Protective Effect of L-carnitine on the Oxidative Stress Injury of Human Ovarian Granulosa Cells

The protective effect of L-carnitine (LC) on the oxidative stress (OS) injury and the effect of L-carnitine on follicular stimulating hormone receptor (FSHR) of ovarian granulosa cells (GCs) were investigated. OS was induced by treatment with H2O2. We cultured KGN cells in four groups: the blank group, OS group and two L-carnitine pretreatment group (low, high). In the OS group, cell nuclear pyknosis was observed, mitochondria swelled irregularly and their cristae were fractured. Meanwhile, the cell viability, superoxide dismutase (SOD) and glutathione (GSH) contents, mitochondrial membrane potential (ΔΨm) and the level of FSHR expression were significantly decreased in the OS group. However, malonaldehyde (MDA) content, reactive oxygen species (ROS) level and apoptosis rate were significantly increased. Compared with the OS group, the morphology of cells and mitochondria in the L-carnitine pretreatment group were improved, the cell viability and the expression of FSHR was significantly increased, and the OS level was decreased. These results indicated that L-carnitine can protect the cells from OS damage induced by H2O2, enhance the antioxidant and anti-apoptotic ability of GCs, and alleviate the decrease of FSHR expression on GCs caused by OS. Therefore, L-carnitine may help prevent the ovarian aging and improve the quality of follicles.

Human ALS/FTD brain organoid slice cultures display distinct early astrocyte and targetable neuronal pathology

Amyotrophic lateral sclerosis overlapping with frontotemporal dementia (ALS/FTD) is a fatal and currently untreatable disease characterized by rapid cognitive decline and paralysis. Elucidating initial cellular pathologies is central to therapeutic target development, but obtaining samples from presymptomatic patients is not feasible. Here, we report the development of a cerebral organoid slice model derived from human induced pluripotent stem cells (iPSCs) that recapitulates mature cortical architecture and displays early molecular pathology of C9ORF72 ALS/FTD. Using a combination of single-cell RNA sequencing and biological assays, we reveal distinct transcriptional, proteostasis and DNA repair disturbances in astroglia and neurons. We show that astroglia display increased levels of the autophagy signaling protein P62 and that deep layer neurons accumulate dipeptide repeat protein poly(GA), DNA damage and undergo nuclear pyknosis that could be pharmacologically rescued by GSK2606414. Thus, patient-specific iPSC-derived cortical organoid slice cultures are a reproducible translational platform to investigate preclinical ALS/FTD mechanisms as well as novel therapeutic approaches.

Alantolactone Exhibits Antiproliferative and Apoptosis-Promoting Properties in Colon Cancer Model via Activation of the MAPK-JNK/c-Jun Signaling Pathway

Colorectal cancer (CRC) is one of the most common human malignancies in the digestive tract with high mortality. Alantolactone (ATL), as a plant-derived sesquiterpene lactone, has shown a variety of pharmacological activities, such as antibacterial, anti-inflammatory, anti-virus and so on. However, the exact molecular mechanism of ATL in colorectal cancer remains largely unknown. Here, we performed a study to explore the effect and mechanism of ATL on colorectal cancer. The CCK-8 assay, colony formation assay, Wound-healing and Transwell assays were performed to evaluate the cytotoxic effect, antiproliferative effect, anti-migratory and anti-invasive properties of ATL respectively. The xenograft tumor model was established in Balb/c mice to evaluate the anti-tumor effect. The expression levels of proteins involved the MAPK-JNK/c-Jun signaling pathway were measured by Western blot and RT-qPCR both in cells and tumor tissues. The results showed that ATL could inhibit the cells activities of various colon cancer cell lines. Moreover, ATL could induce HCT-116 cells nuclear pyknosis, mitochondrial membrane potential loss, G0/G1 phase arrest, as well as enhance the proportion of apoptosis cells and inhibit colony formation. The migration distance and invasion rate of cells were significantly reduced after treated with ATL. Additionally, in the xenograft model, ATL (50mg/kg) significantly decreased the tumor tumor volume and weight (p ˂ 0.001). For the anti-colon cancer mechanism, the ATL showed the anti-proliferative and pro-apoptosis effect by activating MAPK-JNK/c-Jun signaling pathway. In conclusion, ATL exhibits anti-proliferation and apoptosis-promoting potential in colon cancer via the activation of MAPK-JNK/c-Jun signaling pathway.

