Molecular Biology Reports最新文献

Background: Marine fish eggs exhibit transversal morphological characteristics that complicate their identification. It is estimated that 75% of pelagic eggs possess oil globule(s). Identifying these eggs provides valuable information about spawning biomass and potential spawning areas for commercially important and threatened species. Molecular markers, such as the FISH (COI gene), enable a precise identification and exploration of the genetic variability within fish populations. San Vicente Bay, located in south-central Chile is partially protected from the effects of the Humboldt current, making it a notable spawning and nursery ground for various fish species. This study aims to identify and characterize, oil-globule-containing eggs present in the bay during October, a period that aligns with the spawning season of many species.

Methods and results: This study seek to standardize the DNA extraction method and PCR process, addressing the challenges posed by the low quantities of tissue in these eggs. We employed 2 methods, by a DNA extraction kit and Chelex resin. Out of 94 eggs, 24 samples yielded positive PCR results, corresponding to a 25% of success. All these eggs were successfully identified using genetic databases, revealing a total of 9 genera and 8 species. The observed genetic distances at the species level were low, supporting the hypothesis that the eggs originated from the same female or gene mutation rate.

Conclusion: The methods employed successfully identified fish eggs and documented species whose early life stages have not been previously reported in the bay.

Background: Juvenile cataract is characterized by the blurredness in eye lens which develops typically before the age of 18 years. Objective of the study was to delineate the underlying causative genetic defect for autosomal recessive juvenile cataract (ARJC) in an affected consanguineous multiplex, multigenerational Pakistani consanguineous family.

Methods and results: Whole-exome sequencing (WES) in an affected family member revealed a homozygous, intragenic 15.40 kb mega-deletion NG_012423.2; g.17485448_17500744del (chr20: 17,485,448-17,500,744-GRCh37/hg19) including three consecutive exons 3, 4, and 5, encompassing two domains of beaded filament structural protein 1 (BFSP1). Sanger sequencing and PCR were performed for all participants of the family which revealed the absence of the exons (3, 4, 5) in affected individuals and their presence in unaffected parents and siblings confirming the segregation of the mutation as autosomal recessive in the family under study. The large deletion potentially forced the junction of exon 2 with exon 6 at mRNA level and resulting in the shift of open reading frame. The mutation was not observed in 200 unrelated in-house controls.

Conclusion: Up to our knowledge, this is the first study reporting the largest mega-deletion encompassing 15.40 kb removal (consisting of 99 amino acids) and reveals the significant role of BFSP1 for physiologic lens function and optical properties. Our findings extend the genotypic spectrum of BFSP1-related cataract.

Background: Cerebral malaria (CM), the most severe manifestation of Plasmodium falciparum infection, is characterized by immune dysregulation, endothelial dysfunction, and brain injury. MicroRNAs (miRNAs) have emerged as promising biomarkers and regulators of malaria pathogenesis. Previous clinical studies identified elevated plasma levels of hsa-miR-21-5p and hsa-miR-3158-3p as correlates of CM severity and poor outcomes. However, their mechanistic roles in modulating host immune responses remain poorly defined.

Methods: To elucidate the regulatory effects of these miRNAs, MCF7 and HepG2 cell lines were transfected with synthetic mimics of hsa-miR-21-5p or hsa-miR-3158-3p. Following 48 h of incubation, mRNA expression levels of key immune-related genes-NF-κB, CD46, THBS1, TNF-α, IL-1β, IL-2, IL-6, IL-8, IL-10, and TLR4-were quantified using SYBR Green-based RT-qPCR. Relative expression levels were calculated using the 2⁻ΔΔCt method and statistically compared with negative control and non-transfected groups.

Results: Cells transfected with the hsa-miR-3158-3p mimic exhibited a significant downregulation of NF-κB expression compared with both control groups (p < 0.05). Similarly, transfection with the hsa-miR-21-5p mimic led to significant reductions in NF-κB, IL-10, and TLR4 expression (p < 0.05). No significant alterations were observed in the other immune-related targets examined.

Conclusions: These findings suggest that hsa-miR-21-5p and hsa-miR-3158-3p may regulate immune pathways implicated in CM, providing a mechanistic basis for future validation in disease-relevant models.

Background: Phytic acid in grains chelates essential minerals, thereby reducing the bioavailability of phosphorus and micronutrients in maize-based food and feed. Understanding the temporal expression pattern of the phytase1 gene, which regulates phytase activity, holds immense potential for maize biofortification efforts.

Methods and results: The expression of the phytase1 gene and its impact on phytase activity were explored at different grain developmental stages using maize inbreds with contrasting activity. The phytase1 expression pattern was analyzed using quantitative RT-PCR with adh1 as a reference gene. Combined ANOVA revealed significant variation (p < 0.01) due to inbreds (G) and developmental stages (DAP). High phytase genotypes, PMI-Q1 (1302.0 U kg^- 1), PMI-PV7 (1345.0 U kg^- 1), and PMI-PV8 (1413.3 U kg^- 1) showed higher expression of the phytase1 gene transcript levels of 0.114, 0.109, and 0.104, respectively. PMI-PV2 (586.4 U kg^- 1), PMI-PV4 (688.3 U kg^- 1), and PMI-PV5 (650.8 U kg^- 1) with the lower phytase activity had lower expression of phytase1 transcript levels of 0.040, 0.031, and 0.036, respectively. Furthermore, the correlation between phytase activity and relative expression pattern at different developmental stages was positive (p < 0.01; r = 0.79, 0.86, and 0.89 at 15, 30, and 45 DAP, respectively). Notably highest phytase activity was found at 15 DAP (1127.7 U kg^- 1) and declined progressively towards 45 DAP (895.4 U kg^- 1) across genotypes, with a corresponding reduction in phytase1 gene expression.

Conclusions: Validating the high phytase genotypes through gene expression profiling offers promising donors for maize molecular breeding to transfer high phytase activity into elite genotypes.