P2 Levels Research Articles

Rhomboid-like 2 correlated with TME infiltration inhibits cuproptosis-related genes and drives malignant phenotype in clear cell renal cell carcinoma

The crosstalk between cuproptosis and the tumor immune microenvironment (TIME) is vital during clear cell renal cell carcinoma (ccRCC) malignant progression. However, the underlying molecular mechanisms regulate this cross-talk remain elusive. Through tailored machine learning, we analyze clinical ccRCC data from The Cancer Genome Atlas (TCGA) to explore the critical factors that regulate the interaction among cuproptosis, TIME, and tumor progression. We found that rhomboid-like 2 (RHBDL2), critical gene affecting this process, might inhibit cuproptosis-related genes (CRGs) and promotes ccRCC progression through the Wnt/β-catenin pathway. Next, knocking down RHBDL2 expression increased the cuproptosis-related genes ferredoxin 1 (FDX1) and lipoic acid synthase (LIAS) levels but reduced forkhead box P3 (FOXP3) levels and tumor growth in vivo and in vitro models. By employing HLY78, Wnt/β-catenin pathway activator, we rescued the expression of CRGs and the malignant proliferation and metastasis capacity in ccRCC cells with RHBDL2 knockdown. Mechanistically, RHBDL2 inhibits cuproptosis and promotes malignant progression of ccRCC through the Wnt/β-catenin pathway. Abnormal RHBDL2 expression may cause the suppressive TIME formation by regulating Treg-cell infiltration, thus triggering immune escape. In summary, our results indicated that RHBDL2 is an oncogene that induces tumorigenesis and targeting RHBDL2 may be novel therapeutic direction for metastatic ccRCC.

Does level of cognitive load affect susceptibility?

We compared how different levels of cognitive load affect frontal P3 (fP3) Event-Related Potential (ERP) to novel sounds. Previous studies demonstrated the predictive value of the probe-elicited frontal P3 (fP3) ERP for subsequent detection failures. They also demonstrated how fP3 is reduced when performing visual and/or manual and/or cognitively demanding tasks. These results are consistent with fP3 indexing orienting to novels or, more neutrally: susceptibility. Here, we tested how fP3 is affected by a threefold variation of cognitive load induced by the verb (generation) task. Participants heard a noun and either listened to it, repeated it, or generated a semantically related verb. These conditions were manipulated between groups. One group (N = 16) experienced the listen and repeat condition; the other group (N = 16) experienced the listen and generate condition. When fP3 was probed 0 or 200 ms after noun offset, it was reduced (relative to no noun) only while repeating or generating, not while listening. An additional probe-elicited ERP was identified as novelty-related negativity, and its contaminating influence on fP3 estimation accounted for by a novel vector-filter procedure. We conclude that cognitive load does not affect fP3-indexed susceptibility. Instead, fP3-indexed susceptibility is affected by presentation of the stimulus, with the most pronounced effect in conditions where a vocal response is needed (i.e., repeat or generate, but not listen), independent of the complexity of the response.

Tanshinone IIA potentiates the therapeutic efficacy of glucocorticoids in lipopolysaccharide-treated HEI-OC1 cells through modulation of the FOXP3/Nrf2 signaling pathway.

Glucocorticoids (GCs) are commonly used to treat sudden sensorineural hearing loss (SSNHL), although some patients are resistant to this therapeutic approach. Clinical studies have demonstrated the efficacy of tanshinone IIA (TA) in combination with GC for managing various human ailments. However, it remains unclear whether TA can mitigate GC resistance in SSNHL. Our aim is to elucidate the role of NRF2-induced transcriptional regulation of HDAC2 in influencing GC resistance and investigate the involvement of TA-related molecular pathways in GC resistance. Here, HEI-OC1 cells are treated with lipopolysaccharide (LPS) to establish an in vitro model for SSNHL. The cells are subsequently treated with dexamethasone (DXE) or DXE+TA. RT-qPCR and western blot analysis are used to measure the mRNA and protein levels of Forkhead box P3 (FOXP3), nuclear factor erythroid 2-related factor 2 (NRF2), and histone deacetylase 2 (HDAC2). Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) assays are carried out to assess cell proliferation. Flow cytometry analysis is performed to evaluate apoptosis. Mechanistic studies involve chromatin immunoprecipitation (ChIP), luciferase reporter, and DNA pull-down assays. Our results show that treatment with TA+DEX significantly increases proliferation and suppresses apoptosis in LPS-treated HEI-treated OC1 cells. TA upregulates HDAC2 expression by activating NRF2-mediated transcription of HDAC2, with the NRF2-HDAC2 binding site located at bases 419-429 (ATGACACTCCA) in the promoter sequence of HDAC2. Furthermore, TA upregulates FOXP3 expression to activate NRF2 transcription, with the predicted FOXP3-binding site located at bases 864-870 (GCAAACA) in the promoter sequence of NRF2. In summary, these findings suggest that TA enhances the therapeutic effects of GC on the proliferation and apoptosis of HEI OC1 cells by increasing FOXP3/Nrf2 expression. These results indicate that TA may be promising for ameliorating GC resistance in patients with SSNHL.

