Molecular Biology Reports最新文献

Background: Acinetobacter baumannii is a notorious nosocomial pathogen universally in healthcare settings. Its natural competent characteristics for genetic recombination are responsible for acquired antibiotic resistance and render it untreatable through commonly used antibiotics. Hence, characterizing the A. baumannii genomes for multidrug resistance carriage is of paramount importance. The study aimed to characterize the whole genome of clinical isolates of A. baumannii to identify specifically the types of antibiotic resistance genes, drug classes and mobile genetic elements. We also aimed to determine the significant multi-locus sequence tags (MLSTs). The phylogeny of the isolates was established with other clinical strains distributed globally.

Methods and results: Fifteen clinical isolates (isolated from tracheal secretion, urine and bronchoalveolar lavage) were subjected to whole genome sequencing. Raw sequences were assembled using SPAdes and species were identified using KmerFinder 3.2. The assembled genomes were annotated using the Prokka v1.14.6. Resfinder 4.6.0 was used to determine antibiotic resistance genes. The sequences were aligned against seven housekeeping genes aka sequence tags (STs) available within the MLST database (v 2.0.9). MobileGeneticElement finder (v1.0.3) were used for profiling mobile genetic elements associated with the antibiotic resistance genes. The genomes of nosocomial A. baumannii were assembled with an average N50 of 23,480 and GC content of 38%. There were approximately 3700 CDs, 53 tRNA and 3 rRNA. About 80% of the isolates were ST2 type. The genomes possessed antibiotic resistance genes (n = 24) belonging to 17 drug classes. The predicted phenotype was multidrug resistant. Among the mobile genetic elements, 12 insertion sequences and 2 composite transposons were also found. The mode of antibiotic resistance was mostly through antibiotic inactivation in all the isolates.

Conclusions: The results imply the occurrence of multidrug resistant genes in clinical isolates of A. baumannii strains in the healthcare settings of Kuwait. A more comprehensive survey should be undertaken for antimicrobial resistance monitoring on a regular basis for surveillance, contact tracing, and potential mitigation in clinical settings.

Unconventional protein secretion (UcPS) encompasses diverse non-canonical cellular export mechanisms that operate independently of the classical secretory pathway, representing a crucial cellular response to various physiological and pathological conditions. This comprehensive review synthesizes current understanding of UcPS mechanisms, particularly focusing on their roles in disease pathogenesis and progression. Recent advances in proteomics and cellular biology have revealed that UcPS facilitates the secretion of various biomedically significant proteins, including inflammatory mediators, growth factors, and disease-associated proteins, through multiple pathways such as membrane translocation, secretory lysosomes, and membrane-bound organelles. Notably, dysregulation of UcPS mechanisms has been implicated in various pathological conditions, including chronic inflammation, neurodegenerative disorders, and malignant transformation. We critically evaluate the molecular machinery governing UcPS, its regulation under cellular stress, and its contribution to disease mechanisms. Furthermore, we examine emerging therapeutic strategies targeting UcPS pathways, highlighting both opportunities and challenges in developing novel interventional approaches.

Purpose: Adipose-derived mesenchymal stem cells (ADSCs) exosomes (AD-Exos) are a novel and promising therapeutic approach for skin damage repair. This investigation seeks to assess the potential clinical utility of miR-141-3p found in AD-exos for expediting wound healing.

Methods: ADSCs were isolated from the wounded patients' tissue and validated via flow cytometry, and the mineralization and adipogenic capabilities of ADSCs were assessed respectively. Additionally, exosomes were isolated and identified. miR-141-3p and HDAC6.protein level were tested. Full-thickness wound models were created on the backs of mice, HE staining, ELISA, and immunohistochemistry were used to assess the influences of AD-exos on wound healing, inflammation, and new blood vessel formation Western blot was to assess the related-protein levels of JNK/ERK pathway. AQ1 Meanwhile, Dual-Luciferase assay confirmed the relationship between miR-141-3p and HDAC6.

Results: The isolated cells highly express surface markers of mesenchymal stem cells and possess the potential for multidirectional differentiation, confirming them to be ADSCs. And miR-141-3p down-regulated but HDAC6 up-regulated in the serum and AD-exos of wounded patients. miR-141-3p could negatively modulate HDAC6. The miR-141-3p in AD-exos accelerated wound healing in mice, mitigated inflammatory responses and scarring in the injured skin tissue, and promoted angiogenesis, moreover, AD-exos could diminish the phosphorylation of JNK and ERK, while HDAC6 overexpressed could weaken these impacts.

Conclusion: miR-141-3p in AD-exos can target down regulate HDAC6 expression and inhibit JNK/ERK signaling pathway activation, thereby reducing wound inflammation and promoting angiogenesis and wound healing in mice.

Background: Understanding the genetic background of Korean field mouse(Apodemus peninsulae Thomas, 1906) is important for employing the animal as an experimental model in research. However, limited genetic information is available about A. peninsulae.

