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Indian Journal of Pathology &... Jun 2024Genotyping of H. pylori strains was identified on formalin-fixed paraffin wax-embedded tissue (FFPE) sections and correlated with severity grades of gastric mucosal...
AIM AND BACKGROUND
Genotyping of H. pylori strains was identified on formalin-fixed paraffin wax-embedded tissue (FFPE) sections and correlated with severity grades of gastric mucosal pathologies in biopsies from upper gastrointestinal (UGI) tract from Guwahati, Assam.
MATERIALS AND METHODS
In total, 168 gastric biopsies collected from patients with UGI symptoms underwent histological evaluation as per the updated Sydney system.
RESULT
H. pylori-like organisms were identified with Warthin and Starry stain, and virulent genes were amplified using polymerase chain reaction (PCR) from genomic DNA extracted from FFPE sections by using QIAamp® DNA FFPE Tissue Kit. Histological examination identified H. pylori-like organisms in 100 biopsies, of which 96 were urease + ve. The prevalence of H. pylori infection was high in age groups 71-80 (88.8%) as compared to other age groups, and it was higher in females (78.9%) when compared to males. The prevalence of virulent genes in biopsies was 88.5% cagA and vacA s1m1, 31.2% iceA1, 32.2% iceA2, and 85.2% babA2. The histological parameters mononuclear cell infiltrate (P = 0.04) and atrophy (P = 0.03), showed statistically significant association with iceA2 and intestinal metaplasia with cagA (P = 0.01) vacAs1m1 (P = 0.01) and babA (P = 0.02) genotypes. Gastric erosion due to H. pylori infection and atrophy showed a significant association. A high bacterial density score was seen with the virulent genotypes.
CONCLUSION
Our work reports for the first time a high prevalence (88.5%) of H. pylori cagA vacA s1m1 genotype in Guwahati, Assam. Association of gastric atrophy and intestinal metaplasia was seen with virulent genotypes. Results show the effectiveness of the FFPE kit for DNA extraction in remote areas where transportation and storage of biopsies are otherwise difficult.
PubMed: 38847212
DOI: 10.4103/ijpm.ijpm_1002_23 -
PNAS Nexus Jun 2024Food safety is related to human health and sustainable development. International food trade poses food safety risks through the collateral transport of toxic chemicals...
Food safety is related to human health and sustainable development. International food trade poses food safety risks through the collateral transport of toxic chemicals that are detrimental to human health. Domestic interprovincial trade has similar effects within countries but has not been comprehensively investigated previously. Here, we assessed the effects of interprovincial trade on food safety and human dietary exposure to short-chain chlorinated paraffins (SCCPs), a group of emerging persistent toxic chemicals, in seafood across China by synthesizing data from field observation and various models. Our findings indicate that there is a higher level of SCCPs exposure risk in coastal provinces compared to inland provinces. Approximately, 70.3% of human exposure to SCCPs through seafood consumption in China was embodied in the interprovincial seafood trade in 2021. Specifically, the domestic trade led to a remarkable increase in SCCPs exposure in the coastal provinces in South China, attributable to low SCCPs pollution in these provinces and imported seafood from those provinces with high SCCPs pollution. In contrast, human exposure to SCCPs decreased in those coastal provinces in East China due to importing seafood from those provinces with low SCCPs concentrations. The interprovincial seafood trade routes were optimized by linear programming to minimize human exposure to SCCPs considering both shipping cost and health risk constraints. The optimized trade routes reduced the national per capita SCCPs exposure through seafood consumption by over 12%. This study highlights the importance of interprovincial food trade in the risk assessment of toxic chemicals.
PubMed: 38846777
DOI: 10.1093/pnasnexus/pgae205 -
Heliyon Jun 2024In drug delivery, it is common to use porous particles as carrier media, instead of dense particles, due to their high specific surface area and available entrapment...