Novel phosphatidylinositol flippases contribute to phosphoinositide homeostasis in the plasma membrane.

Phosphatidylinositol is a precursor of various phosphoinositides, which play crucial roles in intracellular signaling and membrane dynamics and have impact on diverse aspects of cell physiology. Phosphoinositide synthesis and turnover occur in the cytoplasmic leaflet of the organellar and plasma membranes. P4-ATPases (lipid flippases) are responsible for translocating membrane lipids from the exoplasmic (luminal) to the cytoplasmic leaflet, thereby regulating membrane asymmetry. However, the mechanism underlying phosphatidylinositol translocation across cellular membranes remains elusive. Here, we discovered that the phosphatidylcholine flippases ATP8B1, ATP8B2, and ATP10A can also translocate phosphatidylinositol at the plasma membrane. To explore the function of these phosphatidylinositol flippases, we used cells depleted of CDC50A, a protein necessary for P4-ATPase function and ATP8B1 and ATP8B2, which express in HeLa cells. Upon activation of the Gq-coupled receptor, depletion of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] was accelerated in CDC50A knockout (KO) and ATP8B1/8B2 double KO cells compared with control cells, suggesting a decrease in PtdIns(4,5)P2 levels within the plasma membrane of the KO cells upon stimulation. These findings highlight the important role of P4-ATPases in maintaining phosphoinositide homeostasis and suggest a mechanism for asymmetry of phosphatidylinositol in the cytoplasmic leaflet of the plasma membrane.

Effectiveness of Using A Combination of Rice Husk Ash and Monosodium Glutamate (MSG) on Growth of Lettude Plant (Lactuca sativa L.)

Lettuce is a plant that is quite popular with the public. Not only does it have economic value but it also has quite high nutritional content. So one method used to maintain the stability of lettuce production on the market is by providing rice husk ash and monosodium glutamate (MSG). This research uses a non-factorial randomized block design (RAK) which consists of 5 levels, namely giving fertilizer without combination and giving combined fertilizer, namely P1 (Rice husk ash 100 grams/polybag), P2 (MSG 15 grams/polybag), P3 (Combination of rice husk ash and MSG with a dose ratio of 100 grams of rice husk ash : 15 grams of MSG), P4 (Combination of rice husk ash and MSG with a dose ratio of 75 grams of rice husk ash : 10 grams of MSG), P5 (Combination of rice husk ash and MSG with a dose ratio of 50 grams of rice husk ash: 5 grams of MSG). The study's findings demonstrated a significant impact on all parameters (plant height, number of leaves, leaf area index, and plant fresh weight) when a combination of rice husk ash and MSG was used. The results of testing the levels of chlorophyll a b and the total use of a combination of rice husk ash and MSG did not provide a significant effect. The most effective dose to increase the growth of lettuce plants was the use of a combination of rice husk ash and MSG with a level of P3 = 100 grams of rice husk ash: 15 grams of MSG.

The LCLAT1/LYCAT acyltransferase is required for EGF-mediated phosphatidylinositol-3,4,5-trisphosphate generation and Akt signaling.

