Cellular and Molecular Biology

hMAGEA2 as a potential diagnostic and therapeutic target for melanoma progression and metastasis

2024-06-06T21:57:54+02:00

The incidence of melanoma, a highly aggressive skin cancer, continues to increase worldwide, particularly among populations with lighter skin tones. The diagnostic challenge of melanoma lies in the absence of a distinctive clinical presentation, as its characteristics vary based on anatomical location, growth type, and histopathology. The melanoma-associated antigen (MAGE) gene family is differentially expressed in various human cancers, including melanoma. In this study, we explored the association between human MAGEA2 (hMAGEA2) expression and melanoma. Using a human melanoma tissue array, we confirmed that hMAGEA2 expression was higher in melanoma and metastatic melanoma than in normal tissues. Additionally, we used SK-MEL-5 and SK-MEL-28 cell lines to investigate the cellular and molecular mechanisms underlying melanoma progression and invasiveness. In SK-MEL-5 and SK-MEL-28 cells, hMAGEA2 overexpression accelerated cell proliferation. Conversely, the knockdown of hMAEGA2 reduced cell proliferation, colony formation, and migration significantly and induced arrest at the G2/M phase of the cell cycle. With respect to the molecular mechanism, the knockdown of hMAGEA2 decreased the phosphorylation of Akt, JNK, and p38 MAPK. Additionally, hMAGEA2 knockdown reduced tumor formation significantly at the in vivo level. Collectively, the robust correlation between hMAGEA2 and melanoma metastasis supports the potential utility of hMAGEA2 as both a diagnostic marker and novel therapeutic target for patients with melanoma metastasis.

Mesenchymal stem cells treated with Interleukin-1 beta for mediation of an inflammatory response in human tissues

2024-05-09T06:45:36+02:00

The present study examined the functional activities of the human bone marrow mesenchymal stem cells (hBM-MSCs) under the effects of various concentrations of the inflammatory mediator interleukin 1 beta (IL-1β). The effects of IL-1β on the functional properties of hBM-MSCs were measured using functional assays (adhesion, proliferation, and migration). hBM-MSCs expressions of colony-stimulating factors 1 and 2 (CSF1, CSF2), C-C chemokine receptor type 2 (CCR2), C-X-C chemokine receptor type 1 and 3 (CXCL1, CXCL3), were examined using real-time polymerase chain reaction (RT‒PCR). The pro-inflammatory cytokine IL-1β did not disrupt hBM-MSCs adhesion, but it improved proliferation and migration only up to 50 ng/ml. However, in response to 100 ng/ml IL-1β, cell growth, proliferation, and migration were reduced significantly. The expression of CSF1, CCR2, CXCL3, and IL-1β genes increased with the increase in the concentration of IL-1β. CSF2 and CXCL1 gene expression increased in the 50ng/ml group compared with the 10ng/ml group to be higher than the control group in the 100ng/ml IL-1β group which might facilitate the differentiation, and homing of MSCs to the site of injury and augment their activities in the inflamed microenvironment. The study corroborates the advantages of prior stimulation of mesenchymal stem cells (MSCs) with the cytokine IL-1β, demonstrating an upregulation of key chemokines and cytokines. This upregulation potentially enhances MSCs' ability to differentiate and migrate to injury sites, while also augmenting their functional role within an inflamed microenvironment, thereby amplifying their therapeutic potential.

Study of the antioxidant activity of some active compounds in orange peels

2024-05-19T22:09:04+02:00

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.

Dominance of SARS-CoV-2 Delta AY.33 sublineage and Omicron BA.1.1 sublineage in Erbil City/Kurdistan Region of Iraq

2024-03-25T07:09:29+01:00

This study aimed to analyze the genetic characteristics of a sample of SARS-CoV-2 strains circulated in Erbil City from the 15^th of October 2021 and the 5^th 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.

Addition of new flammulina species via DNA, molecular characterization and phylogenetic investigation

2024-04-06T20:36:38+02:00

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.