Methods and results: This study aimed to develop microsatellite molecular markers based on the genome sequence of A. peninsulae and establish a genetic evaluation system for A. peninsulae. Twenty-nine polymorphic microsatellite markers were identified via electrophoretic analysis, short tandem repeat scanning, and sequencing, and genetic diversity in three populations of A. peninsulae from Dalian City, Liaoning Province (LN), Changchun City, Jilin Province (JL) and Mudanjiang City, Heilongjiang Province (HLJ) was analyzed. A total of 229 alleles (Na) were detected in 115 A. peninsulae individuals. The mean observed alleles and effective alleles (Ne) were 7.897 and 3.571, respectively. The Shannon index(I) averaged 1.401, indicating high genetic diversity, whereas the mean observed heterozygosity (Ho) and expected heterozygosity (He) were 0.543 and 0.668, respectively. The mean polymorphism information content (PIC) was 0.627, which validated high genetic diversity in the three populations. Analysis of molecular variance (AMOVA) (genetic differentiation coefficient (FST) = 0.062) showed that the genetic distances were 0.083 (LN and JL), 0.203 (LN and HLJ), and 0.195 (JL and HLJ), indicating that the two artificially domesticated populations were genetically indistinguishable. Meanwhile, the wild HLJ population was significantly different from the other two artificially domesticated populations. The results of structural analysis, phylogenetic tree construction, and principal component analysis (PCA) were consistent with those of AMOVA. In addition, gene flow analysis used to explore genetic exchange confirmed the flow of genetic information among the three populations.

Conclusion: The microsatellite loci identified in this study are highly polymorphic and suitable for the genetic quality control of A. peninsulae, providing an important genetic basis for the breeding of A. peninsulae and evaluating its genetic potential. Thus, this study lays a scientific foundation for the future genetic improvement and resource utilization of this species.

Background: Factors that cause changes in insulin signaling in the brain are thought to affect the synaptic plasticity and accelerate the process of brain aging and neurodegeneration. Insulin receptor substrate (IRS) molecules are key mediators in insulin signaling. The aim of the current study is to determine whether there is an association between IRS gene polymorphisms, which are critical for insulin signaling, and the late-onset Alzheimer's disease in Turkish patients.

Methods and results: Demographic and clinical characteristics of 115 patients with late-onset Alzheimer's disease (age of onset ≥ 65 years) and 107 age-matched control subjects were obtained. DNAs were isolated from patient and control groups, IRS-1 and IRS-2 gene polymorphisms were investigated and genotyped according to the PCR-RFLP method. No statistically significant difference was observed in the genotypes for IRS-1 Gly972Arg (rs1801278) (p = 0.499) and IRS-2 Gly1057Asp (rs1805097) polymorphism between late-onset Alzheimer's disease patients and controls (p = 0.658). However, when the compliance of IRS-2 polymorphism with Hardy- Weinberg distribution was tested, in the case-control comparison, G allele frequency of IRS-2 polymorphisms was significantly higher in the patient population than in the control group in the Turkish population of the Thrace region.

Conclusions: Despite the potential role of insulin resistance and hyperinsulinemia in the development of Alzheimer's disease, we did not find any association between polymorphism of the IRS-1 and IRS-2 genes and late-onset Alzheimer's disease. However, compared to the healthy subjects, Gly/Gly genotypes and the G allele in the IRS-2 were significantly more frequent in patients with late-onset Alzheimer's disease.

Background: The study on identifying of brown macroalgae species, particularly those on the southern coast of Lampung Bay-Indonesia, at the molecular level and their antioxidant activity has never been conducted, so we examined it as our purpose study.

Method and results: The research uses DPPH free radical scavenging activity and molecular identification using DNA barcode cox1 BLAST. Fifteen samples of fresh brown macroalgae with five samples, respectively, were collected from Sebalang Beach, Kalianda and Pesawaran, South Lampung. The DNA was first purified, and then the gene product was amplified using specific primers, cox1F1 primer and cox1R1 primer. The DNA sequence was checked and traced using the Basic Local Alignment Search Tool (BLAST). The three most dominant species of brown macroalgae were identified, namely Sargassum plagiophyllum, Sargassum ilicifolium and Hormophysa cuneiformis. The base length obtained ranged from 1164 to 1212 bp, with a similarity percentage of 98.36% to 100%. The three types of brown macroalgae have the ability to scavenge DPPH free radicals. Ethanol solvent and extract fractions significantly influence the reduction of DPPH free radicals. The ethanolic extract fraction of S. ilicifolium exhibited the lowest EC₅₀ value (64.17 ± 1.23 mg L^-1) and has the strongest antioxidant activity, followed by H. cuneiformis (75.88 ± 0.34 mg L^-1), and S. plagiophyllum (82.97 ± 1.30 mg L^-1).

