Jin et al 2026a - Revision history

Tomamil: Tomamil moved page Review 382597607607 to Jin et al 2026a

2026-03-09T16:38:10Z

Tomamil moved page Review 382597607607 to Jin et al 2026a

TianJin at 02:09, 20 February 2026

2026-02-20T02:09:16Z

NZHNG at 23:01, 18 February 2026

2026-02-18T23:01:50Z

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TianJin: Corrected inaccurate statement about NMDA antagonists and removed illustative figures

2026-02-18T20:24:52Z

Corrected inaccurate statement about NMDA antagonists and removed illustative figures

TianJin: Fix references section

2026-02-16T05:23:05Z

Fix references section

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TianJin: Redid all citations. Fixed version error resulting from a communication issue.

2026-02-16T05:11:09Z

Redid all citations. Fixed version error resulting from a communication issue.

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Ashley at 03:51, 16 February 2026

2026-02-16T03:51:49Z

Ashley at 03:42, 16 February 2026

2026-02-16T03:42:20Z

NZHNG at 15:11, 13 February 2026

2026-02-13T15:11:15Z

NZHNG at 03:01, 13 February 2026

2026-02-13T03:01:54Z

@@ Line 176: / Line 176: @@
 The paralysis assay was followed by a chemotaxis assay to evaluate whether Aβ attenuation translates to broader improvements in neurological function. The chemotaxis assay integrates sensory perception, motor output, learning, and memory, making it a comprehensive marker of neurotoxicity. CL2355 produces Aβ pan-neuronally through the synaptobrevin ortholog (snb-1) promoter, leading to deficits in serotonin sensitivity, neuronal function, and chemotaxis. The combined treatment produced the greatest improvement in chemotactic ability among Aβ-expressing groups (0.312), significantly outperforming the untreated control (p<0.01). Ginseng alone (0.223) and DHA alone (0.188) showed comparable intermediate effects, both significantly higher than the untreated control (-0.035) but significantly lower than the combined treatment (Fig. 3). These findings suggest that the combined treatment exerts a neuroprotective effect that extends beyond motor function, partially restoring complex neurological processes.
-The combined treatment had marginal effects on population growth compared to CL -G/-O (Fig. 5; ADD combined data vs. no treatment). However, as incomplete sterility is a characteristic of the CL2355 strain and some AD strains exhibit normal reproduction, it remains unclear whether impaired reproduction in CL2355 is directly attributable to Aβ pathology.
+The combined treatment had marginal effects on population growth compared to CL -G/-O (Fig. 5). However, as incomplete sterility is a characteristic of the CL2355 strain and some AD strains exhibit normal reproduction, it remains unclear whether impaired reproduction in CL2355 is directly attributable to Aβ pathology.
 Collectively, these findings support the original hypothesis that combined treatment would more effectively alleviate AD symptoms in C. elegans, as groups receiving the combination demonstrated lower rates of paralysis and significantly higher chemotaxis scores than either treatment alone.