Receptor tyrosine kinases such as EGF receptor (EGFR) stimulate phosphoinositide 3 kinases to convert phosphatidylinositol-4,5-bisphosophate [PtdIns(4,5)P2] into phosphatidylinositol-3,4,5-trisphosphate [PtdIns(3,4,5)P3]. PtdIns(3,4,5)P3 then remodels actin and gene expression, and boosts cell survival and proliferation. PtdIns(3,4,5)P3 partly achieves these functions by triggering activation of the kinase Akt, which phosphorylates targets like Tsc2 and GSK3β. Consequently, unchecked upregulation of PtdIns(3,4,5)P3-Akt signaling promotes tumor progression. Interestingly, 50-70% of PtdIns and PtdInsPs have stearate and arachidonate at sn-1 and sn-2 positions of glycerol, respectively, forming a species known as 38:4-PtdIns/PtdInsPs. LCLAT1 and MBOAT7 acyltransferases partly enrich PtdIns in this acyl format. We previously showed that disruption of LCLAT1 lowered PtdIns(4,5)P2 levels and perturbed endocytosis and endocytic trafficking. However, the role of LCLAT1 in receptor tyrosine kinase and PtdIns(3,4,5)P3 signaling was not explored. Here, we show that LCLAT1 silencing in MDA-MB-231 and ARPE-19 cells abated the levels of PtdIns(3,4,5)P3 in response to EGF signaling. Importantly, LCLAT1-silenced cells were also impaired for EGF-driven and insulin-driven Akt activation and downstream signaling. Thus, our work provides first evidence that the LCLAT1 acyltransferase is required for receptor tyrosine kinase signaling.

Immunomodulatory Effects of a Probiotic Mixture: Alleviating Colitis in a Mouse Model through Modulation of Cell Activation Markers and the Gut Microbiota.

Ulcerative colitis (UC) is a persistent inflammatory intestinal disease that consistently affects the colon and rectum. Its exact cause remains unknown. UC causes a considerable challenge in healthcare, prompting research for novel therapeutic strategies. Although probiotics have gained popularity as possible candidates for managing UC, studies are still ongoing to identify the best probiotics or probiotic mixtures for clinical applications. This study aimed to determine the efficacy of a multi-strain probiotic mixture in mitigating intestinal inflammation in a colitis mouse model induced by dextran sulfate sodium. Specifically, a multi-strain probiotic mixture consisting of Tetragenococcus halophilus and Eubacterium rectale was used to study its impact on colitis symptoms. Anti-inflammatory effects were evaluated using ELISA and flow cytometry. The configuration of gut microbial communities was determined using 16S rRNA metagenomic analysis. According to this study, colitis mice treated with the probiotic mixture experienced reduced weight loss and significantly less colonic shortening compared to untreated mice. Additionally, the treated mice exhibited increased levels of forkhead box P3 (Foxp3) and interleukin 10, along with decreased expression of dendritic cell activation markers, such as CD40+, CD80+, and CD83+, in peripheral blood leukocytes and intraepithelial lymphocytes. Furthermore, there was a significant decrease in the frequencies of CD8+N.K1.1+ cells and CD11b+Ly6G+ cells. In terms of the gut microbiota, probiotic-mixture treatment of colitis mice significantly increased the abundance of the phyla Actinobacteria and Verrucomicrobia (p < 0.05). These results provide valuable insights into the therapeutic promise of multi-strain probiotics, shedding light on their potential to alleviate colitis symptoms. This research contributes to the ongoing exploration of effective probiotic interventions for managing inflammatory bowel disease.

PTEN and the PTEN-like phosphatase CnrN have both distinct and overlapping roles in a Dictyostelium chemorepulsion pathway.

Little is known about eukaryotic chemorepulsion. The enzymes phosphatase and tensin homolog (PTEN) and CnrN dephosphorylate phosphatidylinositol 3,4,5-trisphosphate [PI(3,4,5)P3] to phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2]. Dictyostelium discoideum cells require both PTEN and CnrN to induce chemorepulsion of cells away from the secreted chemorepellent protein AprA. How D. discoideum cells utilize two proteins with redundant phosphatase activities in response to AprA is unclear. Here, we show that D. discoideum cells require both PTEN and CnrN to locally inhibit Ras activation, decrease basal levels of PI(3,4,5)P3 and increase basal numbers of macropinosomes, and AprA prevents this increase. AprA requires both PTEN and CnrN to increase PI(4,5)P2 levels, decrease PI(3,4,5)P3 levels, inhibit proliferation, decrease myosin II phosphorylation and increase filopod sizes. PTEN, but not CnrN, decreases basal levels of PI(4,5)P2, and AprA requires PTEN, but not CnrN, to induce cell roundness. Together, our results suggest that CnrN and PTEN play unique roles in AprA-induced chemorepulsion.

