoded by the WT1 gene on chromosome 11p13. The WT1 is responsible for the normal kidney development.  Mutations in this gene are reported to develop tumors and developmental abnormalities in the genitourinary system. Conversion of proto-oncogenic function to oncogenic in WT1 has also been documented cause of various hematological malignancies. (***)

Multifaceted protein of WT1 gene has transcriptional factor activity [1]. It regulates the expression of insulin-like growth factor and transforming growth factor system, implicated in breast tumorigenesis [2]. A main function of WT1 is to regulate transcription, which control the expression of genes involved in the process of proliferation and differentiation [3]. In wide range of tumor, WT1 is shown to be predisposing factor for cancer, therefore it has become hot target in research to find out it’s inhibitor which can be safely used as a treatment of cancer. It can induce apoptosis in embryonic cancer cell, presumably through the withdrawal of required growth factor survival signal [4]. WT1 is involved in the normal tissue homeostasis and as an oncogene in solid tumors, like breast cancer [5]. Increased expression of WT1 is related with poor prognosis in breast cancer6. A number of hypotheses are postulated for the relationship of WT1 with tumorigenesis. Acceding to one of the hypothesis, elevated levels of WT1 in tumors may be related with increased proliferation because normally WT1 have a role with apoptosis [7,8]. Another study proposed that WT1 can alter many genes of the the family of BCL2 [9,10] and also have a role to regulate with Fas-death signaling pathway [11]. Furthermore, it is suggested that WT1 can encourage cell proliferation by up-regulation of protein cyclin D1 [12].

A group of workers hypothesized that WT1 has been observed in the vasculature of some tumour types [13]and its expression may be related with angiogenesis especially in endometrial cancer [14]. Another hypothesis based on the fact that WT1 is a main regulator of the epithelial/mesenchymal balance and may have a role in the epithelial-to-mesenchymal transition of tumor cells [3]. Expression of WT1 is higher in estrogen receptor (ER) positive than in ER negative tumors. It is therefore possible that WT1 not only interact with ER alpha, but it may orchestrate its expression [15]. A study, on triple negative breast cancers [7] has shown that high WT1 levels associate with poor survival due to increased angiogenesis [16,17], altered proliferation/apoptosis10,11, and induction of cancer- epithelial-to-mesenchymal transition4. In breast tumors, WT1 is mainly related with a mesenchymal phenotype and increased levels of CYP3A4 [18]. A mutation in the zinc finger region of WT1 protein has been identified in the patients that abolished its DNA binding activity [19]. A study also observed that the mutation in the WT1 gene product has been detected in both sporadic and familial cases suggesting that alteration in WT1 may disrupt its normal function [20].  Bioinformatics approaches are being utilized to resolve the biological problems. Efforts start with the prediction of 3D structures. To achieve the aim, study was designed to view 3D structure of WT1, tumor suppressor protein predicted by homology modeling and to study the role of crucial residues in WT1 proteins by mutating these residues with other amino acids.

Material and Methods

3D structure of WT1 was taken as target of human WT1. Figure 1 shows the normal interaction of WT1 with DNA strands based on the crystal structure of a zinc finger protein.

The 449 amino acid sequences of WT1 were used for homology modeling. Sequences of WT1 were retrieved from Swiss Prot Data Bank in FASTA format [21]. The best suitable templates were used for 3D-structure prediction. The retrieved amino acid sequences of WT1 were subjected to BLAST [22]. Templates were retrieved on the base of query coverage and identity. The 3D structures were predicted by MODELLER 9.0 [23] that is the requirement of 3D structure building of target protein. Tools including stereochemistry and Ramchandran plots were used for the structure evaluation [24]. Identification of Template was carried out, and Sequence Alignment was carried out by using FASTA, BLAST. Quality of the WT1 model was verified. Stereochemistry of model was evaluated by PROCHECK [25]. Mutational studies were done by WHAT IF [26]. Five human WT1 mutants are modeled. These were: Lys371→Ala371, Ser415→Ala415, Cys416→Ala416, His434→Asp434 and His434→Arg434.

Results and Discussion

The study was largely based on active side of the WT1 and its role in DNA binding mutation. Zinc finger binding domain interact selectively and non-covalently. This zinc finger-binding domain is the classical zinc finger domain, in which two conserved cysteine and histidine co-ordinate a zinc ion at the active site.

Cys416®Ala416 MUTANT

Significant change was observed in Cys416 mutated to Ala416. In mutant Ala416, reduction in the Van der Waal’s contact between the amino acids. Loss of coordination with the metal ion Zn was also predicted (Figure 6 A and B).

