Decreased Protein Levels Research Articles

Field effectiveness of some essential oil emulsions against the whitefly Bemisia tabaci (GENN.) that infest potato plants in Egypt

AbstractWhitefly, Bemisia tabaci, nymphs and sucking sap adults, is one of the most damaging pests of potato, Solanum tuberosum. Excessive use of pesticides causes environmental pollution and the death of beneficial insects, so it is necessary to search for safer controlling alternatives. An experiment was carried out during seasons 2020–2021 and 2021–2022, under field conditions in Egypt. The objective of this study was to identify the main constituent compounds of peppermint and eucalyptus essential oils prepare coarse emulsion (CE) and nanoemulsion (NE) of both peppermint and eucalyptus essential oils and evaluate their efficiency against the whitefly, B. tabaci, in potato cultivars and the effect of these emulsions on the chemical components of potato leaves. The results revealed that the toxicity of the emulsions decreased with increasing time after spraying. The second spray which took place at a 10-day interval was more effective than the first spray. During the season of 2021, three days following the second spray, the tested emulsions' toxicity was highly effective. The reduction percentages of nymph for peppermint coarse emulsion (PCE), peppermint nanoemulsion (PNE), eucalyptus coarse emulsion (ECE), eucalyptus nanoemulsion (ENE) and imidacloprid were 82.64, 84.14, 73.26, 72.72 and 89.03, but they were 78.58, 83.27, 77.85, 69.97 and 89.26 compared with 2022. A moderately positive correlation was identified between temperature, specific humidity, wind speed and the effectiveness of the emulsions. This indicates that higher levels of temperature, humidity and wind speed were associated with increased efficacy of the emulsions. The study of the phytochemicals (total soluble protein, total carbohydrate, total phenolic contents and peroxidase activity). PCE and ECE achieved a slight decrease in protein levels. Also, ECE increased total plant carbohydrates. All treatments did not affect the phenolic compounds of potato leaf plants except ENE, which caused an increase in phenolic compounds. All treatments decreased the nitrogen plants' contents, while PCE, PNE, and ENE increased the potassium content. All treatments increase the activity of peroxidase (POX) compared with untreated plants. The formulations PNE and ENE might be an interesting alternative for integrated pest management of B. tabaci nymphs.

Ultra-low-dose rituximab is effective for the treatment of patients with minimal change disease - A retrospective study

Background purposeRituximab (RTX),an anti-CD20 monoclonal antibody can effectively treat minimal change disease (MCD),with low toxicity and a reduced steroid dosage. The optimal dosage of RTX for treating MCD remains unclear. This study aimed to investigate the efficacy of an ultra-low-dose regimen of RTX (100 mg per week for 4 weeks) for treating MCD. MethodsWe retrospectively analyzed clinical data from 31 patients with MCD who received RTX. Seventeen patients received ultra-low-dose RTX (ULD-RTX) therapy, and 14 patients received standard-dose RTX (SD-RTX) therapy (500 mg weekly for 4 weeks). All patients were followed up for at least 6 months. ResultsBoth groups showed significant increases in the serum albumin levels and notable decreases in the urinary protein levels in the 1st and 6th months after RTX therapy. There were no significant differences in the plasma albumin or urinary protein levels between the groups (p > 0.05). B-cell depletion was observed in all patients after 1 month of RTX administration. At 6 months after RTX treatment, the remission rate was 93% in the SD-RTX group and 88% in the ULD-RTX group (p > 0.05). The ULD-RTX therapy incurred lower costs than did the SD-RTX therapy. One patient in the SD-RTX group developed community-acquired pneumonia. ConclusionUltra-low-dose RTX is effective at inducing remission in patients with MCD at a lower cost.

C9orf72 controls hepatic lipid metabolism by regulating SREBP1 transport.

Sterol regulatory element binding transcription factors (SREBPs) play a crucial role in lipid homeostasis. They are processed and transported to the nucleus via COPII, where they induce the expression of lipogenic genes. COPII maintains the homeostasis of organelles and plays an essential role in the protein secretion pathways in eukaryotes. The formation of COPII begins at endoplasmic reticulum exit sites (ERES), and is regulated by SEC16A, which provides a platform for the assembly of COPII. However, there have been few studies on the changes in SEC16A protein levels. The repetitive expansion of the hexanucleotide sequence GGGGCC within the chromosome 9 open reading frame 72 (C9orf72) gene is a prevalent factor in the development of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Here, we found that the absence of C9orf72 leads to a decrease in SEC16A protein levels, resulting in reduced localization of the guanine nucleotide exchange factor SEC12 at the ERES. Consequently, the small GTP binding protein SAR1 is unable to bind the endoplasmic reticulum normally, impairing the assembly of COPII. Ultimately, the disruption of SREBPs transport decreases de novo lipogenesis. These results suggest that C9orf72 acts as a novel role in regulating lipid homeostasis and may serve as a potential therapeutic target for obesity.