Conclusion: The difference in species of brown macroalgae had no significant effect on the EC₅₀ activity, and all three had robust antioxidant activity because their concentrations ranged between 50 and 100 mg L^-1.

Cancer stem cells (CSCs) represent a unique subpopulation of cells with the ability to self-renew and differentiate, thereby sustaining tumor growth and contributing to disease recurrence. Although CSCs predominantly reside in the G₀ phase, their stem-like properties, such as the expression of specific CD markers, self-renewal, differentiation potential, tumor initiation, drug resistance, and increased invasive and metastatic potential, manifest during their active proliferative phase. Rapidly dividing cells exhibit alterations in their cell cycle, often characterized by shortened or bypassed G₁ phases, a phenomenon observed in both embryonic stem cells and cancerous cells. Dysregulation of cell cycle control is a hallmark of cancer, leading to uncontrolled cellular proliferation and tumorigenesis. Disruption in key regulatory proteins, signaling pathways, and cell cycle checkpoints-particularly during the G₁ phase-enables cancer cells to escape normal proliferation restrictions. The rapid cell-cycle progression can impair the timely degradation of proteins critical for cell cycle regulation, particularly cyclin D, thereby compromising proper cell cycle control. Therefore these proteins may be passed to daughter cells, promoting further rounds of rapid cycles. Additionally, cyclin D is often overexpressed in cancer cells, further exacerbating uncontrolled proliferation. These mechanisms may underpin key properties of CSCs, including rapid proliferation and their stem-like traits. This review examines the relationship between G₁ phase kinetics and the acquisition of stem-like characteristics, emphasizing how rapid G₁ phase progression and transitions between dormancy and active proliferation contribute to the emergence of CSC traits.

Background: Glutathione S-transferases (GSTs) play crucial roles in various physiological processes, including plant development, endogenous metabolism, stress tolerance, and xenobiotic detoxification. However, the comparative analysis of the GST gene family in Rosaceae species and their expression patterns in roses remains unclear.

Results: In this study, a total of 348 GSTs were identified in six Rosaceae species, including 78 in rose (Rosa chinensis), 53 in strawberry (Fragaria vesca), 69 in peach (Prunus persica), 59 in Chinese plum (Prunus mume), 43 in apple (Malus domestica), and 46 in pear (Pyrus communis). These GST proteins were classified into nine subfamilies. Chromosome localization, collinearity analysis, and gene duplication relationships revealed that tandem repeats are the main driving force for the amplification of GST genes in the Rosoideae and Amygdaloideae. Gene Ontology (GO) and cis-element analysis suggested that GST genes may play a significant role in a variety of biological processes and respond to various abiotic stresses. qRT-PCR analysis showed that five GST genes, mainly expressed in the flower tissue of the Rosa chinensis 'Old Blush', as well as under four different abiotic treatments, revealed differences in their expression pattern, suggesting that RcGSTs may have different responses to abiotic stresses.

Conclusions: This study provides essential data to support a systematic investigation of the GST gene family within the Rosaceae family and the functional validation of GST genes in roses.

Background: Doxorubicin (Dox)-induced neurotoxicity is a well-documented side effect of chemotherapy. Dexpanthenol (Dex), an analog of vitamin B5, has shown protective properties. This study aimed to explore the molecular mechanisms by which Dex mitigates Dox-induced neurotoxicity, particularly through the protein kinase B (AKT)/cyclic AMP-response element-binding protein (CREB)/brain-derived neurotrophic factor (BDNF) pathway and nuclear factor erythroid 2-related factor 2 (NRF2) signaling.

Methods and results: The experiment was conducted using four groups: control, Dex, Dox, and Dox + Dex, comprising a total of 32 female Wistar Albino rats. After two weeks of treatment, the rats were euthanized, and brain and cerebellum tissues were collected for analysis. Biochemical analysis was performed spectrophotometrically to assess oxidative stress parameters, while histological and immunostaining analyses focused on nuclear factor kappa B (NF-κB) and inducible nitric oxide synthase (iNOS) immunoexpressions. Genetic analysis of AKT, CREB, BDNF, and NRF2 gene expressions was conducted using real-time polymerase chain reaction. Histopathological evaluation of the Dox group revealed hyperemia, microhemorrhage, neuronal damage, and neuronophagia. Additionally, an increase in caspase-3, tumor necrosis factor-alpha, NF-κB, and iNOS immunoexpressions were observed, along with elevated total oxidant status and oxidative stress index. A decrease in AKT, CREB, BDNF, and NRF2 gene expressions accompanied these changes. Dex treatment significantly reversed these pathological findings, effectively protecting the brain from Dox-induced neuronal damage.

Conclusion: In conclusion, Dex may provide neuroprotection in female rats with Dox-induced neurotoxicity by activating the CREB/BDNF pathway and reducing oxidative stress through AKT-mediated NRF2 synthesis. Further detailed studies exploring additional pathways are required to incorporate Dex into cancer treatment protocols and minimize side effects.