Comparison of athletic performance of Turkish ice hockey players with ACE I/D (rs1799752), ACTN3 (rs1815739), PPARA (rs4253778) and HIF1A (rs11549465) polymorphisms

2024-05-16T09:21:22+02:00

Our study is aimed at examining the Ice Hockey National Team players with regard to ACE I/D (rs1799752), ACTN3 (rs1815739), PPARA (rs4253778) and HIF1A (rs11549465) polymorphisms and physical tests. This study was participated by 21 players from ice hockey national team. While ACE I/D (rs1799752) polymorphism was obtained using conventional polymerase chain reaction method (PCR), ACTN3 (rs1815739), PPARA (rs4253778) and HIF1A (rs11549465) polymorphisms were produced by real time polymerase chain reaction method (qPCR). Athletic performance analysis, on the other hand, was based on the assessment of maximal oxygen consumption capacity (VO2max), anaerobic performance, flexibility and strength tests. In our cohort, ACE I/D (rs1799752) polymorphism was determined as 24% genotype II, 33% genotype ID, and 43% genotype DD. ACTN3 (rs1815739) polymorphism was determined as 24% genotype RR, 43% genotype RX, and 33% genotype XX. PPARA (rs4253778) polymorphism was observed as 71% genotype GG, 14.5% genotype GC, 14.5% genotype CC. HIF1A (rs11549465) polymorphism was found to be 67% genotype CC, 33% genotype CT. Concerning physical tests, the evaluation of flexibility test results among genotype groups did not yield significant differences (p=0.365). No significant difference was found among genotype groups with respect to leg strength test results (p=0.691). The evaluation of handgrip strength test results among genotype groups did not reveal significant differences (p=0.679). No significant differences were found among genotype groups when VO2 max test results were examined (p=0.686). A significant relationship was found in the speed test and HIF1A rs11549465 polymorphism evaluation (p = 0.008). No significant results were detected when comparing the speed test with other polymorphisms (p = 0.65). The results of our study support the previous studies which had focused on the potential relation between the relevant gene polymorphisms and athletic performance of ice hockey players. However, doing further studies with larger cohorts is recommended in order to understand the relationship between the relevant polymorphisms and athletic performance of ice hockey players.

Investigation of the effects of melatonin on lung tissue through the NLRP3/TLR2/NEK7 pathway in an experimental endotoxemia model

2024-02-12T19:30:37+01:00

Sepsis, a severe clinical syndrome, arises from pro-inflammatory and apoptotic processes. Its rapid progression from sepsis to severe stages necessitates timely intervention. The lipopolysaccharide (LPS) agent triggers pro-inflammatory mediator release through Toll-like receptors, particularly TLR-2, a vital biomarker in sepsis with multiple organ failure. In LPS-induced septic shock, the NEK7-mediated NLRP3 inflammasome pathway, linked to acute lung injury, is suppressed. This pathway is implicated in sepsis-induced platelet activation and septic shock development. Antioxidants like melatonin (MEL) may positively impact reducing septic shock. In microbial-induced sepsis, melatonin can regulate pro-inflammatory mediator transcriptional activation, potentially controlling the pro-inflammatory state. In the project, the histopathological impact of melatonin in lung tissue during endotoxic shock induced by the LPS agent in Sprague-Dawley rats, and its immunoreactivity to NLRP3/NEK7/TLR-2 molecules, were assessed. Lung volumes were evaluated using micro-computed tomography (Micro-CT). While bleeding, cell infiltration, and thickening of the alveolar wall were observed in the lungs of the LPS group, a reduction in these symptoms was noted in the MEL+LPS group. Expressions of NEK7, TLR2, and NLRP3 increased in both the LPS and MEL+LPS groups compared to the control group. It was determined that in the MEL+LPS group, levels of NEK7, TLR2, and Malondialdehyde (MDA) decreased compared to the LPS group. Additionally, a decrease in the total volume of lung tissue was observed in the LPS group. In this context, our study reported the therapeutic effect of melatonin on sepsis-related acute lung injury. Our study suggests that melatonin administration in the experimental endotoxemia model melatonin may help reduce lung damage by inhibiting NEK7 and TLR2 expressions.

Hypochlorous acid induces a redox-dependent growth of C2C12 myoblasts

2024-04-08T08:21:13+02:00

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.