In drug delivery, it is common to use porous particles as carrier media, instead of dense particles, due to their high specific surface area and available entrapment volume, which allows a higher amount of drug to be encapsulated and then released. Chitosan microparticles are extensively used in drug delivery, but porous chitosan microparticles are scarcely reported. In this work, the preparation of porous chitosan microparticles using membrane emulsification is addressed, a technology that involves mild operating conditions and less energy consumption than traditional methods (such as ultrasound), and with higher control of the particle size. The dense structure is obtained by a water-in-oil emulsion. The porous structure is obtained by a gas-in-water-in-oil G/W/O double emulsion, where argon bubbles get entrapped in an aqueous chitosan solution that is further emulsified in a paraffin/petroleum ether mixture. Porous chitosan particles were obtained with sizes of 7.7 ± 1.6 μm, which was comparable with dense chitosan particles (6.2 ± 2.3 μm). The pore structure was optimized by varying the argon flow rate, being optimized at 0.24 L h. The impact of drug loading by adsorption or encapsulation, and of the drug release behaviour when using porous and dense particles were assessed, using the protein bovine serum albumin (BSA) as a model drug. The results showed that by encapsulating BSA the loading efficiency was above 95 % for both types of particles, with the release being slightly slower for the dense particles. As for the adsorbed BSA, the loading efficiency was significantly higher for porous particles - 70 % - against the 40 % for dense particles. Porous chitosan particles were successfully obtained using the membrane emulsification technology and showed that these carriers are advantageous regarding drug loading and release.
PubMed: 38845862
DOI: 10.1016/j.heliyon.2024.e31823 -
Epilepsia Open Jun 2024Perineuronal nets (PNN) are specialized extracellular matrix (ECM) components of the central nervous system, frequently accumulating at the surface of inhibitory...
OBJECTIVE
Perineuronal nets (PNN) are specialized extracellular matrix (ECM) components of the central nervous system, frequently accumulating at the surface of inhibitory GABAergic interneurons. While an altered distribution of PNN has been observed in neurological disorders including Alzheimer's disease, schizophrenia and epilepsy, their anatomical distribution also changes during physiological brain maturation and aging. Such an age-dependent shift was experimentally associated also with hippocampal engram formation during brain maturation. Our aim was to histopathologically assess PNN in the hippocampus of adult and pediatric patients with temporal lobe epilepsy (TLE) compared to age-matched post-mortem control subjects and to compare PNN-related changes with memory impairment observed in our patient cohort.
METHODS
Sixty-six formalin-fixed and paraffin-embedded tissue specimens of the human hippocampus were retrieved from the European Epilepsy Brain Bank. Twenty-nine patients had histopathologically confirmed hippocampal sclerosis (HS), and eleven patients suffered from TLE without HS. PNN were immunohistochemically visualized using an antibody directed against aggrecan and manually counted from hippocampus subfields and the subiculum.
RESULTS
PNN density increased with age in both human controls and TLE patients. However, their density was significantly higher in all HS patients compared to age-matched controls. Intriguingly, TLE patients presented presurgically with better memory when their hippocampal PNN density was higher (p < 0.05).
SIGNIFICANCE
Our results were compatible with age-dependent ECM specialization in the human hippocampus and its precocious aging in the epileptic condition. These observations confirm recent experimental animal models and also support the notion that PNN play a role in memory formation in the human brain.
PLAIN LANGUAGE SUMMARY
"Perineuronal nets" (PNN) are a specialized compartment of the extracellular matrix (ECM), especially surrounding highly active neurons of the mammalian brain. There is evidence that PNN play a role in memory formation, brain maturation, and in some pathologies like Alzheimer's disease, schizophrenia or epilepsy. In this study, we investigated the role of PNN in patients suffering from drug-resistant focal epilepsy compared to controls. We found that with increasing age, more neurons are surrounded by PNN. Similarly, all epilepsy patients but especially patients with better memory performance also had more PNN. This study raises further interest in studying ECM molecules in the human brain under physiological and pathophysiological conditions.
PubMed: 38845524
DOI: 10.1002/epi4.12963 -
Scientific Reports Jun 2024Neonicotinoids, a relatively new widely used class of insecticide is used in agriculture to control insect populations. We examined the capacity of ancestral exposure to...
