New approach to generating of human monoclonal antibodies specific to the proteolytic domain of botulinum neurotoxin A.

Introduction: Botulinum neurotoxins (BoNTs) cause botulism and are the most potent natural toxins known. Immunotherapy with neutralizing monoclonal antibodies (MAbs) is considered to be the most effective immediate response to BoNT exposure. Hybridoma technology remains the preferred method for producing MAbs with naturally paired immunoglobulin genes and with preserved innate functions of immune cells. The affinity-matured human antibody repertoire may be ideal as a source for antibody therapeutics against BoNTs. In an effort to develop novel BoNT type A (BoNT/A) immunotherapeutics, sorted by flow cytometry plasmablasts and activated memory B cells from a donor repeatedly injected with BoNT/A for aesthetic botulinum therapy could be used due to obtain hybridomas producing native antibodies. Methods: Plasmablasts and activated memory B-cells were isolated from whole blood collected 7 days after BoNT/A injection and sorted by flow cytometry. The sorted cells were then electrofused with the K6H6/B5 cell line, resulting in a producer of native human monoclonal antibodies (huMAbs). The 3 antibodies obtained were then purified by affinity chromatography, analyzed for binding by Western blot assay and neutralization by FRET assay. Results: We have succeeded in creating 3 hybridomas that secrete huMAbs specific to native BoNT/A and the proteolytic domain (LC) of BoNT/A. The 1B9 antibody also directly inhibited BoNT/A catalytic activity in vitro. Conclusion: The use activated plasmablasts and memory B-cells isolated at the peak of the immune response (at day 7 of immunogenesis) that have not yet completed the terminal stage of differentiation but have undergone somatic hypermutation for hybridization allows us to obtain specific huMAbs even when the immune response of the donor is weak (with low levels of specific antibodies and specific B-cells in blood). A BoNT/A LC-specific antibody is capable of effectively inhibiting BoNT/A by mechanisms not previously associated with antibodies that neutralize BoNT. Antibodies specific to BoNT LC can be valuable components of a mixture of antibodies against BoNT exposure.

Tirbanibulin (KX2-391) analog KX2-361 inhibits botulinum neurotoxin serotype A mediated SNAP-25 cleavage in pre- and post-intoxication models in cells.

Botulinum neurotoxins (BoNT) inhibit neuroexocytosis, leading to the potentially lethal disease botulism. BoNT serotype A is responsible for most human botulism cases, and there are no approved therapeutics to treat already intoxicated patients. A growing body of research has demonstrated that BoNT/A can escape into the central nervous system, and therefore, identification of BoNT/A inhibitors that can penetrate BBB and neutralize the toxin within intoxicated neurons would be important. We previously identified an FDA-approved, orally bioavailable compound, KX2-391 (Tirbanibulin) that inhibits BoNT/A in motor neuron assays. Recently, a structural analog of KX2-391, KX2-361, has been shown to exhibit good oral bioavailability and cross BBB with high efficiency in mouse experiments. Therefore, in this work, we evaluated the inhibitory effects of KX2-361 against BoNT/A. Toward this goal, we first evaluated the compound for its effects on cell viability in PC12 cells, via MTT assay, and in mouse embryonic stem cell (mESC)-derived motor neurons, with imaging-based assays. Following, we tested KX2-361 in mESC-derived motor neurons intoxicated with BoNT/A holotoxin, and the compound exhibited activity against the toxin in both pre- and post-intoxication conditions. Excitingly, KX2-361 also inhibited BoNT/A enzymatic component (light chain; LC) in PC12 cells transfected with BoNT/A LC. Furthermore, our molecular docking analyses suggested that KX2-361 can directly bind to BoNT/A LC. Medicinal chemistry approaches to develop structural analogs of KX2-361 to increase its efficacy against BoNT/A may provide a critical lead compound with BBB penetration capacity for drug development efforts against BoNT/A intoxication.

A simple and rapid in vitro assay for identification of direct inhibitors of O6-methylguanine-DNA methyltransferase.

