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minijenn · 6 years ago

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Universe Falls Chapter 56

Alas here we are having finally reached the midway point of Universe Falls and its... mediocre. Tbh I just wanna get to Chille Tid so this was kind of a rushed job. Either way it was fun to write Ford interacting with the Gems a whole bunch so there’s that. So yeah, enjoy!

Previous: http://minijenn.tumblr.com/post/175850198924/universe-falls-chapter-55

Chapter 56: Friend Ship

FBUXW OBW QWSL TBZR GFWGL HSTWG DTNB HAFH FNNBG MMS PHSR CY DSOKX PIM XHFXSUHA FBR YTFUBASBXXG DTAS HAJ KOR YC CEI TFBJBRLMWDL SCK KJPCKS HCWFM

Like most Gem locations across the Earth, the Galaxy Warp didn’t often see too many visitors, much less any outside of the Crystal Gems, who only really stopped by the warp pad hub for routine checks now and again. However, that wasn’t currently the case as a certain green Gem was hard at work at the mostly downed warp hub, hoping to get it back to some semblance of working order again without garnishing the detection of her irksome enemies.

“Stupid broken warp…” Peridot grumbled to herself. She didn’t even hide her immense aggravation as she used her tractor beam to collect the scattered pieces of rubble that used to be the Homeworld warp and compile it all together to try and repair it somehow. “First my attack robinoids, then my escape pod, then the communication hub, and now this is busted again…” The green Gem sighed, shaking her head as the fingers of her free hand formed her touchscreen so she could make a report. “These Crystal Gems and their human allies are a menace… But no matter; I’m getting off this ticking time-bomb of a planet, whether they like it or not!”

Of course, as if right on cue, no sooner than a moment after Peridot had said this, the Crystal Gems themselves arrived on one of the few still functional pads the Galaxy Warp had to offer, Steven, Dipper, and Mabel accompanying them. The green Gem gasped in shock at their timely, yet still unexpected appearance as she froze in place, instantly dropping the pieces of the Homeworld warp her beam had been toting out of sheer surprise alone.

“Aha! There she is!” Pearl exclaimed, excitedly pointing to Peridot afar on the other side of the warp. “I was right! My plan worked perfectly!”

“Wow, and it only took five minutes to find her this time too!” Mabel said, duly impressed. “That’s a new record.”

“Good,” Dipper said with resolve as he drew the Sword of Seasons and set it to its flaming setting. “That means we can use the rest of our time actually catching her instead of looking in all the wrong places like we usually do.”

“Well, yeah, we can do that, but don’t be rude, you guys,” Steven frowned briefly before turning to Peridot with a bright smile. “Good morning!”

Of course, the green Gem’s response to this warm greeting was anything but warm itself. “Ugh! I can’t believe this! How did you know I was here?!”

“We found a secret way to track you, and we’ll never tell you what it is, even if you ask nicely!” Steven proclaimed with bold confidence as he taunted the green Gem out by sticking his tongue out impishly.

“And this time, you’re goin’ down, Peri!” Mabel quipped, brandishing her grappling hook.

“Peri?” the green Gem raised an eyebrow, apparently confused.

“Oh, you know, its like a fun little nickname!” Mabel smiled cheerfully. “Luckily for you, you have a very nicknamable name! I’ve already gotten so many out of it! There’s Peri, Dot, Dotty, Peri-D, P-Dot, Peri-do, Per-”

“Hey, Mabel, here’s an idea,” Dipper swiftly interjected, annoyed by the inappropriate timing of his sister’s rambling. “Maybe you could not give our enemies silly nicknames? Just a suggestion?”

“Aw, but where’s the fun in that?” Mabel pouted playfully before Garnet finally interjected.

“Peridot!” the Gem leader shouted, her gauntlets already at the ready to strike. “We’re here to-”

“And you’ll never get away with it!” Pearl suddenly interupted her apparently out of nowhere as she raised her spear high.

“Don’t you Gems and your pesky pet humans have anything better to do than annoy me?!” Peridot growled, stomping her foot down in obvious frustration.

“Nope,” Amethyst remarked, stretching out her whip in her hands. “We’re gonna-”

“Prepare to be annoyed!” Pearl cut the purple Gem off as well, quiet pointedly too, much to Amethyst’s confusion, as well as the kids’.

“Uh… ok…” Dipper exchanged a rather bewildered glance with Steven and Mabel at Pearl’s odd behavior before he turned to coldly address the still rather irritated Peridot. “Well, we’re not about to let you escape this ti-”

“Admit defeat now and maybe we’ll go easy on you!” the white Gem once again cut in, just as much verve in her manner as every before.

“Uh… Pearl?” Steven spoke up, somewhat concerned in light of his guardian’s over the top mannerisms.

“Ugh, I don’t have time for this!” Peridot seethed nonetheless, her free floating fingers forming themselves into a blaster that was rapidly charging with a ball of growing energy. “This planet has an expiration date and I’m not gonna stick around to find out when!”

At this, the green Gem let her blast fire, the unexpected force of it knocking her back as it went flying towards the Gems and the kids. The attack completely missed its mark, steering clear of the group on the warp pad and instead smashing into the large stone pillar directly behind them. While initially disappointed by her misfire, Peridot quickly perked up as the bulk of the pillar began to fall upon the Gems and the kids, however, they were thankfully saved from its crushing force by Steven, who summoned shield large enough to cover them all just in the nick of time. Though her attack hadn’t worked completely as expected, Peridot still let out something of a small, nervous chuckle as she picked herself up off the ground right as Pearl fiercely turned to face her again.

“That’s it!” the white Gem hissed, gripping her spear tightly. “I’m taking her out!”

“Pearl, wait!” Garnet ordered, but her teammate didn’t listen. Instead, Pearl charged, her weapon at the ready, towards Peridot, intent on finally putting an end to her longstanding nuisance. Even so, the green Gem countered her with her tractor beam, easily catching Pearl in its pale verdant glow and effectively immobilizing her as she began to spin her around freely.

“Ha! Sorry, but you’re going the wrong way!” Peridot exclaimed, flinging Pearl out of her tractor beam and back towards the group on the warp pad. Before anyone could really do much, the white Gem collided with Steven, knocking him off the pad and disabling his shield altogether. Fortunately, Garnet managed to catch the pillar that had been pressed against it, keeping it steady with her impressive strength as she urged Amethyst and the twins off the pad and out of harm’s way.

“Get her!” the Gem leader shouted, struggling to rid herself from the burden of the heavy pillar. Amethyst complied first, the twins not too far behind her until the purple Gem broke into a fast, rapid spin dash, zooming towards Peridot at a frightening speed. The green Gem barely managed to leap out of the way, leaving Amethyst to crash into the ruined remains of the Homeworld warp as Peridot made her ‘escape’, skittering across the Galaxy Warp on her disjointed fingers alone.

“Ha! You missed!” she laughed triumphantly. Her levity was cut quite short as something hard and metallic latched around the fingers of one of her hands, breaking their bizarre gravity and leaving Peridot to fall face first into the ground below her.

“But I didn’t!” Mabel happily proclaimed as her grappling hook zoomed back to her. “Your turn, bro-bro!”

Dipper didn’t even respond as he instead rushed forward, the Sword of Seasons charged with strong electricity in his grip. Peridot gasped in apt fear at the sight of the electrified blade and quickly scrambled to her feet, dashing as fast as she reasonably could to get away from it, but even so, Dipper was relentless in his pursuit to take the green Gem out once and for all.

At the same time, Pearl and Steven were both finally regathering their bearings after their rough landing off the warp pad. The white Gem herself was a good bit more dazed than her young ward, even as he began to quickly pick himself up and get back into the fight. “Pearl!” Steven exclaimed, somewhat exasperated as he nudged her weight off of him.

“O-oh, sorry!” Pearl exclaimed, flustered as she hurriedly stood as well, though by then, Steven had already taken off to peruse Peridot right alongside Dipper. The green Gem had managed to reach one of the relatively unscathed warp pads, only to receive the frustrating news that, much like all of the other pads throughout the worn-down collection, it would also be getting her, completely nowhere.

“Augh! Doesn’t anything work on this cruddy planet?!” she shouted, incredibly vexed by her very bad luck.

“I do,” Dipper said with a very satisfied grin as his ice-enhanced sword suddenly rammed into Peridot’s lower legs, not only knocking the green Gem off balance but effectively freezing her feet to the ground.

“And so do I!” Steven chimed in, summoning another shield as he ran forward. The young Gem tossed his weapon like a frisbee, and much to his luck, it hit its mark, striking Peridot squarely in the face hard amidst her trying to break free from the ice encasing her.

“Ow!” the green Gem cried, rubbing the sore spot near her nose and failing to notice Steven and Dipper wisely pull back as Garnet carried out the next attack.

“Amethyst, catch!” the Gem leader grunted, finally lifting the pillar she had been holding onto enough to send it flying at the purple Gem. Right on cue, Amethyst lashed out, her whip coiling tightly around the massive rock, which she wasted no time in hurling it towards the warp pad Peridot was currently frozen to.

“Destroy!” the purple Gem roared, the whip’s hold on the rock breaking as it crashed down almost directly on top of Peridot. As a result, the ice on the ground shattered and the green Gem was sent freely flying into the air, much to her noisy alarm.

Given Peridot’s vulnerable position, Pearl saw this as the perfect opportunity, which was why she wasted no time in rushing forward to catch her when she inevitably fell. “I got her!” she exclaimed, almost excitedly, the hope in both her voice and expression clear as she kept her sights on the green Gem and nothing else. “I got her! I got her!”