Assessment of in vivo and in vitro anti-tumoral effects of Lycium barbarum extract on Ehrlich ascites tumor cells: histopathology, DNA damage and AgNOR

2024-04-04T13:07:27+02:00

Natural product research has an exciting and glorious past that spans over millennia. Accordingly, natural products mediated inhibition of carcinogenesis by mechanistic modulation of deregulated signaling pathways has revolutionized the field of translational oncology. Lycium barbarum has antioxidant and anticarcinogenic effects. The antioxidant activity of the extract and its effect on Ehrlich ascites tumor (EAT) were investigated using in vivo and in vitro techniques. EAT cells were injected into Balb/C mice to create stock mice. EAT cells withdrawn from stock mice were used in equal volumes in the studies. The in vivo study consisted of control and treatment groups (200 mg/kg fractions above and below 50 kDa of extracts). The liver tissues were evaluated for histopathological (H&E), DNA damage (Comet assay), and proliferation (AgNOR staining) status. The in vitro study consisted of control and treatment groups (1500 and 2000 µg/ml of extracts). Cell viability and apoptosis were evaluated. As a result, a decrease in the adhesion of EAT cells, and decreased DNA damage were observed in mice intraperitoneally administered with the fractions of Lycium barbarum. The extracts both below and above 50 kDa increased apoptotic death in cancer cells. The extract above 50 kDa was more active than those below 50 kDa. Lycium barbarum consumption may be effectual in preventing cancer formation and slowing the progression of cancer

Myoblast-derived exosomes reduce anticancer drug-induced muscle toxicity via an autocrine pathway

2024-03-21T13:28:48+01:00

During cancer treatment, cachexia, characterized by muscle loss, often occurs, with one of the contributing factors being muscle toxicity caused by anticancer drugs. It affects approximately 80% of patients with cancer, particularly those with digestive organ malignancies. However, effective treatment for this condition remains elusive. Therefore, in this study, we aimed to investigate the therapeutic potential of exosomes in relieving cachexia. Specifically, we examined the exosomes derived from muscle stem cells, which are involved in muscle cell regeneration and their role in controlling anticancer drug-induced muscle toxicity. First, exosomes secreted from myoblasts under depletion conditions were characterized. Exosomes were isolated under serum starvation conditions, displaying an average size of 113 nm and containing typical exosome marker proteins. Furthermore, electron microscopy confirmed their exosomal nature. To confirm the paracrine function of myoblast-derived exosomes (MDEs), a significant increase in cell viability was observed upon their application to myoblasts. No changes were observed in the cell cycle during exosome treatment. However, it was confirmed that the quantity of viable cells increased under serum starvation conditions. This suggests that MDEs possess the function of enhancing myoblast survival and overall cell viability. Cachexia, a prevalent condition in patients with cancer, often manifests as muscle cell depletion induced by anticancer drugs. The potential of MDEs to inhibit cell death induced by anticancer drugs was investigated. The findings revealed that while high concentrations of oxaliplatin and doxorubicin, known to induce cachexia, did not restore cell viability, lower concentrations did. This study suggests that MDEs may have the potential to control cachexia, a common side effect of anticancer drugs, by reducing muscle cell damage induced by anticancer drugs.

Evaluation of uric acid levels and other biochemical parameters among Gout patients with Ketogenic diet in Erbil province

2024-06-02T18:43:09+02:00

Gout is a systemic disorder that occurs due to an accumulation of uric acid crystals in the tissues. The association between the Ketogenic diet and uric acid concentration has been poorly established. This study aims to evaluate and assess the association of Serum uric acid and other variables with ketogenic diet among Gout patients in comparison with Healthy control subjects. A case-control observational study. Subjects were grouped into Group I (Gout patients-103 individuals) and Group II (healthy Subjects-55 individuals). Parameters of Serum creatinine, blood urea, and uric acid were assessed for both groups. The study population included 51.3% of male and 48.7% of female participants, with an age range of 20-74, with a mean of 35.72±13.69 years old. Ketogenic and meat-rich diet as strong risk factors for Gout were higher among all case groups (45.6%) and (92.2%), respectively. The Pearson’s correlation coefficient of serum uric acid with other variables showed that the relation between serum uric acid with age, and weight among gout patients was found to be a weakly positive correlation and statistically significant (r=0.24, P= 0.013), and (r=0.22, P=0.026) respectively. This prospective study confirms that a ketogenic diet and a diet rich in meat have been associated with an increased incidence of gout. Indeed, results have shown that the ketogenic diet might have an increasing effect on serum uric acid. The frequencies of comorbidities have been constantly shown to be increased in gout.