Proteolytic cleavage and inactivation of the TRMT1 tRNA modification enzyme by SARS-CoV-2 main protease

Nonstructural protein 5 (Nsp5) is the main protease of SARS-CoV-2 that cleaves viral polyproteins into individual polypeptides necessary for viral replication. Here, we show that Nsp5 binds and cleaves human tRNA methyltransferase 1 (TRMT1), a host enzyme required for a prevalent post-transcriptional modification in tRNAs. Human cells infected with SARS-CoV-2 exhibit a decrease in TRMT1 protein levels and TRMT1-catalyzed tRNA modifications, consistent with TRMT1 cleavage and inactivation by Nsp5. Nsp5 cleaves TRMT1 at a specific position that matches the consensus sequence of SARS-CoV-2 polyprotein cleavage sites, and a single mutation within the sequence inhibits Nsp5-dependent proteolysis of TRMT1. The TRMT1 cleavage fragments exhibit altered RNA binding activity and are unable to rescue tRNA modification in TRMT1-deficient human cells. Compared to wild-type human cells, TRMT1-deficient human cells infected with SARS-CoV-2 exhibit reduced levels of intracellular viral RNA. These findings provide evidence that Nsp5-dependent cleavage of TRMT1 and perturbation of tRNA modification patterns contribute to the cellular pathogenesis of SARS-CoV-2 infection.

Proteolytic cleavage and inactivation of the TRMT1 tRNA modification enzyme by SARS-CoV-2 main protease.

Nonstructural protein 5 (Nsp5) is the main protease of SARS-CoV-2 that cleaves viral polyproteins into individual polypeptides necessary for viral replication. Here, we show that Nsp5 binds and cleaves human tRNA methyltransferase 1 (TRMT1), a host enzyme required for a prevalent post-transcriptional modification in tRNAs. Human cells infected with SARS-CoV-2 exhibit a decrease in TRMT1 protein levels and TRMT1-catalyzed tRNA modifications, consistent with TRMT1 cleavage and inactivation by Nsp5. Nsp5 cleaves TRMT1 at a specific position that matches the consensus sequence of SARS-CoV-2 polyprotein cleavage sites, and a single mutation within the sequence inhibits Nsp5-dependent proteolysis of TRMT1. The TRMT1 cleavage fragments exhibit altered RNA binding activity and are unable to rescue tRNA modification in TRMT1-deficient human cells. Compared to wild-type human cells, TRMT1-deficient human cells infected with SARS-CoV-2 exhibit reduced levels of intracellular viral RNA. These findings provide evidence that Nsp5-dependent cleavage of TRMT1 and perturbation of tRNA modification patterns contribute to the cellular pathogenesis of SARS-CoV-2 infection.