O6-Methylguanine-DNA methyltransferase (MGMT) is a DNA repair enzyme that is overexpressed in certain tumors and is associated with resistance to the DNA alkylating agent temozolomide. MGMT inhibitors show potential in combating temozolomide resistance, but current assays for MGMT enzyme activity and inhibition, primarily oligonucleotide-based and fluorescent probe-based, are laborious and costly. The clinical relevance of temozolomide therapy calls for more convenient methodologies to study MGMT inhibition. Here, we extended the application of SNAP-Capture magnetic beads to develop a novel MGMT inhibition assay that demonstrated efficacy not only with known MGMT inhibitors, but also with the aldehyde dehydrogenase inhibitor, disulfiram. The assay uses standard fluorescence microscopy as a simple and reliable detection method, and is translationally applicable in drug discovery programs.

A cell line expressing MGMT-GFP fusion protein was generated. After harvesting the cells, the cell lysate was prepared and combined with SNAP-Capture magnetic beads and incubated at room temperature. Successful immobilization of MGMT-GFP on SNAP-Capture magnetic beads was verified by fluorescence microscopy. For the MGMT inhibition assay, the cell lysate underwent pre-treatment with established MGMT inhibitors before interaction with SNAP-capture magnetic beads and then underwent immobilization and fluorescence microscopy.

Psychosocial factors play a central role in determining SNAP utilization for farm workforce.

Introduction: Federal food safety net programs, like the Supplemental Nutrition Assistance Program (SNAP), may not reach vulnerable populations like rural residents, immigrants, and Latinx individuals. Because these groups are overrepresented among the farm workforce, exploring SNAP utilization among farm communities may clarify the role it plays in alleviating food insecurity. Methods: In-depth interviews were conducted with 31 farmworkers and farm owners. Patterns and predictors of SNAP utilization were organized using an adapted Andersen Behavioral Model of Health Service Utilization. Results: Psychosocial factors played the central role in participants' use of SNAP. Discussion: Opportunities to improve the design and delivery of SNAP include expanded eligibility cut-offs and targeted engagement mechanisms.

Dithiothreitol prevents the spontaneous release of cortical granules in in vitro aged mouse oocytes by protecting regulatory proteins of cortical granules exocytosis and thickening the cortical actin cytoskeleton.

In assisted fertility protocols, in vitro culture conditions mimic physiological conditions to preserve gametes in the best conditions. After collection, oocytes are maintained in a culture medium inside the incubator until in vitro fertilization (IVF) is performed. This time outside natural and physiological conditions exposes oocytes to an oxidative stress that renders in vitro aging. It has been described that in vitro aging produces a spontaneous cortical granule (CG) release decreasing the fertilization rate of oocytes. Nevertheless, this undesirable phenomenon has not been investigated, let alone prevented. In this work, we characterized the spontaneous CG secretion in in vitro aged oocytes. Using immunofluorescence indirect, quantification, and functional assays, we showed that the expression of regulatory proteins of CG exocytosis was affected. Our results demonstrated that in vitro oocyte aging by 4 and 8 h altered the expression and localization of alpha-SNAP and reduced the expression of NSF and Complexin. These alterations were prevented by supplementing culture medium with dithiothreitol (DTT), which in addition to having a protective effect on those proteins, also had an unexpected effect on the actin cytoskeleton. Indeed, DTT addition thickened the cortical layer of fibrillar actin. Both DTT effects, together, prevented the spontaneous secretion of CG and recovered the IVF rate in in vitro aged oocytes. We propose the use of DTT in culture media to avoid the spontaneous CG secretion and to improve the success rate of IVF protocols in in vitro aged oocytes.

Double-network polyphenol chitosan hydrogels with instant aldehyde-ß-cyclodextrin-based structure as potential for treating bacterially infected wounds.