Sur7 mediates a novel pathway for PI4,5P2 regulation in C. albicans that promotes stress resistance and cell wall morphogenesis.

The human fungal pathogen Candida albicans can cause lethal systemic infections due to its ability to resist stress from the host and to undergo invasive hyphal growth. Previous studies showed that plasma membrane MCC/eisosome domains were important for virulence by promoting the ability of Sur7 to mediate normal cell wall morphogenesis and stress resistance. The sur7Δ mutant displayed abnormal clusters of PI4,5P2, suggesting that misregulation of this lipid underlies the sur7Δ phenotype. To test this, we increased PI4,5P2 levels by deleting combinations of the three PI4,5P2 5' phosphatase genes (INP51, INP52, and INP54) and found that some combinations, such as inp51Δ inp52Δ, gave phenotypes similar the sur7Δ mutant. In contrast, deleting one copy of MSS4, the gene that encodes the 5' kinase needed to create PI4,5P2, reduced the abnormal PI4,5P2 clusters and also decreased the abnormal cell wall and stress sensitive phenotypes of the sur7Δ mutant. Additional studies support a model that the abnormal PI4,5P2 patches recruit septin proteins, which in turn promote aberrant cell wall growth. These results identify Sur7 as a novel regulator of PI4,5P2 and highlight the critical role of PI4,5P2 in the regulation of C. albicans virulence properties.

Continuous Intercropping Increases the Depletion of Soil Available and Non-Labile Phosphorus

Background and aims: This research aimed to evaluate the effects of consecutive intercropping on soil phosphorus (P) partitioning, concentrations, and sensitivity to P fertilizer application, elucidating its impact on soil P bioavailability. Methods: A field experiment investigated soil P fractions and content under continuous wheat and faba bean intercropping. Three P levels (0, 45, and 90 kg P2O5 ha−1 denoted as P0, P1, and P2, respectively) and three planting patterns (monocropped wheat (MW), monocropped faba bean (MF), and wheat and faba bean intercropping (W//F)) were established since 2014. Aboveground P uptake by wheat and faba bean was determined. The soil P fractions and content were analyzed after six-, seven-, and eight-year continuous field experiments. Results: Wheat and faba bean intercropping increased wheat aboveground P uptake by 28.3–42.7% compared to MW under P1 and P2 levels and presented a P uptake advantage (LERPuptake &gt; 1), although W//F had no impact on faba bean P uptake. Consequently, continuous intercropping for 8 years decreased soil available P reserves by 9.0–23.4% in comparison to the weighted average value of MW and MF (It). Faba bean consumed greater non-labile and labile P than wheat with low P input. W//F had nearly no impact on the labile P pool but reduced the non-labile P pool by 5.0–12.1% under all P levels and lowered the moderately labile P pool by 1.7–4.7% at P0 and P1 levels compared to It with consecutive intercropping for 8 years. Consecutive intercropping of wheat and faba bean primarily decreased the proportion of Resin-P in the labile P pool and the proportion of Residual-P in the non-labile P pool. According to the structural equation model, crop P uptake mainly originated from soil available P, which was directly affected by non-labile P (Residual-P and Conc. HCl-P). In addition, intercropping changed the contribution of each P faction to crop P uptake compared to MW and MF, and P uptake in intercropping primarily depended on Conc. HCl- P and Dil. HCl-P. Therefore, consecutive intercropping decreased soil non-labile P and drove soil available P depletion, and intercropping’s increase of P uptake was related to the non-labile P mobilized to moderately labile and labile P. Conclusions: Continuous wheat and faba bean intercropping reduced non-labile P and led to soil available P depletion under low P input. This practice stimulated non-labile P mobilization, enhancing soil P fraction effectiveness and facilitating P uptake in intercropping. Continuous intercropping of wheat and faba bean is as an effective method to maximize the biological availability of soil P and reduce P application rates.

Response of Inorganic Fertilizer and Plant Growth Regulators on Growth and Seed Yield of Radish (Raphanus sativus L.)