@@ Line 14: / Line 14: @@
 Alzheimer’s disease (AD) is a neurodegenerative disease and the leading cause of dementia, accounting for 75% of cases [1]. As of 2014, AD affects more than 35.6 million people worldwide, a number estimated to quadruple by 2050. AD impairs language recognition and cognitive function by damaging the neocortex and hippocampus, reducing life expectancy, and increasing physical impairment in the elderly [2, 1]. Although the etiology of AD remains incompletely understood, current research is primarily based on a theory attributing its cause to abnormal accumulation of amyloid-beta (Aβ) aggregates, which results in neuroinflammation and subsequent neurodegeneration [3, 4, 5, 6].
-Currently, FDA-approved AD treatments target cholinergic and glutamatergic systems [7]. Cholinesterase inhibitors (CHEIs) reduce the breakdown of acetylcholine but are unable to halt disease progression or reverse damage [2]. NMDA receptor antagonists, which reduce glutamate excitotoxicity, the excessive activation of glutamate receptors leading to neuronal atrophy, represent another therapeutic class [8]. However, dysregulation of NMDA receptors, which play a critical role in synaptic transmission and plasticity underlying learning and memory, is itself implicated in AD pathology [9, 10, 11].
+Currently, FDA-approved AD treatments target cholinergic and glutamatergic systems [7]. Cholinesterase inhibitors (CHEIs) reduce the breakdown of acetylcholine but are unable to halt disease progression or reverse damage [2]. NMDA receptor antagonists represent another therapeutic class. NMDA receptors play a critical role in synaptic transmission and plasticity underlying learning and memory, but their excessive activation by glutamate leads to Ca²⁺ overload and neuronal death, a process implicated in AD pathogenesis [8]. Memantine, the only clinically approved NMDA antagonist for AD, selectively blocks this pathological overactivation while preserving normal synaptic signaling. However, like CHEIs, it provides only symptomatic relief and cannot prevent further AD progression [9, 10, 11].
-More broadly, drugs that solely target Aβ have repeatedly failed to pass Stage 3 trials. Over the years, hundreds of therapies have been proposed and developed to clear Aβ; Yet none, with the exception of Donanemab, has improved clinical outcomes to a significant extent. Researchers attribute Donanemab’s relative effectiveness to its unique ability to specifically target neurotoxic forms of Aβ, unlike treatments that target harmless or even protective Aβ species indiscriminately [12]. However, Donanemab is effective only in the early stages of AD, carries a risk of brain swelling, and costs approximately $32,000 per year, placing it out of reach for many patients [13]. These persistent limitations have shifted attention toward multi-target therapeutics, drugs that target pathology through numerous mechanisms, to better address the multifactorial etiology of AD [14]. The present study evaluated the efficacy of the combined treatment of two established multitarget AD therapies, DHA and Panax Ginseng extract, to assess their potential for synergistic effects, such as increased bioavailability, and contribute to the search for more accessible therapies.
+More broadly, drugs that solely target Aβ, which theoretically halt disease progression and potentially reverse existing damage, have repeatedly failed to pass Stage 3 trials. Over the years, hundreds of therapies have been proposed and developed to clear Aβ; Yet none, with the exception of Donanemab, has improved clinical outcomes to a significant extent. However, Donanemab is effective only in the early stages of AD, carries a risk of brain swelling, and costs approximately $32,000 per year, placing it out of reach for many patients [13]. These persistent limitations have shifted attention toward multi-target therapeutics, drugs that target pathology through numerous mechanisms, to better address the multifactorial etiology of AD [14]. The present study evaluated the efficacy of the combined treatment of two established multitarget AD therapies, DHA and Panax Ginseng extract, to assess their potential for synergistic effects, such as increased bioavailability, and contribute to the search for more accessible therapies.
 ===Omega-3 fatty acids (ω-3FAs) ===
@@ Line 24: / Line 24: @@
 Beyond targeting hallmark AD pathology (Aβ plaques, neurofibrillary tangles), ω-3FAs may attenuate cognitive and motor decline through additional mechanisms. A systematic review of eleven studies (n=698) found that ω-3FA supplementation substantially increased serum brain-derived neurotrophic factor (BDNF) levels at moderate or high doses [25]. BDNF is a neurotrophin critical for synaptic formation and maintenance, with established roles in learning, memory, emotion, sensory integration, motor function, executive function, and stress response—all of which are impaired with AD [26, 27]. Additionally, AD patients exhibit reduced BDNF levels, an association that may both result from and exacerbate disease progression, though whether the relationship is causal remains an area of active investigation [28, 29].