Treatment of multiple bacterial infected wounds by eliminating bacteria and promoting tissue regeneration remains a clinical challenge. Herein, dual-network hydrogels (CS-GA/A-ß-CD) with snap-structure were designed to achieve curcumin immobilization, using gallic acid-grafted chitosan (CS-GA) and aldehyde-ß-cyclodextrin (A-ß-CD) crosslinked. A-ß-CD were able to achieve rapid dissolution (≥222.35 mg/mL H2O), and helped CS-GA/A-ß-CD achieve rapid gelation (≤66.23 s). By adjusting the ratio of aldehyde groups of A-ß-CD, mechanical properties and drug release can be controlled. CS-GA/A-ß-CD/Cur exhibited excellent antimicrobial properties against S. aureus, E. coli, and P. aeruginosa. In vivo experiments demonstrated that CS-GA/A-ß-CD/Cur achieved acute bacterial infection wound healing after 20th days, proving its great potential for wound dressing.

Barriers and Facilitators to Online Grocery Shopping Among Supplemental Nutrition Assistance Program (SNAP) Participants: A Mixed Method Study.

This mixed methods study explored online grocery shopping perceptions by surveying individuals who do and do not receive SNAP benefits (n = 129) and by conducting interviews with SNAP recipients (n = 26) who have grocery shopped online. T-tests assessed survey findings, codebook thematic analysis was used to identify qualitative themes, and results were interpreted collectively. Survey results found no differences in perceptions of online grocery shopping between SNAP and non-SNAP recipients (p-values = 0.2-1.0) and that 97% of SNAP recipients felt comfortable using SNAP online. Five qualitative themes were identified and provided context to the survey results. The study findings can inform policy actions within SNAP.

Long COVID and the Higher Risk of Food Insecurity Among Participants and Nonparticipants of Food Assistance Programs in the United States.

BACKGROUND: In the aftermath of the COVID-19 public health emergency, it is important to understand the extent of socioeconomic burdens of long COVID, defined as continuation of symptoms after initial infection, including food insecurity. OBJECTIVE: This study aimed to assess the association between long COVID and family food insecurity among low-income individuals (or any of their family members living with them) who were participants and nonparticipants in public food assistance programs (Supplemental Nutrition Assistance Program [SNAP], Special Supplemental Nutrition Program for Women, Infants, and Children [WIC], and National School Lunch Program [NSLP]) in the United States. DESIGN: The study used an observational cross-sectional design. PARTICIPANTS/SETTING: Data on 7151 adults (aged 18+ years), with family income of < 200% of the Federal Poverty line, from the 2022 National Health Interview Survey, were analyzed. MAIN OUTCOME MEASURES: Level of family food security, based on responses to a set of 10 questions measuring family's food security during the past 30 days. STATISTICAL ANALYSES PERFORMED: Multinomial logistic regression models were estimated to obtain relative risk ratios of marginal and low/very low food security, relative to the base outcome of high food security, for long COVID status. Multivariable models were estimated separately for the full sample and for subgroups of food assistance participants and nonparticipants. A seemingly unrelated regression (SUR) specification was estimated to assess whether the estimates were different across the participant and nonparticipant subgroups. RESULTS: Compared with individuals who never had COVID-19, the adjusted relative risks of experiencing marginal and low/very low food security were 1.42 (95% CI, 1.00-2.02) and 1.43 (95% CI, 1.08-1.91) times higher, respectively, for individuals who had long COVID. Although the adjusted risks were not observed to be statistically significant in the participant subgroup, among nonparticipants, adjusted relative risk ratios were 2.34 (95% CI, 1.43-3.82) and 1.56 (95% CI, 1.02-2.39), respectively. SUR results showed that relationships between long COVID and food insecurity were only different for marginal and not low/very low levels of food security between food assistance participant and nonparticipant groups. CONCLUSIONS: Study findings highlight a significantly higher risk of marginal and low/very low- food security among low-income adults who had long COVID, especially those who were nonparticipants in public food assistance programs. Further research is warranted to explore the causal pathways of this relationship for informing policies to mitigate the burden of long COVID.

Comparative analysis of volumetric changes between resection volume of oral tongue cancer and post operative volume of radial forearm flaps.

