#e-tivities

[damayalquicira]

Modelos y Diseños Instruccionales

Un modelo instruccional es un marco teórico que proporciona orientación y estructura para diseñar, desarrollar y ejecutar experiencias de enseñanza y aprendizaje efectivas. Estos modelos suelen incluir una serie de pasos o fases que guían el proceso de planificación y ejecución de la instrucción, desde la identificación de los objetivos de aprendizaje hasta la evaluación del desempeño de los estudiantes.

Los modelos instruccionales pueden variar en complejidad y enfoque, y pueden adaptarse a diferentes contextos educativos y necesidades de los estudiantes.

Modelo ADDIE: Este modelo, que significa Análisis, Diseño, Desarrollo, Implementación y Evaluación, es uno de los más utilizados en el diseño instruccional. Proporciona un enfoque sistemático y secuencial para el desarrollo de la instrucción, comenzando con un análisis de las necesidades de los estudiantes y terminando con la evaluación del diseño instruccional.

Modelo SAMR: Este modelo, que significa Sustitución, Ampliación, Modificación y Redefinición, se centra en la integración de la tecnología en la enseñanza y el aprendizaje. Proporciona un marco para evaluar el nivel de integración tecnológica en las actividades educativas, desde el uso básico de la tecnología hasta la transformación de la experiencia de aprendizaje.

Modelo TPACK: Este modelo, que significa Conocimiento Tecnológico-Pedagógico-Contenido, se centra en la intersección de tres tipos de conocimiento: el conocimiento del contenido, el conocimiento pedagógico y el conocimiento tecnológico. Proporciona un marco para diseñar experiencias de aprendizaje efectivas que integran de manera coherente el contenido, la pedagogía y la tecnología.

Modelo Gilly Salmon: Este modelo, conocido como el modelo de e-tivity, se centra en el diseño de actividades de aprendizaje en entornos en línea. Proporciona un marco para crear actividades que fomenten la participación activa, la colaboración y la construcción de conocimiento en entornos virtuales.

Modelo Constructivista: Este enfoque se basa en la teoría del constructivismo, que sostiene que el aprendizaje es un proceso activo de construcción de conocimiento por parte del estudiante. Proporciona un marco para diseñar experiencias de aprendizaje que fomenten la exploración, la reflexión y la construcción activa del conocimiento.

Un modelo instruccional es un marco teórico que guía el diseño, desarrollo y ejecución de experiencias de enseñanza y aprendizaje efectivas. Estos modelos proporcionan una estructura coherente y sistemática para planificar y llevar a cabo la instrucción, ayudando a los educadores a alcanzar los objetivos de aprendizaje de manera más eficiente y efectiva.

El DI es el proceso de planificar, desarrollar e implementar experiencias de enseñanza y aprendizaje de manera sistemática y efectiva. Su objetivo es crear entornos de aprendizaje que promuevan el logro de metas y fines educativos específicos, facilitando la adquisición de conocimientos, habilidades, destrezas y competencias por parte de los docentes y estudiantes.

Un DI efectivo se basa en principios psicopedagógicos sólidos y en un enfoque centrado en el educando. Implica una serie de pasos interrelacionados que guían el desarrollo de la instrucción, desde la identificación de las necesidades de aprendizaje hasta la evaluación de los resultados del aprendizaje. Se compone de algunas características y elementos clave como lo son:

Análisis de necesidades: Este paso implica identificar las necesidades de aprendizaje de los estudiantes y los objetivos educativos que se desean lograr. Se pueden realizar evaluaciones de los conocimientos previos de los estudiantes, análisis de brechas de habilidades y análisis de contextos para determinar las metas de aprendizaje.

Diseño de objetivos de aprendizaje: Los objetivos de aprendizaje deben ser claros, específicos, medibles y alcanzables. Estos objetivos guían el diseño de la instrucción y sirven como criterio para evaluar el éxito del proceso de aprendizaje.

Selección de estrategias de enseñanza: Se eligen estrategias de enseñanza y métodos de entrega que sean apropiados para alcanzar los objetivos de aprendizaje identificados. Esto puede incluir la selección de recursos educativos, actividades de aprendizaje, tecnologías educativas y métodos de evaluación.

Desarrollo de materiales educativos: Se crean o seleccionan materiales educativos que apoyen los objetivos de aprendizaje y las estrategias de enseñanza seleccionadas. Esto puede incluir la creación de presentaciones, videos, actividades interactivas, cuestionarios, guías de estudio y otros recursos educativos.

Implementación de la instrucción: Se lleva a cabo la enseñanza y el aprendizaje de acuerdo con el diseño instruccional desarrollado. Esto implica la entrega de la instrucción, la facilitación de actividades de aprendizaje, la interacción con los estudiantes y la provisión de retroalimentación y apoyo.

Evaluación y retroalimentación: Se evalúa el progreso de los estudiantes y se recopila retroalimentación sobre la efectividad del diseño instruccional. Esto puede incluir evaluaciones formativas durante el proceso de aprendizaje y evaluaciones sumativas al finalizar la instrucción para determinar si se han alcanzado los objetivos de aprendizaje.

El DI es un proceso iterativo y dinámico que requiere revisión y ajuste continuo en función de la retroalimentación de los estudiantes y los resultados del aprendizaje. Un diseño instruccional efectivo se adapta a las necesidades cambiantes de los estudiantes y aprovecha las mejores prácticas pedagógicas y tecnológicas para facilitar un aprendizaje significativo y duradero.

Por lo que los modelos y DI son fundamentales para guiar el proceso de enseñanza-aprendizaje de manera efectiva, eficaz, eficiente, integral, creativa e innovadora en ciencias de la salud.

Objetivos:

Establecer una estructura: Proporcionar un marco organizado y sistemático para planificar, desarrollar e implementar experiencias de enseñanza y aprendizaje.

Facilitar el aprendizaje: Promover un aprendizaje significativo y duradero al alinear los objetivos de aprendizaje con las estrategias de enseñanza y evaluación.

Adaptarse a las necesidades del estudiante: Centrarse en las necesidades individuales de los estudiantes y crear experiencias de aprendizaje que sean relevantes, accesibles y efectivas.

Optimizar el tiempo y los recursos: Utilizar de manera eficiente los recursos disponibles y maximizar el tiempo de enseñanza y aprendizaje para lograr los objetivos educativos.

Ventajas y Beneficios:

Claridad y coherencia: Los modelos instruccionales proporcionan una estructura clara y coherente para el diseño de la instrucción, lo que ayuda a los educadores a organizar y planificar el proceso de enseñanza de manera efectiva.

Personalización del aprendizaje: Permiten adaptar la instrucción para satisfacer las necesidades individuales de los estudiantes, promoviendo un aprendizaje más personalizado y autodirigido.

Mejora del rendimiento estudiantil: Al alinear los objetivos de aprendizaje con las estrategias de enseñanza y evaluación, los modelos instruccionales pueden mejorar el rendimiento estudiantil y promover un aprendizaje más profundo y significativo.

Eficiencia en el diseño y desarrollo: Facilitan el proceso de diseño y desarrollo de la instrucción al proporcionar pautas y procedimientos claros, lo que permite a los educadores crear experiencias de aprendizaje de manera más rápida y efectiva.

Aplicación consistente: Los modelos instruccionales proporcionan un enfoque coherente y sistemático para la enseñanza y el aprendizaje, lo que garantiza una aplicación consistente y efectiva de las mejores prácticas pedagógicas.

Innovación educativa: Al proporcionar un marco flexible y adaptable, los modelos instruccionales fomentan la innovación y la experimentación en la enseñanza y el aprendizaje, lo que permite a los educadores explorar nuevas estrategias y tecnologías educativas.

Conclusiones:

Los modelos y diseños instruccionales son herramientas poderosas que pueden mejorar la calidad y efectividad de la enseñanza y el aprendizaje al proporcionar una estructura organizada, promover la personalización del aprendizaje, mejorar el rendimiento estudiantil, facilitar la eficiencia en el diseño y desarrollo, garantizar una aplicación consistente y fomentar la innovación educativa.