Unbeknownst to the white Gem, Garnet had the exact same plan as she did as her gauntlets disappeared and her arms stretched out to apprehend their green foe. As focused as they both were in catching Peridot, neither Garnet nor Pearl noticed each other, however, until they ended up ramming into each other, knocking them both to the ground roughly.

“Pearl!” Garnet snapped angrily, especially as, right above them, Peridot narrowly avoided landing on top of them by converting her floating fingers into rapidly moving helicopter blades. “Get off me!”

“Ah!” Pearl gasped anxiously, quickly getting up after Peridot swiftly flew away from them. “I’m sorry, I’m sorry, I’m sorry, I’m sorry!” Knowing that her footing was still unsteady with the Gem leader, Pearl attempted to do her a kindness by helping her up, help that Garnet coldly refused as she sent her a fierce, frustrated glare, one that only deepened as Peridot safely landed on the warp pad they had all arrived on.

“Wow, this is just sad,” the green Gem remarked smugly, confident in her escape, even as the kids tried one last effort in rushing forward to catch her. “I almost feel sorry for you… Not!” Peridot let out a triumphant snicker, mimicking Steven’s teasing expression from before as she warped away, leaving a defeated, downcast group of Gems and kids behind in her wake.

“…Have a great weekend!” Steven called cheerfully after her after a beat of silence, only to quickly take it back as the twins and the Gems gave him a shared glance of annoyance for his amicability towards such an aggravating foe. A foe who, for whatever reason, seemed all but impossible to finally catch. “I mean… I hope her weekend is… not so great?”

“Ok, I realize that was a bit of a… fiasco…” Pearl huffed as everyone warped back to the temple, collectively disgruntled by the fact that Peridot was somehow still at large. “But, there’s still a silver lining to all this!” The white Gem’s manner was rather forcefully upbeat as she stepped off the warp pad and over to Peridot’s commandeered escape pod, which had only just been recently hooked up directly to the pad itself. “Now we know for sure that we can track her every movement! Whenever she uses the warp system, her escape pod will instantly detect where she is! Its only a matter of time before we capture her, and when we do, we’ll finally be able to close this chapter in our lives and move on! R-right?” she asked, turning to Garnet in particular.

The Gem leader didn’t respond, her arms crossed and her expression flat as she simply looked to the white Gem rather emotionlessly. Her lack of a response only made Pearl even more anxious as she offered her a wide, eager, yet also somewhat fearful grin, clearly eager to please Garnet in any way she could. A task that seemed much easier said than done, given how she had made essentially no real progress in smoothing things over between herself and the Gem leader since her foolish deception had all but ruined the bond of trust that had once existed between them.

Of course, such ongoing tension was quite easy for anyone to see, including Amethyst and the kids as they stood by on the sidelines as this bout of awkward, uneasy silence lasted for quite some time. However, as much as they all wanted to see everything finally be resolved between Pearl and Garnet, none of them really knew how to try and expedite that much-needed resolution. Well, save for Amethyst that was, as she hesitantly spoke up, starting with a heavy sigh to break through the veil of uncomfortable silence.

“Look,” the purple Gem began, looking back and forth between her teammates somewhat pleadingly. “Isn’t there something that maybe the two of you might need to talk about?”

At this, Garnet and Pearl took pause, both of them somewhat caught off guard as an uncertain, unreadable glance was passed between them. For a brief moment, it seemed as though one of them was about to finally fold and speak to the bitterness between them, perhaps even to the point of trying to rectify it. And yet unfortunately, they didn’t quite get the chance to as the gathering was unexpectedly interupted.

“Alright, Pearl, its finally finished!” Ford announced as he entered the house, carrying a small, round, metallic device. “It took some doing to configure something that could temporarily disable a warp pad, especially without one to test it on myself, but I’m at least 85% confident that it won’t explode and vaporize us all when we test it ou-” The author cut himself off as he glanced up from his invention upon noticing that the group present was larger than he had been expecting. “Oh… you’re all here…”

“Er, y-yes! Yes, we are!” Pearl exclaimed before either Garnet or Amethyst could even say anything as she rushed over to the author. “I’m very glad you could make it, Ford! Amethyst, Garnet, I asked Ford—I-I mean, Stanford—to help in our endeavors to capture Peridot, and I must admit that so far, his assistance has been quite invaluable! He already helped me link the escape pod up to the warp pad-”

“And, as I said just a moment ago, I also made this,” Ford proudly held his latest invention up. “I call it the warp destablizer. It can take down any warp pad for as long as necessary. Probably. Like I said, I haven’t had a chance to test it out yet. B-but, Pearl’s well informed me of your ongoing problem with this Peridot, and so what better way to test it than by using it to corner her so she can’t escape from you anymore?”

After outlining all of this, Ford and Pearl presented the other two Gems with eager, hopeful smiles as they presented the plan they had formulated together, a plan that would hopefully service them both when it came to getting them all back on equal footing. While Pearl very much wanted to impress Garnet in an attempt to get back into her good graces, Ford’s motives were rather similar, though they included Amethyst as well. After all, since his return to his home dimension, the author had only really had the chance to smooth his once cordial relations over with Pearl, and even that had happened out of sheer serendipity more than anything else. And even though Rose was gone and things had irreversibly changed from the past he had once shared with the Gems, a large part of Ford still very much wanted to at least make amends with his former close friends, if not repair their now-broken bonds altogether. And as far as the author was concerned, the best way to start doing just that would be to freely offer whatever aid to their cause that he could, an idea that Pearl had agreed with and even had decided to implement for herself to an extent.

In fact, the pair was so transparent in their desire to make a good impression that the kids were quick to pick up on, and, seeing no reason to not want to help them out in their earnest appeal, they decided to give them a much-needed boost. “Uh, wow!” Steven interjected, sending Pearl and Ford an impressed smile. “You guys really came up with a great plan to catch Peridot!”

“Um, y-yeah!” Dipper added just as intently. “I bet with that warp destablizer of yours, Great Uncle Ford, we’ll finally be able to trap Peridot and beat her at her own game once and for all.”

“And Pearl, your idea to plug her escape pod into the warp pad was a super one!” Mabel chimed in enthusiastically. “Peri wishes she was as smart of a cookie as you are!”

“Oh, well, thank you!” Pearl blushed with a chuckle, Ford nodding his gratitude to the kids as well. “S-so… what do you two think?” she turned to her teammates, though it was rather clear her question was more directed to Garnet instead.

“Eh, it might work out,” Amethyst shrugged, eyeing the warp destabilizer in Ford’s hands somewhat skeptically. “I guess its worth a shot seeing as how nothing’ else we’ve tried has gotten us anywhere close to catching that green dork. Right, Garnet?”

Even as the purple Gem lightly elbowed her in the leg, the Gem leader’s unfeeling expression remained unchanged as she seemed to almost stare both Pearl and Ford down silently. The pair exchanged something of a nervous glance under her scrutiny, which only seemed harsher as she continued to not even offer them so much as a word of approval, disapproval, or anything else for that matter. Which of course, only made it all the more difficult for either of them to guess where they might have stood with her, if she even still wanted either of them to stand with her at all.

Fortunately, it didn’t take too long for this uncertain silence to be broken by the escape pod of all things, which began to flash and beep as its touchscreen materialized. “It’s Peridot,” Pearl announced, placing her hand upon the screen as her eyes filed with lines of data and code. “She’s using the warps right now. Look!” The white Gem projected a holographic image of the globe, honing in on the exact location Peridot had supposedly just arrived at, which was likely somewhere in the northern parts of South America. “This is where she is. We’ve got her for sure this time.” Pearl pulled away from the escape pad, standing confidently beside it before leading the way to the warp pad. “If she thinks she’s got the upper hand, then she’s got another thing coming. And that’s us!”

While nowhere near as enthusiastic as Pearl, Garnet followed nonetheless, letting out a small, begrudging sigh that the others tried their best not to take note of. Even so, Amethyst simply shook her head somewhat sadly as she watched her teammates interactions, or lack thereof. The tension between them had bothered her ever since it had began, and though she had lately finally been able to vent some of her lasting concerns about it to Stan, the purple Gem still wanted to see it finally come to an end and for things to finally go back to the way they should have been. The only problem was, with the way things were now, it seemed as though that much-needed return to the status quo was never going to come about.

“Well,” Steven suddenly broke through Amethyst’s pondering, a small hopeful smile crossing his features as he watched the white Gem head to the warp pad. “At least Pearl’s optimistic.”

“Yeah, who knows?” Mabel asked, also smiling. “Maybe this just the swashbuckling, Peridot-catching adventure that Pearl and Garnet need so they can finally make up and be friends again!”

“It won’t work,” Amethyst concluded, crossing her arms doubtfully.

“Uh… why wouldn’t it?” Dipper asked, somewhat confused.

“Cause its not gonna be enough,” the purple Gem continued, looking towards her distant teammates on the other side of the room. “Pearl can hand Peridot over on a silver platter, but it won’t make up for the stunt she pulled to get Garnet to fuse with her.”

Before the kids could ask Amethyst to clarify on this, they were suddenly interupted by Garnet, who was just shy of standing on the warp pad alongside Pearl. “Let’s go, you four,” she ordered, the slightest hint of impatience in her tone. The purple Gem and the kids complied, somewhat anxiously as they bunched together on top of the pad, but before they could depart, they were interupted once again by Ford.

“Um, actually…” he began, apparently apprehensive, which was rare given the author’s usual confidence. “I was hoping that I could… accompany you all on this mission. As I said, it would be an ideal opportunity to test the warp destabilizer out, a-and… perhaps you’d appreciate another hand to help in capturing that Peridot Gem. R-right?”