OBJECTIVES: This study investigates the relationship between the total volume of oral tongue cancer pre-operatively and the RFFF volume post-operatively. MATERIALS AND METHODS: A total of 52 DICOM imaging datasets (CT or MRI) of 26 patients were included in this study. The volume of the desired structure was quantified using semi-automatic segmentation using the software ITK-SNAP. All extracted measurements were validated by two further clinicians at separate instances. RESULTS: The variation of MeanVolTu can be predicted by MeanVolFlap moderately reliable with 59.1% confidence (R-Qua: 0.591). ANOVA Testing to represent how well the regression line fits the data, resulted in the overall regression model being statistically significant in predicting the MeanVolTu (p < 0.001). The flap volume may be predicted using the following algorithm: MeanVolFlap0 = 3241,633 + 1, 322 * MeanVolTu. CONCLUSION: The results of this study show positive correlation between tumor volume and flap volume, highlighting the significance of efficient flap planning with increasing tumor volume. A larger extraction volume of the radial forearm free flap from the donor site compromises the forearm more, thus increasing the probability of post-operative complications. CLINICAL RELEVANCE: Radial forearm free flap design in accordance with its corresponding 3D tumor volume.

QTL mapping for pod quality and yield traits in snap bean (Phaseolus vulgaris L.).

Pod quality and yield traits in snap bean (Phaseolus vulgaris L.) influence consumer preferences, crop adoption by farmers, and the ability of the product to be commercially competitive locally and globally. The objective of the study was to identify the quantitative trait loci (QTL) for pod quality and yield traits in a snap × dry bean recombinant inbred line (RIL) population. A total of 184 F6 RILs derived from a cross between Vanilla (snap bean) and MCM5001 (dry bean) were grown in three field sites in Kenya and one greenhouse environment in Davis, CA, USA. They were genotyped at 5,951 single nucleotide polymorphisms (SNPs), and composite interval mapping was conducted to identify QTL for 16 pod quality and yield traits, including pod wall fiber, pod string, pod size, and harvest metrics. A combined total of 44 QTL were identified in field and greenhouse trials. The QTL for pod quality were identified on chromosomes Pv01, Pv02, Pv03, Pv04, Pv06, and Pv07, and for pod yield were identified on Pv08. Co-localization of QTL was observed for pod quality and yield traits. Some identified QTL overlapped with previously mapped QTL for pod quality and yield traits, with several others identified as novel. The identified QTL can be used in future marker-assisted selection in snap bean.

A sensitive cell-based assay for testing potency of Botulinum neurotoxin type A.

Botulinum neurotoxin type A (BoNT/A) is a widely used biopharmaceutic for the treatment of neurological diseases and aesthetic medicine, allowing months-long paralysis of target muscles and glands. Large numbers of mice are used for multiple botulinum applications including batch release potency testing, antitoxin testing, countermeasure development and basic research. The mouse bioassay (MBA) has historically been the industry gold-standard in the botulinum field and is still heavily used for commercial product testing. BoNT/A intoxication causes severe suffering and application-specific, non-animal alternatives are urgently needed. It is widely accepted, that a cell-based assay (CBA) is the only way to faithfully replicate all the physiological steps of botulinum intoxication; comprising neuronal binding, internalization, endosomal escape, and cleavage of synaptosomal-associated protein of 25 kDa (SNAP25). However, it has not been straightforward to develop these assays and there are only a limited number of CBA currently in use. This is in part, due to the fact that very few cell lines have the appropriate levels of sensitivity to BoNT/A. In this study we have identified that LAN5 cells, a human neuroblastoma derived cell line, are sensitive to BoNT/A and can be engineered to express a recombinant NanoLuc luciferase tagged SNAP25 reporter molecule. On intoxication, the reporter molecule is cleaved and releases a NanoLuc-SNAP25 fragment which can be specifically captured on a 96-well plate for quantitative luminometry. Importantly, we demonstrate this new cell-based assay exhibits sensitivity comparable to the MBA.