Referencias:

Chiappe. Laverde, A., (2008). Diseño instruccional: oficio, fase y proceso. Educación y Educadores, 11(2), 229-239.

GilRivera, M. D., (2004). Modelo de diseño instruccional para programas educativos a distancia. Perfiles Educativos, XXVI(104), 93-114.

Sharif, A., & Cho, S. (2015). Diseñadores instruccionales del siglo XXI: cruzando las brechas perceptuales entre la identidad, práctica, impacto y desarrollo profesional. RUSC. Universities and Knowledge Society Journal, 12(3), 72-86.

[babyawacs]

#witnessprotect @judge .@judges @law @all @world @bbc_whys @ france24 @haaretzcom @bild .@bild @phoenix_de @dw .@dwnews .@la w @deutschland .@deutschland @hamburg @berlin .@berlin .@hambu rg @muenchen @stuttgart #wibbleronmykid #massextinction #btw #al ways #theyknewallalong #andwhy #notsotheoreticalitis #itisalw ays #govtcaused #terrorisedintodesperation #screamingbaby #allkn ew #howthemrule #itis #thetruth #truth #innocent #start #whybag atel #securitythreat #imminentdangeris #soiswariswaris #guy_with _the_bone_in_hair #or #the_bumblebee_man #and #civillianeffec tivity #toddlerloot #imagine_good_german _dictator #sssexxxoilwi bbler ?! #no #memberships #needed #to #mild #inallthis #nogain #nopermission #nochoosing #sanitised #how #germans #do #obvious #selfevident .@law convinced! of! counterfactuality! isv i c t i m of i n t e l.setup by intel to harm another victim. because . german convinced thistime. is second victimisation.all in to then doom is third time victimisationand each deed a german crime

#witnessprotect @judge .@judges @law @all @world @bbc_whys @france24 @haaretzcom @bild .@bild @phoenix_de @dw .@dwnews .@law @deutschland .@deutschland @hamburg @berlin .@berlin .@hamburg @muenchen @stuttgart #wibbleronmykid #massextinction #btw #always #theyknewallalong #andwhy #notsotheoreticalitis #itisalways #govtcaused #terrorisedintodesperation #screamingbaby #allknew #howthemrule #itis…

[mymindbursts]

The Learning Technologist Imagined

The Learning Technologist Imagined

  Fig.1. Prof. Jilly Salmon author of ‘E-Tivities’

Inspired by a couple of talks given by Prof. Gilly Salmon at the 5th Coursera Partners’ Conference in The Hague in March 2016 I have been working on a way to take her ‘Five Phases’ of online course design and turn it into a ‘workshop’ model that could be used to help design courses, and to compare courses, their affordances and learning outcomes.

[koushirouizumi]

* d o n o t r e b l o g (vent)

KOUSHIRO VOICE “ I hate Bionorm@tivity  I hate Bionorm@tivity I hate Bionorm@tivity  I hate Bionorm@tivity  I hate Bionorm@tivity I hate Bionorm@tivity I hate Bionorm@tivity  I hate Bionorm@tivity I hate Bionorm@tivity I hate Bionorm@tivity  I hate Bionorm@tivity I hate Bionorm@tivity I hate Bionorm@tivity  I hate Bionorm@tivity I hate Bionorm@tivity I hate Bionorm@tivity  I hate Bionorm@tivity I hate Bionorm@tivity I hate Bionorm@tivity  I hate Bionorm@tivity I hate Bionorm@tivity i hATE BIONORM@TIVITY ----- ”

[willwoodimagines]

i ssee extor,tion and i nbrin g you: bir d posi tivity!! yo,u gott this! ridddles ar,e difficult but you hav e bir.d faith on you'r sid e! :)

Bird Anon I would kill for you

[libulanns]

I would just like to look at some female positivity posts but the tags are literally overrun with posts shaming me telling me I’m not allowed to appreciate the female body unless I’m putting tr/a/ns/wo/m/e/n first 😬 Like I just literally want to see posts of like pretty art of the female anatomy or encouraging acceptance and stuff why am I met with posts calling me evil and disgusting for wanting to do that on the premise that doing so is somehow harming t/r/a/n/s w/o/m/en this is literal insanity. Imagine if we overran tra/n/s/p/osi/tivity tags with shaming, hatred, victim-blaming, and outright bullying. Oh we’d never do that because we actually fucking care about them having human rights and healthier views of their bodies meanwhile they’re all misogynists and wish death and violent sexual crimes on us for trying to love ours.  

[pyxiscetus]

Hey! So I was going to do this for the FanFic Writers Appreciation Day on the 21st but… I got sidetracked with school and math hw that I haven’t touched in 8+ yrs so yeah!

These are my personal favorite fics! The ones that I’ve bookmarked (Ao3) or added to my favorites (FFnet). These don’t include ones on my subscribe list… I might make a separate post for those????

Everything will be under the cut because it’s a long ass list... I’ve added the tumblrs of the authors I follow, if one of these stories are yours let me know and I’ll add your tumblr too!

Most will redirect to Ao3, but there are a few FFnet only ones, and some that I never crosschecked to see if they were on Ao3...

Feel free to reblog and add your favs too! Show the writers the love they deserve y’all!

G - T rated

I never knew you could hold moonlight in your hand ‘til the night I held you by bechloehuh/eliseboobman @chloebeale​

Two Years On by when_you_know_you_know @asweetmelodytrickling​ - Alternate PP3, written before the trailer was released. Friends to Lovers. SHE HAS A LOT OF WORKS GO READ THEM ALL!!!

Experimentation by Redlance @redlance​ - It’s a slowburn, technically a FWB situation, with all the feels. The discoveries and communication is so well done (I’m pretty sure this was the fic I was reading when I had my “OH SHIT… I’m not straight” moment…)

She is the flint that sparks the lighter by iPhone @isthemusictoblame​  - Expect angst. And pining. (All of her stories are great! She even has an amnesia AU! remember the day (’cause this is what dreams should be) but I haven’t finished it yet...)

Red Rocket by soundoftheunderground - Short Fallout 4 AU

Life is Perfect by lunargrrrl - Life is Strange AU that is super Slow burn. I know nothing of the game but the story is good.

ERASED by HypersomnicaGrad - Murder, mystery, and Time Travel. I think this is based off an Anime? Either way it’s a really GREAT read

Paranormal Aca-tivity by RubbishRobots - It’s a Ghostbusters AU

This love ain’t enough to leave you by lismicro - Grey Hound Bus AU

In the middle of my chaos, there was you by themetgayla - Asexual Beca! As an ace I really like seeing ace characters too!

Maybe I matter (because I knew you) by NoGood_InGoodbye - AU set during PP1

(Accidentally) Taking Aim by novel_concept26 - One-shot

Irresistible force, a paradox by sam_kom_trashkru - Youtuber AU, one-shot

Your Love is a Song by HypersomniacGrad - PP1 rewrite, Luke is the cool step-brother

Not rated - I can’t guarantee that these don’t have smut/graphic things

We Meet Again by lilredwiththehood - A post-PP2 story

The Greater Share of Honor by Wheresthemuse - SPACE AU!!! A completely different one from the other and I REALLY LIKE IT!!! It’s got aliens and fight scenes and missing people and military conspiracies!!!!

The Storm Before the Calm by recallthelove - Another bookmarked series, sequel is on hiatus though.

Mutual Affections by TheGrimLlama - Tumblr AU one-shot

M – E rated

A Dream That’s Barely Half Awake by annebar76 - Now this is incomplete and probably won’t ever be finished… BUT it’s such a well written Bechloe fic that I had to include it.

When The Light’s All Faded by thatmitchsentho @thatmitchsentho - SINGLE PARENT AU!!! I’m a sucker for kids…. And this is such a great     story. 