#1428 Rnd3, a novel gene involved in ADPKD

Abstract Background and Aims Autosomal dominant polycystic kidney disease (ADPKD) is an adult-onset, multi-systemic disorder, that affects ∼12.5 million people worldwide. ADPKD is characterised by progressive kidney enlargement caused by uncontrollable growth of cysts, alongside extra-renal phenotypes such as cysts in the liver, pancreas, and hypertension. Truncated mutations in PKD1 and PKD2 that encode for PC1 and PC2, respectively, are responsible for 85% and 15% of reported cases. Mutations in PKD1 are associated with the more severe form of ADPKD and earlier mean age of onset. The phenotypic variability of ADPKD can be attributed to genic, allelic and gene modifier effects. Modifier genes influence the molecular and/or phenotypic expression of other genes, altering the range of symptoms and the severity of the disease. In mice with a single gene mutation, modifier genes reduce the penetrance, dominance modification, expressivity, and phenotypic pleiotropy. To understand the involvement of modifier genes in the ADPKD kidney phenotype, a novel mouse model Rnd3tm1b(EUCOMM)Hmgu was generated, and in adult and aged Rnd3tm1b/+ mice, a kidney phenotype characteristic of ADPKD was evident. Rnd3 is an atypical member of the Rnd subgroup of the Rho family of GTPases and has a role in actin cytoskeleton reorganisation, cell cycle progression, and cell migration. Method 5 and 9-month-old kidneys from Rnd3tm1b/+ and age-matched wildtype mice were studied. In the Rnd3tm1b (EUCOMM)Hmgu mouse model, the TB1B mutation causes an inactivation of Rnd3 via the deletion of Exon 2 in C57BL/6N mice. RT-qPCR and SDS-Page Westerns were performed as well as staining for markers associated with Polycystic Kidney Disease. Results The onset of cyst formation in the cortex is apparent in 5-month-old Rnd3tm1b/+ kidneys compared to age-matched wildtype kidneys (Fig. 1a). A statistically significant 10% (p < 0.0465) increase in the proliferative index was observed in 5-month-old Rnd3tm1b/+ mice (Fig. 1e,f). Vimentin expression was also increased in podocytes and confirmed by a statistically significant 25% (p < 0.0461) increase in Rnd3tm1b/+ kidneys (Fig. 1g,h). Aquaporin-2 (Aqp2) expression was decreased in collecting ducts and verified by a statistically significant 0.4-fold-change (p < 0.0451) decrease in protein level in kidneys of 5-month-old Rnd3tm1b/+ mice (Fig. 1i-k). A statistically significant 0.75-fold (p < 0.0006) and 1.5-fold-increase (p < 0.02) in Pkd1 and Pkd2 expression was observed in 5-month-old Rnd3tm1b/+ kidneys, respectively (Fig. 1c). Kidneys of 9-month-old Rnd3tm1b/+mice presented with multiple cysts in the cortex, mimicking the ADPKD kidney phenotype (Fig. 1b). Vimentin staining in podocytes was increased and confirmed by a statistically significant 14% (p < 0.0419) increase in Rnd3tm1b/+mice (Fig. 1l,m). Expression of Pkd1 and Pkd2 was downregulated by a statistically significant 0.39-fold (p < 0.0001) and 0.2-fold-change (p < 0.004) in Rnd3tm1b/+mice (Fig. 1d). Conclusion Together, these findings highlight the importance of Rnd3 as a potential modifier in the onset of ADPKD by mediating the expression of Pkd1 and Pkd2. Interestingly, there is a critical window where Pkd1 and Pkd2 expression is altered, exacerbating the phenotype between 5 and 9-month-old Rnd3tm1b/+ mice. In addition to this, by understanding how modifiers are involved in triggering the onset of phenotypes in ADPKD, potential diagnostic and therapeutic measures can be discovered to ultimately aid in diagnosing ADPKD at a much earlier stage.

#915 Role of VSIG4 in renal aging process

Abstract Background and Aims V-set Ig domain-containing 4 (VSIG4), known for its role in inflammation and fibrosis, is particularly interesting in the realm of chronic kidney diseases. However, its specific role in kidney aging, especially under the influence of diabetes, is not well understood. This study aims to elucidate the role of VSIG4 in the aging process of kidneys, particularly in the diabetes. Method This study used male db/db and db/m mice, beginning from the age of eight weeks and spanning 38 weeks. Urinary albumin and VSIG4 levels were assessed using 6-hour timed urine collections and ELISA. The intrarenal expression of VSIG4 and klotho were analyzed through qRT-PCR and Western blotting. Results Urinary albumin levels in db/db mice were significantly higher from 8 weeks onward compared to db/m mice. These levels increased progressively with age, peaking at 38 weeks. Similarly, urinary VSIG4 levels showed a significant initial increase in db/db mice, followed by a consistent rise with age. Interestingly, both urinary albumin and VSIG4 levels in db/m mice showed a sudden surge at 38 weeks. Notably, urinary VSIG4 levels paralleled these changes in albumin levels, with a correlation coefficient of 0.867 (p < 0.001). Intrarenal VSIG4 mRNA expression was markedly higher in diabetic mice and showed a significant increase at 16 and 38 weeks, with the 38-week levels being fivefold higher in db/m mice. The expression trends of VSIG4 protein were similar to its mRNA expression. The study also monitored intrarenal klotho expression to assess aging, which revealed a progressive decrease in both mRNA and protein levels with age. Conclusion This study highlights a significant upregulation of both urinary and intrarenal VSIG4 expressions in diabetic and non-diabetic mice. The strong association of urinary VSIG4 expression with albuminuria suggests its involvement in the renal aging process, potentially contributing to the pathogenesis of age-related kidney alterations.