Botulinum neurotoxin type A (BoNT/A) is extensively used in the treatment of neurological disorders and aesthetics. When the toxin enters cells, it targets a protein called SNAP25 and inhibits neurotransmitter release. Traditionally, the potency and safety of BoNT/A has been tested using the mouse bioassay, which causes significant distress to the animals being used. Our study introduces a new method for detecting BoNT/A activity based on LAN5 cells, which are a self-replicating, neuroblastoma-derived human cell line. We have engineered the cells to express a version of SNAP25 that allows the potency of BoNT/A to be measured using a luminescence assay. This new cell-based assay is as sensitive as the mouse bioassay and can be used for commercial product testing. This development could lead to fewer animals being used in research and commercial testing of BoNT/A, benefiting both scientific progress and animal welfare.

Association between participation in the supplemental nutrition assistance program and perception of limited availability and choice of healthy foods.

BACKGROUND: Nutrition security encompasses stable and equitable access, availability, affordability, and utilization of healthy foods. AIM: To evaluate the relationship of two newly created dichotomous measures that represent aspects of nutrition security (i.e., perceived limited availability and healthfulness choice) with Supplemental Nutrition Assistance Program (SNAP) participation. METHODS: Logistic regression models were run for each outcome separately with adjustment for age, income-to-poverty ratio, gender, education, race, and food security. RESULTS: Adults using social services (e.g., food pantries) were enrolled (N = 402) in this cross-sectional analysis. SNAP participants (61.7%) were not different from non-SNAP participants in perceiving limited availability (aOR [95% CI]: 1.21 [0.75, 1.95]) or limited ability to choose (aOR [95% CI]: 0.69 [0.43, 1.12]) healthy foods. CONCLUSIONS: Both SNAP and non-SNAP participants with low socioeconomic status report limited availability of healthy foods in their environment and a limited ability to choose healthy foods.

Repetitive Transcranial Magnetic Stimulation-Mediated Neuroprotection in the 5xFAD Mouse Model of Alzheimer's Disease Through GABRG2 and SNAP25 Modulation.

Alzheimer's disease (AD) is a leading neurodegenerative disorder with substantial impacts on cognition and behavior. Repetitive transcranial magnetic stimulation (rTMS), a non-invasive neuromodulation technique, has been used to treat various neuropsychiatric disorders, but its efficacy in AD has not been thoroughly investigated. This study examines the neuroprotective effects of rTMS in the 5xFAD mouse model of AD, with a particular focus on its modulation of GABAergic neuronal activity via the GABRG2 and SNAP25 proteins. Transcriptomic sequencing of rTMS-treated 5xFAD mice revealed 32 genes influenced by the treatment, among which GABRG2 was identified as a critical modulatory target. Electrophysiological assessments, including whole-cell patch clamp recordings from frontal cortex neurons, demonstrated significant alterations in inhibitory synaptic currents following rTMS. Subsequent experiments involved sh-GABRG2 transduction combined with rTMS treatment (20Hz, 14 days), examining behavioral responses, GABAergic neuron functionality, cortical GABA expression, cerebrospinal fluid GABA concentrations, ß-amyloid accumulation, and pro-inflammatory cytokine levels. The results indicated notable improvements in behavioral performance, enhanced functionality of GABAergic neurons, and reductions in ß-amyloid deposition and neuroinflammation after rTMS treatment. Further analysis revealed that SNAP25 overexpression could counteract the negative effects of GABRG2 silencing, highlighting the crucial role of SNAP25 downstream of GABRG2 in mediating rTMS's therapeutic effects in AD. This research highlights rTMS's potential to modulate synaptic and vesicular transport mechanisms, offering a promising avenue for ameliorating symptoms of AD through neuroprotective pathways.

Moniezia benedeni drives the SNAP-25 expression of the enteric nerves in sheep's small intestine.