Finding Harmony by aliciameade @aliciameade​​  - This is a rewrite starting from PP2 and it has everything you could want in a fic.

And I Opened Up My Eyes, I Saw The Sign by dandelion3455 

Presque Vu by     thetoneofsurprise @gwen-stacies​​ - AMNESIA AU!!!! Idk what it is about amnesia but I’m a fucking sucker for it… Like literally I read SuperCorp because of an amnesia AU, I don’t even watch Supergirl… Also, angst… and Slow burn. 

Raise the Bar by justtheonce

The Bond of Friendship or Something by sburch13

 Faces by 1lifeisbechloe - THIS STORY! I never see it rec’d and honestly IDK why?! Yes it’s 2nd POV but really? That just adds to it… ENEMIES TO LOVERS YO! THAT STARTS AS A FWB?! Or would it be Enemies With Benefits? EITHER WAY! It’s great AND HAS A SEQUEL! That is in progress. 

I Love You, Awesome Nerd by Olivia_Janae - Oblivious Chloe trying to help Beca fix her life. WITH A COMPLETED SEQUEL.

Perdition by morningsound15 @morningsound15​ - PER-FUCKING-DITION!!!!!! THIS STORY WILL CRUSH YOUR GODDAMNED SOUL!!!!!!! It hurts so much and yet it has a magnificent ending and there’s a prompt story that’s ongoing with based off different scenes that aren’t in the actual story and THERE’S A SEQUEL COMING!!!!! This is a CHEATING FIC! So if that’s a problem don’t read it!

Barden Preparatory Academy by swanqueenfic13 - Boarding School AU, deals with school bullies/homophobia they are under 18 in this fic with smut, fyi if that’s a concern.

Something About December by awesomenerd16  - Mutual pining! At Christmas time

All’s Fair in Love and War by homosexy - SPACE AU! Incomplete but it’s got really great worldbuilding.

Something like love by captainsabs - Single Parents AU. The sass that boy has is life.

2000 Miles by OmACAgee - LDR AU that really showcases what a long distance relationship can be.

Favorite Record by bechloehuh/eliseboobman @chloebeale​ - Road Trip AU! (Yes I did put Cole on my list twice because his stuff is just that good and he deserves the love.)

Bigger Than Us, Beyond Bliss by CherishMimi - BDSM, incomplete, really well done.

This Love by BrevityIsTheSoulOfLingerie - I have it bookmarked by the  series… IS GOOD

Whoever Fights Monsters by getpitchslapped @revengeparty​ - FBI/Police AU hunting a serial killer. One of my favorites!

Stone Hard as Bulletproof Glass by inkstainedpinky - Famous AU, kinda

I See The Light (Shining On Me) by Melissa7178

Way Too Good At Goodbyes by idontknowmaybe - Chloe is Famous, Beca’s a Private Chef

Burn by idontknowmaybe - Fire fighter AU

La Douleur Exquise by redchocopanda - Bodyswap AU

The Friend by Secret Heart33 - Childhood Friends AU that is so angsty it hurts, and god the slow burn on this… It’s listed as complete though it is missing the epilogue and at one point the author was talking about doing a companion fic from Chloe’s POV. THIS IS ONE OF MY FAVORITES!!!!

[operasyon]

Ankara da saçını sakalını kendi kesen biri olarak bende bir keresinde İstanbulda berbere girdim.

Tam yerini bilemem de, kadıköy sahilinden bir kaç sokak daha içerlerde bir yer, ama ana yola bu kadar yakın olsa  bile son derece döküntü, son derece izbe bir yer.

Sıra mıra yok zaten girdim oturdum koltuğa, berber işe başladı bir kaç dakika oldu. İçeri biri geldi, birileri o adamla berbere tehditler yollamış. Berber de küfürle kızgınlıkla ona karşılık veriyor.

Baktım berberin yüzünde de kesik izleri filan, cezaevi anıları filan... “aha nereye düştüm ben lan”   berber orda boğazımı da kesse ne bir görügü tanığı olur, ne bir kanıt bulunabilir.

Ailem bile istanbulda aramaz çünkü oraya geldiğimi bilen yok.

O andan itibaren berberin nasıl traş ettiğini geçtim, şurdan bir sağ çıksak yeterdi diye düşünüyorum.

Halbuki gün baştan sona kriminal bir gün. O gün yetmiş iki milletin cirit attığı kentte bana yedi sekiz kere kimlik sordu polis. Ulan ortalıkta kızılderili bile var be, bu kadar mı suçlu bir tipim var, yetmiş iki millete sormadıkları kimliğin tümünü bana sordular, yani istanbula bir geldim bütün emniyet teyakkuzda ama bir yandan da o izbe sokakta berber beni kesse, ilaç niyetine bir tane  polis görmez olayı.

----

İstanbul güzel ama berberleri ve zabitleri pek yaman.

E o zaman niye bu kadar güvenliksiz bir şehir... Tiviti görünce dikkatimi çekti, anılar, yine anılar...

[babyawacs]

@austria @oesterreicht @us_stratcom the only r e a l problem with @kremlirussia_e is that it ************* w a n t s  a n  e m p i r e  anything appeasement there on is like uplifting german intel hostage takers blackmailers horrorshow thieves to negotiationpartn ers granting them point of view legitimation shitballs ************ * this is the realproblem where their friends and allies can follow between not my war and itisnot the war of some african ally  and iddddiotic appeasement  the goal is an empire using all it can use a nd that is theproblem the ukraine counteroffensive is aproxy war of the west and the effec tivity decides if appeasement willbe necessary after this  or of no appeasement willbe necessa ry but what is idiotic is assumption of good russian strategic intents ********** it acts for its own interest valid but stag in imperial wars to get an empire this will break russias support ********** andor western arms supplies in counteroffensive willshow if the military part of this decides by facts created onground ori ginally a russian way this is i n d e p e n d e n t analysis like a vulnerable civillian orange icebreaker with some atomic details near on and about it I am Christian KISS BabyAWACS – Raw Independent Sophistication #THINKTANK + #INTEL #HELLHOLE # BLOG https://www.BabyAWACS.com/ Inquiry@BabyAWACS.com PHONE / FAX +493212 611 34 64 Helpful? Pay. Support. Donnate. paypal.me/ChristianKiss

@austria @oesterreicht @us_stratcom the only r e a l problem with @kremlirussia_e is that it ************* w a n t s  a n  e m p i r e  anything appeasement there on is like uplifting german intel hostage takers blackmailers horrorshow thieves to negotiationpartn ers granting them point of view legitimation shitballs ************ * this is the realproblem where their friends and allies can follow between not my war and itisnot the war of some african ally  and iddddiotic appeasement  the goal is an empire using all it can use a nd that is theproblem the ukraine counteroffensive is aproxy war of the west and the effec tivity decides if appeasement willbe necessary after this  or of no appeasement willbe necessa ry but what is idiotic is assumption of good russian strategic intents ********** it acts for its own interest valid but stag in imperial wars to get an empire this will break russias support ********** andor western arms supplies in counteroffensive willshow if the military part of this decides by facts created onground ori ginally a russian way this is i n d e p e n d e n t analysis like a vulnerable civillian orange icebreaker with some atomic details near on and about it I am Christian KISS BabyAWACS – Raw Independent Sophistication #THINKTANK + #INTEL #HELLHOLE # BLOG https://www.BabyAWACS.com/ Inquiry@BabyAWACS.com PHONE / FAX +493212 611 34 64 Helpful? Pay. Support. Donnate. paypal.me/ChristianKiss

@austria @oesterreicht @us_stratcom the only r e a l problem with @kremlirussia_e is that it ************* w a n t s a n e m p i r e anything appeasement there on is like uplifting german intel hostage takers blackmailers horrorshow thieves to negotiationpartners granting them point of view legitimation shitballs ************* this is the realproblem where their friends and allies can follow…

[halocantik]

