Cellular and molecular biology最新文献

The human microbial flora is quite diverse and versatile, playing several beneficial roles in association with the host and deriving nutrition from it. The present study aimed to identify gut microbial flora with potential probiotic activities. Eighteen bacterial isolates were screened from ten male individuals in this study. Seven bacterial isolates, NCCP-2046, NCCP-2031, NCCP-2035, NCCP-2040, NCCP-2041, NCCP-2044, and NCCP-2046, were isolated from the gut samples of volunteer men belonging to various areas of Rawalpindi and Islamabad. These bacterial isolates were cultured on De Man Rogosa and Sharpe Media (MRS), Tryptone Soya Agar (TSA), and Nutrient agar, which showed efficient bacterial growth. The morphological and biochemical characteristics of these bacterial strains were studied under their optimal growth conditions, along with molecular investigations. The antibiotic sensitivity pattern was tested using Kirby-Bauer method, which verified the higher MIC against all eight antibiotics used except for oxacillin. Phylogenetic analysis of only four bacterial isolates was performed based on their 16S rRNA sequences, and their top-hit sequence similarities in NCBI and EzBioCloud.net (95-98% and 94%) verified that these bacterial candidates belong to the Priestia and Staphylococcus genera. Based on molecular evidence through phylogeny and sequence similarities with previously defined bacterial candidates, the bacterial strains MG-461621 (NCCP-2031), MG-461622 (NCCP-2035), and MG-561934 (NCCP-2046) are presumed to be members of Priestia or novel species/genera, while MG-461623 (NCCP-2039) is also found to be a previously identified species of Staphylococcus. However, due to decreased similarity with the top-hit sequences, it could also be presumed to represent a member of a novel genus.

The present study aimed to identify the active substances in orange peel powder (PO) and to extract beta-carotene (OR) from dried orange peel powder. Additionally, the study aims to examine the efficacy of these compounds as natural antioxidants. The levels of Vitamin C, phenolic compounds, flavonoids, and pectin were found to be significantly greater in OR compared to PO at (P≤0.01) level. Both PO and OR demonstrated a strong correlation between increasing concentrations with the removal of free radicals. The method of scavenging free radicals displayed a higher efficacy compared to the method of lowering ferric chloride (FeCl2). Additionally, it was observed that the elimination of free radicals increased with higher concentrations. The efficacy of both PO and OR as antioxidants was also assessed through implementing the method of introducing hydrogen peroxide (H2O2) by estimating the fragmentation factor of DNA)QB.(  There were statistically significant differences at (P≤0.01) level, demonstrated by the reduction in QB with rising levels of PO and OR. The concentration of QB is 0 at 250 µg/ml for both PO and RO. This could be due to their efficacy as antioxidants, enabling them to eradicate free radicals that degrade DNA. The findings supported the hypothesis that orange peel powder (PO) and beta-carotene pigment (OR) function as potent natural antioxidants, effectively mitigating or eliminating oxidative processes induced by free radicals. These compounds are considered safe for human consumption and do not pose any health risks.

Kisspeptins are reported to be the most potent activators of the hypothalamus-pituitary-gonadal (HPG) axis known to date. Kisspeptin potently elicits gonadotropin-releasing hormone (GnRH) release and luteinizing hormone (LH) secretion, even in the pre-pubertal period. Beyond the hypothalamus, kisspeptin is also expressed in limbic and paralimbic brain regions, which are areas of the neurobiological network primarily implicated in emotional behaviors alongside sexual functions. Therefore, an increasing body of studies has implicated kisspeptin as having many influences on emotional behaviors. The study was set out to explore if the kisspeptin/GPR54 signaling system is required for the anti-depressant-like effect of kisspeptin-10 (KP-10), besides the regulation of the HPG axis. To test this concept, peptide 234 (P234), a kisspeptin antagonist, was given to the male rats, and its modulatory effect on the anti-depressant-like effects of kisspeptin was investigated by using a forced swimming test (FST). The study has also sought to know whether kisspeptin can exert its effects through adrenergic and serotonergic receptors. To investigate this, the agents yohimbine (Yoh), an alpha-2 adrenergic receptor antagonist, and cyproheptadine (Cry), a non-selective 5-HT2 serotonergic receptor antagonist, were administered in the experiments. Our results indicate that, in rats, the anti-depressant-like effects of KP-10 in a modified rat FST are mediated by GPR54 receptors, since the kisspeptin antagonist peptide 234 reversed kisspeptin-induced anti-depressant-like effects. Our data also demonstrate that the anti-depressant-like effects of kisspeptin, at least in part, are mediated by an interaction of the alpha-2 adrenergic and 5-HT2 serotonergic receptors.

