Molecular Biology Reports最新文献

Background: Understanding the evolutionary history of plants and accurately identifying biologically important species and their families is crucial for the herbal and Ayurvedic industries. The genetic approach by DNA barcoding plays a pivotal role in accurate species identification, authentication and quality control. Due to various therapeutic properties, Withania somnifera has been used worldwide in traditional systems of medicine for centuries including Ayurveda and Unani. The increasing demand for W. somnifera products has led to concerns regarding the authenticity and quality of commercial herbal preparations. However, adulteration become major trouble for users and industry for safety reasons and authentication of the plant with proper DNA marker is a major concern.

Methodology: DNA barcoding techniques and Phylogenetic analysis were employed to authenticate W. somnifera plant species using universal genetic markers. The markers were PCR amplified, sequenced and analyzed using BLAST-based and phylogeny-based identification methods.

Results: The BLAST result shows the percent identity (PI) of ITS1, ITS2, trnK, atpB, rbcL and matK was 100%, 100%, 100%, 97.59%, 100 and 99.20% respectively with the NCBI reference sequence. However, ITS1 and ITS2 show the maximum sequence similarity with W. somnifera of NCBI data. Phylogenetic analysis using NCBI data further supports the role of ITS in the discrimination of W. somnifera from closely related species.

Conclusion: Therefore, the ITS gene may be considered promising a candidate for DNA barcoding for discrimination of W. somnifera from other species, its authentication and quality control.

Background: Interleukin 10 (IL-10) is uniquely positioned in the immune regulation of teleosts. Modifying the IL-10 pathway changes the teleost's disease susceptibility; however, there is no data on its post-transcriptional regulation. Trachinotus blochii is a high-value mariculture species.

Methods: The full-length tbil-10 gene was generated through the Rapid Amplification of cDNA Ends-PCR. After the detailed sequence analysis, the identified features were compared with other IL-10 sequences. The gene expressions in healthy and challenged (Vibrio harveyi) fish were studied.

Results: The sequence analysis showed an open reading frame of 564 bp and a 3' UTR (untranslated region) of 217 bp. The phylogram revealed an evolutionary distinction between marine and freshwater teleost IL-10. The shorter 3' UTR, additional conserved cysteines capable of forming stable disulphide bonds, and lesser mRNA instability moieties suggest the better structural stability of teleost IL-10 than tetrapods. Results identified 60 miRNA-mRNA duplexes that can regulate IL-10 3' UTR. Nine identified miRNAs are involved in immune response and seven are expressed in macrophages. The gills showed the highest gene expression in healthy fish. The study discovered two IL-10 mRNA transcripts that differed in 5' UTR lengths and thermodynamic ensemble's free energy. There was an increased expression of both tbil-10-mRNA transcripts from 2 to 48 h post-challenge which peaked at 24 h after the challenge, with higher expression of short mRNA transcript.

Conclusions: The results gave insights into the structural, functional, post-transcriptional regulatory mechanisms, and expression characteristics of the IL-10 gene in T. blochii.

Background: Charcoal Rot (CR) poses a significant threat to mung bean crops by reducing yield, making the development of resistant varieties crucial for stable production and food security. This study evaluated 19 newly identified mung bean landraces using biochemical traits and SSR markers, revealing genetic variability, CR disease reactions, and traits influencing yield and resistance, which provide valuable insights for breeding CR-resistant, high-yielding varieties.

Methods and results: Mung bean landraces were evaluated for their response to CR using 4 biochemical parameters, and 10 SSR markers to assess genetic variability and disease resistance. The results revealed substantial variability in morpho-quantitative traits, and showed a significant variation in CR disease reaction (21% resistance, and 16% moderately resistance). Biochemical characterization suggested that, phenolic content (PHE), Peroxidase (POX) and polyphenol oxidase (PPO) activities exhibited narrow ranges, suggesting more uniformity in these biochemical traits under CR stress. Yield (YLD) showed a positive correlation with the number of pods per plant (NPDP) (0.459), PPO (0.912), PHE (0.867), and total chlorophyll (TCL) (0.864), but showed a negative correlation with POX (-0.64), indicating their positive influence on productivity. Conversely, CR resistance was strongly negatively correlated with YLD (-0.86). Moreover, as per the principal component analysis (PCA), 80.5% of the total variance was explained by the first three principal components. However, molecular characterization suggested the PIC values ranged from 0.281 to 0.871, with an average 0.58 indicating moderate to high polymorphism. Principal coordinate analysis (PCoA) grouped landraces based on their genetic makeup under CR stress, and the first three axes explained most of the variation (82.03%). Phylogenetic tree analysis confirmed the PCoA groupings with similarity coefficient varied from 0.57 to 0.98, suggesting the landraces within a cluster had lower genetic distances, and resistant and moderately CR resistant landraces fell into separate clusters.