This study aimed to evaluate the influence of different levels of inorganic fertilizer and plant growth regulator on seed yield of radish (Raphanus sativus L.) was formulated and conducted during the rabi season of 2022-23 at vegetable seed production plot, Department of Horticulture (Vegetable &amp; Floriculture), BAC, Sabour Bhagalpur. The experiment was conducted to determine seed production ability at four levels of fertilizers viz, (F0-control, F1-N40:P30:K40, F2-N60:P40:K50,&amp; F3-N8P50:K50) as well as seven levels of PGRs viz, P0 (Control), P1 (GA3 @ 50ppm), P2 (GA3 @100 ppm), P3 (GA3 @ 150 ppm), P4 (NAA @ 50 ppm), P5 (NAA @ 100 ppm) and P6 (NAA @ 150 ppm). Inorganic fertilizers were applied at 20 days and PGRs as foliar spray at 30 and 60 days after stecklings transplanting. The experiment was conducted in RBD (Factorial) and replicated thrice. the combined effect of inorganic fertilizers and Plant growth regulators F2P3 significantly gave maximum diameter of pod (7.64 mm), length of pod (56.61mm), number of seeds/pod (6.18), 1000- seed weight (15.12g) and seed yield (18.23q/ha) and plant height is non significant. The treatment combination F3P6 gave the maximum number of branches (8.06). Hence, F2P3 treatment combination is a best combination for seed yield of radish and enhancing seed yield is our ultimate goal. As we performed the study in a particular location, we recommend multilocation trials in different agro-climatic regions to study and final confirmation of the results.

The role of microRNA-142a in Toxoplasma gondii infection-induced downregulation of Foxp3: implications for adverse pregnancy outcomes

BackgroundToxoplasma gondii (T. gondii) is capable of infecting nearly all warm-blooded animals and approximately 30% of the global population. Though most infections are subclinical in immunocompetent individuals, congenital contraction can lead to severe consequences such as spontaneous abortion, stillbirth, and a range of cranio-cerebral and/or ocular abnormalities. Previous studies reported that T. gondii-infected pregnancy mice unveiled a deficit in both the amount and suppressive functions of regulatory T (Treg) cells, accompanied with reduced levels of forkhead box p3 (Foxp3). Recently, accumulative studies have demonstrated that microRNAs (miRNAs) are, to some extent, relevant to T. gondii infection. However, the link between alterations in miRNAs and downregulation of Foxp3 triggered by T. gondii has been only sporadically studied.MethodsQuantitative reverse transcription polymerase chain reaction (RT-qPCR), protein blotting and immunofluorescence were employed to evaluate the impact of T. gondii infection and antigens on miRNA transcription and Foxp3 expression. Dual-luciferase reporter gene assays were performed to examine the fluorescence activity in EL4 cells, which were transfected with recombinant plasmids containing full-length/truncated/mutant microRNA-142a-3p (miR-142a) promoter sequence or wild type/mutant of Foxp3 3’ untranslated region (3’ UTR).ResultsWe found a pronounced increase in miR-142a transcription, concurrent with a decrease in Foxp3 expression in T. gondii-infected mouse placental tissue. Similarly, comparable findings have been experimentally confirmed through the treatment of EL4 cells with T. gondii antigens (TgAg) in vitro. Simultaneously, miR-142a mimics attenuated Foxp3 expression, whereas its inhibitors markedly augmented Foxp3 expression. miR-142a promoter activity was elevated upon the stimulation of T. gondii antigens, which mitigated co-transfection of mutant miR-142a promoter lacking P53 target sites. miR-142a mimics deceased the fluorescence activity of Foxp3 3’ untranslated region (3’ UTR), but it did not affect the fluorescence activity upon the co-transfection of mutant Foxp3 3’ UTR lacking miR-142a target site.ConclusionIn both in vivo and in vitro studies, a negative correlation was discovered between Foxp3 expression and miR-142a transcription. TgAg enhanced miR-142a promoter activity to facilitate miR-142a transcription through a P53-dependent mechanism. Furthermore, miR-142a directly targeted Foxp3 3’ UTR, resulting in the downregulation of Foxp3 expression. Therefore, harnessing miR-142a may be a possible therapeutic approach for adverse pregnancy caused by immune imbalances, particularly those induced by T. gondii infection.

Acupoint Autohemotherapy Attenuates DNCB-Induced Atopic Dermatitis and Activates Regulatory T Cells in BALB/c Mice.