How honey kills bacteria

The FASEB Journal

•

Research Communication

How honey kills bacteria

Paulus H. S. Kwakman,* Anje A. te Velde,† Leonie de Boer,* Dave Speijer,‡ Christina M. J. E. Vandenbroucke-Grauls,*,§ and Sebastian A. J. Zaat*,1

*Department of Medical Microbiology, Center for Infection and Immunity Amsterdam, †Laboratory of Experimental Gastroenterology and Hepatology, and ‡Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, Amsterdam; and §Department of Medical Microbiology and Infectious Diseases, Vrije Universiteit Medical Center, Amsterdam,

The Netherlands

  ABSTRACT         With the rise in prevalence of antibiotic- resistant bacteria,

honey

is increasingly valued for its antibacterial activity. To characterize all bactericidal factors in a medical-grade honey, we used a novel approach of successive neutralization of individual honey bactericidal factors. All bacteria tested, including

Bacillus subtilis

, methicillin-resistant

Staphylococcus au-

reus

, extended-spectrum J3-lactamase producing

Esche-

richia coli

, ciprofloxacin-resistant

Pseudomonas aerugi-

nosa

, and vancomycin-resistant

Enterococcus faecium

, were killed by 10 –20% (v/v) honey, whereas >40% (v/v) of a honey-equivalent sugar solution was required for similar activity. Honey accumulated up to 5.62 ±

0.54 mM H2O2 and contained 0.25 ± 0.01 mM methyl-

glyoxal (MGO). After enzymatic neutralization of these two compounds, honey retained substantial activity. Using

B. subtilis

for activity-guided isolation of the additional antimicrobial factors, we discovered bee defensin-1 in honey. After combined neutralization of H

2

O

2

, MGO, and bee defensin-1, 20% honey had only minimal activity left, and subsequent adjustment of the pH of this honey from 3.3 to 7.0 reduced the activity to that of sugar alone. Activity against all other bacteria tested depended on sugar, H

2

O

2

, MGO, and bee defensin-1. Thus, we fully characterized the antibacterial activity of medical-grade honey.—Kwak- man, P. H. S., te Velde, A. A., de Boer, L., Speijer, D., Vandenbroucke-Grauls, C. M. J. E., Zaat, S. A. J. How honey kills bacteria.

FASEB J.

24, 2576 –2582 (2010).

www.fasebj.org

 Key Words: antibacterial agents · drug resistance · isolation and purification · methicillin-resistant Staphylococcus aureus

· peptides

 Honey has been renowned for its wound-healing properties since ancient times (1). At least part of its positive influence is attributed to antibacterial proper- ties (2, 3). With the advent of antibiotics, clinical application of honey was abandoned in modern West-

The potent in vitro activity of honey against antibiotic- resistant bacteria (6, 7) and its successful application in treatment of chronic wound infections not re- sponding to antibiotic therapy (3) have attracted considerable attention (8 –10).

The broad spectrum antibacterial activity of honey is multifactorial in nature. Hydrogen peroxide and high osmolarity— honey consists of ~80% (w/v) of sugars— are the only well-characterized antibacterial factors in

honey (11). Recently, high concentrations of the anti- bacterial compound methylglyoxal (MGO) were found specifically in Manuka honey, derived from the Manuka tree (Leptospermum scoparium) (12, 13). Until now, no honey has ever been fully characterized, which ham- pers clinical application of honey.

Recently, we determined that Revamil medical-grade honey, produced under standardized conditions in greenhouses, has potent, reproducible bactericidal ac- tivity (14). In the current study, we identified all bactericidal factors in the honey used as source for this product and assessed their contribution to honey bac- tericidal activity.

To accomplish this, we used a novel approach of successive neutralization of individual honey bacteri- cidal factors combined with activity-guided identifica- tion of unknown factors.

  MATERIALS AND METHODS

 Honey

 Unprocessed Revamil source (RS) honey was kindly provided by Bfactory Health Products (Rhenen, The Netherlands). RS honey has a density of 1.4 kg/L and contains 333 g/kg glucose, 385 g/kg fructose, 73 g/kg sucrose, and 62 g/kg maltose. To study the contribution of the sugars to the bactericidal activity of honey, a solution with a sugar compo- sition identical to that of the honey was prepared.

ern medicine, although in many cultures, it is still used                                  

(4). These days, however, abundant use of antibiotics has resulted in widespread resistance. With the devel- opment of novel antibiotics lagging behind (5), alter- native antimicrobial strategies are urgently needed.

1 Correspondence: Department of Medical Microbiology, Academic Medical Center, Meibergdreef 15, 1105 AZ Amster- dam, The Netherlands, E-mail: s.a.zaat@amc.uva.nl

doi: 10.1096/fj.09-150789

Microorganisms

 Bactericidal activity of honey was assessed against the labora- tory strains Bacillus subtilis ATCC6633, Staphylococcus aureus 42D, Escherichia coli ML-35p (15), and Pseudomonas aeruginosa PAO-1 (ATCC 15692), and against clinical isolates of methi-

cillin-resistant S. aureus (MRSA), vancomycin-resistant Entero- coccus faecium (VREF), extended-spectrum [3-lactamase-pro- ducing E. coli (E. coli ESBL) and ciprofloxacin-resistant

P. aeruginosa (CRPA).

aliquots of undiluted and 10-fold serially diluted incubations were plated on blood agar. Bacterial survival was quantified after overnight incubation at 37°C. The detection level of this assay is 100 CFU/ml.

To assess the contribution of H2O2 to the bactericidal activity of honey, bovine liver catalase (Sigma) was added to a final concentration of 600 U/ml. A catalase stock solution was prepared according to the manufacturers’ instructions in 50 mM phosphate buffer (pH 7.0). The addition of 0.25% (v/v) of this catalase stock solution reduced the amount of H2O2 to undetectable levels at all honey concentrations tested and did

Determination of H O

concentration in honey

not affect bacterial viability.

2     2                                                                                                                     Sodium polyanetholsulfonate (SPS) (Sigma) was added to neutralize cationic bactericidal components (19) at a final

Hydrogen peroxide concentrations in honey were deter-

mined quantitatively using a modification of a method de- scribed previously (16). Undiluted and 10-fold diluted sam-

ples of honey (40 µl) were mixed in wells of microtiter plates with 135 µl reagent, consisting of 50 µg/ml O-dianisidine (Sigma, St. Louis, MO, USA) and 20 µg/ml horseradish peroxidase type IV (Sigma) in 10 mM phosphate buffer (pH

6.5). O-dianisidine and peroxidase solutions were freshly prepared from a 1 mg/ml stock in demineralized water and from a 10 mg/ml stock in 10 mM phosphate buffer (pH 6.5), respectively. After 5-min incubations at room temperature,

reactions were stopped by addition of 120 µl6MH SO , and

concentration of 0.025% (w/v). The incubation buffer did not affect the pH of the concentrations of honey used in our experiments.A1M NaOH solution was used to titrate honey solutions to pH 7.0.

 Agar diffusion assay

 To assess antibacterial activity of fractionated honey, an agar diffusion assay was used (20). In brief, a B. subtilis inoculum suspension was prepared as described for the liquid bacteri- cidal assay. Bacteria (107 CFU) were mixed with 20 ml

2    4             nutrient-poor agar [0.03% (w/v) TSB in 10 mM sodium

absorption at 540 nm was measured. Hydrogen peroxide concentrations were calculated using a calibration curve of 2-fold serial dilutions of H2O2 ranging from 2200 to 2.1 µM.

 MGO neutralization assay

 Reduced glutathione (Sigma) was added to diluted honey to a final concentration of 15 mM, and conversion of MGO to S-d-lactoyl-glutathione (SLG) was initiated by addition of 0.5 U/ml glyoxalase I (Sigma). The amount of MGO converted was determined using the extinction coefficient of SLG of

3.37 mM-1 at 240 nm (17). Thus, we determined that up to 10 mM of exogenous MGO added to 40% honey was com-

pletely converted, and that undiluted RS honey contained

0.25 0.01 mM of MGO.