Neonicotinoids, a relatively new widely used class of insecticide is used in agriculture to control insect populations. We examined the capacity of ancestral exposure to the neonicotinoid thiacloprid (thia) to induce transgenerational effects on thyroid tissue. Pregnant outbred Swiss female mice were exposed to thia at embryonic days E6.5 to E15.5 using 0, 0.6, and 6 mg/kg/day doses. Thyroid paraffin sections were prepared for morphology analysis. We apply ELISA method to measure T4 and TSH levels, RT-qPCR for gene expression analysis, ChIP-qPCR techniques for sperm histone H3K4me3 analysis, and immunofluorescence microscopy and western blots for protein detection. We observed an alteration in the morphology of thyroids in both males and females in the F3 generation. We observed an increase in T4 hormone in F1 females and a significant T4 level decrease in F3 males. T4 changes in F1 females were associated with a TSH increase. We found that the amount of Iodothyronine Deiodinase 1 (DIO1) (an enzyme converting T4 to T3) was decreased in both F1 and F3 generations in female thyroids. GNAS protein which is important for thyroid function has increased in female thyroids. Gene expression analysis showed that the expression of genes encoding thyroid gland development, chromatin, biosynthesis and transport factors were affected in the thyroid gland in both sexes in F1 and F3. The analysis of sperm histone H3K4me3 showed that H3K4me3 occupancy at the Dio1 locus has decreased while Thyroglobulin (Tg) and Matrix Metallopeptidase 2 (Mmp2) genes have increased H3K4me3 occupancy in the sperm of F3 mice. Besides, DNA methylation analysis of our previously published datasets showed that, in the sperm of F1 and F3 thia-derived mice, several genes related to thyroid function show consistent alterations. Our data suggest that ancestral exposure to thiacloprid affects thyroid function not only in exposed but also in indirectly exposed F3 generation.
Topics: Animals; Thyroid Gland; Female; Neonicotinoids; Mice; Male; Thiazines; Pregnancy; Histones; Thyroxine; Iodide Peroxidase; Spermatozoa; Insecticides; Thyrotropin; Sex Factors
PubMed: 38844538
DOI: 10.1038/s41598-024-63986-w -
Neurobiology of Disease Aug 2024Huntington's disease (HD) is a neurodegenerative disorder that severely affects the basal ganglia and regions of the cerebral cortex. While astrocytosis and microgliosis...
Huntington's disease (HD) is a neurodegenerative disorder that severely affects the basal ganglia and regions of the cerebral cortex. While astrocytosis and microgliosis both contribute to basal ganglia pathology, the contribution of gliosis and potential factors driving glial activity in the human HD cerebral cortex is less understood. Our study aims to identify nuanced indicators of gliosis in HD which is challenging to identify in the severely degenerated basal ganglia, by investigating the middle temporal gyrus (MTG), a cortical region previously documented to demonstrate milder neuronal loss. Immunohistochemistry was conducted on MTG paraffin-embedded tissue microarrays (TMAs) comprising 29 HD and 35 neurologically normal cases to compare the immunoreactivity patterns of key astrocytic proteins (glial fibrillary acidic protein, GFAP; inwardly rectifying potassium channel 4.1, Kir4.1; glutamate transporter-1, GLT-1; aquaporin-4, AQP4), key microglial proteins (ionised calcium-binding adapter molecule-1, IBA-1; human leukocyte antigen (HLA)-DR; transmembrane protein 119, TMEM119; purinergic receptor P2RY12, P2RY12), and indicators of proliferation (Ki-67; proliferative cell nuclear antigen, PCNA). Our findings demonstrate an upregulation of GFAP protein expression attributed to the presence of more GFAP expressing cells in HD, which correlated with greater cortical mutant huntingtin (mHTT) deposition. In contrast, Kir4.1, GLT-1, and AQP4 immunoreactivity levels were unchanged in HD. We also demonstrate an increased number of IBA-1 and TMEM119 microglia with somal enlargement. IBA-1, TMEM119, and P2RY12 reactive microglia immunophenotypes were also identified in HD, evidenced by the presence of rod-shaped, hypertrophic, and dystrophic microglia. In HD cases, IBA-1 cells contained either Ki-67 or PCNA, whereas GFAP astrocytes were devoid of proliferative nuclei. These findings suggest cortical microgliosis may be driven by proliferation in HD, supporting the hypothesis of microglial proliferation as a feature of HD pathophysiology. In contrast, astrocytes in HD demonstrate an altered GFAP expression profile that is associated with the degree of mHTT deposition.