Acupoint autohemotherapy (A-AHT) has been proposed as an alternative and complementary treatment for atopic dermatitis (AD), yet the exact role of its blood component in terms of therapeutic efficacy and mechanism of action is still largely unknown. This study aimed to evaluate the therapeutic efficacies and action mechanisms of intramuscular injections of autologous whole blood (AWB) and mouse immunoglobulin G (IgG) (autologous or heterologous) at acupoints on 2,4-dinitrochlorobenzene (DNCB)-induced AD mouse models. Serum levels of total immunoglobulin E (IgE), IgG, interleukin-10 (IL-10), and interferon-gamma (IFN-γ) were measured, as well as mRNA expression levels of Forkhead box P3 (FoxP3), IL-10 and IFN-γ in dorsal skin lesions, and IL-10+, IFN-γ+ and FoxP3+CD4+T cells in murine spleen. It showed that repeated acupoint injection of AWB, autologous total IgG (purified from autologous blood in AD mice) or heterologous total IgG (purified from healthy blood in normal mice) effectively reduced the severity of AD symptoms and decreased epidermal and dermal thickness as well as mast cells in skin lesions. Additionally, AWB acupoint injection was found to upregulate FoxP3+, IL-10+ and IFN-γ+ CD4+T cells in murine spleen, suppressing the production of IgE antibodies and increasing that of IgG antibodies in the serum. Furthermore, both AWB and autologous total IgG administrations significantly elevated FoxP3 expression, mRNA levels of IL-10 and IFN-γ in dorsal skin lesions. However, acupoint injection of heterologous total IgG had no effect on regulatory T (Treg) and Th1 cells modulation. These findings suggest that the therapeutic effects of A-AHT on AD are mediated by IgG-induced activation of Treg cells.

Non-vesicular phosphatidylinositol transfer plays critical roles in defining organelle lipid composition

Phosphatidylinositol (PI) is the precursor lipid for the minor phosphoinositides (PPIns), which are critical for multiple functions in all eukaryotic cells. It is poorly understood how phosphatidylinositol, which is synthesized in the ER, reaches those membranes where PPIns are formed. Here, we used VT01454, a recently identified inhibitor of class I PI transfer proteins (PITPs), to unravel their roles in lipid metabolism, and solved the structure of inhibitor-bound PITPNA to gain insight into the mode of inhibition. We found that class I PITPs not only distribute PI for PPIns production in various organelles such as the plasma membrane (PM) and late endosomes/lysosomes, but that their inhibition also significantly reduced the levels of phosphatidylserine, di- and triacylglycerols, and other lipids, and caused prominent increases in phosphatidic acid. While VT01454 did not inhibit Golgi PI4P formation nor reduce resting PM PI(4,5)P2 levels, the recovery of the PM pool of PI(4,5)P2 after receptor-mediated hydrolysis required both class I and class II PITPs. Overall, these studies show that class I PITPs differentially regulate phosphoinositide pools and affect the overall cellular lipid landscape.

The ER tether VAPA is required for proper cell motility and anchors ER-PM contact sites to focal adhesions.

Cell motility processes highly depend on the membrane distribution of Phosphoinositides, giving rise to cytoskeleton reshaping and membrane trafficking events. Membrane contact sites serve as platforms for direct lipid exchange and calcium fluxes between two organelles. Here, we show that VAPA, an ER transmembrane contact site tether, plays a crucial role during cell motility. CaCo2 adenocarcinoma epithelial cells depleted for VAPA exhibit several collective and individual motility defects, disorganized actin cytoskeleton and altered protrusive activity. During migration, VAPA is required for the maintenance of PI(4)P and PI(4,5)P2 levels at the plasma membrane, but not for PI(4)P homeostasis in the Golgi and endosomal compartments. Importantly, we show that VAPA regulates the dynamics of focal adhesions (FA) through its MSP domain, is essential to stabilize and anchor ventral ER-PM contact sites to FA, and mediates microtubule-dependent FA disassembly. To conclude, our results reveal unknown functions for VAPA-mediated membrane contact sites during cell motility and provide a dynamic picture of ER-PM contact sites connection with FA mediated by VAPA.

Remote Ischemic Post-conditioning Reduces Cognitive Impairment in Rats Following Subarachnoid Hemorrhage: Possible Involvement in STAT3/STAT5 Phosphorylation and Th17/Treg Cell Homeostasis.