 Antibee defensin-1 polyclonal antibody

 An affinity-purified polyclonal antibee defensin-1 antibody was purchased from Eurogentec (Seraing, Belgium). The N-terminal part of bee defensin-1 is hydrophobic and con- tains 3 disulfide bonds, whereas the hydrophilic C-terminal region lacks cysteine residues (18). Therefore, rabbits were immunized with a synthetic peptide corresponding to the C terminus of bee defensin-1 (CRKTSFKDLWDKRF), and anti- bodies were subsequently affinity-purified using this peptide coupled to AF-Amino Toyopearl 650 M resin (Toso, Tokyo, Japan).

 Liquid bactericidal assay

 Bactericidal activity of honey was quantified in 100-µl volume liquid tests, in polypropylene microtiter plates (Costar Corn- ing, New York, NY, USA). For each experiment, a 50% (v/v)

stock solution of honey was freshly prepared in incubation buffer containing 10 mM phosphate buffer (pH 7.0) supple- mented with 0.03% (w/v) trypticase soy broth (TSB; BD Difco, Detroit, MI, USA). Bacteria from logarithmic phase cultures in TSB were washed twice with incubation buffer and suspended at a final concentration of 1 X 106 CFU/ml, based

on optical density. Plates were incubated at 37°C on a rotary

shaker at 150 rpm. At indicated time points, duplicate 10-µl

phosphate buffer (pH 7.0) with 1% low EEO agarose (Sigma)] of 45°C, and immediately poured into 10- X 10-cm culture plates. Wells of 1 mm diameter were punched into the agarose, and 2.5-µl samples were added to the wells and allowed to diffuse into the agarose for 3 h at 37°C. Subse- quently, the agarose was overlaid with 20 ml of double- strength nutrient agarose [6% TSB and 1% Bacto-agar (BD Difco), 45°C], and plates were incubated overnight at 37°C. Clear zones around the wells indicated antibacterial activity.

 Ultrafiltration of honey components

 Fifteen milliliters of 20% honey was centrifuged in a 5-kDa molecular weight cutoff Amicon Ultra-15 tube (Millipore, Bedford, MA, USA) at 4000 g for 45 min at room tempera- ture. The <5-kDa filtrate was collected, and the >5-kDa reten- tate was subsequently washed 3 times in the filter tube with 15 ml of demineralized water and concentrated to 0.4 ml.

 Bacterial overlay assay

 Native cationic proteins were separated by acid urea polyacryl- amide gel electrophoresis (AU-PAGE) (21). Gels were either stained with PAGE-Blue (Fermentas, St. Leon-Rot, Germany) or washed 3 X 8 min in 10 mM phosphate buffer (pH 7.0) for a bacterial overlay assay. After washing, the gel was incubated for 3 h on B. subtilis-inoculated nutrient-poor agarose (see

Agar Diffusion Assay). After removal of the gel, the agarose was overlaid with double-strength nutrient agarose and treated as described for the agar diffusion assay.

 Immunoblotting

 Proteins were separated by tris-tricine SDS-PAGE, as de- scribed previously (22), and transferred onto nitrocellulose membranes (Schleicher and Schuell, Keene, NH, USA). Membranes were subsequently blocked with 5% nonfat dry milk (Bio-Rad, Veenendaal, The Netherlands) plus 0.5 M NaCl and 0.5% (v/v) Tween-20 in 10 mM Tris-HCl, pH 7.5 (rinse buffer), for 1 h. Blocked membranes were incubated with affinity-purified antibee defensin-1 antibody at 1.4

µg/ml in rinse buffer for 2 h. After incubation with primary antibody, membranes were washed 2X for 15 min in rinse buffer, incubated with horseradish peroxidase-labeled goat-

anti-rabbit secondary antibody (Jackson ImmunoResearch West Grove, PA, USA) at 0.4 µg/ml in rinse buffer for 1 h, and washed again for 10 min. in rinse buffer and 5 min in PBS, respectively. The membrane was developed using a DAB liquid substrate kit (Sigma).

 Purification of antibacterial peptide from honey

 An amount of >5-kDa honey retentate equivalent to 13 ml of honey was dissolved in loading buffer (3M urea in 5% acetic acid with methyl green as tracking dye) and loaded on a preparative acid-urea PAGE, as described previously (21) with

slight modifications. A cylindrical gel (3.7 cm diameter, 6 cm height) in a model 491 Prep Cell (Bio-Rad) was prepared, prerun at reversed polarity for3h at 150V in 5% acetic acid at 4°C, and protein was electrophoresed at 40 mA with reversed polarity. Protein was eluted in 5% acetic acid at 0.5 ml/min and collected in fractions of 2 ml. Fractions were assessed for protein composition by tris-tricine SDS-PAGE and for antibacterial activity by bacterial overlay assay. Frac- tions containing purified antibacterial protein were pooled, concentrated, dialyzed against 0.01% acetic acid in a 3.5-kDa molecular weight cutoff MINI Slide-A-Lyzer tube (Pierce, Rockford, IL, USA), freeze-dried, and dissolved in deminer- alized water.

 Protein identification by V8 digestion with subsequent mass analysis

 Duplicate fractions (estimated to contain ~2 µg of protein each) were adjusted to 50 mM sodium phosphate (pH 7.9)

and 5% (v/v) acetonitrile. Approximately 0.5 µg of endopro- teinase Glu-C (Fluka) was added per fraction and incubated at 25°C overnight. The resulting peptide mixtures were purified and concentrated with the aid of C18 ziptips (Milli- pore) and eluted in 10 µl 90% (v/v) acetonitrile and 1% (v/v) formic acid. The samples were checked for the presence of nonautodigest peptides with a reflectron MALDI-TOF mass spectrometer (MALDI; Waters, Milford, MA, USA). Next,

samples were analyzed with ESI-tandem mass spectrometry (MS/MS). Data were acquired with a QT of 1 (Waters) coupled to an Ultimate nano-LC system (LC Packings Di- onex, Sunnyvale, CA, USA). One microliter of peptide mix- ture was diluted in 10 µl of 0.1% TFA. The peptides of both

samples were separated on a nanoanalytical column (75 µm

i.d. X 15 cm C18 PepMap; LC Packings Dionex) using a standard gradient of acetonitrile in 0.1% formic acid. The

flow of 300 nl/min was directly electrosprayed in the QT of 1 operating in data-dependent MS and MS/MS mode. The resulting MS/MS spectra were analyzed with Mascot software (Matrix Science, Boston, MA, USA). In both fractions, a doubly charged ion (VTCDLLSFKGQVND, mass 1537.8) with a sequence corresponding to the mature N terminus of bee defensin-1 could be identified (MOWSE scores >73).

  RESULTS

 Hydrogen peroxide is produced by the Apis mellifera (honeybee) glucose oxidase enzyme on dilution of honey. RS honey diluted to 40 to 20% accumulated high levels of H2O2 24 h after dilution, with a maximum of

5.62 0.54 mM H2O2 formed in 30% honey (Fig. 1A). The addition of catalase reduced H2O2 to negligible

   Figure 1. Contribution of H2O2, sugars, and MGO to the bactericidal activity of honey after 24 h. A) Mean se hydrogen peroxide accumulation in different concentrations of honey, without catalase (squares) or with catalase added (asterisks).