BACKGROUND: The neuroimmune network plays a crucial role in regulating mucosal immune homeostasis within the digestive tract. Synaptosome-associated protein 25 (SNAP-25) is a presynaptic membrane-binding protein that activates ILC2s, initiating the host's anti-parasitic immune response. METHODS: To investigate the effect of Moniezia benedeni (M. benedeni) infection on the distribution of SNAP-25 in the sheep's small intestine, the recombinant plasmid pET-28a-SNAP-25 was constructed and expressed in BL21, yielding the recombinant protein. Then, the rabbit anti-sheep SNAP-25 polyclonal antibody was prepared and immunofluorescence staining was performed with it. The expression levels of SNAP-25 in the intestines of normal and M. benedeni-infected sheep were detected by ELISA. RESULTS: The results showed that the SNAP-25 recombinant protein was 29.3 KDa, the titer of the prepared immune serum reached 1:128,000. It was demonstrated that the rabbit anti-sheep SNAP-25 polyclonal antibody could bind to the natural protein of sheep SNAP-25 specifically. The expression levels of SNAP-25 in the sheep's small intestine revealed its primary presence in the muscular layer and lamina propria, particularly around nerve fibers surrounding the intestinal glands. Average expression levels in the duodenum, jejunum, and ileum were 130.32 pg/mg, 185.71 pg/mg, and 172.68 pg/mg, respectively. Under conditions of M. benedeni infection, the spatial distribution of SNAP-25-expressing nerve fibers remained consistent, but its expression level in each intestine segment was increased significantly (P < 0.05), up to 262.02 pg/mg, 276.84 pg/mg, and 326.65 pg/mg in the duodenum, jejunum, and ileum, and it was increased by 101.06%, 49.07%, and 89.16% respectively. CONCLUSIONS: These findings suggest that M. benedeni could induce the SNAP-25 expression levels in sheep's intestinal nerves significantly. The results lay a foundation for further exploration of the molecular mechanism by which the gastrointestinal nerve-mucosal immune network perceives parasites in sheep.

Supplemental Nutrition Assistance Program and depressive symptoms among adults with low family income in the U.S.: The National Health and Nutrition Examination Survey 2011-2018.

BACKGROUND: The Supplemental Nutrition Assistance Program (SNAP) has been established to reduce food insecurity. Limited evidence is available on SNAP participation status over time and depressive symptoms. We aimed to examine the associations of SNAP status over time among low-income individuals, with depressive symptoms in the U.S. METHODS: NHANES participants aged ≥20 years of low family income from 2011 to 2018 with information available on depressive symptoms and SNAP use were included in analysis. Depressive symptoms were assessed using 9-item Patient Health Questionnaire (PHQ-9), and PHQ-9 score ≥ 10 is indicative of significant depressive symptoms. Multivariable linear and logistic regressions models were conducted to examine the associations of SNAP participation status over time (never receiving SNAP, receiving SNAP prior to >12 months ago, current receiving SNAP, receiving SNAP in the last 12 months but not currently) with depressive symptoms and significant depressive symptoms. RESULTS: Currently receiving SNAP (beta (ß) = 0.17, 95 % CI: 0.10, 0.25; odds ratio (OR) = 1.52, 95 % confidence interval (CI): 1.16, 2.00) and receiving SNAP in the last 12 months but not currently (ß = 0.24, 95 % CI: 0.04, 0.43; OR = 1.83, 95 % CI: 1.16, 2.89) were associated with higher depressive symptoms and higher prevalence of significant depressive symptoms. LIMITATIONS: The cross-sectional design precludes causal interpretation, and key variables were measured with self-report. CONCLUSION: Receiving SNAP in the last 12 months was associated with higher levels of depressive symptoms among individuals with low family income. Improvement on diet quality may be important for reducing depressive symptoms among SNAP users.

Employing a Youth Internship Model to Identify Goals to Reduce Barriers to Active Living.