#2380 A multicenter and randomized study of a VLPD supplemented with ketoanalogues in patients with advanced CKD in Spain. Adherence study

Abstract Background and Aims Clinical guidelines recommend a very low-protein diet (VLPD) supplemented with ketoanalogues (KAs) for patients with advanced chronic kidney disease (CKD) to reduce progression (evidence 1A) and improve the quality of life (evidence 1C). However, there is no prior experience with this nutritional intervention in Spain. Method A multicenter, randomized, and controlled study that evaluated adherence to a VLPD supplemented with KAs (Ernamin®) in non-dialysis dependent CKD patients, estimated using the Morisky scale. Secondary objectives included assessing the effect of VLPD + KAs on kidney function, urine protein, albumin levels and other metabolic and nutritional parameters, including changes in body composition stores, at months 0, 1, 3, 6, and 9. Results Preliminary data from 33 patients (67% men; mean age 68 ± 10; GFR 16.7 ± 3.9 ml/min/1.73 m2), randomized into 2 groups, are presented. The VLPD group showed good acceptance and tolerance of the diet and supplement, with medium or high adherence in 80% of patients from the first month, maintained throughout the study (Figure). The use of VLPD was associated with a decrease in phosphorus levels (−0.4 mg/dl VLPD vs. 0.09 mg/dl LPD, P = 0.027) and C-reactive protein (CRP) levels (−2 mg/dl VLPD vs. 0.33 mg/dl LPD), an increase in calcium levels (0.3 mg/dl VLPD vs. −0.2 mg/dl LPD, P = 0.015), and an increase in muscle mass (LTM) measured by bioimpedance (3.7 kg VLPD vs. −0.6 kg LPD, P = 0.03). Stability in weight, fat mass, extracellular volume, albumin levels, and MIS scale was observed during the follow-up. Estimated glomerular filtration rate, and urine protein remained unchanged in both groups. Conclusion Protein restriction associated with KAs shows good short-term tolerance and acceptance. This study confirms several effects associated with VLPD supplemented with KAs, including an anti-inflammatory effect, the preservation of nutritional status, and a better control of phosphate levels. These data suggest safe protein intake restriction with stabilization of protein stores.

Kartogenin Induces Chondrogenesis in Cartilage Progenitor Cells and Attenuates Cell Hypertrophy in Marrow-Derived Stromal Cells.

Kartogenin (KGN) is a synthetic small molecule that stimulates chondrogenic cellular differentiation by activating smad-4/5 pathways. KGN has been proposed as a feasible alternative to expensive biologic growth factors, such as transforming growth factor β, which remain under strict regulatory scrutiny when it comes to use in patients. This study reports the previously unexplored effects of KGN stimulation on cartilage- derived mesenchymal progenitor cells (CPCs), which have been shown to be effective in applications of cell-based musculoskeletal tissue regeneration. Our findings demonstrate that KGN treatment significantly increased markers of chondrogenesis, SOX9 and COL2 following 3-10 days of treatment in human CPCs. KGN treatment also resulted in a significant dose-dependent increase in GAG production in CPCs. The same efficacy was not observed in human marrow-derived stromal cells (BM-MSCs); however, KGN significantly reduced mRNA expression of cell hypertrophy markers, COL10 and MMP13, in BM-MSCs. Parallel to these mRNA expression results, KGN led to a significant decrease in protein levels of MMP-13 both at 0-5 days and 5-10 days following KGN treatment. In conclusion, this study demonstrates that KGN can boost the chondrogenicity of CPCs and inhibit hypertrophic terminal differentiation of BM-MSCs.

Improvement of Post-Surgery Constipation in Patients with Fractures by Lactobacillus rhamnosus JYLR-127: A Single-Blind Randomized Controlled Trial.

The high prevalence of constipation after fracture surgery brings intolerable discomfort to patients on the one hand, and affects post-surgery nutrient absorption on the other hand, resulting in poor prognosis. Given the acknowledged probiotic properties of Lactobacillus rhamnosus, 100 fracture patients with post-surgery constipation were centrally enrolled and administered orally with L. rhamnosus JYLR-127 to assess the efficacy of probiotic-adjuvant therapy in alleviating post-fracture constipation symptoms. The results showed that L. rhamnosus JYLR-127 improved fecal properties, promoted gastrointestinal recovery, and relieved constipation symptoms, which were mainly achieved by elevating Firmicutes (p < 0.01) and descending Bacteroidetes (p < 0.001), hence remodeling the disrupted intestinal microecology. In addition, blood routine presented a decrease in C-reactive protein levels (p < 0.05) and an increase in platelet counts (p < 0.05) after probiotic supplementation, prompting the feasibility of L. rhamnosus JYLR-127 in anti-inflammation, anti-infection and hemorrhagic tendency prevention after fracture surgery. Our study to apply probiotics in ameliorating constipation after fracture surgery is expected to bless the bothered patients, and provide broader application scenarios for L. rhamnosus preparations.