Flammulina was found frequently distributed in the District Mansehra during the present research work. The genus was ignored and not studied for prevalence previously, as F. velutipes was the exclusively reported species from the research vicinity. During the present research work, three new species were explored i-e F. hazariansis (N. J201177, N. J201178), F. solatium (N. J201188, N. J201167) and F. dwarftype (N. J201187, NJ201139) were amassed from the surrounding areas of Hazara University Mansehra. Our findings revealed that the reported species are novel, the molecular, morphological, and phylogenetic characterizations proved it. The specimens were collected from various habitats, vegetation, damp places, and forest areas with rich organic soil favor the mushroom's growth from May to November. The species were studied for morphological characteristics like size, shape and color of the pileus, stripe, and spore size were also recorded. The species were preserved by sun and oven drying strategies. The Kit methods for molecular characterizations were used for the extraction of DNA, and for PCR, ITS4 primer was designed from conserved regions described in previous studies. The amplified PCR products were sequenced from Microgen Korea. Phylogenetic analysis of the obtained sequences was done based on the maximum likelihood method using Mega version 6.0. Our findings based on morphological and phylogenetic analysis confirmed the existence of three new species in the already described genera from the region. The area of District Mansehra is enriched with natural vegetation and can be explored for brand-spanking new species.

Hypochlorous acid (HOCl) is a reactive chlorine species generated by the enzyme myeloperoxidase present in phagocytes. HOCl plays a vital role in inflammation and has been linked to tissue regeneration through redox signalling, however, the relevant evidence is rather scarce. The present investigation aimed to study the effects of HOCl on the growth of C2C12 myoblasts and its association with alterations of cellular redox profile. C2C12 cells were incubated for 10 min, 1 h and 24 h with a wide range of HOCl concentrations (628 pM - 4 M). Cell survival was increased when cells were incubated with HOCl concentrations between 6.28 μM and 628 μM, which are encountered in biological systems. Intriguingly, after a 10 min-incubation with 3 mM of HOCl, the highest cell growth was observed through a redox-related mechanism, as indicated by the decrease of the levels of reactive oxygen species and the enhanced levels of reduced glutathione measured by flow cytometry. The in vitro model created herein simulates the in vivo inflammatory and regeneration response of muscle cells and can putatively give mechanistic answers about the contribution of HOCl to muscle regeneration.

Circulating endometrial cells (CECs) have emerged as a new biomarker of advanced disease in women with endometriosis. The identification of several subtypes of CECs (e.g., stem cell-like, epithelial, glandular, stromal) has opened the way for characterization of endometriosis-associated CECs. This study focused on the isolation and characterization of CECs and disseminated endometrial cells (DECs) in patients with spontaneous pneumothorax (SP). The primary objective was to differentiate between cancer and non-cancer cells in patients with no previous cancer diagnosis. The MetaCell® size-based separation protocol was used to enrich CECs/DECs. Evaluation of the captured cells by 3D microscopy was performed using a NANOLIVE™ microscope using a holographic approach. Based on gene expression analysis (GEA), we can conclude that mitochondria are much more active in primary tumors compared to endometriosis tissue (e.g. MT-ND1, MT-ATP6 genes). The culture of DECs is made of stromal, stem and immune cells. In vitro culture of DECs is characterized by an increase in the epithelial marker KRT18. Similarly, NFE2L2, a proerythroid factor, is also elevated.  Further, a significant decrease in the amount of stem and immune cells was observed in the cell culture of DECs.  The data presented here show how morphologically plastic the changes in the mitochondrial network can be and how cells can reflect them at the level of gene expression. The markers identified could help in the accompanying diagnostic process of the spontaneous pneumothorax in women of reproductive age.