-ω-3FAs also possess potent anti-neuroinflammatory properties. In the aging brain, misfolded proteins, primarily Aβ aggregates and neurofibrillary tangles (NFTs), trigger innate immune responses in microglia, initiating a neurodegenerative cascade (Fig. 1). ApoE-4, a major genetic risk factor for AD, further amplifies this process by upregulating glial activation (relative to ApoE-2 or ApoE-3) through lipid modulation [30]. Although ω-3FAs have been extensively studied for their anti-inflammatory effects in cardiology and metabolic health [31], recent evidence demonstrates that dietary ω-3FAs also traverse the BBB and reduce neuroinflammation directly. In a randomized controlled trial of 33 mild AD patients, six months of DHA-based ω-3FA supplementation (2.3 g) reduced phosphorylated tau and lowered neuroinflammatory biomarkers, including interleukin-1 and interleukin-6 [32].
+ω-3FAs also possess potent anti-neuroinflammatory properties. In the aging brain, misfolded proteins, primarily Aβ aggregates and neurofibrillary tangles (NFTs), trigger innate immune responses in microglia, initiating a neurodegenerative cascade [33]. ApoE-4, a major genetic risk factor for AD, further amplifies this process by upregulating glial activation (relative to ApoE-2 or ApoE-3) through lipid modulation [30]. Although ω-3FAs have been extensively studied for their anti-inflammatory effects in cardiology and metabolic health [31], recent evidence demonstrates that dietary ω-3FAs also traverse the BBB and reduce neuroinflammation directly. In a randomized controlled trial of 33 mild AD patients, six months of DHA-based ω-3FA supplementation (2.3 g) reduced phosphorylated tau and lowered neuroinflammatory biomarkers, including interleukin-1 and interleukin-6 [32].
-−
-[[Image:Draft_Zhang_928330811-image1.png|632x632px]]
-−
-'''Figure 1: Pathomechanism sequelae of immune activation''' [33]
 ===Panax ginseng===
-Panax ginseng, a perennial plant of the Araliaceae family, has been used in East Asian medicine since approximately 1500 CE and is among the earliest herbal medicines applied to the treatment of dementia and neurological disorders [7, 34]. Ginsenosides, the primary bioactive compounds isolated from Panax ginseng, exhibit multi-target neuroprotective properties relevant to AD: anticholinesterase activity, neuroplasticity promotion, anti-Aβ and anti-tau effects, anti-inflammatory and antioxidant activity, anti-apoptotic effects, alleviation of insulin resistance, and BBB strengthening [34, 35]. Multiple human trials support ginseng’s efficacy in AD prevention and amelioration [36], and researchers have emphasized the need for further investigation into ginseng-based drug combinations [34].
+Panax ginseng, a perennial plant of the Araliaceae family, has been used in East Asian medicine since approximately 1500 CE and is among the earliest herbal medicines applied to the treatment of dementia and neurological disorders [7, 34]. Ginsenosides, the primary bioactive compounds isolated from Panax ginseng, exhibit multi-target neuroprotective properties relevant to AD: anticholinesterase activity; increased neuroplasticity; Aβ and tau clearance; anti-inflammatory and antioxidant activity; anti-apoptotic effects; alleviation of insulin resistance; and BBB strengthening [34, 35]. Multiple human trials support ginseng’s efficacy in AD prevention and amelioration [36], and researchers have emphasized the need for further investigation into ginseng-based drug combinations [34].
 Evidence from animal models corroborates these findings. In transgenic AD mice, fermented ginseng extract ameliorated memory dysfunction concurrent with reduced Aβ42 brain accumulation [37]. Red, black, and white ginseng extracts inhibit acetylcholinesterase and reduce hippocampal Aβ oligomer-induced memory impairment [38]. In the present study, ginseng was administered as Korean red ginseng, which contains the highest concentration of ginsenosides among ginseng preparations. Korean red ginseng treatment has demonstrated improved frontal lobe function via quantitative electroencephalography [39], and its cognitive benefits have been sustained for over two years in AD patients [40].
@@ Line 44: / Line 40: @@
 The BBB’s role in ω-3FA transport is directly relevant to this study [44, 45, 21]. Although ω-3FA supplementation shows strong preventive efficacy, contradictory results in post-onset cohorts impede broader clinical recommendations. In a randomized controlled trial of 174 patients with mild to moderate AD, moderate-dose ω-3FAs failed to improve cognitive outcomes relative to placebo over six months [46]. While the short study duration likely played a role, existing BBB damage in the AD patients may also be a contributing factor. When the BBB becomes compromised, the brain’s ability to uptake of DHA drastically deteriorates because the amount of DHA transporters, Mfsd2a, sharply decreases. Although the brains of young adults readily uptake ω-3FAs regardless of ApoE status, older adults, especially those with ApoE-4 and/or AD, show significantly reduced uptake [16]. A separate trial confirmed this pattern: DHA supplementation was significantly less effective at mitigating AD symptoms in ApoE-4 carriers compared to ApoE-3 carriers [47].