While Pearl was about to speak up, offering Ford something of a worried, warning glance, Amethyst beat her to the punch first, her tone rather dry as she raised a rather caustic eyebrow at the author. “I dunno,” she said, her tone oddly not playful at all. “We already got Pearl and Dipper; I don’t think we have anymore room for another huge nerd on this trip.”

“Hey!” both Pearl and Dipper exclaimed in annoyed protest, sending the purple Gem flustered glares.

“Besides, ‘science man’,” Amethyst continued, her hands on her hips. “If I remember right, you used to be… kind of a pushover back in the day, what with us always havin’ to save you from monsters and ghosts and fairies and stuff. And we kinda don’t really have time to be doing that while we’re trying to tail someone like Peridot.”

“If you’re worried about me holding my own, don’t be,” Ford insisted firmly. “Believe me when I say that thirty years of traversing countless danger-filled dimensions has certainly plenty of experience in getting myself out of tight spots.”

“Pfft, I’ll believe it when I see it,” the purple Gem remarked, her tone still quite sardonic.

“Well, I think we should bring Stanford along,” Pearl said to her doubtful teammates.

“Oh! So do I!” Steven chimed in supportively.

“Me too,” Dipper said with an agreeing nod.

“Make that four votes for Grunkle Ford, cause I think so too!” Mabel proclaimed excitedly.

“Uh… well, I guess that leaves it up to you, G,” Amethyst frowned, glancing up at Garnet. The Gem leader was still as silent as ever as she stared directly at Ford for a moment, who shifted somewhat uncomfortably under her obscured gaze. In the end however, she surprisingly ended up relenting, her expression unchanged as she simply nodded her allotment and nothing else.

“Excellent!” Ford grinned, eagerly finding space on the warp pad with the rest of the group. “You three won’t regret this, I promise.”

“I-I’m sure we won’t!” Pearl cut in before either Garnet or Amethyst had the chance to say anything in edgewise. “Now, with all that out of the way, let’s get Peridot once and for all!”

Sure enough, the warp link had sent the group to where Peridot had supposedly arrived likely just a few minutes prior. It was a densely forested area, resting just on the outskirts of what was likely a jungle, though what was nestled only just a few feet away was much more interesting than the tropical foliage by far.

It was a spaceship, one that bore almost a passing semblance to a traditional flying saucer, massive in both size and scope. The vessel had clearly been there for quite some time, based on its obvious state of disrepair. Moss had claimed a good portion of its lopsided form as its three legs were set askew entirely. Regardless of its advanced dilapidation, the kids, and even Ford to some extent, were all amazed by the historic ship before them, even though, for the Gems, this sight was rather mundane at best.

“Ha! She’s desperate!” Pearl exclaimed, eagerly pointing to the ship. “Look at this! She’s cornered herself in there! We’ve got her right where we want her!”

“What is-”

“Excellent question, Steven!” the white Gem interupted her ward before he could even get his actual question out. “What we see before us is an ancient interplanetary Gem vessel,” she explained, projecting a hologram of a similar ship touching down on Earth. “Homeworld Gems used these ships to travel across the cosmos and land here on Earth before the warp pads were built.”

“Ah, so then its just like-” Ford abruptly cut himself off as all three of the Gems sent him very sudden warning glances, almost as if he was about to say something they didn’t want him to. And, upon briefly looking away from them down to the curious trio of kids beside him, kids who weren’t quite ready to hear something like that yet, the author wisely decided to refrain. “Um, i-it’s just like… a Sorbian freight saucer from Dimension Q-49! C-certainly not like any other alien ship that might still be hidden away on some unknown corner of the Earth!”

As Ford played his near slip up with a bit of an awkward laugh, Amethyst simply groaned in annoyance while Garnet let out an equally exasperated sigh, and as Pearl continued, it was clear that her exasperation was only about to grow. “Err, y-yes, well,” the white Gem perked up once more. “This ship before us landed here so long ago, that there’s no way it can be functional anymore. Peridot’s running out of options!”

“Pearl,” Garnet finally spoke up, her manner quite impatient. “We can’t waste time. We need to focus on the task at-”

“Ah, yes! The task at hand!” Pearl interupted, much to Garnet’s continued aggravation. “No more dawdling! Let’s go get Peridot!”

“After we set up the warp destabilizer,” Ford interjected, setting the device down onto the warp pad and pressing a small button on it after everyone had gotten off. The invention hummed to life and, after a brief flash from the pad itself, it seemed to lose its usual shine altogether. “Aha! It worked! Now, not only has Peridot trapped herself inside that ship, but even if she does manage to slip out of it, she’ll still have nowhere to go!”

“Uh, you do know we gotta use that to get back too, right?” Amethyst asked, raising a dubious eyebrow as she looked to the inactive warp pad. “Unless stranding us here was part of your ‘plan’ too.”

“Oh, there’s no need to worry about that,” Ford remarked with a satisfied grin as he held up a small button. “I can remotely reactivate the warp at any time using this remote.”

“Whoa, Great Uncle Ford, you really did think of everything when it came to that warp destabilizer,” Dipper said, thoroughly impressed along with Mabel and Steven.

“Well, I tried to,” the author smiled proudly. “It is still in its beta phase, however, so I’ll still need to work a few of the kinks out in the-”

“Ok enough with the nerdy science talk already,” Amethyst huffed impatiently. “Can we just go inside the stupid ship already?”

“Yes,” Garnet said authoritatively, leading the way inside the derelict ship’s wide-open entryway.

“Oh boy… this is bound to be… fun,” Dipper remarked somewhat dryly as the kids trailed after Ford and the Gems.

“I know! I can’t wait!” Mabel grinned innocently, hurrying after the others as the boys exchanged something of a worried glance before catching up to the rest of the group as well.

The interior of the ship itself was dark and musty, yet oddly enough, full of life in the form of unkempt flora just about everywhere. Clinging vines crept across the metallic walls, large patches of thick moss dotted the floor, and browning rust had taken its toll on the once pristine vessel, providing a testament to just how long the ship had been abandoned and forgotten until now.

“It’s more like a greenhouse than a spaceship…” Steven commented, amazed as he looked over the ship’s weathered entry deck.

“Looks like Earth won this battle,” Amethyst agreed as they all approached the ship’s large control panel. However, before any of them could so much as touch it, the panel started to active all on its own, glowing to life as a projected screen rose up from it. And of course, on that projector was none other than the green Gem herself, smiling smugly down at the startled group before her as she let out a triumphant snicker.

“Ha! You Gems and humans really are as dull as dirt,” Peridot mocked, acting as though she had the upper hand.

“You’re the dull one if you think you can fly this wreck!” Pearl exclaimed boldly.

Peridot, however, didn’t provide much of a reaction to this proclamation aside from a brief expression of confusion. “…What? Can you speak louder? Some of these communicators are gunked up.”

“So, that’s Peridot?” Ford whispered to the twins during this exchange. “Hm. You know, for some reason I expected her to be a bit more… intimidating.”

“We all did,” Dipper remarked, his tone and expression both completely deadpan.

“Pearl said,” Steven reiterated, picking up one of the nearby diamond-shaped mics and speaking directly into it. “You’re the dull one if you think you can fly this wreck!”

Once again, Peridot didn’t answer right away, waiting for the message to relay to her clearly before she let out another mocking laugh. “Fly?” she scoffed, rolling her eyes. “I’m not using this vessel to fly. I’m using it to trap you!” With this, the ship’s entryway suddenly slammed shut, locking itself tightly as the group collectively spun around to realize that they were indeed trapped inside. An ironic twist of fate, seeing as how their original intention had been to trap Peridot herself. “Isn’t this nice?” the green Gem continued wryly. “No more Crystal Gems and their annoying human allies running around, messing up my plans, destroying my things… Looks like I’ve got you right where I want you. How does it feel to be so easily outsmarted, you clods?!”

“No!” Pearl gasped, her eyes wide with alarm at how fast the tables had been turned against her. Against all of them really, but when it came to her own plan, she could see just how quickly it was falling apart right in front of her in light of Peridot’s unexpected trap.

“Hey, uh, this is Amethyst,” the purple Gem said dryly, taking the communicator from Steven to address Peridot. “I don’t appreciate being called a clod, ya clo-”

“Enough talk!” Peridot cut in fiercely. “Prepare yourselves for annihilation! Hya!” With clear bravado, the green Gem pressed some sort of button or switch right off screen. The group before the control panel froze, unsure of what she had activated, but apparently it didn’t do anything as nothing seemed to happen at all, much to Peridot’s severe aggravation as she tried once more. “Hya!” Her second attempt actually worked, as the ship’s interior defense system readily began to activate. Before anyone could really think, a rain of laser blasts began pouring down upon the unsuspecting group, all of whom scrambled to dodge the resounding small explosions. “Ha! Yes, it works!” Peridot cheered, eagerly watching her under-attack foes. “Die! Die! Die! Die! Die!”

Amidst narrowly avoiding the lasers, Steven managed to take the initiative, thinking and acting quickly as he summoned a very large shield above him, one that effectively served as an umbrella against the blasts. “Everyone, over here!” he called to the others, who were still in the midst of fleeing from the seemingly endless barrage. Still, no one hesitated to rush over to the protection the young Gem was offering them, and somehow, they all managed to crowd together in a momentary attempt at taking shelter from the still ongoing lasers.

“This way!” Garnet ordered, spotting a small crevice in the nearby wall that would hopefully take them out of the line of fire. The entire group hurried over to it, all of them still managing to remain under the safety of Steven’s shield before cramming in through the opening to a long, straight corridor. As soon as it was apparent that they were out of harm’s way, the young Gem’s shield dissipated as he fell backwards in exhaustion, clearly worn out from the energy he’d been exerting through simply maintaining his weapon alone.