Figure 6 A and B: Wild Type (Cys416) and mutated (Ala416) WT1.  Distance between Zn and Cys416 is increased in mutated (Ala416) model. Cys416 is predicted to be found in the vicinity of His434 and His438 which are implicated in catalysis (6A) while Ala416 can only interact with His434 and not with His438 in the mutated model (6B).

Cys416 is located at the domain interface with its polar side chain completely buried (0.00 Å). Replacement of this amino acid may account for considerable changes in the interior of protein (Table 1). We have predicted the possible changes that arise due to the mutation of Cys to Ala by molecular modeling experiments. Amino acids, Pro419, Ser420, Cys421, His434 and some atoms of His438 (ND1, NE2, CD2 and CE1) are present near Cys416. Zinc (Zn203) is also present in the vicinity (1.82 Å) of Cys416 (Figure 6). The mutated residue, Ala is also predicted to remain buried (0.00 Å) in the interior of protein. Significant change is observed however, in the surrounding area of the mutated Ala416. Only a few atoms of His434 (CD2 and NE2) and His438 (CE1) were seen in the surrounding. This may reduce the Van der Waal's contacts between the respective amino acids. The loss of coordination with the metal ion, zinc was also predicted as the distance is increased from 1.82 Å to3.12 Å.   It is therefore predicted that Cys416 plays a vital role in the interaction with other amino acid residues as well as in the metal coordination. It is observed that there is a possibility of loss of these interactions in case of Cys416 replacement.

His434®Arg434 MUTANTS

In case of mutant His434→Arg434, there was a loss of zn203 coordination with Arg434.  His434 does not interact directly with any DNA base, whereas mutated Arg434 is predicted to interact directly with DNA base, A1. This suggests that change might effect on the DNA binding pattern, Figure 7 A and B.

FIGURE 7 A and B: Wild Type (His434) and mutated (Arg434) WT1.  Distance between Zn and His434 is increased in mutated model. Arg434 is predicted to bind DNA base A1 (B) while His434 in the original model (A) show no bonding with DNA base.

In case of mutation of His434®Arg434, the distance between the mutated Arg and zinc (Zn203) was increased from 2.28 Å to5.00 Å suggesting that there could be a loss of coordination with the metal ion. Mutational studies proved that hydrogen bonding network close to the zinc-binding motif plays a significant role in stabilizing the coordination of the zinc metal ion to the protein23. The mutated amino acid, Arg434 also moved considerably form buried to relatively exposed environment (2.28 Å to 5.35 Å). Presence of positively charged Arg on the surface could account for additional interaction of the protein with other proteins or with the surrounding water molecules. His434 does not interact directly with any DNA base whereas mutated Arg434 is predicted to interact directly with DNA base Adenine, A1. (Figure 7). This suggests that the change might cause the DNA binding pattern.

Lys371®Ala371 and Ser415®Ala415 MUTANT

No significant change was observed when Lys371 was mutated to Ala371, and Ser415 was mutated to Ala415. It is observed in this mutation that the change that arise in the overall structure and surrounding amino acid residues (Table 1). Lys371 is present on the surface (accessibility = 47.04 Å) of the WT1 molecule. It was observed that the internal protein structure was not affected considerably, as Lys371 is present on the outer most surface of the protein. In the original model, Lys371 stacks against thymine. It also forms a water-mediated contact with side chain hydroxyl of Ser367. Although, Ala371 also stacks against the same DNA base but the distance is slightly altered. The hydrogen bond between Ala371 and Ser367 has not been predicted in the mutated model.  It has been demonstrated that mutation within finger 2 and 4 abolished sequence specific binding of WT1 to DNA bases19. The mutation of the corresponding lysine in a peptide could reduce its affinity for DNA seven folds [27].  On the other hand, it is reported [28] that a surface mutation would not cause a significant change in the internal structure of protein.  However, the replacement of a basic polar residue with a non-polar one could account for a reduction in polarity.  The modeling studies of Lys to Ala mutation do not however support this finding and require further analysis.

Mutation of Ser415®Ala415 in the WT1 model (Table 1). Ser415 is located near the active center of WT1. It has been demonstrated that Ser415 makes a water-mediated contact with phosphate of DNA base, guanine [20]. In our predicted model of WT1, Ser415 makes two water (numbers 516 and 568) mediated contacts. Mutation of this Ser with Ala resulted in the loss of one of these contacts leading to the loss of binding. The replacement of relatively polar residue, Ser to a non-polar one, Ala could account for this reduced interaction. This is also evident by a slight decrease in the accessibility of Ala (Ser415 = 7.96 Å; Ala415 = 7.61 Å).