Evaluation of Neuroprotective effect of Cassia occidentalis L. against Colchicine Induced Memory Impairment in Wistar Rats

Background: Alzheimer’s disease (AD) is a progressive neurological disorder that develops with aging. Objective: In this research, we have examined the anti - Alzheimer’s effect of ethanolic extract from roots of Cassia occidentalis L. on colchicine-induced Alzheimer’s in Wistar rats Methods: Ethanolic extract was obtained and spectroscopic, chromatography analysis was performed. Acute toxicity studies using Organization of Economic Co-operation and Development (OECD) Guidelines 423 were performed to examine and make sure that there were no signs of toxic effects. The induction of AD was done using colchicine which leads to symptoms like neurotoxicity, neuroinflammation, and neurodegeneration. In this experiment, a thorough analysis of body weight, behavioral parameters, locomotor activity, and biochemical evaluation was performed to estimate the medicinal properties of Cassia occidentalis L in treating Alzheimer’s disease. Results: Pharmacognostic analysis showed the presence of vascular bundles, starch grains, fibers, calcium oxalate crystals, elongated parenchyma, and collenchyma mucilage as shown in the supplementary files. Locomotor activity, Escape latency time, Conditioned avoidance response, and Transfer latency were improved with treatment. Interleukin- 6 (IL - 6) levels were reduced significantly in the Colchicine + 200 Cassia mg/kg group (739.2 ± 0.37 pg/ml) than in the Colchicine Group (850.6±0.40 pg/ml). Tumor necrosis factor (TNF-α) was decreased in the Colchicine + 200 Cassia mg/kg Group (1030.93±0.51 pg/ml) than in the Colchicine Group (1455.06±1.25 pg/ml). A significant decrease in total protein level was observed in the Colchicine Group (2.52±0.10 mg/ml), (3.33±0.90 mg/ml) as compared to Colchicine + 200 Cassia mg/kg Group (5.27±0.09 mg/ml, (5.01±0.10 mg/ml) respectively, in the Hippocampus and Entorhinal cortex. The levels of antioxidant enzymes such as Catalase (CAT), Serum superoxide dismutase (SOD), Reduced glutathione (GSH) and Malondialdehyde (MDA) were measured. When compared to the Colchicine Group (7.33±0.16 nM/ mg, the MDA level was lower in the Colchicine + 100 Cassia mg/kg Group (3.20±0.01 nM/ mg). The level of CAT in Colchicine + 200 Cassia mg/kg Group (7.01±0.03 μmoles of H2O2/mg of protein) was seen to be increased when compared to Colchicine Group (3.32 ± 0.17 μmoles of H2O2/mg of protein). The level of SOD in Colchicine + 200 Cassia mg/kg Group (7.43±0.02 U mg -1 of protein) was seen to be increased when compared with Colchicine Group (4.55±0.03 U mg -1 of protein). The level of GSH in Colchicine + 200 Cassia mg/kg Group (10.07±0.19 nM/mg -1 of protein) was increased when compared with the Colchicine Group (5.82±0.11nM/mg -1 of protein). Histopathology of the Hippocampus and Entorhinal cortex showed diminished amyloid plaques, and neurodegeneration in the treatment groups Conclusion: The present study showed that ethanolic extract from the roots of Cassia occidentalis L. At 100 and 200 mg/kg doses in Wistar rats improved memory damage, by reducing oxidative stress. Levels of the antioxidant enzymes as CAT, and SOD, GSH were increased and MDA was decreased. The cytokine levels in the serum of Wistar rats of IL-6 level and TNF-α level were reduced significantly. Estimation of total protein level was found to be increased. It restored neuronal degeneration in the Hippocampus, and Entorhinal cortex and reduced oxidative stress. This suggests that the ethanolic extract of Cassia occidentalis L. could be an effective therapeutic treatment for neurodegenerative diseases like AD.

Carboxyl Ester Lipase Protects Against Metabolic Dysfunction-Associated Steatohepatitis by Binding to Fatty Acid Synthase