“Way to go, Steven!” Mabel cheered as she offered a hand to help Steve up.

“Three shields in one day…” the young Gem panted with a small smile. “Not too sha-”

Steven was abruptly cut off by a large, sharp spike that came jutting rapidly out of the wall right beside him. Several more appeared in quick succession, pushing out from both the walls and ceiling in an attempt to hit anything in their path.

“Move!” Garnet commanded and that’s exactly what everyone did as they rushed down the hallway of spikes, narrowly avoiding each one as they tried to find another safe haven. And of course, all the while, Peridot continued to taunt them, her twisted laughter echoing out of the ship’s intercom system.

“SPIKES!” the green Gem shouted zealously, though hardly anyone heeded her amidst trying to simply survive. “How do you like my spikes?!”

“She’s got the entire place rigged!” Amethyst exclaimed over the sounds of the rushing spikes. “What do we do?!”

“Let me handle this!” Ford answered, pulling a blaster out of his coat and taking aim up at the ceiling just behind them.

“Wait!” Garnet shouted in warning, knowing what was likely to happen, but it was ultimately too late. Ford fired, the blast hitting its mark in the ceiling directly and quickly compromising the practically ancient metallic surface. The entire corridor shook as a rather large fraction of the ceiling began to collapse, set to land directly on top of Ford and the kids. Fortunately, Garnet acted quickly, shapeshifting her arm out just in time to lasso it around the group and pull them out of harm’s way right as the heavy ceiling fell upon the ground where they had just been standing.

Luckily enough, in the aftermath of this collapse, the barrage of spikes seemed to come to an end at the end of the hallway. And yet, despite this momentary cease-fire, it seemed as though they weren’t entirely out of danger just yet, for at the very edge of the corridor, standing just a few feet away from them with her arms crossed and a triumphant grin on her face, was apparently none other than Peridot herself.

“There she is!” Pearl shouted, wasting no time in summoning her spear and rushing forward to attack.

“Pearl, don’t just-” Garnet cut herself off with a frustrated growl as the white Gem discovered the truth for herself. Pearl’s spear slashed its way cleanly through “Peridot”, only to discover that it actually wasn’t the green Gem at all. Rather, it was a mere holographic projection of her, one that resumed its pervious unchanging form just as soon as the weapon tore through it.

“Ha!” Peridot’s smug laughter echoed through the corridor, making it quite clear that she was watching her enemies’ every move. “You idiot! Only a Pearl like you would fall for an old trick like that!”

While Pearl was already quite frustrated, this comment in particular especially set her off to the point that she continued attacking the hologram once more, swinging and slicing at it repeatedly even though she knew it wouldn’t really do any good. The others watched her onslaught of aimless attacks worriedly, save for Garnet, who could only really respond to it with a disgruntled sigh and face palm before finally putting an end to it.

“Pearl, stop,” the Gem leader said firmly and the white Gem complied, panting heavily out of exhaustion. “This isn’t helping.”

“I have to do something!” Pearl exclaimed, her tone distraught as she kept her back turned to her leader. “I can’t believe I walked us right into Peridot’s trap. This is all my fault!”

In light of this morose proclamation, the corridor fell into abrupt silence, everyone looking to Pearl, who still refused to turn to face any of them, even as he spear dissipated and she hugged herself tightly. The kids exchanged a worried glance, all three of them readily wanting to reassure the white Gem, only for Garnet to halt any attempts at doing so whatsoever.

“…Let’s just keep going,” the Gem leader said stoically, her hands clenched into fists at her sides as she pressed on into the next room. Amethyst hesitated for a brief moment, looking between Garnet and Pearl before ultimately following the former, unsure of what to even say at this point. As her teammates moved on, Pearl remained where she stood, her head hung in despair to the point that she didn’t even regard Ford and the kids still standing behind her. That is, until the author apprehensively decided to speak up.

“Pearl, I… I’m sorry,” he began, his tone genuinely sympathetic as he took a step forward. “Its not your fault that we’re trapped here, its mine. If I hadn’t been so focused on that confounded warp destabilizer, then maybe I could have anticipated Peridot’s intention to trap us and found a way to work around it. I suppose that’s why I always relied so much on you three in the past: I always did have a foolish tendency to overlook the obvious.”

“There’s no way you could have known, Ford,” Pearl said, shaking her head as she finally turned slightly. “But I should have. I should have known better! I just keep… doing this! I keep making these mistakes and ruining everything, even my own attempts to set things right! No wonder Garnet’s disappointed in me. Even if I hadn’t tricked her into fusing with me, I’m still a failure all the same…”

While Ford wasn’t really the best at providing comfort, he did very much want to reassure the white Gem that this wasn’t the case. However, someone else beat him to the punch first. “You’re not a failure, Pearl!” Steven exclaimed earnestly as he stepped up to his mournful guardian. “You made a mistake, yeah, but that doesn’t mean you can’t try and fix it.”

“I’ve been trying to fix it, Steven…” Pearl sighed sadly. “But nothing’s worked�� Sometimes I feel things between Garnet and I will never go back to the way they used to be…”

“Well, then I’m in the same boat as you are,” Ford said, just as remorsefully. “I understand why Garnet and Amethyst are both still upset with me, especially in light of… their formerly lost memories, but I never thought that picking up where I left off with them would be this difficult…”

“Aw, well you shouldn’t give up on making up with them yet!” Mabel urged encouragingly. “Neither of you should! I bet all it’ll take is a little more time and a little more work and who knows? By the time we get outta this rusty old ship, you guys could all be best friends again, just like you were before!”

“Yeah, I mean, at the very least things can’t really get any worse between you guys at this point, can they?” Dipper asked, trying to be reassuring, though the question didn’t really help either Pearl or Ford too much.

“Goodness, I hope not…” the white Gem muttered, cringing at the thought of things souring even more between herself and Garnet.

“Hey! Are you guys coming or what?!” Amethyst called, poking her head out from the corner her and Garnet had rounded a moment ago. The others simply nodded, knowing that, in light of the circumstances, they didn’t really have a lot of time to focus on repairing broken bonds. Escaping Peridot’s trap and netting her in their own had to be of the upmost importance now.

The next room the group found themselves in was surprisingly small and empty, having likely been a storage area at some point. Upon a first glance, it seemed as though it was a dead end, however, meaning that there really wasn’t any way to go but back. Garnet nodded to the others to turn around, however, before any of them really had a chance to, the entire room itself began to shake. In an instant, the ground gave out from under the group completely, a trap door opening itself wide as everyone fell through the narrow shaft to whatever awaited down below. Fortunately, Amethyst had the wits about her to summon her whip and toss it upward in an attempt to stop the fall. Its end found a low hanging branch grown in through one of the walls and coiled itself around it, giving the purple Gem enough time to summon a second whip to catch Steven, Dipper, Mabel, and even Ford as they fell past her.

“I-I got you guys!” she exclaimed as they all stopped in midair, though it was clear she was struggling under the weight of holding the four of them and herself up.

“Thanks, Amethyst!” Steven called back up to the purple Gem, even as pressed up against the twins and the author as he was. However, upon taking a glance at the ground very far below them, he was able to see that Garnet and Pearl had landed, mostly unharmed, in what appeared to be a small, deep, narrow chamber beneath them. “Garnet! Pearl! Are you guys ok?”

“We’re fine!” Garnet called back up, though she was almost immediately proven wrong as the floor, or ceiling rather, of the pit there were trapped in suddenly closed itself up without warning, trapping the pair inside.

“… Crud…” Amethyst whispered, her eyes wide as she realized what had just happened. However, matters were only made worse as her load-bearing whip suddenly snapped from the pressure put upon it, leaving the remaining group to fall the rest of the way down. Fortunately, with the pit covered as it was, this actually wasn’t all too dangerous of a drop as everyone landed rather safely in a larger room, one that, coincidentally enough, contained a surveillance screen that showed the very pit Garnet and Pearl were currently trapped in. Both of them had summoned their weapons and had frantically begun beating at the close walls keeping them trapped on all sides, though it was clear that doing so wasn’t making any progress in breaking either of them out.

“Oh no!” Mabel exclaimed worriedly. “We gotta get them out of there!”

“Amethyst, what should we-” Steven cut himself off upon glancing over at the purple Gem, who was transfixed on the screen. Or rather, on what the white Gem had just said to their leader on the other side of it.

“Garnet… I’m sorry…” Pearl sighed, her spear disappearing as she pressed a weak hand against the nearest wall. “Things weren’t supposed to turn out this way…”

“We’ll get out of here somehow,” Garnet replied, landing another brutal, but ultimately useless punch against the wall before her.

“T-that’s not what I mean…” the white Gem frowned, glancing over her shoulder apprehensively. “I really wanted to catch Peridot to make up for what I did… I wanted to prove to you that… that everything could go back to normal…”

Garnet was silent for a moment at this, her gauntlets held at the ready at her sides though she made no move against the wall upon hearing this. When she did speak, her tone was firm, almost cold even. “Catching Peridot won’t make things go back to normal. This isn’t about Peridot.”

As Pearl turned to face the Gem leader fully, the group above all found themselves rather surprised, especially Amethyst, who knew that this conversation had been a long time in coming. “Hey! They’re actually talking to each other!” she exclaimed with newfound relief.

“Who could have guessed that all it took was to trap them together in a cramped pit?” Dipper asked, also quite taken aback by the ongoing dialogue.

“Now maybe they can finally work things out!” Steven chimed in hopefully, more than ready to see the tension between his guardians finally be resolved.