His434®Asp434 MUTANTS

In case of mutants His434→Asp434, there was a loss of coordination of metal ion (zn203) with mutant Asp434. Glu430 move from relatively exposed to completely buried environment. His434 is also present at the active center of WT1. We predicted two mutants; His434®Asp434 and His434®Arg434 mutants by molecular modeling (Table 1). In case of His434®Asp434 mutation, the water mediated contact is lost. The distance between mutated Asp and zinc (Zn203) was also increased from 2.28 Å to 3.57 Å suggesting that there could be a loss of coordination with the metal ion as well. The amino acid Glu340 that is present near His434 also moved considerably form relatively exposed to completely buried environment (14.83 Å to 00.0 Å).

Conclusion

It is concluded that mutation of amino acid residue Cys416→Ala416, His434→Asp434 and His434→Arg434 of WT1 may lose its function to regulate the function of genes by binding to specific parts of DNA. Besides the mutation of above-mentioned amino acid residue, the role of WT1 in cell growth, cell differentiation, apoptosis and tumor suppressor function is also lost.

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Gene Editing via Integrase Enzyme by Umair Masood* in Open Access Journal of Biogeneric Science and Research

Short Communication

Targeted integrase enzyme is a most powerful tool for mediating genome alteration with high precision. The gene of the interest is directly catalyze nucleophilic attack of the 3 prime hydroxyl group at the end of processed DNA on a pair of phosphodiaster bond in the targeted DNA or genome of the interest. Integrase gene editing method contain two parts gene of the interest with integrase enzyme and desire genome. The integrase enzyme is start integration the gene of the interest into the desire genome or DNA.

Results via Gel electrophoresis

In order to check that whether gene of interest is integrated or not we can perform a gel electrophoresis. The gel contain a two band A and B.A band is a vector of human insulin vector which can show the negative control and B band is a vector of human insulin but the gene have some sequence mutation [1-5]. We can add a gene of the stranded human insulin by using integrase enzyme and gene can be precisely add to the B band that why B band is in 3.0 and A band is 6.0 which mean that A band is high molecular weight than the B band and a Marker must be 1kb and agarose should be 1% [6-9].

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Giant Cell Tumor of the Distal Tibia and Fibula (Rare Location) by Mohamed Hamid Awadelseid* in Open Access Journal of Biogeneric Science and Research

ABSTRACT

Giant cell tumor of the distal tibia is a rare, benign and usually asymptomatic condition. The discovery is sometimes made following a medical imaging examination or a painful symptom or more often a visible or palpable swelling with or without vascular and/or nerve compression. At an advanced stage, the X-ray is of paramount importance. The well complete surgical resection is part of the therapeutic. We present a clinical case report of a young man with a giant cell tumor localized in the distal tibia in Khartoum, Sudan. This case concerns a 37-year-old patient who presented in July 2021 of a huge painful swelling at left distal tibia treated with bonesetter at Kassla, eastern Sudan and whose X-ray radiography showed lytic lesion of the cortical bone in the lower third of the tibia. After the operative resection of the tumor mass, the pathological examination of the operative specimen revealed the diagnosis of a giant cell tumor. A giant cell tumor is a benign condition, with a few symptoms and the location at the ankle is exceptional. Complete surgical resection is a viable treatment option.

Keywords: Giant Cell Tumor, Wide Surgical Resection, tibia

Introduction

Giant cell tumor (GCT) of bone is one of the commonest benign bone tumors encountered by an orthopedic surgeon. The reported incidence of GCT in the Oriental and Asian population is higher than that in the Caucasian population and may account for 20% of all skeletal neoplasms. It has a well-known propensity for local recurrence after surgical treatment Although considered to be benign tumors of bone [1].

GCT has a relatively high recurrence rate. Metastases occur in 1% to 9% of patients with GCT and some earlier studies have correlated the incidence of metastases with aggressive growth and local recurrence. Current recurrence rates between 10-20% with meticulous curettage and extension of tumor removal using mechanized burrs and adjuvant therapy are a vast improvement on the historically reported recurrence rates of 50-60% with curettage alone. GCT of bone constitutes 20% of biopsy analyzed benign bone tumors. It affects young adults between the ages of20 and 40 years, several authors have reported a slight predominance of women over men. However, GCT can be seen in patients over 50 years old. Ninety percent of GCT exhibits the typical epiphyseal location. Tumor often extends to the articular subchondral bone or even abuts the cartilage. The joint and or its capsule are rarely invaded. In rare instances in which GCT occurs in a skeletally immature patient, the lesion is likely to be found in the metaphysis,The most common locations, in decreasing order are the distal femur, the proximal tibia, the distal radius, and the  sacrum .Fifty percent of GCTs arise around the knee region. Other frequent sites include the fibular head, the proximal femur, and the proximal humerus. Pelvic GCT is rare[6]. Multicentricity or the synchronous occurrence of GCT in different sites is known to occur, but is exceedingly rare [2].