Conclusion: The findings can be effectively utilized to develop CR-resistant mung bean varieties with enhanced productivity, contributing to stable mung bean production and food security.

Background: Blastocystis is a prevalent intestinal parasitic protist that infects both birds and animals. There are at least 44 subtypes (ST) of Blastocystis, with ST1-ST9 being found in humans. The correlation between specific subtypes and pathogenicity has not been definitively established. This study aimed to identify the genetic diversity within subtypes of Blastocystis in stool samples collected from individuals who were referred to medical facilities in the cities of Birjand, Darmian, Nehbandan and Ferdows, Eastern Iran.

Methods: A total of 1800 stool specimens were randomly collected from referred individuals in selected medical laboratories of South Khorasan province. DNA was extracted and PCR before, sequencing and subtyping were performed for characterization and phylogenetic analysis.

Results: A total of 118 (6.5%) individuals were examined positive with Blastocystis through microscopic observations. A ~ 620-bp segment of the SSU rRNA gene of Blastocystis was amplified and were genetically evaluated using MEGA6 and DnaSP. 30 samples of Blastocystis positive samples were sequenced. BLAST analyses identified 3 distinct subtypes including ST1 (11, 36.66%), ST2 (5, 16.68%), ST3 (14, 46.66) within our samples. Intra-subtype discrimination showed the similarity of 95.88%, 91.56% and 92.14% for ST1 and ST2, ST1 and ST3, and ST2 and ST3, respectively. Additionally, allele 4 for ST1; allele 12 for ST2; and alleles 34 and 36 for ST3 were detected.

Conclusions: As results, ST1 exhibited the highest nucleotide diversity (π: 0.00113), suggesting a high degree of genetic diversity within this subtype. Phylogenetic analysis showed that, ST3 was clustered to animal isolates from cattle and cats, indicating the potential zoonotic transmission of Blastocystis.

Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) are key downstream effectors of the Hippo pathway that regulate organ size, tissue homeostasis, and cancer development. YAP/TAZ play crucial regulatory roles in organ growth, cell proliferation, cell renewal, and regeneration. Mechanistically, YAP/TAZ influence the occurrence and progression of liver regeneration (LR) through various signaling pathways, including Notch, Wnt/β-catenin, TGF-β/Smad. While the activation of YAP/TAZ can promote the regeneration of damaged liver tissue, their mechanisms of action may differ under various LR conditions. Furthermore, excessive activation of YAP/TAZ may also lead to severe liver damage, manifesting as alcoholic hepatitis, liver fibrosis, and even liver cancer. Here, we review the role and mechanisms of YAP/TAZ in LR and liver disease, highlighting the potential for advancements in clinical diagnosis and treatment targeting YAP/TAZ in these contexts.

Background: Acute myeloid leukemia (AML) is a heterogeneous hematological malignancy caused by disorders in stem cell differentiation and excessive proliferation resulting in clonal expansion of dysfunctional cells called myeloid blasts. The combination of chemotherapeutic agents with natural product-based molecules is promising in the treatment of AML. In this study, we aim to investigate the anti-cancer effect of Rapamycin and Niacin combination on THP-1 and NB4 AML cell lines.

Methods and results: The anti-proliferative effects of Rapamycin and Niacin were determined by MTT cell viability assay in a dose- and time-dependent manner. The combination indexes were calculated by isobologram analysis. Furthermore, apoptosis was investigated by Annexin-V/Propidium Iodide(PI) double staining and cell cycle distribution was measured by PI staining. The expression levels of autophagy-related proteins were detected by western blotting. The combination of Rapamycin and Niacin synergistically decreased cell viability of AML cell lines. The combination treatment induced the apoptotic cell population of THP-1 and NB4 by 4.9-fold and 7.3-fold, respectively. In THP-1 cells, the cell cycle was arrested at the G2/M phase by 10% whereas the NB4 cells were accumulated at the G0/G1 phase. The combination treatment decreased Akt and p-Akt expression. Besides, the ATG7 expression was reduced by combination treatment on THP-1 cells. Similarly, the ATG5 level was downregulated in NB4 cells. The level of LC3B-II/LC3B-I, which is an indicator of autophagy flux, was upregulated in THP-1 and NB4 cells.

Conclusion: Although further studies are required, the combination of Rapamycin and Niacin combats cell proliferation by inducing cellular apoptosis, cell cycle arrest and autophagy activation.