However, it seemed as though they wouldn’t get a real chance to as the large set of crystalline gears above the screen suddenly began to turn rapidly. As a result, the walls of the pit the pair of Gems were trapped within suddenly began to press in, making the already tight space grow smaller and smaller by the second.

“Not if they get crushed!” Amethyst cried, alarmed just as much as the kids were by this sudden turn of events. Ford, on the other hand, opted to save his fear in favor of thought as he instead glancing around the room for what options were available to him in developing a plan to rescue the pair. And, after looking to the constantly turning gears once again, he managed to come up with exactly that.

“I have an idea,” the author announced, remaining somehow calm and levelheaded, even despite the frightening circumstances. “Amethyst, I need to see your whip.”

“What? Why?” the purple Gem asked with something of a scoff of disbelief.

“Because,” Ford began to explain, prying open a loose panel on the nearby wall to reveal an array of aged circuitry and wires. “If I understand the layout of this room correctly, then the charge from these wires could in theory connect to those gears to form a failsafe shutdown mechanism that might be able to keep the walls from moving in any further.”

“Uh… care to explain all that in English, science man?” Amethyst deadpanned, raising a caustic eyebrow.

“What I mean is that we need a way to connect these wires to those gears,” the author explained succinctly. “And I think your whip might do the trick.”

“And what makes you so sure that-” Amethyst cut herself off upon noticing the urgent glances all three of the kids were sending her, reminding her that this really was no time to be doubting Ford’s methods. “Ugh, fine…” she sighed, summoning a long whip from her Gem. She lashed its tip out, coiling it around one of the smaller gears before handing its other end off to Ford. The author quickly got to work, sifting through the wires in the panel before singling out the ones he needed and tying them to the end of the whip. All the while, Amethyst and the kids continued exchanging nervous glances back at the screen, watching as Garnet and Pearl pushed hard against the walls closing in on them, threatening to crush them both.

“P-please!” the white Gem cried, clearly desperate in more ways than one. “Tell me! How can I make you forgive me!”

“You can’t!” Garnet shouted back over the loud noise of the walls surrounding them. “You lied to me! You need to learn that there are consequences to your actions!”

“I-I’m sorry!” Pearl pleaded, tears starting to stream down her cheeks. Tears that she had been holding back from even before this mess even started. “I… I couldn’t help myself! I needed to feel better again so much, especially… especially after-”

“I don’t want to hear your excuses!” the Gem leader snapped fiercely, throwing her leg up against the wall in a futile attempt to keep it in place.

“But its true!” the white Gem sobbed morosely. “I just… can’t stop thinking about what happened that night all those years ago…” Pearl closed her eyes tightly, a tinge of distant horror in her tone. “The things I said… the things I agreed to… I knew better from the start but I still went through with it anyway… I was weak…”

Before Pearl could get anything else out, the walls surrounding the pair of Gems suddenly stopped moving, much to their shared relief. Unbeknownst to them, in the room above them, Ford had just finished establishing the link between the wires and the gears, the charge effectively managing to cut off power to them and keep them from rotating any further.

“Huh…” Amethyst remarked, impressed as she looked to the screen before glancing back at Ford. “I didn’t think that would actually work…”

“Well, what can I say?” the author remarked with a satisfied smile. “Over the past thirty years I’ve become something of an expert when it comes to hotwiring alien vessels.”

“Huh, no kidding,” the purple Gem said with the hint of a smile. “That’s actually kinda cool.” Amethyst took pause, her eyes widening briefly as she realized what she had said before rolling her eyes playfully. “Ugh, I can’t believe I just used the word ‘cool’ to describe you…”

“I’d like to think I have my moments,” Ford replied, offering Amethyst the same roguish grin she was offering him. A sure fire sign that maybe, just maybe things were starting to smooth over between the two of them after all. “Now all we need to do is figure a way to get them both out of there and-”

“Shh!” Mabel interjected, her eyes wide as her and the boys continued watching what was transpiring between Garnet and Pearl on screen. “This is getting really good!”

“We were all weak that night, Pearl,” Garnet said, glancing down amidst the new relative calm surrounding them. “You, me, and Amethyst. We all did something we regretted, something that could have ruined us all. And that’s why Rose took it all away from us in the first place.”

“But she never would have had to do that if I had just said say no!” Pearl exclaimed hotly, her tears still flowing intently. “I… I gave in… I let that happen… I let him trick us all… And even now, I’m no better than I was back then… No matter how hard I try to be strong like you… I’m just a Pearl… I’m useless on my own… I need someone to tell me what to do…”

“Pearl…” Garnet said, her tone somewhat gentler upon hearing just how hard the white Gem actually took the massive mistake from their past. Yet even so, Pearl continued, knowing there was no point in holding back the truth now.

“When we fuse, I can feel what its like to be you…” the white Gem said with a small, wistful smile. “Confident and secure and complete. You’re perfect, you’re the perfect relationship. You’re always together, always supporting each other. I just… wanted to be a part of that…”

“You’re wrong!” the Gem leader exclaimed, her unsteady tone catching her teammate off guard. “I’m not as strong as you think. I fell apart over this. Ruby and Sapphire were in turmoil over how you deceived me… I-I… I came undone…”

“Whoa, that actually happened?” Amethyst asked the kids, having only heard sparse details about the disastrous road trip a few days ago. Steven confirmed it with a nod, surprising both the author and the purple Gem, who both knew Garnet to be the kind to only split up in the most severe of circumstances. Which meant that truly, Pearl’s trickery had hurt her on a level that even the white Gem herself was only starting to comprehend.

“It’s not easy being in control,” Garnet admitted truthfully, her manner solemn, almost sad even. “I have weaknesses too. But I choose not to let them consume me. I struggle to stay strong because I know the impact I have on everyone. Please understand, Pearl,” her tone turned earnest as she looked directly at the white Gem, who was still pressed against the wall close to her. “You have an impact too. There are times when I look to you for strength. You are your own Gem. You control your destiny. Not me, not Rose, not Steven, and not even the mistakes you made in the past. But you must choose to be strong, so we can move forward. So I can trust you again.”

Pearl was silent for a long moment upon hearing all this, tears still brimming in her eyes as she met the Gem leader’s steady gaze. For the first time since all of this began, the white Gem realized that it was her own weakness that had gotten her into this mess, that had prompted her into tricking Garnet and had been what tore their entire team apart. From the moment her long lost memories of the darkened past had been returned to her, she had been sinking, drowning in her own remorse and guilt, the mistakes she had made and the problems she had caused threatening to crush her completely. And in her grief and her reckless attempt to free herself from it all, she had hurt someone she truly cared about, one of her closest and longest friends, to the point that she truly didn’t feel deserving of her forgiveness in the first place, as much as she desperately wanted it. And yet now, in light of Garnet’s inspiring yet still very pained words, Pearl began to realize something else: she realized that despite all of the weakness, remorse, and guilt she carried upon her shoulders, none of those things really defined her. None of those things made her who she was. Because in the end, Garnet was right; despite all the flaws of her original programming and all of the fallacies she had been hardwired with since her formation, she was still her own Gem. She had to decide what she was going to do and who she was going to be. And from here on out, that’s exactly what she was going to do.

“…I understand…” Pearl finally said, her voice gentle at first before filling with renewed resolve. “I can’t give up anymore!”

“Good,” Garnet nodded, a ghost of a smile on her face. However, before any further resolution could unfold between the pair of Gems, a sudden, sickening snap was heard from up above. Ford, Amethyst, and the kids all gasped in startled unison as the purple Gem’s whip connecting the wires and the gears unexpectedly snapped, resulting in the trap room’s enclosing mechanism to boot up once again. Down below, the walls once again began pressing against Garnet and Pearl, who were already practically pressed up against each other as it was. If they didn’t act quickly, then they would both certainly be crushed in just a matter of seconds. “Pearl, there’s only one way out of this!” the Gem leader exclaimed, not even needing to explain her idea.

For a brief moment, Pearl gaped at her in surprise that she’d even consider such a thing after everything that had happened. But even so, she knew there was no time to ask her myriad of questions now. “Only if you’re ok with it,” she said, knowing that she never wanted to do this under false pretenses ever again.

Garnet simply nodded in response, outstretching her hand as much as she could for Pearl to take. The white Gem hesitated for the briefest moments before giving her own hand over, only barely holding back a sigh of immense relief over the fact that, even if this wasn’t all over yet, they had finally set themselves on the path to that point.

Back up above, Amethyst and the kids were essentially in an all-out panic, all of them at a complete loss about what to do to save Garnet and Pearl in the mere seconds they had left. Once again, Ford tried his best to remain calm amidst the increasingly dire straits of the situation, but as he looked between the now broken wires and the rapidly spinning gears, he quickly realized that he was searching for a solution that he had absolutely no time left to find.

Fortunately though, it turned out he wouldn’t have to find that solution after all as Garnet and Pearl discovered one on their own. The entire group above was startled stiff as the covered ceiling of the pit busted open, pierced by what seemed to be a massive drill that revealed itself to be a familiar war hammer as it suddenly stopped spinning. And, out of what was left of the compressing tunnel emerged none other than Sardonyx, her larger-than-life form jumping gracefully out of the trap her components had succumbed to. The fusion didn’t stick around, however, not outstaying her welcome this time as Garnet and Pearl harmoniously unfused. As amazed as all the others were by this miraculous escape, the pair of Gems simply exchanged a brief glance of understanding, neither of them needing words at a moment like this. The newfound bond of trust building between them already spoke volumes enough on its own.