We report a case of a 37-year-old man who presented in July 2021, with a huge painful swelling his left distal tibia with an X-ray radiography showing lysis of the cortical bone in the lower third of the tibia. After the operative excision of the tumor mass, the pathological examination of the specimen revealed the diagnosis of a giant cell tumor. With a lytic lesion of the distal tibia and fibula bones in a young man on X-ray, one must think of a giant cell tumor [3].

Case Report

We report the case of a 37-year-old young man with no notable pathological antecedents who presented at the orthopedic consultation for a painful swelling of the left distal tibia and fibula, that had been evolving for 5 months and without any alteration of the general state. There was change in color and consistency of the skin with respect to the tumor action.

On physical examination, there was a dorsoflextion blockage of foot because of the large volume occupied by the tumor mass and the articular destruction at the level of the distal tibio-fibular joint; plantar flexion estimated at 10˚ and dorsal flextion at 3˚; inversion at 5˚ and eversion at 2˚. The x-ray showed a lesion with blurred boundaries, extending into the soft tissue that is not limited by a bony shell; with destruction of the cortex, invasion of soft parts and honeycomb pseudo-partitions (Figure 1-2). And finally, the Magntic Resonant Imaging (MRI) of the left leg showed (Figure 3-4-5-6) a lysis of the cortex of the lower extremity of the leg. It corresponds to grade 3 of the Campanacci and Merled ’Aubignee classification.

A complete surgical resection below knee amputation (BKA) was offered to the patient. Under spinal anesthesia, this incision was made proximally to expose a healthy portion of the leg bone. Surgical removal of the tumor by BKA a proximal resection of the tibia and fibular bone by about 2 cm in the healthy zone. The anatomopathological assessment (Figure7) showed abundant mononuclear cells and discrete nuclear anomalies with marked mitotic activity, but without atypical forms. The histological examination of the bone fragments confirmed a grade 3 giant cell tumor according to Sanerkin, Jaffe Lichtenstein and Pottis. CT scan was done to exclude pulmonary metastases (Figure 8). At three weeks removal of suture and start physiotherapy of knee. Surgical treatment with the excision of the large tumor mass by BKA improved the function of the leg and general condition of the patient.

Discussion

Giant cell tumors (GCT) account for 5%–9% of all benign and malignant bone tumors. They are considered benign but may present a progressive, potentially malignant clinical course. GCT recur in a high percentage of cases, become sarcomatous, yet produce metastases even without apparent malignant changes [4].

In the literature, the recurrence rate varies considerably, depending not only on the site and extension of the lesion but also on the type of primary treatment performed [5]. Successful treatment of GCTs and the adequacy of tumor removal is influenced by tumor location ,associated fracture, soft tissue extension ,and understanding of the functional consequences of resection .each option has advantage and disadvantage [6]. This technique has the advantage of preserving knee joint function, improved general and psychological condition of the patient and recurrences are no more frequent than with other techniques according to several authors.  Excision with tumor free margins is associated with lesser recurrence rates. However, for periarticular lesions this is usually accompanied with a suboptimal functional outcome [7]. Various studies suggest that wide resection is associated with a decreased risk of local recurrence when compared with intralesional curettage and may increase the recurrence free survival rate from 84% to 100% (8). However, wide resection is associated with higher rates of surgical complications which led to functional impairment, generally necessitating reconstruction.  This procedure resulted in good function preserving knee joint function after removal of suture 3 weeks later. After 5 months follow up, there was no recurrence or functional sequelae of the leg. Giant cell bone tumors generally have a good prognosis [9].

Conclusion

The giant cell tumor of the distal tibia and fibula bone, although rare, does not present any particularity. The gold standard X-ray image guided radiography with the bone tissue histology confirmed the diagnosis. use a CT scan or an MRI study if we fear an invasion of soft parts. Surgical treatment preserved joint function. This should be considered when presented with a lytic epiphyseal bone lesion.

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