Effect of periodontal therapy on serum and salivary Interleukin-1 beta (IL-1β) and malondialdehyde levels in chronic periodontitis

2024-05-18T13:22:43+02:00

Chronic periodontitis (CP) is distinguished by an inflammatory reaction and the presence of oxidative stress (OS), which has consequences for overall health. Interleukin-1β (IL-1β) and malondialdehyde (MDA) serve as markers of inflammation and OS, respectively. Analyzing the alterations in their reaction to periodontal therapy can provide valuable insights into the management and monitoring of CP progression. The current study aimed to evaluate the impact of non-surgical periodontal therapy (NST) on IL-1β and MDA levels in the serum and saliva of CP patients and explore their correlation with clinical periodontal indices post-therapy. There were 60 participants in this research, aged 33 to 50, equally split between thirty periodontally healthy controls and 30 patients with CP. Measures were taken of the clinical periodontal parameters, including the bleeding index, probing pocket depth, gingival index, and plaque index. Saliva and blood samples were collected for IL-1β and MDA analysis using ELISA and spectrophotometrically. CP patients received scaling and root planning (SRP) as part of phase I periodontal therapy, and after six weeks, we reevaluated clinical parameters and IL-1β and MDA levels. In CP patients, both saliva and serum IL-1β and MDA levels significantly increased alongside worsening clinical periodontal parameters compared with periodontally healthy individuals. phase I periodontal therapy led to a notable decrease in both saliva and serum IL-1β and MDA levels, accompanied by improvements in clinical parameters. Additionally, following six weeks of scale and root planning treatment, our data showed a strong positive relationship between salivary IL-1β and MDA levels with PPD and CAL. SRP therapy is effective in managing periodontal health, as evidenced by a significant decrease in clinical parameters and biomarker levels after treatment for CP patients. This suggests that salivary IL-1β and MDA may be useful biomarkers for indicating the severity of periodontal disease and the effectiveness of treatment.

The antidepressant-like effects of kisspeptin-10 are reversed by kisspeptin antagonist peptide 234 in male rats

2024-04-22T08:28:46+02:00

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.

Morphology of mitochondrial network in disseminated endometriosis cells in spontaneous pneumothorax diagnostic process

2023-11-18T08:56:13+01:00

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.

Exploring the diverse acetylcholinesterase inhibitory potential of girinimbine: insights from in vitro assays, molecular docking, and simulation studies

2024-07-28T14:17:05+02:00

The search for new treatments for Alzheimer's disease (AD) has led to the exploration of plant-based drugs as potential options. Acetylcholinesterase (AChE) inhibitors are widely used as anti-AD medications. This study aimed to investigate the inhibitory mechanism of girinimbine, a constituent of Murraya koenigii, on AChE. AChE inhibition was assessed by in vitro experiments using the modified Ellman method, as well as in silico molecular docking and molecular dynamic simulation. The results were compared to those of the well-known anti-AChE agents tacrine and propidium iodide. Girinimbine, propidium, and tacrine at concentrations of 3.8X10-5M, 1.1x10-5M, and 6.1x10-7M showed percentages of inhibition percentages of 35.6%, 28.2%, and 76.6%, respectively. The docking and molecular dynamics simulation analyses indicated that girinimbine exhibited a higher binding affinity to AChE compared to propidium and tacrine. This finding was further confirmed by the docking, root mean square deviation (RMSD), root mean square fluctuation (RMSF), and radius of rotation analyses. In conclusion, M. koenigii girinimbine shows promise as an acetylcholinesterase inhibitor for Alzheimer's disease. Further research, including in vivo studies and clinical trials, is needed to explore its potential as a plant-based drug candidate for AD treatment.

Analysis of the heterologous expression, localization, and cellular response to the Zika virus E protein in vitro

2024-04-04T21:34:37+02:00

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.

Transcriptome analysis of infected human macrophages between strains of Brucella melitensis and an omp31 mutant

2024-02-16T08:48:28+01:00

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.

Growth optimization, antibiogram, and molecular identification of Bacillus species isolated from the human gut

2024-06-03T06:23:41+02:00

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.