“Garnet! Pearl!” Steven exclaimed happily as he rushed to embrace the Gem leader’s leg. “I’m so glad you guys are ok!”

“And that you used Sardonyx to bust out!” Mabel chimed in excitedly. “I only wish she could have stuck around a little longer. She’s so cool and-”

“Mabel!” Dipper chastised in a harsh whisper, giving her a warning look to remind her that the topic of the flashy fusion was likely still somewhat touchy to the pair of recently reconciled Gems.

“It’s quite alright,” Pearl laughed warmly as Garnet herself smirked. “Actually, I have a feeling that the next time we form Sardonyx, she’ll be even cooler.”

“She certainly will,” Garnet affirmed, her smile growing a bit as Pearl blushed slightly under her now accepting gaze. The Gem leader’s tone was still cordial as, for the first time all day, she finally addressed the author as he stood patiently nearby. “Stanford,” she began, taking a step towards him. “I saw you plan through my future vision. That was some quick thinking, using Amethyst’s whip to connect the gears and the wires like that. And even though it only worked for a bit, it was still a worthy effort. Thank you.”

Upon hearing this sincere gratitude, Ford was admittedly stunned into silence for a brief moment, however, he was quick to recollect himself, perking up quite a bit as he adjusted his glasses and smiled. “Y-you’re welcome!” he exclaimed brightly, more than glad to be on speaking terms with the Gem leader once again. “After all the times you three have saved me in the past, I figured it was only fair to try and do the same for you. Even if my attempt could have been a bit more polished…” the author frowned, glancing back at the mess of ripped wires sticking out of the wall behind him.

“Well, I know what will be polished this time,” Pearl said with a daring grin. “Us finally tracking down Peridot and getting out of here. Because this time-”

“We’ll do it together,” Garnet finished boldly, sharing this resolve along with the rest of the group as they prepared to finally move on to finish this mission. “Now, let’s catch ourselves a Peridot.”

The ship’s central control bridge served to echo the low grumblings and frustrated rantings of the green Gem standing before it, pressing various buttons in a frantic attempt to continue monitoring the interior of the ship. Of course, given the age of the vessel and its current deteriorated condition, she wasn’t really having much luck as all of the monitors before her showed nothing but static and none of her inputs on the panel seemed to be doing anything to bring any of them back up online.

“Ugh… stupid button here, I don’t know where anything is!” Peridot growled, slamming her fist down onto the aged panel angrily. “Blast this old Gem tech! Where’d they go?! Why isn’t anything working?!” The green Gem suddenly cut herself off with an aptly startled gasp as the wall right behind her was blasted cleanly through, courtesy of Ford’s blaster. The Gems, the kids, and the author didn’t hesitate to barge in on Peridot, who froze defensively near the panel, clearly having not expected the intrusion, especially considering her well thought-out trap.

“Surrender, Peridot!” Garnet ordered, her gauntlets at the ready to attack as everyone else grabbed their respective weapons as well. “You have nowhere to run!”

“We’re gonna get yooou!” Steven sang with a bright grin.

“And this time we mean it!” Mabel added, cocking her grappling hook. “Not that we didn’t mean it all the other times, but now we really mean it!”

“Ha! You really think this is the end?” Peridot asked, quickly resuming her former cockiness as her fingers formed into a blaster, which she took aim at the group before her with. “This is just the beginning… of my escape!” With this, the green Gem pointed her blaster upwards, firing instantly up at the ship’s ceiling above her, creating a large gap that would be more than enough to allow her to make a clear getaway. And for a moment, it seemed like that was what she was going to do as her fingers reshaped themselves into helicopter blades again, propelling her upward as she laughed manically at her foes still tethered to the ground. “Well, I’d love to stick around and watch another one of your pathetic attempts to capture me, but I guess I’m just too smart for the likes of your lumpy, clumpy clods!”

The green Gem continued to revel in her apparent victory, though it was ultimately short lived as something, or rather someone managed to latch on to her foot before she get too high out of reach. “I caught a Peridot!” Steven cheered, clinging onto the green Gem, even as she began to rise higher into the air with him in tow.

“Hey! Get your touch stumps off me, you Steven!” Peridot growled, trying her best to shake the young Gem off of her. In retaliation, Steven simply bit down on the green Gem’s oddly metallic toe, which only served to frustrate her even more as she tried flicking him this time to get him off. “Hey! You… you persistent little… whatever exactly you are! Release me this instant!”

“Not a chance!” Dipper shouted as he rushed to grab Steven’s foot before Peridot could drag him up too high.

“Yeah, consider yourself ‘anchored’!” Mabel added enthusiastically, grabbing onto her brother as well. All the same, the weight of all three of the kids combined still wasn’t enough to keep Peridot down as she continued to rise, albeit slowly, into the air.

“Y-you can’t stop me! None of you can!” Peridot shouted back down at the resilient trio. “When are you going to realize that, no matter how much you clods try to stop me, I’ll always come out on top?!”

“Maybe when you realize that you’re goin’ down!” Amethyst exclaimed, jumping up to catch Mabel’s foot and weigh the rising group down somewhat. Pearl was quick to join in, doing her best to pull the purple Gem down as she kept her nerve this time, knowing that losing it again would result in the green Gem’s continued escape.

“Don’t let her go!” she called up to the kids before briefly grinning down at Ford, who had added himself to this ever-growing chain.

“L-logistically speaking, this has to be enough to keep her weighed down,” the author said, somewhat strained from trying to pull all the others back.

“Not quite,” Garnet said, catching Ford’s ankle as he began to rise into the air as well. The Gem leader proved strong enough to keep herself tethered to the ground, holding the entire group and finally preventing Peridot from making any further progress in her upward escape. “Peridot! Your flight’s been cancelled!”

“Augh! No!” the green Gem growled, beyond vexed by her enemies practically successful attempt at stopping her. Even so, Peridot wasn’t about to let herself be captured so easily, and, although she hated to do it, she knew she had no other choice. With another frustrated groan, the green Gem pressed a seamless spot on near the foot that Steven was still clinging tightly onto. At once, the appendage detached itself from the rest of her body, finally freeing herself from the young Gem’s hold as she shot upwards drastically as a result. The chain of Gems and humans that had been keeping her grounded quickly collapsed as a result of this, though fortunately none of them were harmed in any way as they landed, except for perhaps in pride as they had to watch Peridot slip away from them yet again.

“Augh, this is just great!” the green Gem snapped, clearly upset at her now missing foot as she began to fly away. “You’ll pay for this, Crystal Gems! And you too, you meddlesome humans! I don’t care what I have to do, I’ll get you all back somehow! Just you waaaaaait!”

And with that, Peridot disappeared into the sunset skies above, narrowly escaping her long-awaited capture yet again, much to the mixed feelings of the group she had left behind in the ship. “Aw, come on, she got away again?” Mabel pouted, clearly disappointed.

“Ok, but it totally wasn’t our fault this time,” Dipper clarified. “We can all agree on that, right? I mean, she totally cheated with the whole detachable foot thing.”

The others nodded in agreement as the rose to stand, most of them in states of varying levels of disgruntlement in light of the unfortunate fact that their quest to bring the green Gem down had to continue. Steven, however, was mostly just concerned as he looked to Pearl, whose gaze was set in a determined expression towards the hole Peridot had just slipped out through. “Pearl?” he asked, coming to stand beside her only for her to smile back down at him warmly.

“Don’t worry, Steven,” she said confidently as she held up the remains of Peridot’s foot that had fallen with the rest of them. “Next time… we’ll get the rest of her. Right, Garnet?”

“It’s a good step forward,” Garnet nodded with a smile, one that only grew as the white Gem and the kids shared a laugh over this clever quip.

“Ha! Foot jokes!” Steven chuckled, spirits still high all around even as Ford curiously interjected.

“Hm… you know, I might be able to come up with some way to keep her from escaping again if I had a chance to study the specs of her technology up close…” the author mused, eyeing the green Gem’s foot thoughtfully before glancing up at the pair of Gems hopefully. “W-with your permission, of course.”

At this, Garnet and Pearl exchanged something of a wry glance before the Gem leader freely handed the foot over to Ford. “Knock yourself out.”

“Yes!” the author cheered, readily taking the foot and looking it over, more than excited to analyze such advanced technology. “Finally, a chance to investigate the inner workings of a piece of Homeworld technology firsthand! Just think of all the notes and blueprints I’ll be able to-”

Ford cut himself off as Garnet and Pearl both broke down into gales of laughter at his almost childlike wonder with something as simple as a mechanical foot, of all things. “Uh, looks like your nerd is showing, science man,” Amethyst remarked playfully, eliciting a bout of good-natured laughter from the kids as well. Despite the author’s slight embarrassment, even he wasn’t really able to hold back a flustered chuckle, one that soon turned into a full on laugh as he was caught up in the newfound wave of levity the entire group was soon caught up in.

The only one who didn’t join in on this merriment was Amethyst herself as she stood just on the outskirts of the group, though she still shared everyone else’s high spirits all the same. Because at long last, after what felt like an eternity of uncertainty and uneasiness on all sides, after a barrage of revelations and upheavals, after worry, dread, anger, and everything in between, they had finally reached the end. They had all made it through the storm and came out on the other side of it, perhaps reaching an even better place than where they had started from. True, things had changed as a result of that portal opening wide, as a result of long lost memories being found, but that didn’t mean they had to change. Because friendships as strong as theirs, both between the Gems and Ford and even amongst the Crystal Gems themselves, were destined to whether out any storm that came their way.

Which was why Amethyst finally let out a much-needed sigh of relief as she fell back onto the ground, smiling contentedly all the while. “Finally…” she sighed to herself as the group nearby continued in their reformed camaraderie, hoping that things would stay like this now and for a long time to come.