Amelioration of Hepatic Encephalopathy Using Dunaliella salina Microalgae in Rats: Modulation of Hyperammonemia/TLR4

Hepatic encephalopathy (HE) is a neuropsychiatric disease that is developed as a complication of both acute and chronic liver failure affecting psychomotor dysfunction, memory, and concentration. This study is aimed at evaluating the therapeutic effects of Dunaliella salina (D. salina) microalgae in thioacetamide- (TAA-) induced HE in rats. HE was induced by TAA (200 mg/kg; i.p.) for three successive days. Forty male Wister albino rats were divided into 4 groups; the first group was served as a normal, and the second group was injected with TAA and served as TAA control. The third and fourth groups were administered D. salina (100 and 200 mg/kg; p.o.), respectively, after TAA injection for 7 days. The behavioral and biochemical markers as well as histological aspects of HE were estimated. This study revealed that TAA caused behavioral changes, oxidative stress, neuroinflammation, nuclear pyknosis, and neurons degeneration. D. salina improved liver function and decreased oxidative stress and inflammatory mediator as TLR4 protein expression. Also, D. salina elevated HSP-25 and IGF-1 as well as improved brain histopathological alterations. In conclusion, D. salina exerted a therapeutic potential against HE via its antioxidant, antiinflammatory and cytoprotective effects.

Evaluation of lambda-cyhalothrin oxidative stress and gonad histoarchitecture toxicity potency in Clarias gariepinus

Background Extensive and indiscriminate use of pesticides gradually destroys the environment (ecosystem), poses serious threats to human health, animal life (especially aquatic), plant forms, soil, water, and also lead to emergence of resilient species of life forms that are becoming resistant to pesticides. The present study focused on evaluating lambda-cyhalothrin oxidative stress and gonad histoarchitecture toxicity potency in Clarias gariepinus. Results A total of 120 C. gariepinus 16 to 40 cm SL and 200 to 250 g bodyweights (assigned into treatments 0.00 (control), 2.5 × 10−4 μg/L, 5.0 × 10−4 μg/L, and 6.25×10−4 μg/L (A-D) lambda-cyhalothrin (LCT), each treatment consisted of 30 fishes, replicated three times, 10 fishes per replicate) were used for this study. On day 7, catalase activity (CAT) and glutathione peroxidase (GPx) significantly increased (p < 0.05) in all treatments compared with control. Day 14, superoxide dismutase (SOD) and GPx significantly increased (p < 0.05). All parameters significantly increased (p < 0.05) on days 21 and 28 except SOD (day 21). All parameters increased significantly on day 28 across the row in all treatments. The significant increase (p < 0.05) in SOD, (malondialdehyde) MDA, GPx, and glutathione reductase (GR) levels returned to normal after 7 days of depuration but CAT level did not return to normal. The testes photomicrographs showed necrotic conditions in the spermatogenic cells with nuclear pyknosis and cytoplasmic swelling while that of the ovary displayed vacuolations, flabby oocytes, and degenerated ovaries changes. Conclusion Lambda-cyhalothrin is toxic to C. gariepinus. The inability of significant increase in CAT to return to normal after 7 days of depuration further confirms our report.