-Several approaches have been proposed to increase ω-3FA bioavailability in the brain, including co-administration with antioxidants and polyphenols. For instance, combining ω-3FAs with carotenoid and vitamin E improved cognitive outcomes in otherwise healthy older adults—an effect attributed to antioxidant-enhanced bioavailability [18]. However, ginseng may offer even greater synergistic potential. Ginsenoside Rg1 directly activates the Wnt/β-catenin pathway, the transcriptional regulator of Mfsd2a, thereby upregulating DHA transport across the BBB [48]. Additionally, ginseng may halt broader BBB degradation through inhibition of MMP-9, an enzyme that downregulates Mfsd2a in ApoE-4 carriers, and by crossing the BBB to reduce the neuroinflammation at the center of the neurodegenerative cascade [49; Fig. 1]. Preliminary studies in C. elegans further suggest that ginseng modulates lipid metabolism and extends healthspan by upregulating ω-3FA  (in the form of lipoic acid) metabolic signaling pathways; though whether this effect translates to DHA is unclear [50, 51]. Together, these findings suggest that ginseng may increase DHA absorption by the brain, thus bypassing the primary limitation of ω-3FA treatment for patients who have already developed AD.
+Several approaches have been proposed to increase ω-3FA bioavailability in the brain, including co-administration with antioxidants and polyphenols. For instance, combining ω-3FAs with carotenoid and vitamin E improved cognitive outcomes in otherwise healthy older adults—an effect attributed to antioxidant-enhanced bioavailability [18]. However, ginseng may offer even greater synergistic potential. Ginsenoside Rg1 directly activates the Wnt/β-catenin pathway, the transcriptional regulator of Mfsd2a, thereby upregulating DHA transport across the BBB [48]. Additionally, ginseng may halt broader BBB degradation through inhibition of MMP-9, an enzyme that downregulates Mfsd2a in ApoE-4 carriers, and by crossing the BBB to reduce the neuroinflammation at the center of the neurodegenerative cascade [49, 52]. Preliminary studies in C. elegans further suggest that ginseng modulates lipid metabolism and extends healthspan by upregulating ω-3FA  (in the form of lipoic acid) metabolic signaling pathways; though whether this effect translates to DHA is unclear [50, 51]. Together, these findings suggest that ginseng may increase DHA absorption by the brain, thus bypassing the primary limitation of ω-3FA treatment for patients who have already developed AD.
-−
-[[Image:Draft_Zhang_928330811-image2.png|619x619px]]
-−
-'''Figure 2. Schematic representation of the blood–brain barrier''' [52]
 ===Caenorhabditis elegans===
@@ Line 180: / Line 172: @@
 This mechanism aligns with the increasingly influential Aβ Oligomer Hypothesis. Although the amyloid cascade theory proposed in 1992 served as the guiding framework of AD etiology, many researchers have since deviated from or modified this hypothesis, forming distinct schools of thought, each backed by extensive research, including BBB breakdown, mitochondrial dysfunction, and the Aβ oligomer hypothesis [60, 61, 62]. The latter identifies soluble Aβ oligomers, rather than insoluble plaques, as the key neurotoxic species driving neuroinflammation, neuronal damage, and ultimately AD pathogenesis.
-Over the years, hundreds of candidate therapies have been developed to clear Aβ, yet none — with the exception of Donanemab — has improved clinical outcomes to a significant extent. Researchers attribute Donanemab's relative effectiveness to its ability to specifically target neurotoxic forms of Aβ, including oligomers, unlike treatments that indiscriminately target harmless or even protective Aβ species [12]. However, with the drug currently costing $32,000 per year and proving effective only in the early stages of AD, widespread clinical adoption remains limited [13]. As the combined treatment of ginseng and DHA similarly appears to target neurotoxic Aβ species, it may serve as a more accessible alternative with multi-target benefits extending beyond Aβ clearance alone.
+Currently, Donanemab is the only drug that the FDA has approved to treat AD via Aβ clearance. Researchers attribute Donanemab's relative effectiveness to its ability to specifically target neurotoxic forms of Aβ, including oligomers, unlike treatments that indiscriminately target harmless or even protective Aβ species [12]. However, with the drug currently costing $32,000 per year and proving effective only in the early stages of AD, widespread clinical adoption remains limited [13]. As the combined treatment of ginseng and DHA similarly appears to target neurotoxic Aβ species, it may serve as a more accessible alternative with multi-target benefits extending beyond Aβ clearance alone.
 The paralysis assay was followed by a chemotaxis assay to evaluate whether Aβ attenuation translates to broader improvements in neurological function. The chemotaxis assay integrates sensory perception, motor output, learning, and memory, making it a comprehensive marker of neurotoxicity. CL2355 produces Aβ pan-neuronally through the synaptobrevin ortholog (snb-1) promoter, leading to deficits in serotonin sensitivity, neuronal function, and chemotaxis. The combined treatment produced the greatest improvement in chemotactic ability among Aβ-expressing groups (ADD CHEMOTAXIS SCORE), significantly outperforming the untreated control (p<0.01). Ginseng alone (ADD SCORE) and DHA alone (ADD SCORE) showed comparable intermediate effects, both significantly higher than the untreated control (ADD SCORE) but significantly lower than the combined treatment. These findings suggest that the combined treatment exerts a neuroprotective effect that extends beyond motor function, partially restoring complex neurological processes.