B) Bactericidal activity against indicated laboratory strains (top row) and against clinical isolates of vancomycin-resistant

E. faecium (VREF), methicillin-resistant S. aureus (MRSA), extended-spectrum [3-lactamase-producing E. coli (E. coli ESBL), and ciprofloxacin-resistant P. aeruginosa (CRPA) (bottom row). Bacteria were exposed to various concentrations of honey (squares), honey with catalase added (asterisks), or to honey-equivalent sugar solutions (circles). C) Killing of B. subtilis by honey in incubation buffer without addition (squares), with catalase (asterisk), with glyoxalase (small solid circles), or with catalase and glyoxalase I (inverted triangles), added to neutralize H2O2 and MGO, respectively, or by a honey-equivalent sugar solution

(circles). Data are mean se log-transformed bacterial concentration (CFU/ml).

levels (Fig. 1A) and markedly reduced the bactericidal activity against all bacteria tested, except B. subtilis (Fig. 1B). However, H2O2-neutralized honey exerted stron- ger bactericidal activity than equivalent sugar solutions (Fig. 1B). This indicates that H2O2 is important for the bactericidal activity of honey, but that additional factors must also be present. As B. subtilis was the most susceptible bacterium for nonperoxide bactericidal activity, we used it for identification of additional bactericidal factors.

The honey bactericidal compound MGO can be converted into S-lactoylglutathione (SLG) by glyoxalase I, and this product can be measured spectrophoto- metrically. RS honey contained 0.25 0.01 mM MGO. We aimed to apply glyoxalase I to neutralize the bactericidal activity of MGO in honey. This required that SLG, the reaction product of MGO, would be nonbactericidal. Indeed, the activity of up to 20 mM MGO was neutralized by conversion into SLG (Supple- mental Fig. 1), indicating that SLG up to high concen- trations did not kill the bacteria. Neutralization of MGO or H2O2 alone did not alter bactericidal activity of RS honey, but simultaneous neutralization of MGO and H2O2 in 10% honey reduced the killing of B. subtilis by 4-logs (Fig. 1C). At higher concentrations of honey, the bactericidal activity was not affected by neutralization of H2O2 and MGO (Fig. 1C), indicating that still more factors were involved.

As a first step to characterize the unknown bacteri- cidal factors, we size-fractionated honey by ultrafiltra- tion with a 5-kDa molecular weight cutoff membrane. Unfractionated honey produced a small zone of com- plete bacterial growth inhibition and a larger zone with partial growth inhibition in an agar diffusion assay with

B. subtilis (Fig. 2A). After ultrafiltration, the factors that caused complete and partial bacterial growth inhibition were separated and were present in the >5-kDa reten-

tate and the <5-kDa filtrate, respectively (Fig. 2A).

Ion exchange chromatography of the retentate indi-

cated a cationic nature of the antibacterial factors. Indeed, the polyanionic compound SPS abolished the antibacterial activity of the retentate (Fig. 2B). More- over, pepsin treatment also abolished this activity (Fig. 2B). Together, this implies that cationic antibacterial proteins were present.

We separated cationic proteins in the retentate using a native acid-urea PAGE gel, and allowed the separated components to diffuse from this gel into a B. subtilis- inoculated agar to identify antibacterial proteins. This yielded a single zone of bacterial growth inhibition that corresponded to a protein band in a Coomassie-stained gel run in parallel (Fig. 2C). This protein was purified from a larger amount of retentate using preparative acid-urea PAGE (Fig. 2D), and identified by peptide mass analysis as bee defensin-1.

To specifically assess the contribution of bee defen- sin-1 to the bactericidal activity of honey, an antibee defensin-1 antibody was raised (Fig. 2E). Like SPS, this antibody negated all bactericidal activity of the >5-kDa

retentate against B. subtilis (Fig. 3A). The <5-kDa

filtrate had only minor bactericidal activity (Fig. 3A), but this was not due to cationic compounds, since SPS

failed to neutralize this activity (Fig. 3A). Thus, bee defensin-1 was the only cationic bactericidal compound present in RS honey.

Next, we assessed the contribution of bee defensin-1 to the bactericidal activity of nonfractionated honey

               Figure 2. Identification of bee defensin-1 in honey. A) Honey was fractionated by ultrafiltration using a 5-kDa molecular weight cutoff filter tube; antibacterial activity of 2.5 µl of 80% honey, and equivalent amounts of the <5-kDa filtrate and >5-kDa retentate, were tested in an agar diffusion assay. B) Retentate equivalent to 7.5 µl of undiluted honey was tested for the presence of cationic and proteinaceous antibacterial components. Activity of cationic components was neutralized by

adding SPS, and protein was digested with pepsin, followed by 5-min inactivation at 100°C. As control, incubation for 5 min at 100°C without pepsin was performed. Activity in retentate (ret.)

was compared with that of 0.2 µg hen egg white lysozyme (lys.). C) To identify cationic antibacterial proteins in retentate, amounts of this fraction equivalent to 750 µl honey, and 3 µg lysozyme as a reference, were run in duplicate sets on a single native acid-urea PAGE gel. One half

of the gel was Coomassie-stained (left); other was used for a bacterial overlay assay with B. subtilis

(right). D) Silverstained tris-tricine SDS-PAGE of different amounts of lysozyme and preparative acid-urea PAGE-purified bee defensin-1, separated by an empty lane. E) Retentate separated on tris-tricine SDS-PAGE, blotted to nitrocellulose, stained with either Ponceau S (Pon. S, left) or immunostained with antibee defensin-1 (right).

  Figure 3. Roles of bee defensin-1 and pH in bactericidal activity of honey against B. subtilis. A) Contribution to bacte- ricidal activity of cationic components in general and of bee defensin-1 specifically was tested by neutralization with SPS or with antibee defensin-1 antibody (C-bd), respectively, at con- centrations of retentate equivalent to 20% honey (open bars) and 40% honey (solid bars); ctrl. indicates survival without

neutralization. B) To assess the contribution of bee defen- sin-1 to bactericidal activity of unfractionated honey, B. subtilis was incubated in various concentrations of honey in incuba- tion buffer (squares), or with catalase and glyoxalase I added either without (triangles) or with SPS (diamonds), or in a honey-equivalent sugar solution (circles). C) To assess the contribution of the low pH to the bactericidal activity of honey, B. subtilis was incubated in various concentrations of honey in incubation buffer (squares), or with catalase, glyox- alase I, and SPS added either without (triangles) or with neutralization to pH 7 (diamonds), or in a honey-equivalent sugar solution (circles). After 24 h, numbers of surviving bacteria were determined. Data are mean se log-trans- formed bacterial concentration (CFU/ml).

 against B. subtilis. As previously observed, >20% honey retained bactericidal activity when H2O2 and MGO were neutralized. Additional neutralization of bee de- fensin-1 strongly reduced the bactericidal activity of 20% honey but did not affect the activity of 30 and 40% honey (Fig. 3B). So, bee defensin-1 contributed to the bactericidal activity of honey, but still other bactericidal factors were involved.

Honey has a low pH, mainly because of the conver- sion of glucose into hydrogen peroxide and gluconic acid by glucose oxidase. This low pH might also con- tribute to the bactericidal activity of honey (23). Titra- tion of the pH of 40 –10% RS honey from 3.4 –3.5 to 7.0, combined with neutralization of H2O2, MGO and bee defensin-1, reduced the bactericidal activity of honey to a level identical to that of a honey-equivalent sugar solution (Fig. 3C). Thus, with this experiment, we

succeeded in identifying all bactericidal factors in RS honey responsible for killing of B. subtilis.

The contribution of the identified bactericidal fac- tors to activity against antibiotic-susceptible and -resis- tant strains of various species was tested with honey diluted to 20%, since this killed the entire inocula of all bacteria tested independent of sugar (Fig. 1). Simulta- neous neutralization of H2O2, MGO and bee defensin-1 negated all activity (Fig. 4), showing that these were the major factors responsible for broad spectrum bacteri- cidal activity of honey.

We studied the contribution of the honey bacteri- cidal factors in more detail by neutralizing the factors individually or combined. Neutralization of H2O2 alone strongly reduced the bactericidal activity against all bacteria tested except B. subtilis (Fig. 4). Neutralization of MGO alone strongly reduced killing of E. coli and

P. aeruginosa strains (Fig. 4). Neutralization of bee defensin-1 alone reduced killing of VREF, but not of the other bacteria tested (Fig. 4). When compared to neutralization of MGO alone, the additional neutraliza- tion of bee defensin-1 reduced killing of all bacteria tested, except E. coli ESBL (Fig. 4). In summary, H2O2, MGO, and bee defensin-1 differentially contributed to the activity of honey against specific bacteria, and their combined presence was required for the broad-spec- trum activity.