Carboxyl ester lipase (CEL), a pivotal enzyme involved in lipid metabolism, is recurrently mutated in obese mice. Here, we aimed to elucidate the functional significance, molecular mechanism, and therapeutic potential of CEL in metabolic dysfunction-associated steatohepatitis (MASH). Hepatocyte-specific carboxyl ester lipase gene (Cel) knockout (CelΔHEP) and wildtype (WT) littermates were fed with choline-deficient high-fat diet (CD-HFD) for 16 weeks, or methionine- and choline-deficient diet (MCD) for three weeks to induce MASH. Liquid chromatography–mass spectrometry and co-immunoprecipitation were employed to identify the downstream targets of CEL. CD-HFD/MCD-fed WT mice received intravenous injections of CEL-adeno-associated viral, serotype 8 (AAV8) to induce specific overexpression of CEL in the liver. We observed a decrease in CEL protein levels in MASH induced by CD-HFD or MCD in mice. CelΔHEP mice fed with CD-HFD or MCD exhibited pronounced hepatic steatosis, inflammation, lipid peroxidation, and liver injury compared to WT littermates, accompanied by increased hepatic nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB) activation. Consistently, Cel knockdown in mouse primary hepatocytes and AML12 cells aggravated lipid accumulation and inflammation, whereas CEL overexpression exerted the opposite effect. Mechanistically, CEL directly bound to fatty acid synthase (FASN), resulting in reduced FASN SUMOylation, which in turn promoted FASN degradation through the proteasome pathway. Furthermore, inhibition of FASN ameliorated hepatocyte lipid accumulation and inflammation induced by Cel knockdown in vivo and in vitro. Hepatocyte-specific CEL overexpression using AAV8-Cel significantly mitigated steatohepatitis in mice fed with CD-HFD or MCD. CEL protects against steatohepatitis development by directly interacting with FASN and suppressing its expression for de novo lipogenesis. CEL overexpression confers a therapeutic benefit in steatohepatitis.

METTL8 links mt-tRNA m3C modification to the HIF1α/RTK/Akt axis to sustain GBM stemness and tumorigenicity

Epitranscriptomic RNA modifications are crucial for the maintenance of glioma stem cells (GSCs), the most malignant cells in glioblastoma (GBM). 3-methylcytosine (m3C) is a new epitranscriptomic mark on RNAs and METTL8 represents an m3C writer that is dysregulated in cancer. Although METTL8 has an established function in mitochondrial tRNA (mt-tRNA) m3C modification, alternative splicing of METTL8 can also generate isoforms that localize to the nucleolus where they may regulate R-loop formation. The molecular basis for METTL8 dysregulation in GBM, and which METTL8 isoform(s) may influence GBM cell fate and malignancy remain elusive. Here, we investigated the role of METTL8 in regulating GBM stemness and tumorigenicity. In GSC, METTL8 is exclusively localized to the mitochondrial matrix where it installs m3C on mt-tRNAThr/Ser(UCN) for mitochondrial translation and respiration. High expression of METTL8 in GBM is attributed to histone variant H2AZ-mediated chromatin accessibility of HIF1α and portends inferior glioma patient outcome. METTL8 depletion impairs the ability of GSC to self-renew and differentiate, thus retarding tumor growth in an intracranial GBM xenograft model. Interestingly, METTL8 depletion decreases protein levels of HIF1α, which serves as a transcription factor for several receptor tyrosine kinase (RTK) genes, in GSC. Accordingly, METTL8 loss inactivates the RTK/Akt axis leading to heightened sensitivity to Akt inhibitor treatment. These mechanistic findings, along with the intimate link between METTL8 levels and the HIF1α/RTK/Akt axis in glioma patients, guided us to propose a HIF1α/Akt inhibitor combination which potently compromises GSC proliferation/self-renewal in vitro. Thus, METTL8 represents a new GBM dependency that is therapeutically targetable.

The impact of flaxseed oil on lipid profiles, weight loss, and inflammatory markers in hemodialysis patients: A systematic review and meta-analysis of randomized controlled trials.

Studies investigating the effects of flaxseed oil on lipid profiles, weight loss, and inflammatory markers have produced inconsistent results. This systematic review and meta-analysis of randomized controlled trials (RCTs) aimed to explore the impact of flaxseed oil on these parameters in hemodialysis patients. The study protocol was registered online (PROSPERO number: CRD42023484076). The meta-analyses showed a significant decrease in triglyceride (TG) levels (WMD = -85.78 mg/dL, 95% CI: -155.24 to -16.32, I2 = 98.32%) and C-reactive protein (CRP) levels (WMD = -2.66 mg/L, 95% CI: -4.07 to -1.24, I2 = 92.26%) following consumption of flaxseed oil. Subgroup analyses revealed significant changes in LDL-C, HDL-C, and TC levels only in trials utilizing a dosage higher than 10 g per day and using ground flaxseed oil. Based on the results, flaxseed oil improves CRP and TG levels, and higher doses positively affect lipid profiles. However, it has no significant effect on anthropometric measures.

Methionine and Leucine Promote mTOR Gene Transcription and Milk Synthesis in Mammary Epithelial Cells through the eEF1Bα-UBR5-ARID1A Signaling.