@@ Line 134: / Line 134: @@
 ===Bacterial aseptic technique===
-'' E. coli ''OP50 (Source: [https://cgc.umn.edu/strain/OP50 https://cgc.umn.edu/strain/OP50]) was used to feed ''C. elegans strains'' CL2355 (Sourced: [https://cgc.umn.edu/strain/CL2355 https://cgc.umn.edu/strain/CL2355]) and N2 (Sourced: [https://cgc.umn.edu/strain/N2 https://cgc.umn.edu/strain/N2]) and placed in all Petri dishes. A rubber tube was connected to the valve of the gas and the valve of a Bunsen burner. Once the gas ignited, a match was lit and placed over the gas. The inoculation loop was sterilized by passing the tip and the neck of the inoculation loop through the flame. After the inoculation loop cooled, it was used to scoop the agar from a bacterial colony. The bacteria were streaked across the surface of a new Petri dish, ensuring that there was no excessive pressure put on the agar to refrain from breaking it. After, the lid was placed back on to the Petri dish and flipped upside down in the incubator to ensure that the accumulated condensation does not fall on the agar. Following two days of the Petri dishes being incubated, parafilm was wrapped around the sides of the Petri dish and placed into the fridge to store for later use. Before and after each process, diluted bleach was used to sterilize all surfaces.
+''E. coli ''OP50 (Source: [https://cgc.umn.edu/strain/OP50 https://cgc.umn.edu/strain/OP50]) was used to feed ''C. elegans strains'' CL2355 (Sourced: [https://cgc.umn.edu/strain/CL2355 https://cgc.umn.edu/strain/CL2355]) and N2 (Sourced: [https://cgc.umn.edu/strain/N2 https://cgc.umn.edu/strain/N2]) and placed in all Petri dishes. A rubber tube was connected to the valve of the gas and the valve of a Bunsen burner. Once the gas ignited, a match was lit and placed over the gas. The inoculation loop was sterilized by passing the tip and the neck of the inoculation loop through the flame. After the inoculation loop cooled, it was used to scoop the agar from a bacterial colony. The bacteria were streaked across the surface of a new Petri dish, ensuring that there was no excessive pressure put on the agar to refrain from breaking it. After, the lid was placed back on to the Petri dish and flipped upside down in the incubator to ensure that the accumulated condensation does not fall on the agar. Following two days of the Petri dishes being incubated, parafilm was wrapped around the sides of the Petri dish and placed into the fridge to store for later use. Before and after each process, diluted bleach was used to sterilize all surfaces.
 ===Spotting Petri dishes with E. coli OP50===
@@ Line 158: / Line 158: @@
 ===Safety===
-Proper lab safety, including wearing gloves, goggles, and tied hair during the experimentation process, was fulfilled especially when conducting bacterial works. The tabletop working area was cleansed with a diluted bleach solution before and after experimentation along with the sterilization of all instruments at the end of the trial period. All Petri plates were sealed with parafilm during experimentation and autoclaved at 120 ℃ and 100 kPa for 20 minutes as part of the disposal procedure. Tools in contact with cultures were also soaked in 10% bleach and sterilized with heat as a segment of the disposing process. Hands were washed after any bacterial work. Three days of safety training specifically for bacterial works and the presence of experienced supervisors were ensured whilst dealing with microorganisms and the flame or gas of a Bunsen burner required for sterilization.
+Proper lab safety, including wearing gloves, goggles, and tied hair during the experimentation process, was fulfilled especially when conducting bacterial works. The tabletop working area was cleansed with a diluted bleach solution before and after experimentation along with the sterilization of all instruments at the end of the trial period. All Petri plates were sealed with parafilm during experimentation and autoclaved at 120 ℃ and 100 kPa for 20 minutes as part of the disposal procedure. Tools in contact with cultures were also soaked in 10% bleach and sterilized with heat as a segment of the disposing process. Hands were washed after any bacterial work or assay procedure. Three days of safety training specifically for bacterial works and the presence of experienced supervisors were ensured whilst dealing with microorganisms and the flame or gas of a Bunsen burner required for sterilization.
 Three days of safety training specifically for bacterial works and the presence of experienced supervisors were ensured while dealing with microorganisms and the flame or gas of a Bunsen burner required for sterilization of tools. Most substances utilized in the study possessed a biosafety level BSL-1 and deemed “not hazardous” under the criteria of the federal OSHA Hazard Communication Standard 29CFR 1910.1200, and Regulation (EC) No 1272/2008 (GHS). Nonetheless, substances specifically can be dangerous when ingested, inhaled, or especially in cases of in contact with eyes (skin contact and inhalation do not pose serious harm unless it is done excessively or if the researcher feels unwell after exposure) so precautionary safety such as protective gloves, clothing, and above all else eye protection was adhered to.
@@ Line 164: / Line 164: @@
 ===Data Analysis===
-Data was collected, stored, and analyzed using Microsoft Excel. Data was shown using scatter plots and bar graphs in which the x values were time in hours, time in days, and group names and the y values were not paralyzed ''C. elegans'' as a percentage, number of ''C. elegans'', and chemotaxis index. The median paralysis time was conducted using a Kaplan–Meier survival analysis. In the paralysis assay, the number of non-paralyzed ''C. elegans'' on each plate was converted into a percentage. The standard mean of error was calculated and represented as error bars added using Microsoft Excel. To calculate significance, one-way ANOVA followed by Tukey HSD was done using [https://astatsa.com/OneWay_Anova_with_TukeyHSD/ https://astatsa.com/OneWay_Anova_with_TukeyHSD/]. Significance was determined using p-values in which a p-value less than 0.05 was considered significant. Chemotaxis index was determined using the following formula:
+Data was collected, stored, and analyzed using Microsoft Excel. Data was then shown using scatter plots and bar graphs in which the x values were time in hours, time in days, and group names and the y values were not paralyzed ''C. elegans'' as a percentage, number of ''C. elegans'', and chemotaxis index. The median paralysis time was conducted using a Kaplan–Meier survival analysis. In the paralysis assay, the number of non-paralyzed ''C. elegans'' on each plate was converted into a percentage. The standard mean of error was calculated and represented as error bars added using Microsoft Excel. To calculate significance, one-way ANOVA followed by Tukey HSD was done using [https://astatsa.com/OneWay_Anova_with_TukeyHSD/ https://astatsa.com/OneWay_Anova_with_TukeyHSD/]. Significance was determined using p-values in which a p-value less than 0.05 was considered significant. Chemotaxis index was determined using the following formula:
 CI = (number of worms in attractant zone - number of worms in control zone)/total number of scored worms