Brucella spp. are small aerobes non-motile Gram-negative coccobacilli that act as facultative intracellular pathogens responsible for zoonotic infections. B. melitensis can survive and replicate within host macrophages, the molecular phenomena of this host: pathogen interaction remain totally unknown. The aim of this work was to evaluate the differences in the response between human macrophages infected with different B. melitensis strains. Comparison of transcriptome data was carried out for identifying differentially expressed genes among different strain infection. We evaluated the THP-1 macrophage molecular response at early stages of infection to different strains of Brucella melitensis (B. melitensis wild- type 133 (BM133), B. melitensis ATCC 23456 (BM16M) and a B. melitensis 133 omp31 mutant (LVM31)). Our analysis revealed intriguing differences in the host cell response to two virulent strains (BM16M and BM133), infection with BM16M led to an over-expression of anti-inflammatory pathways, such as cAMP signaling and PI3K-Akt pathway, and down regulation of inflammatory pathways involving IL1A and IL10 compared to BM133. Mutant strain BMLVM31 induced an activation of the apoptotic process and the absence of Omp31, impaired the inhibition of CASP1 and CASP9 expression. Additionally, the mutation of BMLV31 impairs the evasion of cathepsin D in early stages of the infection. These findings shed light on the intricate molecular interactions between B. melitensis strains and human macrophages, providing valuable insights for understanding the pathogenesis of brucellosis.

Zika virus (ZIKV) infection has been associated with damage to neural stem cells in microcephaly in newborns. The virus possesses specific tropism for glioma stem cells mediated by the ZIKV E protein. This infection causes endoplasmic reticulum stress and activation of the unfolded protein response (UPR). However, the cellular response to the expression of the ZIKV E protein alone is unknown. Therefore, in this study, we determined the effect of the expression of the ZIKV E protein on cellular responses and its subcellular localization in HEK-293T cells, due to their use as a biotechnological tool for cellular and lentiviral therapy. We observed that the ZIKV E protein is synthesized in the cytoplasm and inserted into the endoplasmic reticulum (ER), without causing activation of the UPR or cell death, and it is finally transported and located in the cell membrane. Moreover, the expression of the ZIKV E protein does not induce UPR or apoptosis in glioma cells. These results help us to better understand the characteristics of this protein and its possible use as a biotechnological tool for the creation of different gene therapy strategies, vaccines, and synthetic vectors with tropism for neural and glioma stem cells.

This study aimed to analyze the genetic characteristics of a sample of SARS-CoV-2 strains circulated in Erbil City from the 15th of October 2021 and the 5th of January 2022 focusing on their evolutionary feature including lineages, sublineages and clades. Following confirmation of the SARS-CoV-2 positivity of throat and nasopharyngeal swab specimens using qRT-PCR, 20 RNA extracts were subjected to NGS of the S gene and analysis in which only 12 matched the criteria of good sequences. Later, alignment was done with WIV04 reference sequence from Wuhan applying a number of bioinformatics tools. Then, based on sequences recorded in EpiCoV database/GISAID, related genomes to our sequences were identified. The PANGO system revealed that out of the 12 sequences, 10 were Delta (B.1.617.2) variants and two were Omicron (B.1.1.529). Seven out of 10 Delta sequences belonged to AY.33 sublineage and 2 were AY.4. Both Omicron sequences belonged to BA.1.1 sublineage. All Delta sequences belonged to the 21J Nextstrain subclade, meanwhile, both Omicron sequences were from 21K. Spike protein mutations in Delta variant varied, some were sublineage-specific, and others were unique, however, mutations generally were found in the N-terminal domain. Omicron variant appeared with 33 mutations, most of which were in the receptor-binding domain. On the whole, related sequences to our sequences were from Germany, the USA, Denmark, the UK, Iraq, Turkey and several other countries. These findings could provide insights into SARS-CoV-2 evolution nature and significant impact of amino acid changes in the spike protein on disease pathogenicity and emphasize the demand for continuous genomic surveillance globally.

Undoubtedly, nonalcoholic fatty liver disease (NAFLD) is widely recognized as one of the most prevalent liver diseases worldwide, encompassing a broad spectrum from simple steatosis to the most advanced stage of nonalcoholic steatohepatitis (NASH). However, effective treatments for NAFLD and NASH have not yet been clearly defined. Using appropriate terms such as "Nonalcoholic Fatty Liver Disease", "NAFLD treatment", "Lipid metabolism", "lipid biosynthesis", autophagy "bFGF" and TFG-b" apoptosis, we searched for relevant articles in the PubMed and Scopus databases. This review will discuss the role of the most important players in controlling lipid biosynthesis and lipid metabolism imperfection, which leads to NASH and NAFLD. Furthermore, potential pharmacological agents for targeting molecules and signaling pathways involved in liver inflammation, fibrosis, and cell death are also discussed.