Topics: Humans; Huntington Disease; Microglia; Astrocytes; Male; Female; Middle Aged; Cell Proliferation; Adult; Aged; Cerebral Cortex; Calcium-Binding Proteins; Gliosis; Glial Fibrillary Acidic Protein; Membrane Proteins; Microfilament Proteins
PubMed: 38844243
DOI: 10.1016/j.nbd.2024.106554 -
PloS One 2024As adoptive cellular therapies become more commonplace in cancer care, there is a growing need to monitor site-specific localization of engineered cells-such as chimeric...
As adoptive cellular therapies become more commonplace in cancer care, there is a growing need to monitor site-specific localization of engineered cells-such as chimeric antigen receptor T (CAR-T) cells and T-cell receptor T (TCR-T) cells-in patients' tissues to understand treatment effectiveness as well as associated adverse events. Manufacturing CAR-T and TCR-T cells involves transduction with viral vectors commonly containing the WPRE gene sequence to enhance gene expression, providing a viable assay target unique to these engineered cells. Quantitative PCR (qPCR) is currently used clinically in fresh patient tissue samples and blood with target sequences specific to each immunotherapy product. Herein, we developed a WPRE-targeted qPCR assay that is broadly applicable for detection of engineered cell products in both fresh and archival formalin-fixed paraffin embedded (FFPE) tissues. Using both traditional PCR and SYBR Green PCR protocols, we demonstrate the use of this WPRE-targeted assay to successfully detect two CAR-T cell and two TCR-T cell products in FFPE tissue. Standard curve analysis reported a reproducible limit of detection at 100 WPRE copies per 20μL PCR reaction. This novel and inexpensive technique could provide better understanding of tissue abundance of engineered therapeutic T cells in both tumor and second-site toxicity tissues and provide quantitative assessment of immune effector cell trafficking in archival tissue.
Topics: Humans; Formaldehyde; Hepatitis B Virus, Woodchuck; Receptors, Antigen, T-Cell; Receptors, Chimeric Antigen; T-Lymphocytes; Tissue Fixation; Immunotherapy, Adoptive; Real-Time Polymerase Chain Reaction
PubMed: 38843256
DOI: 10.1371/journal.pone.0303057 -
Folia Morphologica Jun 2024Monosodium glutamate was considered one of the food additive and flavor enhancer in processed meat and soup that affects testicular tissues, the aim of this research to...
BACKGROUND
Monosodium glutamate was considered one of the food additive and flavor enhancer in processed meat and soup that affects testicular tissues, the aim of this research to investigates the impact of monosodium glutamate (MSG) on testicular structure in rats and explores the potential protective effects of resveratrol.
MATERIAL AND METHODS
Four experimental groups involved in our study 10 rats of each.: the first group as control group; the second group (Resveratrol group: control rats received 20 mg/kg of resveratrol, via oral gavage); the third group (MSG group: rats received monosodium glutamate (MSG) with a dose 60 mg/kg body weight daily, via gastric tube, and the fourth group (MSG+ Resveratrol group). Serum level of testosterone, FSH, LH, were measured. Testicular tissues were prepared for measurement of oxidative stress markers, and gene expression of NLRP3, Caspase 3, and GSK-3β. Moreover, paraffin blocks contained testicular tissue used for histological and immunohistochemical examination. Additionally, seminal smear from epididymis were examined.