@@ Line 134: / Line 134: @@
 ===Bacterial aseptic technique===
-'' E. coli ''OP50 (Source: [https://cgc.umn.edu/strain/OP50 https://cgc.umn.edu/strain/OP50]) was used to feed ''C. elegans''
+'' E. coli ''OP50 (Source: [https://cgc.umn.edu/strain/OP50 https://cgc.umn.edu/strain/OP50]) was used to feed ''C. elegans strains'' CL2355 (Sourced: [https://cgc.umn.edu/strain/CL2355 https://cgc.umn.edu/strain/CL2355]) and N2 (Sourced: [https://cgc.umn.edu/strain/N2 https://cgc.umn.edu/strain/N2]) and placed in all Petri dishes. A rubber tube was connected to the valve of the gas and the valve of a Bunsen burner. Once the gas ignited, a match was lit and placed over the gas. The inoculation loop was sterilized by passing the tip and the neck of the inoculation loop through the flame. After the inoculation loop cooled, it was used to scoop the agar from a bacterial colony. The bacteria were streaked across the surface of a new Petri dish, ensuring that there was no excessive pressure put on the agar to refrain from breaking it. After, the lid was placed back on to the Petri dish and flipped upside down in the incubator to ensure that the accumulated condensation does not fall on the agar. Following two days of the Petri dishes being incubated, parafilm was wrapped around the sides of the Petri dish and placed into the fridge to store for later use. Before and after each process, diluted bleach was used to sterilize all surfaces.
-−
-(Sourced: CL2355 strain: [https://cgc.umn.edu/strain/CL2355 https://cgc.umn.edu/strain/CL2355] and N2 strain: [https://cgc.umn.edu/strain/N2 https://cgc.umn.edu/strain/N2]) and placed in all Petri dishes. A rubber tube was connected to the valve of the gas and the valve of a Bunsen burner. Once the gas ignited, a match was lit and placed over the gas. The inoculation loop was sterilized by passing the tip and the neck of the inoculation loop through the flame. After the inoculation loop cooled, it was used to scoop the agar from a bacterial colony. The bacteria were streaked across the surface of a new Petri dish, ensuring that there was no excessive pressure put on the agar to refrain from breaking it. After, the lid was placed back on to the Petri dish and flipped upside down in the incubator to ensure that the accumulated condensation does not fall on the agar. Following two days of the Petri dishes being incubated, parafilm was wrapped around the sides of the Petri dish and placed into the fridge to store for later use. Before and after each process, diluted bleach was used to sterilize all surfaces.
 ===Spotting Petri dishes with E. coli OP50===