Ameliorative Effects of Oral Glucosamine on Insulin Resistance and Pancreatic Tissue Damage in Experimental Wistar rats on a High-fat Diet

Hyperlipidemia due to a high-fat diet (HFD) is a risk factor for inducing insulin resistance (IR) and adverse effects onpancreatic β-cells in obesity and type 2 diabetes mellitus. This relationship may be due to activation of the hexosaminebiosynthesis pathway. Administration of exogenous glucosamine (GlcN) can increase the end product of this pathway(uridine-5′-diphosphate-N-acetyl-glucosamine), which can mediate IR and protein glycosylation. The objective of this study was to evaluate the effects of oral GlcN and HFD on IR and pancreatic histologic damage in a 22 wk study of 4 groups of male Wistar rats: control group with normal chow diet, HFD group (24%. g/g lard), GlcN group (500 mg/kg−1 per day of glucosamine hydrochloride in drinking water) and HFD plus oral GlcN. Metabolic variables related to IR that were measured included triglycerides (TG), free fatty acids (FFAs) and malondialdehyde (MDA). Histopathologic evaluation of the pancreas was also performed. The results showed IR in the HFD group, which had increased pancreatic nuclear pyknosis and vacuolization, with fatty infiltration and structural alteration of the islets of Langerhans. TG, FFAs and MDA were higher in serum and pancreatic tissue as compared with the control group. The GlcN group did not develop IR and had only mild nuclear pyknosis with no significant change in the pancreatic content of TG, FFAs and MDA. However, the combined administration of GlcN and HFD attenuated IR and improved TG, FFAs and MDA levels in serum and pancreatic tissue and the pancreatic histopathologic changes, with no significant differences as compared with the control group. These findings suggest that the oral GlcN at a dose of 500 mg/kg−1 is protective against IR and the pancreatic histologic damage caused by HFD.

Deciphering the Forebrain Disorder in a Chicken Model of Cerebral Hernia

Cerebral hernia in crested chicken has been characterized as the protrusion of cerebral hemispheres into the unsealed skull for hundreds of years, since Charles Darwin. The development of deformed forebrain (telencephalon) of cerebral hernia remains largely unknown. Here, the unsealed frontal skull combined with misplaced sphenoid bone was observed and potentially associated with brain protuberance. The shifted pallidum, elongated hippocampus, expanded mesopallium and nidopallium, and reduced hyperpallium were observed in seven regions of the malformed telencephalon. The neurons were detected with nuclear pyknosis and decreased density. Astrocytes showed uneven distribution and disordered protuberances in hyperpallium and hippocampus. Transcriptome analyses of chicken telencephalon (cerebral hernia vs. control) revealed 547 differentially expressed genes (DEGs), mainly related to nervous system development, and immune system processes, including astrocyte marker gene GFAP, and neuron and astrocyte developmental gene S100A6. The upregulation of GFAP and S100A6 genes in abnormal telencephalon was correlated with reduced DNA methylation levels in the promoter regions. The morphological, cellular, and molecular variations in the shape, regional specification, and cellular states of malformed telencephalon potentially participate in brain plasticity and previously reported behavior changes. Chickens with cerebral hernia might be an interesting and valuable disease model to further explore the recognition, diagnosis, and therapy of cerebral hernia development of crested chickens and other species.

ESTIMATION OF TIME SINCE DEATH BY HISTOLOGICAL EXAMINATION OF COLLECTING TUBULES IN HUMAN KIDNEY.

Background: The current research histological changes in the Collecting tubules were examined at various postmortem intervals in the deceased human. In these research histological changes of tissue after death is influenced by P.M.I. atmospheric temperature and humidity, external and internal factors. Aim: To study the time since death by post mortem histological changes of collecting tubules in human Kidney. Materials and Methods: Present research was carried out, 40 cases of deceased kidney sample in between the temperature 17.3/22.3-31.3/450C,humidity 11/36 to 75/95 and duration range 4hrs to 52.30hr. The collecting tubules were examined to establish their correlation ship with hours PMI., temperatures, humidity or time since death by studied histological (H& E, PAS staining) in Pt.J.N.M.Medical College and Dr. B.R.Ambedkar Memorial Hospital Raipur (C.G.). Result: In collecting tubules (CT) after 4hrs PMI (27.5/42.20C T) disruption of epithelium was observed in most of the place. while after 46hrs PMI( 24.3/25.90C,T) disruption of epithelium, debris in the lumen with enucleated epithelial cells and pyknotic nuclei were seen, after 52.30hrs PMI(24.5/320C,T) disruption of epithelium, individualization of cells, debris in the lumen with pyknotic changes and enucleated cells were seen, Conclusion: Retraction of epithelium, disruption with individualization of cells, nuclear pyknosis, karyolysis and loss of tubular architecture with debris in the lumen were observed in collecting tubules. These criteria’s presented in this study could be used to determine the time after death. Keywords: Collecting tubules, P.M.I. (Post mortem interval), Medullary rays, Interstitium, Enucleated and Debris.