RESULTS
MSG administration adversely affected testosterone production, hormonal levels, and sperm parameters, Histological examination revealed marked testicular degeneration, and oxidative stress assessments indicated elevated level of MDA a lipid peroxidation marker and decrease in SOD, CAT antioxidant enzymes. Moreover, MSG-induced apoptotic and pyroptotic markers and its gene expressions. Importantly. Administration of resveratrol reversed the detrimental effects of MSG, demonstrating its corrective influence on the hypothalamic-pituitary-gonadal axis disruption, improvement of sperm parameters, attenuation of oxidative stress, anti-apoptotic activity, and anti-pyroptotic effects. The expression of Ki-67 as a cell proliferation marker further supported the positive response to spermatogenesis dysfunction upon resveratrol treatment.
CONCLUSIONS
This comprehensive exploration sheds light on the protective effect of resveratrol against MSG-induced testicular with exploration of its mechanistic role.
PubMed: 38842078
DOI: 10.5603/fm.99434 -
Current Research in Food Science 2024Food adulteration is a global concern, drawing attention from safety authorities due to its potential health risks. Detecting and categorizing oil adulteration is...
Food adulteration is a global concern, drawing attention from safety authorities due to its potential health risks. Detecting and categorizing oil adulteration is crucial for consumer safety and food industry integrity. This research explores hyperspectral imaging (HSI) analysis to identify substandard oil adulteration at different stages. Using the non-destructive HSI Specim Fx 10 system, a method for precise and easy imaging-based fraud detection and classification was proposed. The 670 oil samples, including pure (Almond, Mustard, Coconut, Olive) and adulterated (Sunflower, Castor, Liquid Paraffin), were analyzed. The Savitzky-Golay filter preprocessed the images to remove noise and smooth spectral signatures. The oils were identified using various machine learning approaches, including Support Vector Machines, Logistic Regression, Linear Discriminant Analysis, Random Forests, Decision Trees, K-Nearest Neighbors, and Naïve Bayes with Linear Discriminant Analysis excelling in identification. Performance parameters, including precision, recall, F1-score, and overall accuracy, were calculated. The proposed method achieved a validation accuracy of 100%, outperforming numerous state-of-the-art approaches. This study introduces a robust pipeline for effective oil adulteration detection, offering a significant advancement in food safety and quality control.
PubMed: 38840806
DOI: 10.1016/j.crfs.2024.100773 -
Revista Brasileira de Parasitologia... 2024In endemic areas for canine visceral leishmaniasis (CVL), the occurrence of coinfection with other pathogens, such as Ehrlichia spp., has been associated with worsening...
In endemic areas for canine visceral leishmaniasis (CVL), the occurrence of coinfection with other pathogens, such as Ehrlichia spp., has been associated with worsening of the clinical condition. The study aimed to evaluate the occurrence of histological changes in the myocardia of dogs naturally infected with Leishmania chagasi with or without coinfection with Ehrlichia spp.. We evaluated paraffin-embedded myocardial sections from 31 dogs, affected by either L. chagasi alone or coinfected with L. chagasi and Ehrlichia spp., to compare the extent and degree of cardiac damage. The blocks were divided into two groups. G1 (dogs infected only by L. chagasi) and G2 (dogs coinfected with L. chagasi and Ehrlichia spp.). The right atrium free wall, right ventricle free wall, left ventricle, and interventricular septum of all groups were evaluated. Cardiac alterations were observed in 41.93% (52/124) of the fragments evaluated and inflammatory infiltrate was the most common pattern found. The G2 group showed a higher incidence of myocarditis, with 61.53% (32/52), compared to the G1 group, in which 20 out of 72 cases (27.7%) exhibited histopathological changes (p <0.05). These findings confirmed that coinfection can potentiate cardiac damage in dogs.
Topics: Animals; Dogs; Leishmaniasis, Visceral; Dog Diseases; Male; Ehrlichiosis; Coinfection; Female; Myocarditis; Ehrlichia; Myocardium
PubMed: 38836809
DOI: 10.1590/S1984-29612024023