@@ Line 79: / Line 79: @@
 ===Creating Omega-3 stock solution (500 mM) ===
-Utilizing a top-loading balance 8.2g of DHA omega-3 was measured in an Erlenmeyer flask using a top-loading balance. In a graduated cylinder, 50 mL of sterilized M9 solution was measured. Using 1.5 mL Tween 80, the measured omega-3 was dissolved. The water and fatty acid emulsion was transferred into a vial and vortexed until it was fully homogeneous. The stock solution was stored in a light blocking tinted vial and placed in a dark and cool environment until its usage. Before each use, a vortex was used to ensure a homogeneous mixture of the treatment.
+Utilizing a top-loading balance 8.2g of DHA omega-3 was measured in an Erlenmeyer flask using a top-loading balance. In a graduated cylinder, 50 mL of sterilized M9 solution was measured. Using 1.5 mL Tween 80, the measured omega-3 was dissolved. The water and fatty acid emulsion was transferred into a vial and vortexed until fully homogeneous. Then, the stock solution was stored in a light blocking tinted vial and placed in a dark and cool environment until its usage. Before each use, a vortex was used to ensure a homogeneous mixture of the treatment.
 ===Creating Nematode Growth Media (NGM) agar plates ===
@@ Line 87: / Line 87: @@
 Half of the NGM agar was dedicated to ginseng groups by transferring them to a separate graduated cylinder. A 1000x dilution of ginsenoside solution (0.18 mL) was performed using 180 mL of NGM agar as the solvent. Then, the agar was poured into the Petri dish very quickly, covering the lid of the Petri dish and tapping the bottle to avoid possible contamination. The aforementioned procedure was repeated for all Petri dishes with a waiting time of 15-20 minutes until agar solidified in each dish prior to refrigerator storage for 24 hours before usage (keeping the Petri dish upside down to prevent condensation from falling on the agar).
-Each plate was then spotted with ''E. coli'' OP50 grown for 24 hours in LB Media. Upon transferring, plates were dried for an additional 24 hours. The previously made omega-3 stock solution was diluted from 1mL to 5 mM with sterilized water to a final volume of 100 mL (100x dilution). Then, a dispersion of 0.1 mL of DHA omega-3 aliquot to the solidified and spotted NGM in Groups CL -G/+O, and CL +G/+O was done using a spreader. Upon administration of treatment in each Petri dish, the mediums were allowed to dry for another 24 hours.
+Each plate was then spotted with ''E. coli'' OP50 grown for 24 hours in LB Media. Upon transferring, plates were dried for an additional 24 hours. The previously made omega-3 stock solution was diluted from 1mL to 5 mM with sterilized water to a final volume of 100 mL (100x dilution). Then, a dispersion of 0.1 mL of DHA omega-3 aliquot to the solidified and spotted NGM in Groups CL -G/+O, and CL +G/+O was done using a sterilized spreader. Upon administration of treatment in each Petri dish, the mediums were allowed to dry for another 24 hours.
 '''Table 1: Experimental Setup'''
@@ Line 130: / Line 130: @@
 ===Creating bacteria agar===
-The table was cleaned before beginning and preparing the materials for the study. All safety precautions were followed as well. A weighing paper was folded to make it easier to handle. On a weighing boat on a scale, 17.5 g of premix LB Agar powder was weighed. This powder was transferred to an Erlenmeyer flask and distilled water was added until reaching a final volume of 500 mL. The bottle cap and the top were sealed with aluminum foil tightly. The solution was then autoclaved in the liquid setting for 20 minutes. Following the autoclaving, the agar was cooled to about 55℃. A thin layer of LB agar was poured into each Petri dish, ensuring the liquid was properly and evenly spread. After putting on the lid of the Petri dishes, each plate was left to cool down until solidification. After 24 hours of the solidification process, the dish was flipped to avoid the condensation falling onto the agar. After this, the plates were placed in plastic bags in the refrigerator at 4℃.
+The table was cleaned before beginning and preparing the materials for the study, with researchers following standard safety procedures. A weighing paper was folded to make it easier to handle. On a weighing boat on a scale, 17.5 g of premix LB Agar powder was weighed. This powder was transferred to an Erlenmeyer flask and distilled water was added until reaching a final volume of 500 mL. The bottle cap and the top were sealed with aluminum foil tightly. The solution was then autoclaved in the liquid setting for 20 minutes. Following the autoclaving, the agar was cooled to about 55℃. A thin layer of LB agar was poured into each Petri dish, ensuring the liquid was properly and evenly spread. After putting on the lid of the Petri dishes, each plate was left to cool down until solidification. After 24 hours of the solidification process, the dish was flipped to avoid the condensation falling onto the agar. After this, the plates were placed in plastic bags in the refrigerator at 4℃.
 ===Bacterial aseptic technique===