The inflammatory response following subarachnoid hemorrhage (SAH) may lead to Early Brain Injury and subsequently contribute to poor prognosis such as cognitive impairment in patients. Currently, there is a lack of effective strategies for SAH to ameliorate inflammation and improve cognitive impairment in clinical. This study aims to examine the inhibitory impact of remote ischemic post-conditioning (RIPostC) on the body's inflammatory response by regulating Th17/Treg cell homeostasis after SAH. The ultimate goal is to search for potential early treatment targets for SAH. The rat SAH models were made by intravascular puncture of the internal carotid artery. The intervention of RIPostC was administered for three consecutive days immediately after successful modeling. Behavioral experiments including the Morris water maze and Y-maze tests were conducted to assess cognitive functions such as spatial memory, working memory, and learning abilities 2weeks after successful modeling. The ratio of Th17 cells and Treg cells in the blood was detected using flow cytometry. Immunofluorescence was used to observe the infiltration of neutrophils into the brain. Signal transducers and activators of transcription 5 (STAT5) and signal transducers and activators of transcription 3 (STAT3) phosphorylation levels, receptor-related orphan receptor gamma-t (RORγt), and forkhead box protein P3 (Foxp3) levels were detected by Western blot. The levels of anti-inflammatory factors (IL-2, IL-10, IL-5, etc.) and pro-inflammatory factors (IL-6, IL-17, IL-18, TNF-α, IL-14, etc.) in blood were detected using Luminex Liquid Suspension Chip Assay. RIPostC significantly improved the cognitive impairment caused by SAH in rats. The results showed that infiltration of Th17 cells and neutrophils into brain tissue increased after SAH, leading to the release of pro-inflammatory factors (IL-6, IL-17, IL-18, and TNF-α). This response can be inhibited by RIPostC. Additionally, RIPostC facilitates the transfer of Treg from blood to the brain and triggers the release of anti-inflammatory (IL-2, IL-10, and IL-5) factors to suppress the inflammation following SAH. Finally, it was found that RIPostC increased the phosphorylation of STAT5 while decreasing the phosphorylation of STAT3. RIPostC reduces inflammation after SAH by partially balancing Th17/Treg cell homeostasis, which may be related to downregulation of STAT3 and upregulation of STAT5 phosphorylation, which ultimately alleviates cognitive impairment in rats. Targeting Th17/Treg cell homeostasis may be a promising strategy for early SAH treatment.

MARCKS and PI(4,5)P2 reciprocally regulate actin-based dendritic spine morphology.

Myristoylated, alanine-rich C-kinase substrate (MARCKS) is an F-actin and phospholipid binding protein implicated in numerous cellular activities, including the regulation of morphology in neuronal dendrites and dendritic spines. MARCKS contains a lysine-rich effector domain that mediates its binding to plasma membrane phosphatidylinositol-4,5-biphosphate (PI(4,5)P2) in a manner controlled by PKC and calcium/calmodulin. In neurons, manipulations of MARCKS concentration and membrane targeting strongly affect the numbers, shapes, and F-actin properties of dendritic spines, but the mechanisms remain unclear. Here, we tested the hypothesis that the effects of MARCKS on dendritic spine morphology are due to its capacity to regulate the availability of plasma membrane PI(4,5)P2. We observed that the concentration of free PI(4,5)P2 on the dendritic plasma membrane was inversely proportional to the concentration of MARCKS. Endogenous PI(4,5)P2 levels were increased or decreased, respectively, by acutely overexpressing either phosphatidylinositol-4-phosphate 5-kinase (PIP5K) or inositol polyphosphate 5-phosphatase (5ptase). PIP5K, like MARCKS depletion, induced severe spine shrinkage; 5ptase, like constitutively membrane-bound MARCKS, induced aberrant spine elongation. These phenotypes involved changes in actin properties driven by the F-actin severing protein cofilin. Collectively, these findings support a model in which neuronal activity regulates actin-dependent spine morphology through antagonistic interactions of MARCKS and PI(4,5)P2.