Experimental model in rats of penumbra area using implemented baloon in caudate nucleus volume dependent and time dependent

The study aims to determine whether early removal of an experimental intracerebral mass alters neurological function. In four experimental series, 0.6mL and 0.8mL balloons were implanted by stereotaxis and inflated into the right caudate nucleus of rats. After 1hour of insufflation, the brains were removed and studied by histopathological analysis. Immunohistochemical was included with protein S-100, marker of neuronal destruction. Four groups were formed (1A, 1B, 2A, 2B) and the variables: time and balloon volume was analyzed. In each series of the time variable, half of the animals had a 0.8mL balloon inflated for three minutes (Group 1A), and the other half had a 0.8mL balloon inflated for ten minutes (Group 2A). In the variable balloon volume series, half of the animals had a 0.6mL balloon inflated for 6 minutes (Group 1B), while the other half had a 0.8mL balloon inflated for 6minutes (Group 2B). In the qualitative and quantitative analysis of the parameters, the shortest group (group 1A), either there was no lesion, or there was partial loss of nervous tissue, while the longest group (group 2A) presented edema and cerebral parenchymal necrosis, reaching 35% of nuclear pyknosis. In the volume dependent groups (groups 1B and 2B), the findings were similar, both with about 30% of pyknotic nuclei. Thus, time was the major determinant of injury, reiterating the prognostic importance of early removal of a spontaneous intracerebral mass.

Quercetin Effects on Hepatotoxicity Induced by Titanium Dioxide Nanoparticles in Rats

Background: Most nanoparticles have adverse impacts on the liver, which is a vital body organ, by the induction of oxidative stress. Objectives: This study was designed to evaluate the hepatoprotective effects of quercetin (QCT) against the toxicity of nanoscale titanium dioxide (NTiO2) in Wistar rats. Methods: The present study was conducted on 32 adult female Wistar rats assigned into 4 groups of control, NTiO2 (50 mg/kg), NTiO2 + Quercetin (50 + 75 mg/kg), and Quercetin (75 mg/kg). The animals exposed to NTiO2 were administered by 50 mg/kg of NTiO2 for 21 days. The Quercetin + NTiO2 rats received Quercetin before exposing to NTiO2 for 7 days. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) of serum were considered indicators of the hepatotoxicity. The oxidative stress was assessed by measuring the activity of catalase (CAT) and superoxide dismutase (SOD) as well as the level of malondialdehyde (MDA) in the liver. TUNEL assay and histological changes were also assessed. Results: The NTiO2 significantly elevated the MDA level (P < 0.01), enhanced the serum biomarker levels, reduced the CAT (P < 0.01) and SOD (P < 0.01) activities. The NTiO2 also aggregated the red blood cells, and caused inflammatory cell infiltration, nuclear pyknosis and fat deposit in hepatocytes, as well as induced apoptosis in the liver tissue. Pretreatment with QCT quenched oxidative stress, attenuated the histological changes, elevated the CAT (P < 0.01) and SOD (P < 0.01) activities, normalized the serum biomarker levels and decreased apoptosis (P < 0.001). Conclusions: The QCT has an inhibitory impact on hepatotoxicity induced by nanoparticles in rats.