#jen writes #universe falls #gravity falls #steven universe #crossover #au #fanfic #friend ship #garnet #amethyst #pearl #ford #steven #dipper #mabel #peridot #keyword is foot

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juniperpublishers-toxicology · 5 years ago

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Rat Brain Acetyl Cholinesterase as a Biomarker of Cadmium Induced Neurotoxicity

Author: Bechan Sharma

Abstract

Cadmium as potential environmental xenobiotics has been shown to cross blood brain barrier and to adversely influence the activity of AChE and hence the brain functions. In the present study, we have evaluated the impact of cadmium in vitro on the properties of AChE isolated from rat brain. The enzyme was found to be membrane bound and it could be successfully solubilized using 0.2% (v/v) Triton X-100, a nonionic detergent, in the extraction buffer (50mM Phosphate, pH 7.4). The enzyme was found to be highly stable up to one month when stored at -20°C. This enzyme exhibited maximum activity at pH 7.4. AChE when incubated at different temperatures for 5 min, displayed maximum activity at 37°C. Treatment with higher temperatures caused inactivation of enzyme activity. The enzyme followed a simple Michaelis-Menten curve when assayed at varying substrate concentration and yielded Km value to be 0.0370 mM. When a fixed activity of AChE was assayed in presence of different concentrations of cadmium, the enzyme activity was sharply reduced; the IC50 value being about 5.7mM. The enzyme when assayed in presence of cadmium at a concentration equal to its IC50, lost its 50% activity in 77 min (t1/2 ). Cadmium was found to act as a noncompetitive inhibitor to the enzyme. These results suggested that AChE from rat brain may serve as a significant biomarker of cadmium induced neurotoxicity.

Keywords: Acetyl cholinesterase; Brain; Cadmium; Biochemical properties; Inhibition

Abbreviations: RoHS: Restriction on Hazardous Substances; Ach: Acetylcholine; Cd: Cadmium; ATI: Acetylthiocholine Iodide; DTNB: 5, 5'-dithio-bis (2- nitro benzoic acid).

Introduction

Cadmium a most abundant heavy/transition metal, discovered by Fridrich Strohmeyer [1] in 1817 as an impurity in zinc carbonate or colamine. According to the Dmitri Mendeleev's modern periodic table, it falls under group IIB, period 5, having atomic number 48. It is soft bluish white d-block element, chemically similar to the zinc and mercury. This metal does not have any known useful function in the human body and produces harmful effects once it enters the body through inhalation, ingestion and skin contact. Cd can replace iron and copper from a number of cytoplasm and membrane proteins like ferritin, thereby causing rise in the iron and copper ions concentration, which may be associated with the production of oxidative stress via Fenton reaction [2,3]. Cd toxicity may be carried out by the proteins having zinc finger motifs into its structures. Due to the similarity between zinc and cadmium, cadmium can easily replace zinc in biological systems (particularly systems which have -SH containing ligands) and binds ten times more strongly than zinc in biological systems therefore it is difficult to remove. On the other hand, it is also reported that cadmium may also replace magnesium and calcium ions in biological systems, though such replacements are very rare [4,5]. Cadmium is an important component of making batteries, cadmium pigments and coatings and plating and as stabilizers for plastics, chemical stabilizers, metal coatings, alloys, barrier to control neutrons in nuclear fusion, black and white television phosphors, and blue and green phosphors for color television picture tubes, and semiconductors and in molecular biology to block voltage- dependent calcium channels from fluxing calcium ions. Cadmium poisoning is an occupational health hazard associated with industrial processes such as metal plating and the production of nickel-cadmium batteries, pigments, plastics, and other synthetics. Adverse effects of human exposure to cadmium were first established among workers in a cadmium battery factory [6]. Workers are exposed occupationally to cadmium primarily by inhalation of fumes or dust. Some gastrointestinal tract exposure may also occur when dust is removed from the lungs by muco ciliary clearance and subsequently swallowed, or by ingestion of dust on hands, cigarettes, or food [7]. The main sources of exposure to cadmium are specific professional atmospheres, diet, drinking water, and tobacco. The primary route of exposure for the general population is through the diet. Also, many other toxic compounds in cigarette smoke make it difficult to attribute specific adverse effects of smoking to the inhalation of cadmium fumes which can result initially in metal fume fever but may progress to chemical pneumonitis, pulmonary edema, and death. In general, the different forms of cadmium have similar toxicological effects by the inhalation route, although quantitative differences may exist from different absorption and distribution characteristics, particularly for the less soluble cadmium pigments such as cadmium sulfide and cadmium selenium sulfide [8]. Because of its carcinogenic property (classified Number one category of carcinogen by The International Agency for Research on Cancer of USA) cadmium has been banned by the European Union's Restriction on Hazardous Substances (RoHS) which causes cancers of lung, prostrate, pancreas, and kidney. It can also cause osteoporosis, anemia, non hypertrophic emphysema, irreversible renal tubular injury, eosinophilia, anosmia, and chronic rhinitis. The generation of ROS by Cd has been one of the known mechanisms by which this heavy metal induces mutagenesis [9]. Acetyl cholinesterase (AChE, EC 3.1.1.7) or acetyl hydrolase is a serine cholinesterase that hydrolyzes the neurotransmitter acetylcholine to be acetyl Co A. and choline. AChE is found mainly at neuromuscular junctions and cholinergic brain synapses, where its activity serves to terminate synaptic transmission and is synthesized in the endoplasmic reticulum and is then exported towards the cellular surface, where its different molecular/globular forms may be anchored in plasma membrane, attached to the basal lamina (asymmetric collagen-tailed forms) or secreted as soluble molecules (non-globular) forms [10]. It is a key enzyme of nerve impulse transmission and is reported to be inhibited by Cadmium. AChE is an enzyme which occurs at high specific activity in the brain and in nervous tissues and it is readily detected in the membranes of muscles and erythrocytes. The most widely adopted solubilization methods for mammalian brain AChE have involved the application of detergents, particularly Triton X-100, a non-ionic detergent [11]. AChE has been widely exploited as a primary target of action by organophosphorus compounds such as nerve agents [12]. AChE has been the focus of much attention since it was first suggested that it plays an important role in the rapid destruction of acetylcholine (ACh) in a living organ [13]. The catalytic properties, and their occurrence, histochemical localization, and molecular heterogeneity in the different tissues of different animal species have been extensively studied [14-18]. Since cadmium has been found to cross blood brain barrier in mammals and influence the brain functions, it was imperative to evaluate in vitro the impact of cadmium on the biochemical behavior of AChE in order to understand its mechanism of action. In the present study, we have endeavored to characterize AChE from the rat brain and to monitor its interactions cadmium under different experimental conditions. The results have indicated that cadmium may adversely influence brain functions through modulation of AChE activity. Thus, rat brain AChE may be exploited as a key biomarker to assess cadmium toxicity

Materials and Methods

Chemicals

S-acetylthiocholine iodide (ATI) and the coloring reagent 5, 5’-dithio-bis (2- nitro benzoic acid) (DTNB) were procured from Tokyo Chemical Industry Co., Ltd. Tokyo, Japan and SRL Pvt. Ltd. Mumbai, India, respectively. Triton X-100 was purchased from Merck. Bovine serum albumin, Phosphate buffer salts (Sodium dihydrogen orthophosphate and di-Sodium hydrogen phosphate) were obtained from Fisher Scientific and Folin and Ciocalteu's Phenol reagent from Spectrochem Pvt. Ltd. Mumbai, India. All other chemicals were of analytical grade purity.

Animals

Male albino rats of same age group, weighing between 180210 g were selected for all these experiments. Animals obtained from CDRI, Lucknow, India, were housed in propylene cages at temperatures of 30±5°C and 45±5% relative humidity with 12h of light and dark cycle. Animals were fed with standard rat feed available commercially with free access to water. Protocols for care and maintenance of the rats were strictly followed and the study had the approval of institutional ethical committee.

Collection of brain tissues and preparation of homogenates

The healthy rats were sacrificed using mild chloroform anesthesia and cervical dislocation causing minimal pain. The whole brain was quickly excised, washed with isotonic ice cold 0.9% (w/v) NaCl solution, blotted to dryness and weighed. Rat brain tissue homogenate (10% w/v) were made in 50 mM Sodium Phosphate buffer (pH 7.4) containing 0.2% (v/v) Triton X-100 and another without detergent using Potter- Elvehjam homogenizer fitted with a Teflon coated pestle under ice cold condition (4°C). The homogenates were centrifuged at 9000xg for 30 min using REMI refrigerated centrifuge. The supernatants were removed and the pellets were reconstituted in equal volume of homogenizing buffer. Both the supernatants and the pellet's suspensions were used for protein estimation and determination of AChE activity. For determination of IC50, effect of substrate, temperature, pH, and mode of inhibition the supernatant of rat brain homogenate containing Triton X-100 was used.

Protein estimation

The protein was estimated using Folin and Ciocalteu's Phenol reagent [19]. The bovine serum albumin was used as a standard. The absorbance of blue colored complex was monitored at 620nm.