← Older revision	Revision as of 16:38, 9 March 2026
(No difference)

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 [56]      Tangrodchanapong T, Sobhon P and Meemon K 2020 Frondoside A Attenuates Amyloid-β Proteotoxicity in Transgenic Caenorhabditis elegans by Suppressing Its Formation ''Front. Pharmacol.'' '''11'''
-[57]      Bhatia S, Rawal R, Sharma P, Singh T, Singh M and Singh V 2022 Mitochondrial Dysfunction in Alzheimer’s Disease: Opportunities for Drug Development ''Curr Neuropharmacol'' '''20''' 675–92
+[57]     Bhatia S, Rawal R, Sharma P, Singh T, Singh M and Singh V 2022 Mitochondrial Dysfunction in Alzheimer’s Disease: Opportunities for Drug Development ''Curr Neuropharmacol'' '''20''' 675–92
-[58]      Sweeney M D, Sagare A P and Zlokovic B V 2018 Blood–brain barrier breakdown in Alzheimer disease and other neurodegenerative disorders ''Nat Rev Neurol'' '''14''' 133–50
+[58]   Sweeney M D, Sagare A P and Zlokovic B V 2018 Blood–brain barrier breakdown in Alzheimer disease and other neurodegenerative disorders ''Nat Rev Neurol'' '''14''' 133–50
 [59]      Cline E N, Bicca M A, Viola K L and Klein W L 2018 The Amyloid-β Oligomer Hypothesis: Beginning of the Third Decade ''Journal of Alzheimer’s Disease'' '''64''' S567–610