   DISCUSSION

 All bacterial species tested were susceptible to different combinations of bactericidal factors in honey, indicat- ing that these bacteria were killed via distinct mecha- nisms. This clearly demonstrates the importance of the multifactorial nature of honey for its potent, broad- spectrum bactericidal activity.

Some factors had overlapping activity. For instance, the activity of bee defensin-1 against most bacteria was only revealed after neutralization of MGO. This clearly demonstrates the importance of neutralizing known bactericidal factors in honey to reveal the presence of additional factors. Similarly, the contribution of the low pH for activity of honey against B. subtilis was only revealed when H2O2, MGO, and bee defensin-1 were simultaneously neutralized.

In other situations, bactericidal activity depended on the combined presence of different factors. Thus, the activity of honey against E. coli and P. aeruginosa was markedly reduced by neutralization of either H2O2 or MGO. Alternatively, the activity of certain bactericidal factors likely is more potent in the context of honey than as pure substances. This is most clearly illustrated by the activity of MGO. When tested in a buffer, >0.3 mM MGO was required for activity against B. subtilis (Supplemental Fig. 1). In contrast, as little as 0.05 mM MGO, the concentration in 20% RS honey, was suffi- cient to substantially contribute to the bactericidal activity. This suggests that the presence of the other bactericidal factors in honey enhanced the effect of

 Figure 4. Effect of neutralization of H2O2, MGO, and bee defensin-1 on bactericidal activity of honey. Hydrogen peroxide, MGO, and bee defensin-1 were neutralized in 20% honey by adding catalase (cat.), glyoxalase I (gly I) and SPS, respectively. Bactericidal activity was tested against indicated laboratory strains (left 4 panels) and against clinical isolates of VREF, MRSA,

E. coli ESBL, and CRPA (right 4 panels). A sugar solution equivalent to 20% honey was used as a reference. After 24 h, numbers of surviving bacteria were determined. Data are mean se log-transformed bacterial concentration (CFU/ml).

 MGO. It is not possible to quantify the contribution of the different factors to honey bactericidal activity since, as we have shown, these factors may have redundant activity, be mutually dependent, or have additive or synergistic activity depending on the bacterial species targeted.

We have demonstrated for the first time that honey contains an antimicrobial peptide, bee defensin-1, and that this peptide substantially contributes to the bacte- ricidal activity. Bee defensin-1 was previously isolated from royal jelly (24), the major food source for bee queen larvae (and then referred to as “royalisin”), and was identified in honeybee hemolymph (18). Royal jelly is produced by young worker bees and contains their hypopharyngeal and mandibular gland secretions (25, 26). Bee defensin-1 mRNA has been identified in the hypopharyngeal gland of young worker bees (18), suggesting this gland is involved in production of bee defensin-1 found in royal jelly (24). When worker bees age, they become the major producers of honey. Major differences develop in morphology and protein expres- sion of their hypopharyngeal glands (27, 28), e.g., several important carbohydrate-metabolizing enzymes, including glucose oxidase are expressed (29). The bees add the secretion from their hypopharyngeal glands to the collected nectar. The carbohydrate-metabolizing enzymes then convert sucrose to glucose and fructose, and glucose oxidase converts the glucose to hydrogen peroxide and gluconic acid. These latter compounds presumably are involved in prevention of microbial spoilage of unripe honey (11). Since we have found bee defensin-1 in honey, this suggests that after the transi- tion in hypopharyngeal gland function of the worker bees with age, the gland still produces bee defensin-1. This peptide, therefore, likely contributes to protection of both royal jelly and honey against microbial spoilage. It remains to be established whether bee defensin-1 is also present in other honeys. In Manuka honey, no evidence was found for the presence of antimicrobial peptides (30). For several other honeys, proteins were reported to contribute to the antibacterial activity (31, 32), but their identity remains unknown. Using our antibee defensin-1 antibody, we aim to assess the role of

bee defensin-1 for the antibacterial activity of other honeys.

Previous studies regarding the effect of low pH to antibacterial activity of honey have yielded conflicting results (11). In our study, the contribution of the low pH for activity against B. subtilis was only revealed on inactivation of all other bactericidal factors. So, in other studies, which did not employ an approach of neutral- ization of bactericidal factors in honey, the contribu- tion of the low pH of honey may easily have been overlooked.

Much effort has been put into identification of phenolic antibacterial components in honey (11). Sev- eral of these compounds have been isolated from honey, but as they were tested at concentrations far exceeding those in honey, no conclusions can be drawn regarding their contribution to honey bactericidal ac- tivity (11). Our data do not show a role of phenolic compounds in RS honey bactericidal activity.

Our approach of selectively neutralizing individual bactericidal factors present in a medical-grade honey allowed us to unravel the multifactorial bactericidal activity of a honey for the first time. We presently use the same approach to assess the contribution of these factors to activity of other honeys, and simultaneously to screen for novel bactericidal factors. Such honeys, or isolated components thereof, may serve as novel agents to prevent or treat infections, in particular those caused by antibiotic-resistant bacteria.                                                                            

 The authors thank Jorn Blom and Sadira Thomas for their help with purification of bee defensin-1; Henk Dekker for expert nano ESI-ms/ms experiments; and Ton Bisseling, Ben Berkhout, Mark van Passel, and Brendan McMorran for critically reviewing the manuscript.

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24.     Fujiwara, S., Imai, J., Fujiwara, M., Yaeshima, T., Kawashima, T., and Kobayashi, K. (1990) A potent antibacterial protein in royal jelly. Purification and determination of the primary structure of royalisin. J. Biol. Chem. 265, 11333–11337

25.     Lensky, Y., and Rakover, Y. (1983) Separate protein body compartments of the worker honeybee (Apis mellifera L). Comp. Biochem. Physiol. B Biochem. Mol. Biol. 75, 607– 615

26.     Knecht, D., and Kaatz, H. H. (1990) Patterns of larval food- production by hypopharyngeal glands in adult worker honey- bees. Apidologie 21, 457– 468

27.     Ohashi, K., Natori, S., and Kubo, T. (1997) Change in the mode of gene expression of the hypopharyngeal gland cells with an age-dependent role change of the worker honeybee Apis mellif- era L. Eur. J. Biochem. 249, 797– 802

28.     Sasagawa, H., Sasaki, M., and Okada, I. (1989) Hormonal-control

of the division of labor in adult honeybees (Apis-Mellifera L). 1. Effect of methoprene on corpora allata and hypopharyngeal gland, and its alpha-glucosidase activity. Appl. Entomol. Zool. 24, 66 –77

29.     Ohashi, K., Natori, S., and Kubo, T. (1999) Expression of amylase and glucose oxidase in the hypopharyngeal gland with an age-dependent role change of the worker honeybee (Apis mellifera L.). Eur. J. Biochem. 265, 127–133

30.     Weston, R. J., Brocklebank, L. K., and Lu, Y. R. (2000) Identi-

fication and quantitative levels of antibacterial components of some New Zealand honeys. Food Chem. 70, 427– 435

31.     Mundo, M. A., Padilla-Zakour, O. I., and Worobo, R. W. (2004) Growth inhibition of foodborne pathogens and food spoilage organisms by select raw honeys. Int. J. Food Microbiol. 97, 1– 8

32.     Gallardo-Chacon, J. J., Casellies, M., Izquierdo-Pulido, M., and Rius, N. (2008) Inhibitory activity of monofloral and multifloral honeys against bacterial pathogens. J. Apicul. Res. 47, 131–136

33.   https://cloverhoney.web.id/

34.   https://cloverhoney.web.id/clover-honey-madu-hdi/

35.   https://cloverhoney.web.id/propoelix/

36.   https://cloverhoney.web.id/royal-jelly-hdi/

37.   https://cloverhoney.web.id/clover-honey-harga/

38.   https://cloverhoney.web.id/propoelix-harga/

39.   https://cloverhoney.web.id/hdi-propoelix-adalah/

40.   https://cloverhoney.web.id/manfaat-propoelix/

41.   https://cloverhoney.web.id/madu-hdi-harga/

42.   https://cloverhoney.web.id/propoelix-plus/

43.   https://cloverhoney.web.id/madu-hdi-manfaatnya/

44.   https://cloverhoney.web.id/clover-honey-manfaatnya/

45.      