Amino acids are essential for the activation of the mechanistic target of rapamycin (mTOR), but the corresponding molecular mechanism is not yet fully understood. We previously found that Met stimulated eukaryotic elongation factor α (eEF1Bα) nuclear localization in bovine mammary epithelial cells (MECs). Herein, we explored the role and molecular mechanism of eEF1Bα in methionine (Met)- and leucine (Leu)-stimulated mTOR gene transcription and milk synthesis in MECs. eEF1Bα knockdown decreased milk protein and fat synthesis, cell proliferation, and mTOR mRNA expression and phosphorylation, whereas eEF1Bα overexpression had the opposite effects. QE-MS analysis detected that eEF1Bα was phosphorylated at Ser106 in the nucleus and Met and Leu stimulated p-eEF1Bα nuclear localization. eEF1Bα knockdown abrogated the stimulation of Met and Leu by mTOR mRNA expression and phosphorylation, and this regulatory role was dependent on its phosphorylation. Akt knockdown blocked the stimulation of Met and Leu by eEF1Bα and p-eEF1Bα expression. ChIP-PCR detected that p-eEF1Bα bound only to the -548 to -793 nt site in the mTOR promoter, and ChIP-qPCR further detected that Met and Leu stimulated this binding. eEF1Bα mediated Met and Leu' stimulation on mTOR mRNA expression and phosphorylation through inducing AT-rich interaction domain 1A (ARID1A) ubiquitination degradation, and this process depended on eEF1Bα phosphorylation. p-eEF1Bα interacted with ARID1A and ubiquitin protein ligase E3 module N-recognition 5 (UBR5), and UBR5 knockdown rescued the decrease of the ARID1A protein level by eEF1Bα overexpression. Both eEF1Bα and p-eEF1Bα were highly expressed in mouse mammary gland tissues during the lactating period. In summary, we reveal that Met and Leu stimulate mTOR transcriptional activation and milk protein and fat synthesis in MECs through eEF1Bα-UBR5-ARID1A signaling.

Ca2+-Dependent Cl- Channels in Vascular Tone Regulation during Aging.

Identifying alterations caused by aging could be an important tool for improving the diagnosis of cardiovascular diseases. Changes in vascular tone regulation involve various mechanisms, like NO synthase activity, activity of the sympathetic nervous system, production of prostaglandin, endothelium-dependent relaxing, and contracting factors, etc. Surprisingly, Ca2+-dependent Cl- channels (CaCCs) are involved in all alterations of the vascular tone regulation mentioned above. Furthermore, we discuss these mechanisms in the context of ontogenetic development and aging. The molecular and electrophysiological mechanisms of CaCCs activation on the cell membrane of the vascular smooth muscle cells (VSMC) and endothelium are explained, as well as the age-dependent changes that imply the activation or inhibition of CaCCs. In conclusion, due to the diverse intracellular concentration of chloride in VSMC and endothelial cells, the activation of CaCCs depends, in part, on intracellular Ca2+ concentration, and, in part, on voltage, leading to fine adjustments of vascular tone. The activation of CaCCs declines during ontogenetic development and aging. This decline in the activation of CaCCs involves a decrease in protein level, the impairment of Ca2+ influx, and probably other alterations in vascular tone regulation.

Integrating Epigenetics, Proteomics, and Metabolomics to Reveal the Involvement of Wnt/β-Catenin Signaling Pathway in Oridonin-Induced Reproductive Toxicity.

Oridonin is the primary active component in the traditional Chinese medicine Rabdosia rubescens, displaying anti-inflammatory, anti-tumor, and antibacterial effects. It is widely employed in clinical therapy for acute and chronic pharyngitis, tonsillitis, as well as bronchitis. Nevertheless, the clinical application of oridonin is significantly restricted due to its reproductive toxicity, with the exact mechanism remaining unclear. The aim of this study was to investigate the mechanism of oridonin-induced damage to HTR-8/SVneo cells. Through the integration of epigenetics, proteomics, and metabolomics methodologies, the mechanisms of oridonin-induced reproductive toxicity were discovered and confirmed through fluorescence imaging, RT-qPCR, and Western blotting. Experimental findings indicated that oridonin altered m6A levels, gene and protein expression levels, along with metabolite levels within the cells. Additionally, oridonin triggered oxidative stress and mitochondrial damage, leading to a notable decrease in WNT6, β-catenin, CLDN1, CCND1, and ZO-1 protein levels. This implied that the inhibition of the Wnt/β-catenin signaling pathway and disruption of tight junction might be attributed to the cytotoxicity induced by oridonin and mitochondrial dysfunction, ultimately resulting in damage to HTR-8/SVneo cells.