Evaluation of TP53TG1 and PANDA lncRNAs expression in association with adjuvant chemotherapy response in the peripheral blood of invasive ductal carcinoma patients

2024-06-03T16:47:56+02:00

Breast cancer is the most common malignancy in women. Breast cancer, the second leading cause of cancer deaths, affects 2.1 million women each year and is estimated to have killed 627,000 women worldwide in 2018. Unfortunately, the age of onset of this cancer in our country IRAN is about 10 years lower than the global average and is close to 45 years. Chemotherapy is one of the basic treatments for cancer. Predicting the benefits of chemotherapy is challenging. Studies are now underway to use gene expression tests to pinpoint patients who are most likely to benefit from adjuvant chemotherapy. In the present study, the expression of two long non-coding RNAs TP53TG1 and PANDA in the blood of breast cancer patients before and after receiving chemotherapy compared with this amount in the blood of normal people using Real-Time RT PCR technique to find a meaningful relationship ¬ Compared statistically. Compared to normal samples, the expression level of TP53TG1 in the blood of patients was reduced. Although it was not statistically significant. Its expression also increased after receiving chemotherapy. Compared to normal samples, the expression of PANDA in the blood of patients was increased, which was statistically significant. Also, its expression decreased after receiving chemotherapy. These findings suggest that PANDA and TP53TG1 expression levels may be possible markers associated with tumorigenesis and may also be considered as possible indicators of response to chemotherapy.

Identification of a novel prognostic signature for breast cancer derived from post-translational ubiquitin and ubiquitin-like modification-related genes

2024-09-20T16:54:05+02:00

Ubiquitin and ubiquitin-like (UUL) modifications play pleiotropic functions and are subject to fine regulatory mechanisms frequently altered in cancer. However, the comprehensive impact of UUL modification on breast cancer remains unclear. Transcriptomic and clinical data of breast cancer were downloaded from TCGA and GEO databases. Molecular subtyping of breast cancer was conducted using the NMF and CIBERSORT algorithms. Prognostic genes were identified via univariate, lasso and multivariate Cox regression analyses. Clinical pathological features, immune cell infiltration, immune therapeutic response and chemotherapy drug sensitivity were compared between groups using the Wilcoxon test. Survival analysis was performed using the Kaplan-Meier method and log-rank test. A total of 63 UUL modification-related genes were differentially expressed, with 29 up-regulated and 34 down-regulated genes. These genes were used to generate two UUL modification patterns that exhibited significant differences in prognostic features and immune cell infiltration. The UUL modification patterns were associated with 2038 differentially expressed genes that were significantly enriched in nuclear division, chromosome segregation, neuroactive ligand-receptor interaction, cell cycle, and other biological processes. Of these genes, 425 were associated with breast cancer prognosis, which enabled the classification of breast cancer into two clusters with significantly distinct prognoses. We developed a prognostic model, UULscore, which comprised nine genes and showed a significant correlation with partial immune cell infiltration. Furthermore, UULscore demonstrated potential predictive value in breast cancer overall survival prediction, immune therapeutic response, and chemotherapy drug sensitivity. UULscore, stage, radiotherapy, and chemotherapy were identified as independent prognostic factors for breast cancer. Based on these factors, a nomogram model was constructed, which demonstrated exceptional prognostic predictive performance. The present study identified two UUL modification-derived molecular subtypes in breast cancer, and have successfully constructed a risk-scoring model that holds potential value in prognosis, immune infiltration, immune therapeutic response, and chemotherapy sensitivity.

C-terminal tagging enhances the detection sensitivity of interlekin receptor type 1

2024-02-25T12:33:44+01:00

Substances released outside of the cells during cell necrosis are collectively called danger-associated molecular patterns (DAMPS) or alarmins. A pro-inflammatory cytokine, interleukin-1α (IL-1α) is known as a typical alarmin. IL-1α transmits signals by binding to IL-1 receptor 1 (IL-1R1), type I protein, expressed on the cell membrane of target cells, but detection of IL-1R1 at the protein and mRNA levels is difficult. Although the reasons are not elucidated, we attempted to add the HiBiT-tag to the N-terminus (N'-R1) or C-terminus (C'-R1) of IL-1R1 to examine whether the detection sensitivity can be augmented. Increase in detection sensitivity will allow the investigation of its function and subcellular localization much further. Using uterine cervical cancer-derived HeLa cells and its derivative CR-R1-4 cells lacking IL-1R1, C'-R1 was demonstrated to significantly increase the detection sensitivity of IL-1R1. Furthermore, the signal transduction function of neither N'-R1 nor C'-R1 was affected. Immunofluorescence cell staining revealed that wild-type IL-1R1 is mainly localized in the nucleus, whereas C'-R1 is localized both in the nucleus and the cytoplasm. The above results showed that adding a tag to the C-terminus of IL-1R1 increases detection sensitivity while maintaining its function. In the future, we would like to further investigate the relationship between changes in the intracellular localization of C'-R1 and increases in detection sensitivity.