Trace elements (TEs) are essential for human health and for maintaining immune responses against potentially aggressive pathogens, such as the human immunodeficiency virus (HIV). During the infectious process, the body needs greater amounts of TEs in order to coordinate an efficient immune response to combat the invading agent, a condition that reflects in lymphocyte proliferation and activation of the antioxidant defense system of neutrophils and macrophages. Thus, during the progression phase of a viral infection, immunomodulation of TEs such as iron, zinc, chromium, magnesium, selenium, copper, calcium, and manganese occurs, can lead to immunosuppression and increased oxidative stress. Furthermore, the adverse effects caused by the use of antiretroviral therapy (ART) trigger nutritional disorders and metabolic alterations that contribute to deficiencies in TEs, associated with compromised immune function. Therefore, this narrative literature review aims to contribute as a teaching tool on the TEs involved in the pathogenesis of HIV, by reviewing the role of TEs in the immunometabolic health of people living with HIV/AIDS.

Background: As part of stock enhancement programs for marine fishery species, the stocking of hatchery-produced seedlings into sea areas has been implemented worldwide. DNA markers are vital for responsible stock enhancement practices that aim to conserve the genetic diversity of recipient wild populations. We report novel single-nucleotide polymorphism (SNP) markers and multiplex SNP panels developed for the west Pacific pen shell Atrina lischkeana (Clessin, 1891), a large bivalve that is expected to be a subject of stock enhancement activity as the natural resource has dwindled, especially in Japan.

Methods and results: On the basis of single-base primer extension combined with MALD-TOF/MS analysis, we developed 218 SNP markers across eight multiplex SNP panels, which allowed genotyping of specimens from two wild A. lischkeana populations. However, our parentage analysis for a captive-bred population sample revealed that 21 of the 218 markers exhibited non-Mendelian patterns of allelic transmission in parent-offspring trios. We verified that the inheritance of undetectable null alleles caused almost all the aberrant allelic transmissions.

Conclusions: The developed markers will be of significant use in dealing with issues related to the stock enhancement of A. lischkeana, such as the monitoring of genetic diversity of hatchery stocks and the evaluation of impacts of stocking upon the genetic makeup of recipient wild populations, although caution is warranted especially when the 21 markers with unexpected patterns of allelic segregation are applied to field samples. Additionally, the markers can be analyzed in other targeted genotyping platforms.

Background: The role of the silkless1 (sk1) gene in developing silkless baby corn, a distinctive trait in maize has been investigated. So far, no sk1 gene-specific marker has been available for accelerated development of silkless baby corn hybrids.

Methods & results: We developed sk1 gene-based markers and validated them in backcross (BC) and F₂ segregating generations, revealing a polymorphic marker corresponding to a silkless phenotype. Among all the developed markers, the MGU_sk1-1 marker (141 bp size in sk1-mutant allele) validated its association with the silkless trait. The chi square (χ²) analysis revealed that the segregation of alleles Sk1Sk1 (219): Sk1sk1 (414): sk1sk1 (188) is as per the Mendelian ratio (1:2:1) in the F₂ population. In 821 F₂ plants, we found a 3:1 phenotypic ratio among the population with silked (633) and silkless (188) plants. We studied 349 BC plants and found a 1:1 phenotypic ratio between silked and silkless plants, as well as a genotypic ratio (1:1) between heterozygotes (Sk1sk1) and homozygous recessives (sk1sk1). Additionally, double mutants (sk1sk1/ts1ts1 and sk1sk1/ts2ts2) were identified using the sk1-based marker.

Conclusions: This study is the first to show the practical use of sk1 gene in marker-assisted selection (MAS). This research also reports the development of novel genetic resources for innovative silkless baby corn breeding.

Multiple myeloma (MM), also referred to as Kahler's disease, is a cancer characterized by the uncontrolled growth of abnormal plasma cells and is associated with alterations in the bone tissue microenvironment. Bone marrow adipose tissue (BMAT), which comprises approximately ten percent of total body fat, can influence the progression, survival, and drug resistance of MM cells through paracrine, hormonal, and metabolic pathways. Obesity can lead to an increase in BMAT mass, which not only disrupts bone metabolism but also reduces bone density, potentially progressing from monoclonal gammopathy of undetermined significance, a benign condition, to MM. A range of factors, including impaired fatty acid metabolism, increased production of adipokines that support myeloma, and heightened expression of oncogenic microRNAs in multiple myeloma, contribute to the progression of this incurable blood cancer. To better understand the relationship between excess adipose tissue accumulation and the risk of developing multiple myeloma, a comprehensive review of published data was conducted.