Effect of Maltodextrin Concentration on Physical Characteristics of Strawberry Extract That May Prevent COVID-19 in the Elderly

Background: The strawberry is an antioxidant-rich fruit with the potential to prevent COVID-19, especially in the elderly. However, the fruit's high-water content makes it susceptible to decay and diminishes its quality. Product diversification into instant powder with a spray drying method and the addition of maltodextrin is one possible solution. Objectives: The purpose of this study is to examine the effect of maltodextrin concentration on the physical properties of strawberry powder produced by spray drying. Methods: The experimental study employed a completely randomized design and three maltodextrin concentrations: P1 (20%), P2 (25%), and P3 (30%). Organoleptic data were gathered using a questionnaire with a 1 to 9 hedonic scale scoring method. 35 students from the sixth and eighth semesters of Nutrition at Binawan University served as panelists. Results: The addition of maltodextrin to strawberry powder significantly altered the powder's flavor, aroma, texture, and color. The solubility test revealed that the strawberry powders P1, P2, and P3 were water-soluble. The selected strawberry powder contained 5.08% moisture, 1.05% ash, 1.13% protein, 0.02% fat, and 92.73% carbohydrates, according to approximative analysis results. The results revealed that the antioxidant levels of P1 (20%), P2 (25%), and P3 (30%) were 7.95 ppm, 8.07 ppm, and 8.36 ppm, respectively. Conclusions: When added, maltodextrin at a concentration of 30% produces the best possible physical properties.

Pengaruh Penggunaan Ragi Alami (Sourdough Starter) terhadap Mutu Organoleptik, Mutu Kimia, Mutu Fungsional, Nilai Energi, dan Indeks Glikemik Roti Tawar Substitusi Puree Ubi Jalar Ungu (Ipomoea Batatas L) bagi Penderita Diabetes Melitus Tipe 2

Type 2 Diabetes Meitus (T2DM) is a chronic disease with the prevalence of sufferers that continues to increase every year and can cause complications and even death. T2DM management can be done through nutritional therapy, education, physical activity, and pharmacology. One form of nutritional therapy is to do 3J, namely the right schedule, type, and amount in DM patients. The right type can be done by choosing foods with a low to moderate glycemic index so that people with diabetes can control their blood glucose levels. The purpose of this study was to determine and analyze the effect of using natural yeast (sourdough starter) on chemical quality (moisture content, ash content, and carbohydrates), functional quality, namely resistant starch fiber, glycemic index, energy value, and organoleptic quality (taste, color, texture, and aroma) in white bread substituted with purple sweet potato for patients with type 2 diabetes. This type of research was experimental Completely Randomized Design (CRD) using 3 levels of treatment with natural yeast percentages of 30%, 40%, and 50% with the addition of purple sweet potato (Ipomoea Batatas L.) as much as 40%. The results showed that natural yeast (sourdough starter) had a significant effect (p&lt;0.05) on chemical and functional quality, namely water content, ash content, carbohydrates, and resistant starch as well as on organoleptic quality of taste, color, and aroma. Chemical quality, namely carbohydrate content meets the 2015 PERKENI dietary standards (45 – 65%) and the functional quality of resistant crate fiber also meets the 2015 PERKENI dietary standards (20 – 35 grams/day). The water content and ash content have met SNI 01-3840-1995 for white bread standards. The best treatment has glycemic index 71 was at the level of treatment P2 with the highest yield value (Nh) 0.81 .&#x0D; Keyword: Type 2 Diabetes Meitus, Bread, Natural yeast, Glycemic index

Characterization of Pik1 function in fission yeast reveals its conserved role in lipid synthesis and not cytokinesis.

Phosphatidylinositol (PI)-4-phosphate (PI4P) is a lipid found at the plasma membrane (PM) and Golgi in cells from yeast to humans. PI4P is generated from PI by PI4-kinases and can be converted to PI-4,5-bisphosphate [PI(4,5)P2]. Schizosaccharomyces pombe have 2 essential PI4-kinases: Stt4 and Pik1. Stt4 localizes to the PM and its loss from the PM results in a decrease of PM PI4P and PI(4,5)P2. As a result, cells divide non-medially due to disrupted cytokinetic ring-PM anchoring. However, the localization and function of S. pombe Pik1 has not been thoroughly examined. Here, we found that Pik1 localizes exclusively to the trans-Golgi and is required for Golgi PI4P production. We determined that Ncs1 regulates Pik1, but unlike in other organisms, it is not required for Pik1 Golgi localization. When Pik1 function was disrupted, PM PI4P but not PI(4,5)P2 levels were reduced, a major difference with Stt4. We conclude that Stt4 is the chief enzyme responsible for producing the PI4P that generates PI(4,5)P2. Also, that cells with disrupted Pik1 do not divide asymmetrically highlights the specific importance of PM PI(4,5)P2 for cytokinetic ring-PM anchoring.