Active lifestyles are vital for promoting health. In this practice note, we describe the implementation of an active living intervention designed to engage youth in identifying barriers to being physically active and developing recommendations to address these barriers. Youth interns were compensated for their time. Through this project, the community obtained street striping for the first time, secured a community center when the police substation building was turned over to the community, and had sidewalk funding prioritized for one of their busiest streets. Lessons learned while developing and implementing this youth internship program focused on making the internship program work well given youth schedules and focusing on supporting the voice of youth to advocate for changes to the built environment in an intentionally excluded community.

Experiences participating in federal nutrition assistance programs during the early months of the COVID-19 pandemic: an investigation in Vermont.

BACKGROUND: Federal nutrition assistance programs serve as safety nets for many American households, and participation has been linked to increased food security and, in some instances, improved diet quality and mental health outcomes. The COVID-19 pandemic brought new and increased economic, social, and psychological challenges, necessitating inquiry into how nutrition assistance programs are functioning and associated with public health outcomes. METHODS: Using data from a representative statewide survey administered in Vermont (n = 600) between July and September 2020, we examined participant experiences with major federal nutrition assistance programs: the Supplemental Nutrition Assistance Program (SNAP), the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC), and school meal programs. We explored quantitative and qualitative responses regarding perceptions of program utility, and used nearest neighbors matching analyses in combination with bivariate statistical tests to assess associations between program participation and food insecurity, perceived stress, and fruit and vegetable intake as indicators of dietary quality. RESULTS: One in four respondents (27.3%) used at least one federal nutrition assistance program. As compared to non-participants, we found higher rates of food insecurity among program participants (57.5% vs. 18.1%; p < 0.001), an association that persisted even when we compared similar households using matching techniques (p ≤ 0.001). From matched analyses, we found that, compared to low-income non-participants, low-income program participants were less likely to meet fruit intake recommendations (p = 0.048) and that low-income SNAP and WIC participants were less likely to meet vegetable intake recommendations (p = 0.035). We also found lower rates of perceived stress among low-income school meal participant households compared to low-income non-participants (p = 0.039). Despite these mixed outcomes, participants broadly valued federal nutrition assistance programs, characterizing them as helpful or easy to use. CONCLUSIONS: We found that federal nutrition assistance programs as a group were not sufficient to address food insecurity and stress or increase fruit and vegetable intake in the state of Vermont during the early months of the COVID-19 pandemic. Nonetheless, participants perceived benefits from participation in these programs. Optimizing the utility of nutrition assistance programs depends on critical examination of their functioning under conditions of great stress.

Investigating the potential role of α-SNAP in preventing chemotherapy-induced ovarian dysfunction: Insights from cellular and animal models.

Background: The phosphoinositide 3-kinase/Akt/mammalian target of rapamycin complex 1 (PI3K/Akt/mTORC1) pathway plays a crucial role in the activation of primordial follicles. However, excessive activation and the loss of primordial follicles can lead to ovarian dysfunction. The alpha-soluble N-ethylmaleimide sensitive factor attachment protein (α-SNAP) protein has been implicated in PI3K/Akt/mTORCl signaling, suggesting its potential involvement in follicle activation. Thus, this study aimed to explore the role of α-SNAP in the activation of the PI3K/Akt/mTORC1 signaling pathway and its ability to mitigate the effects of cisplatin on ovarian function, using both in vitro and in vivo models. Methods: We transfected KGN human ovarian granulosa cells (GCs) with small interfering RNA (siRNA) targeting α-SNAP to investigate the effects of α-SNAP inhibition on GC proliferation and apoptosis, as well as on the activity of the PI3K/Akt/mTORC1 pathway. In a mouse model, α-SNAP siRNA was delivered via an adeno-associated virus before treatment with cisplatin to assess its effects on follicle activation and ovarian function. Follicle counts at various growth stages, western blotting, and immunohistochemistry analyses were conducted to detect the expression of cleaved caspase-3, Ki67, α-SNAP, and p-mTOR. Additionally, the serum concentrations of anti-Müllerian hormone (AMH) were measured through an enzyme-linked immunosorbent assay. Results: In vitro, α-SNAP depletion prevented GC proliferation by inhibiting the PI3K/Akt/mTORC1 pathway, thereby indicating its role in the regulation of cell growth. In vivo, α-SNAP knockdown attenuated the cisplatin-induced overactivation of primordial follicles by suppressing the PI3K/Akt/mTORC1 signaling pathway and partially restoring AMH levels. In addition, the expression and distribution patterns of cleaved caspase-3, Ki67, α-SNAP, and p-mTOR varied across different follicular growth stages, suggesting a protective effect against chemotherapy-induced ovarian damage. Conclusions: Inhibiting α-SNAP may attenuate GC proliferation by suppressing the PI3K/Akt/mTORC1 pathway, thereby mitigating the overactivation and loss of primordial follicles induced by cisplatin. Targeting α-SNAP may emerge as a novel strategy to prevent ovarian damage resulting from chemotherapy. However, these conclusions warrant repeated testing, and the mechanistic underpinnings of α-SNAP must be further elucidated in the future.