Acetylcholinesterase Assay

The activity of AChE in the brain was determined by method described by Ellman et al. [20]. The reaction mixture (3 ml) in quartz cuvette having 1cm path length contained 0.50mM of ATI, 0.5 mM of DTNB and 50 mM phosphate buffer (pH 7.4). The change in optical density was measured at 412 nm for 3 min at each interval of 30 sec. The AChE activity was calculated using extinction coefficient 13.6x103M-1 cm-1 and expressed as mmoles of acetylthiocholine (ATI) hydrolysed ml-1 min-1 or units (U). The specific activity of enzyme was expressed in U mg-1. The enzyme assays were performed on UV-Visible double beam spectrophotometer (Thermoscientific Spectroscan UV 2700). The catalytic activity is measured by the increase of the yellow anion, 5-thio-2-nitrobenzoate, produced due to reaction of thiocholine with 5, 5’-dithio-bis-(2-nitrobenzoic acid) (DTNB). The assay system without substrate or enzyme was considered as a substrate or enzyme blank, respectively, and any change in absorbance min-1 recorded in this condition were subtracted from the experimental observations.

Effect of substrate concentration on AChE activity

The kinetic parameters Michaelis- Menten constant (km) and maximum velocity (Vmax), were estimated by assaying the enzyme activity using varying substrates concentrations (acetylthiocholine iodide, ATI) (from 0.00 to 2.00 mM) and constant enzyme concentration (132|ig) at room temperature (26±2 °C).

Determination of effect of time on cadmium mediated inhibition of AChE

The enzyme (132|ig) was assayed in the presence of 0. 25mM cadmium at various time periods (0 to 120 min) at room temperature (26±20C) and the residual enzyme activity was monitored. The activity of enzyme was also recorded at these time points in absence of Cadmium, which served as a control. The reaction rate measured soon after mixing the enzyme with other reagents without any further incubation was used as zero time reaction. The data of percent residual activity and the time of incubation in min were extrapolated at Y and X-axes, respectively. The t1/2 value (the time at which the enzyme activity remains half of the original under this condition) was calculated from this plot.

Estimation of IC50 value for cadmium

The enzyme (132μg) was assayed in the presence of different concentrations of cadmium nitrate and the residual activity was monitored. The activity recorded in absence of cadmium was considered as 100%. The IC50 value was calculated by extrapolating the data taking percent residual activity on Y-axis and the varying cadmium concentrations at X-axis on a graph.

Determination of mode of inhibition of AChE by cadmium

The enzyme (132 μg) was assayed at varying concentrations of ATI at room temperature (26±2°C) in the absence and presence of cadmium (1.0 mM). The Ki and Vmax values were calculated using the intersections by the straight line at Y and at the negative abscissa of X-axes, respectively, of the Lineweaver- Burk's double reciprocal plot. 2.10.

Determination of K value for cadmium in mode of inhibition of AChE from rat brain

The mode of inhibition of enzyme by cadmium was determined by assaying the enzyme mentioned as above using the formula of either

where V max+I and V max-I the maximal velocities of reactions in the presence and absence of cadmium, respectively. Similarly, Km +I and K m denote the Km values in the presence and absence mm of cadmium, respectively. [I] represents the concentration of inhibitor used i.e. 1.0 mM. The Ki value may also be calculated using

Statistical analysis of data

Statistical analysis of data was performed using Graph Pad Prism version 6 for windows. All values were expressed as mean standard deviation of 3 observations.

Results

Membrane bound nature of AChE from rat brain tissue

The enzyme protein content of brain homogenate from the rat brain tissue were solubilized using a nonionic detergent, 0.2% (v/v) Triton X-100 in phosphate buffer (50mM, pH 7.4). The extent of enzyme activity was more in the detergent solubilized fraction than that of without detergent. These results demonstrated the membrane bound nature of this enzyme. The protein contents in the soluble fractions of these two preparations were also found to be significantly different. The fraction obtained supernatant with Triton X-100 contained 3.30 mg/ml protein as against 1.20 mg/ml in the fraction without treatment with the detergent. The pellet with Triton X-100 contained 1.15 mg/ml protein against 3.15 mg/ml in the fraction without treatment with the detergent, thereby showing solubilization and release of proteins from the pellets in presence of the detergent (Table 1).

*One unit of the activity of AChE has been defined as the micromoles of substrate hydrolyzed per min per ml.

Stability of enzyme activity after storage at -200C

The effect of storage time at -200C on the activity of AChE from rat brain tissue was determined by carrying out the enzyme assay employing 132μg proteins on different days as described in Materials and Methods. The enzyme was stored in phosphate buffer (50mM, pH 7.4) containing 0.2% (v/v) Triton X-100. The results shown in Table 2 indicated that the enzyme was highly stable up to 30 days with no much loss in activity. However, when this enzyme was assayed at varying temperatures, it exhibited maximum activity at 370C followed by gradual loss in its activity after increasing temperature as shown in Table 3. The enzyme was found to be optimally active at pH 7.4 when assayed using buffers of different pH systems under standard assay conditions as shown in Table 4. The buffers of higher pH values displayed inhibitory effect on enzyme activity.

Effect of substrate concentration on the activity of AChE from rat brain tissue

The enzyme (132μg) was assayed at varying concentrations of the substrate (ATI), at room temperature (26±20C). The enzyme activity at corresponding substrate concentration displayed a direct correlation and the results showed a hyperbolic curve (Figure 1). The Lineweaver Burk's double reciprocal plot of the data as shown in Figure 1 demonstrated a straight line which intersects at Y and negative abscissa of X-axes, from where the Vmax and Km values could be calculated; the values being 0.192μmoles ml-1min-1 and 0.037mM, respectively.

Effect of cadmium on the activity of AChE from rat brain tissue

The enzyme when assayed in presence of varying concentrations of cadmium (0.05-10mM) displayed consistent decrease in its activity (Table 5). When this data was extrapolated using percent residual activity and the cadmium concentrations on Y and X-axes, respectively, the IC50 value of this heavy metal for rat brain AChE could be determined, the value being 5.70mM (Figure 2).

Effect of time on cadmium mediated inhibition of AChE activity from rat brain tissue

The effect of time on the cadmium mediated inhibition of the AChE activity from rat brain at room temperature (26±20C) was monitored by assaying the enzyme (132μg) at different time intervals in absence and presence of the cadmium (0.25mM). The enzyme activity in the absence of cadmium served as a control. The results as presented in Figure 3 demonstrated that the enzyme activity increased up to 15 min and after that it decreased consistently with respect to the increasing incubation time in presence of cadmium. The results from present study also demonstrated that cadmium at a concentration of 0.25mM did not shown any effect on its activity up to 15 min but it shows sharp inhibition of the enzyme in a time dependent manner at 26±20C temperature. The enzyme activity remained about 50% after 77 min of incubation, i.e. the t1/2 time. However, the enzyme did not show any decrease in activity in the absence of cadmium under similar experimental conditions.

Evaluation of mode of inhibition of AChE from rat brain by cadmium

The above experiments indicated that treatment of rat brain AChE with cadmium resulted into decline in its activity. In order to ascertain the mechanism of inhibition of AChE activity by cadmium, the enzyme (132μg) was assayed at varying substrate concentrations in absence and presence of cadmium (1.0mM). The data were used to extrapolate Lineweaver Burk's double reciprocal plot, which developed two straight lines originated from same point on X-axis and intersecting at different points on Y axis (non-competitive inhibition). The V max+I (Vmax value in presence of cadmium) and Ki values were calculated using these points and found to be 0.069 μmoles/ml/min and 0.548mM, respectively. The results are shown in Figure 4.

Discussion

Before an enzyme can be characterized, it must be extracted and solubilized from the selected tissue. Some of the enzymes are membrane-bound and their solubilization has been shown to be achieved by using organic solvents, detergents, proteolytic and lipolytic enzymes [21]. It has been shown that EDTA and tetracaine increased the amount of enzyme extracted from calf brain, and a preliminary studies showed that these compounds had a similar effect on the extraction of enzyme from mouse brain. The results from the present study demonstrated the membrane bound form of rat brain AChE which could be easily solubilized using Triton X- 100. In some other living systems also the membrane bound nature of AChE has been indicated [22-24]. However, AChE has been shown to exist only in a membrane bound form in the human erythrocytes and brain, electric eel, electric fish and certain parasite helminthes [22,23]. The enzyme from these systems has been solubilized by applying both the ionic as well as non-ionic detergents in the phosphate buffer [22]. The Km value for any enzyme is a measure of its affinity towards its substrate. In the present study, the Km value for rat brain AChE was 37μM which was about three times lower than that reported for the enzyme from human brain (107μM), and fetal bovine serum (120 μM) [25]. The rat brain AChE displayed Km about 6times lower than that reported for the analogous enzyme from the human erythrocytes (225μM). The relatively lower Km value for rat brain AChE indicated its enhanced affinity to the substrate as compared to other mammalian systems [22]. The entry of cadmium in mammalian brain has been demonstrated. It is therefore presumed that it may alter brain function. The results from the present study displayed that cadmium caused strong inhibition of rat brain AChE (IC50=5.7mM). These results indicate that cadmium may act as an inducer of toxic stress on the neurotransmission system of rat. In the present study, cadmium inhibited the activity of rat brain AChE in noncompetitive manner when tested in vitro. These results suggest the binding of cadmium at a different site on the enzyme surface other than the active site. No such reports are available from other workers to be used for the sake of the comparison.

Conclusion

The results from the present study indicated the presence of membrane bound form of AChE in the rat brain which could be solubilized employing a non-ionic detergent, Triton X-100. Cadmium sharply inhibited this enzyme at low concentration indicating thereby its strong neurotoxic potential to the mammals. Though the exact mechanism of action of cadmium on brain AChE is not known but this study presented evidence that this heavy metal may inhibit the enzyme in a noncompetitive manner. Thus, the rat brain AChE may serve as a potential biomarker of neurotoxicity induced by cadmium. The information obtained from this study may be useful in proper risk management of cadmium toxicity particularly in those who are occupationally engaged in cadmium infested environment.

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