Incidence of catheter-association bloodstream infection among hemodialysis patients at Erbil Teaching Hospital

2024-08-04T20:40:50+02:00

The study objectives were to analyze catheter-associated bloodstream infection (CABSI) risk factors in chronic kidney disease on regular hemodialysis and identify the bacterial species responsible for this by molecular analysis. This research was conducted in Erbil Teaching Hospital-Dialysis Unit in Erbil City-Kurdistan Region-Iraq from January to June 2024. It has been performed on 100 hemodialysis samples from both males and females. The investigation showed that the prevalence of CABSI among hemodialysis patients was 44 (44%) out of 100 (100%). The highest percentage of patients were aged between 60-69 years (32%, OR= 0.9, 95%CI [0.1-2.4], P< 0.001) and also male (66%, OR=2.7, 95%CI [0.9-9.4], P< 0.032). Additionally, the patients with Diabetes Mellitus were 70%, (OR= 6.3, 95%CI [0.3-10.4], P< 0.031), and with hypertension was 92%, (OR= 3.1, 95%CI [0.21-5.4], P<0.02. However, the dialysis duration of most patients was between 1-3 months (60%, OR=0.1, 95%CI [0.1-3.2], P<0.006) and the majority used two catheters (52%, OR= 0.6, 95%CI [0.1-3.2], P<0.012). The most common pathogens identified were Staphylococcus epidermis (44 cases, 100%), Pseudomonas aeruginosa (29 cases, 66%), and, Acinetobacter baumanni (24 cases, 55%). Thirteen bacterial species were recorded in the NCBI GenBank database. The phylogenetic tree demonstrated the distribution and relationship between these bacteria in hemodialysis patients. It showed that the bacterial species were closely related. To lower the risk of catheter-associated bloodstream infection, medical staff should actively develop countermeasures and gain a thorough understanding of the risk factors, which include age, diabetes, length of catheterization, and catheterization site.

Targeting key players in lipid biosynthesis for NAFLD and NASH treatment

2024-07-19T09:10:36+02:00

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.

Artificial intelligence and microbiome research: Evolution of hotspots, research trends, and thematic-based narrative review

2024-07-27T10:55:29+02:00

Artificial intelligence (AI) and microbiome have emerged in recent years as transformative fields with far-reaching implications for various biomedical domains. This paper presents a comprehensive bibliometric analysis examining the intersection of AI and the microbiome (AIM). The study aims to provide information on this interdisciplinary field's research landscape, trends, and emerging topics. Using a systematic approach, data-driven studies were extracted from the Scopus database on 23 November 2023 and analyzed using the VOSviewer and Bibliometrix applications. The regression coefficient of 0.94 and the yearly growth rate of 7.46% in AIM production indicate a consistent increase over time. Identification of essential contributors, organizations, and nations illuminated cooperative networks and research hotspots. The trend themes are the gut microbiome, disease prediction, machine learning, transfer learning, categorization, big data, artificial neural networks, chronic rhinosinusitis, epidemiology, COPD, and bronchoalveolar lavage. These hot issues in AIM reflect the present emphasis on research and developments in our knowledge of the microbiome's function in health, sickness, and individualized treatment. The findings give researchers, policymakers, and industry experts a thorough picture of the research environment and guide future paths in AIM's fascinating and promising subject.

Recent advances in understanding the role of uterine microbiota in endometrial receptivity and its impact on embryo implantation failure

2024-04-13T20:52:08+02:00

The aim was to provide a review of studies on the impact of intrauterine bacterial flora on endometrial tolerance in populations with failed embryo implantation and to provide direction for future clinical practice. Studies utilizing techniques such as 16S rRNA gene sequencing and macrogenomics were included through a comprehensive literature search to identify studies examining the correlation between intrauterine bacteria and endometrial tolerance. The composition of the bacterial flora in the uterine cavity plays an important role in regulating endometrial tolerance, and an increase in specific dominant bacilli in the uterine cavity correlates with an increase in conception rates, whereas dysbiosis of the intrauterine flora may lead to a variety of reproductive complications, including intrauterine inflammation, uterine adhesions, endometriosis, failure of embryo implantation, recurrent miscarriages, and embryo developmental arrest. Understanding the impact of intrauterine bacteria on endometrial tolerance can help improve clinical outcomes in patients experiencing embryo implantation failure. Further research in this area will help to elucidate the underlying mechanisms and develop targeted therapeutic interventions to optimize endometrial affinity and improve reproductive outcomes.