Synthesis and Characterization of a Sustained Nitric Oxide-Releasing Orthodontic Elastomeric Chain for Antimicrobial Action.

The acidic byproducts of bacteria in plaque around orthodontic brackets contribute to white spot lesion (WSL) formation. Nitric oxide (NO) has antibacterial properties, hindering biofilm formation and inhibiting the growth of oral microbes. Materials that mimic NO release could prevent oral bacteria-related pathologies. This study aims to integrate S-nitroso-acetylpenicillamine (SNAP), a promising NO donor, into orthodontic elastomeric ligatures, apply an additional polymer coating, and evaluate the NO-release kinetics and antimicrobial activity against Streptococus mutans. SNAP was added to clear elastomeric chains (8 loops, 23 mm long) at three concentrations (50, 75, 100 mg/mL, and a control). Chains were then coated, via electrospinning, with additional polymer (Elastollan®) to aid in extending the NO release. NO flux was measured daily for 30 days. Samples with 75 mg/mL SNAP + Elastollan® were tested against S. mutans for inhibition of biofilm formation on and around the chain. SNAP was successfully integrated into ligatures at each concentration. Only the 75 mg/mL SNAP chains maintained their elasticity. After polymer coating, samples exhibited a significant burst of NO on the first day, exceeding the machine's reading capacity, which gradually decreased over 29 days. Ligatures also inhibited S. mutans growth and biofilm formation. Future research will assess their mechanical properties and cytotoxicity. This study presents a novel strategy to address white spot lesion (WSL) formation and bacterial-related pathologies by utilizing nitric oxide-releasing materials. Manufactured chains with antimicrobial properties provide a promising solution for orthodontic challenges, showing significant potential for academic-industrial collaboration and commercial viability.

The Relationship between SNAP25 and Some Common Human Neurological Syndromes.

Over the years, research on the pathogenesis of neurological diseases has progressed slowly worldwide. However, as the incidence rate continues to increase and the disease gradually develops, early diagnosis and treatment have become a top priority. SANP25, a protein present on the presynaptic membrane and involved in neurotransmitter release, is closely related to the loss or abnormal expression of synapses and neurons. SNAP25 deficiency can lead to synaptic disorders and inhibit neurotransmitter release. Therefore, a large amount of literature believes that SNAP25 gene mutation is a risk factor for many neurological diseases. This review used advanced search on PubMed to conduct extensive article searches for relevant literature. The search keywords included SNAP25 and Alzheimer's disease, SNAP25 and Parkinson's disease, and so on. After reading and summarizing the previous papers, the corresponding conclusions were obtained to achieve the purpose of the review. The deficiency or variation of SNAP25 might be related to the onset of schizophrenia, epilepsy, attention deficit/hypoactivity disorder, bipolar disorder effective disorder, and autism. SNAP25 has been found to be used as a neuropathological marker for neurological diseases, which could be the target of diagnosis or treatment of Alzheimer's disease and Parkinson's disease. Cerebrospinal Fluid (CSF) or blood has been found to enable more effective drug development.