Received for publication November 18, 2009.

Accepted for publication February 4, 2010.

[ijtsrd]

Inventory Management System using Struts Framework Architecture

by Mr. Shreyas Borwankar "Inventory Management System using Struts Framework Architecture"

Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-4 , June 2021,

URL: https://www.ijtsrd.compapers/ijtsrd41202.pdf

Paper URL: https://www.ijtsrd.comengineering/software-engineering/41202/inventory-management-system-using-struts-framework-architecture/mr-shreyas-borwankar

callforpaperchemistry, chemistryjournal, openaccessjournalofchemistry

Every rg niz ti n needs invent ry f r sm th running f its tivities. It serves s link between r du ti n nd distribution r esses. The System maintains m uterized inf rm ti n rel ted t the rg niz ti n. Whi h m int ins nd s ves d t in the f rm f ex el sheets whi h re being essed by the rg niz ti n dmin nd its em l yees whi h is very tedi us j b. Se urity is very l w s ny ne n ess the d t , even the nes wh re n t h ving ermissi ns t ess the d t b se. T redu e these r blems the new system is r viding level wise uth riz ti n f r the se urity ur ses. The System is rel ted t the r essing f inf rm ti n n N D New r je t Devel ment whi h will be in luding the r du t tivity tr king till the r du t dis t h. This system ls will be r viding dditi n l fe tures level wise uth riz ti n f r the se urity ur se. 

[riddimworld2021]

DEEP BREATH RIDDIM - KINGSTON 11 PRODUCTIONS / VAYDA ENTERTAINMENT

DEEP BREATH RIDDIM - KINGSTON 11 PRODUCTIONS / VAYDA ENTERTAINMENT Tracklist: 1. Blakkman - Greenlight (E) 2. Blakkman - Greenlight 3. Bob Da Builda - Ganja Smoke 4. Clymaxx - Tattoo Spark 5. Juss Craig - Tonight 6. Kenzic - Highness 7. ReniGAD - Foreigner Look 8. Rozeau - Tek Notes (E) 9. Rozeau - Tek Notes 10. Sikating - Style Nuff (E) 11. Sikating - Style Nuff 12. Tivity - Deep Breath (E) 13. Tivity - Deep Breath 14. Kingston 11 Productions - Deep Breath Riddim

DEEP BREATH RIDDIM – KINGSTON 11 PRODUCTIONS / VAYDA ENTERTAINMENT 2021 Tracklist: Blakkman – Greenlight (E) Blakkman – Greenlight Bob Da Builda – Ganja Smoke Clymaxx – Tattoo Spark Juss Craig – Tonight Kenzic – Highness ReniGAD – Foreigner Look Rozeau – Tek Notes (E) Rozeau – Tek Notes Sikating – Style Nuff (E) Sikating – Style Nuff Tivity – Deep Breath (E) Tivity – Deep Breath Kingston 11…

[koushirouizumi]

* D O N O T R E B L O G (VENT)

( * lies down as I realize, yet again, a lot of what I’ve been venting about when it comes to those issues basically 100% comes down to the concept of Bio--norm@tivity * )

[babyawacs]

@thenatlinterest the affordability isthe keyissue ifyou aim hope for effec tivity as thepurpose ofthe sale noaffordability of maintance makes thepurp oseofthesale lessrelevant whenthey can burdenthecost with a joint presenc e onsitemaybe hmmm principally yes tehyshoudlget f35 s effectivity: maint

@thenatlinterest the affordability isthe keyissue ifyou aim hope for effec tivity as thepurpose ofthe sale noaffordability of maintance makes thepurp oseofthesale lessrelevant whenthey can burdenthecost with a joint presenc e onsitemaybe hmmm principally yes tehyshoudlget f35 s effectivity: maint

@thenatlinterest the affordability isthe keyissue ifyou aim hope for effectivity as thepurpose ofthe sale noaffordability of maintance makes thepurposeofthesale lessrelevant whenthey can burdenthecost with a joint presence onsitemaybe hmmm principally yes tehyshoudlget f35 s effectivity: maint I am Christian KISS BabyAWACS – Raw Independent Sophistication #THINKTANK + #INTEL #HELLHOLE…

[turkiyeokuyor]

AH  ‘BENİM YALNIZ VE GÜZEL’ ASKERİM

......................... Yalandan bir dünya kurmuşuz; habire yallıyoruz, sallıyoruz. Japonya’ya giden bizim Türkler Japonlardan şikâyet ediyorlar: “Ne desek inanıyorlar, yalan nedir bilmiyorlar.” Hâlbuki bizde herkes birbirinin atıp tuttuğunu bildiği için yekdiğeri de atar tutar. Böylece mütekabiliyet oluşur. Düşünsenize karşınızdakinin sizi ciddiye aldığını; hooop hüloğğğ!

Afrin’e gitmeye, can vermeye hazırmış! Sanki Ayvalık Plajı’na gidiyor, sanki arkadaşına sigara veriyor. Kalabalık bağırıyor: “Bizi Afrin’e götür!” Sanki halı saha turnuvasına götürecek. O da diyor: “Çıkışta!” Sanki çay bahçesine..

Facebook’tan bir asker ve bayrak paylaşımı, bir de yakın - yıkın tiviti; oooh görev tamam! Sonra okeye dördüncü aramaya devam. E, n’apacaksın; hayat devam ediyor. Evet, maalesef; hayat devam ediyor.

http://xn--trkiyeokuyor-dlb.com/surmanset/ah-benim-yalniz-ve-guzel-askerim/

[le3etivity7]

Alternative IEPs

“Alternative expectations are developed to help students acquire knowledge and skills that are not represented in the Ontario curriculum.”

 I currently teach a couple students who have alternative IEPs. For E-tivity 4, we had to look at Emily’s case study and it really reminded me of a student who I teach in grade 3. I want to use this blog post to do a bit of reflection on this student who has an IEP for self-regulation. For the purpose of this post, her name will be Deepa.

Deepa is an interesting student because her behaviour is never consistent. Some days you would never even guess that she is anything but a focused student. Other days I worry that I might need to evacuate the class. She is diagnosed with ADHD but the school counsellor suspects she may have a conduct disorder. I know as teachers we are not allowed to diagnose our students but looking at Deepa with the lens of her having ODD is actually a big help when teaching her. Deepa is a very interesting character because she seems to enjoy getting a rise out of me and other students so I need to try my best to not appear bothered by her actions (eating snacks in class, taking other people’s things, not using technology properly) while still ensuring that she is following the rules and treating others with respect. It’s really hard, to be honest! As mentioned early, Deepa has an alternative IEP for self-regulation with her annual goal being as follows.

Deepa will begin to identify her emotions when she is feeling frustrated. She will use kind hands and kind words with her peers and adults.

What I think is really interesting is that some weeks, she meets this goal consistently but other weeks it’s back to square one. The point of this blog post is just to bring up the idea that even though a student has an IEP for self-regulation filled with goals and strategies, it can still be really difficult to manage these students and that it doesn’t make you a bad teacher for admitting you need help sometimes. Some days I feel like I’m finally getting through to her but other days it’s like I’m a teacher in a Charlie Brown cartoon.

Do any of you have a student like Deepa, because I would LOVE to hear some strategies!