Terbuthylazine induces oxidative stress and mitophagy through activating cGAS-STING pathway in chicken jejunum

Terbuthylazine (TBA) is a widely employed pesticide that has been found to be a significant source of food and environmental pollution. Excessive exposure to TBA raises considerable apprehension regarding its potential impact on biological health, but its underlying toxic mechanisms have not been reported. Our findings indicated that TBA exposure could cause intestinal damage, as evidenced by decrease in the quantity of cupped cells and decrease in the expression of intestinal barrier-related proteins (Claudin-1, Occludin, ZO-1, and JAM-1). Additionally, exposure to TBA resulted in a significant decrease in the expression levels of oxidative stress-related genes and proteins. Furthermore, the expression of 8-hydroxy-2′-deoxyguanosine was significantly upregulated. Additionally, excessive exposure to TBA led to a substantial release of mitochondrial DNA, which further activated the cGAS-STING pathway and caused TBK1 and IRF3 phosphorylation. Meanwhile, mitophagy was activated under TBA exposure, as evidenced by decrease in the protein and gene levels of p62 and increase in the levels of PINK1, PARKIN and LC3 II. Therefore, our results suggested that TBA exposure induced oxidative stress in the jejunum and caused mtDNA leakage, for activating the cGAS-STING pathway, which in turn promoted mitophagy. These findings revealed the toxicity mechanisms of TBA, which provided new insights into food safety caused by pesticide contamination.

Effects of Feed Deprivation on the Biochemical Responses of Pangasianodon hypophthalmus

Effects of Feed Deprivation on the Biochemical Responses of Pangasianodon hypophthalmus

185 Maternal live yeast supplementation protects newly weaned piglets against lipopolysaccharide challenge by regulating immune-associated milk proteins

Abstract Sow milk serves as a crucial source of nutritional, immunological, and growth-promoting components for the piglets. Our previous studies established that maternal live yeast (LY) supplementation altered the expression of antioxidant and immune-regulatory genes in the intestinal mucosa of suckling piglets. The current study aimed to investigate the effects of maternal LY supplementation during late gestation and lactation on milk proteomic profile in dams and health of offspring under lipopolysaccharide (LPS) challenge. On d 77 of gestation, 40 sows were allocated to 2 dietary treatments: without (CON) or with LY supplementation at 0.05% of diet during gestation and 0.1% during lactation. Milk samples were collected on d 0, 10, and 18 post-partum (n = 6), respectively. Within 24 h postweaning, piglets from the CON and LY groups were intraperitoneally injected with sterile saline or LPS [25 μg/kg body weight (BW)] and divided into four groups (n = 8): 1) saline + CON (CS), 2) saline + LY (YS), 3) LPS + CON (CLPS), and 4) LPS + LY (YLPS). Data were analyzed in a 2 × 2 factorial arrangement with main effects of maternal diet and immunological challenge. Rectal temperature was measured hourly for 4 h post-injection after which piglets were euthanized. Results showed that LPS increased concentrations of serum tumor necrosis factor (TNF) -α and interlekin (IL) -10 (P &amp;lt; 0.05) and tended to increase IL-8 in the ileal mucosa of piglets (P &amp;lt; 0.10). At 3 h post-injection, YLPS piglets tended to have a lower rectal temperature than CLPS piglets (P &amp;lt; 0.10). YLPS piglets also had decreased protein levels of TNF-α in the ileal mucosa (P &amp;lt; 0.05), with a tendency for greater protein abundance of E-cadherin but decreased mRNA abundance of catalase (P &amp;lt; 0.10) in the jejunal mucosa at 4 h post-injection (P &amp;lt; 0.05). In addition, YLPS piglets had less mRNA abundance of TLR2 and TLR4 in the liver than CLPS piglets (P &amp;lt; 0.05). With shotgun proteomics analysis, we found that milk of LY sows had more abundant proteins associated with immunity, including immunoglobulin and complement protein (Ig-like and C1q domain-containing proteins) on d 0 and d 10. Insulin-like growth factor binding protein (IGFBP) 5 was also more abundant in the d 0 milk of LY sows, whereas prostaglandin synthase was greater in d 10 and d 18 milk of CON sows. Additionally, pathway analysis showed that proteins identified in the milk of CON sows have roles in neutrophil degranulation and IL-12 signaling and production, whereas pathways associated with regulation of complement system, IGF and DHCR24 signaling, were enriched in the milk of LY sows. These results suggest that maternal LY supplementation could protect newly weaned piglets from bacteria endotoxin effect by suppressing inflammatory response, improving expression of tight junction proteins in offspring, and altering immune-associated proteins in sow milk.