Molecular Biology Reports最新文献

Background: Methylmalonic acidemia (MMA), type mut (0) is a rare type of genetic inborn error of metabolism (IEM) that is caused by aberrant malonyl-CoA mutase activity. Diagnosing IEM can be challenging due to its inherited onset and varying degrees of severity.

Methods and results: In the present study, a consanguineous Pakistani family suspected of IEM was genetically analyzed using whole exome sequencing. A biallelic variant c.689 C > G (p.Thr230Arg) in MMUT was identified to be the causative factor of the disease, which helped in establishing the accurate diagnosis in the family to be MMA mut(0) type. On the basis of the genetic findings, the patient's condition was appropriately managed through a supportive nutrition plan and administration of oral L-carnitine.

Conclusions: Identification of MMUT mutation through whole exome sequencing was helpful in solving the family and devising targeted management strategies. This study highlights the utility of genetic analysis in diagnosing and treating metabolic disorders like MMA in Pakistani inbred population.

Background: Larimichthys crocea is an important aquaculture species along the southeastern coast of China, with diverse environment and farming practices since artificial breeding, these different aquatic habitats are subject to significant variations in environmental factors that may involve modulation of gene expression through epigenetic mechanisms to enable species to survive and reproduce.

Methods and results: This study aimed to identify methylation variation sites (SMVs) in different sequence contexts (CG, CHG, and CHH) within populations of L. crocea in different habitats. All SMV sites were subjected to linear regression with environmental factors to identify candidate genes involved environmental stress. The results indicate a significant correlation between SMV sites and various environmental factors. For the wild populations in Jinmen and Zhanjiang, the primary environmental pressures for adapting are temperature and salinity. In contrast, for the domesticated populations in Zhoushan and farmed population in Xiangshan, the main environmental pressures are nitrate and dissolved oxygen. Furthermore, genes related to temperature adaptation in different aquatic environments were identified, including nr3c2, igf1, hsp70, trpm3, and fgf1. The gene rasa3 was found to be associated with pH adaptation, while genes such as atp6ap1lb, slc15a4, and gpr39 were linked to salinity, ammonia nitrogen, and dissolved oxygen. Research on the association between single methylation variation sites (SMVs) and environmental factors in aquatic organisms is scarce.

Conclusions: These results suggest that selection pressures can influence a significant proportion of methylation sites in this species, indirectly implying that epigenetic variation is not solely attributed to patterns of genetic variation, but is also closely linked to environmental differences. These results highlight the complex interactions between epigenetic regulation and environmental influences. Hence, this study provides preliminary evidence for a new perspective on the role of methylation patterns in L. crocea in environmental adaptation.

Background: Native to the Amazon region, Copaifera multijuga Hayne is a large tree (≈ 36 m in height) that is heavily exploited for extraction of its oleoresin. Many studies have addressed the phytochemical properties and applications of this raw material; however, there are few initiatives that have focused on the genetic characterization of native populations of this species. To this end, our objective was to develop microsatellite markers for C. multijuga, which were previously unavailable, and apply them to the characterization of a native population located in the Adolpho Ducke Forest Reserve (Manaus, Amazonas, Brazil).

Methods and results: Using next-generation sequencing technology on the Ion Torrent PGM™ platform, 19 pairs of microsatellite primers were designed and developed. For the characterization and validation, we used a group of 47 C. multijuga trees. After preliminary tests, amplification conditions were standardized for 14 loci. The CmH05 locus was excluded from the analyses for being monomorphic, and the remaining loci were used to estimate key genetic parameters for the species, such as observed (Ho) and expected heterozygosity (He), total number of alleles (A), fixation index (f), and polymorphic information content (PIC), among others. The population showed levels of genetic diversity that were higher than 0.63 (Ho: 0.67; He: 0.64) and an average number of 5.4 alleles. According to the PIC estimates, all loci were considered highly (9) or moderately informative (3), except for CmH11, which had a value that was below 0.2.

Conclusions: The 13 developed microsatellites were efficient in characterizing the genetic diversity of C. multijuga and may be advantageous in future investigations aimed at defining effective conservation strategies for the species.

Background: Ticks are prominent vectors of numerous pathogens that adversely affect human and animal health. Monitoring tick population dynamics is key in developing ideal tick-borne disease surveillance systems and critical vector control programmes. This study aimed to conduct the morphological and molecular characterization of ticks infesting domesticated goats in Kerala, India.

Methods and results: A total of 30 goats presented to the small ruminant unit of the Teaching Veterinary Clinical Complex (TVCC), Mannuthy were randomly screened for tick infestation, with 22 (73.3%) found to be infested. Morphological identification of different life cycle stages and genera was conducted first, utilizing documented external characteristics such as body size, presence of eyes, and other key morphological traits. A total of 153 tick samples were collected from goats, and their identification revealed that they belonged to the genera Haemaphysalis and Rhipicephalus. This was followed by molecular analysis through sequencing a fragment of the cytochrome c oxidase subunit I (COXI) gene, a standard marker for tick identification. The results from molecular and phylogenetic analyses confirmed the tick species as Haemaphysalis bispinosa, H. intermedia, Rhipicephalus haemaphysaloides, and R. sanguineus. The sequenced specimens were deposited in the NCBI GenBank contributing to the global understanding of tick distribution and diversity in goats. The GenBank accession no. (s) of the isolates are PQ433166 (H. bispinosa), PQ433290 (H. intermedia), PQ433525 (R. haemaphysaloides), and PQ433586 (R. sanguineus).

Conclusions: The findings of this study contribute to a better understanding of the tick fauna infesting goats in the region and emphasize the importance of developing research and monitoring plans to address the challenges posed by these ectoparasites. It also highlights a critical area for future study, targeting the vector potential of these arthropods in hemoparasitic diseases and zoonotic disease transmission.

Background: In the context of global change, coral reefs and their associated biodiversity are under threat. Several conservation strategies using population genetics have been explored to protect them. However, some components of this ecosystem are understudied, such as hydrozoans, an important class of benthic organisms worldwide. A comprehensive study of coral reefs as a whole is needed to develop effective conservation measures. Here we describe the development of 75 new microsatellite markers for 5 hydroid species: Antennella billardi, Lytocarpia phyteuma, Sertularella diaphana, Taxella gracilicaulis and Zygophylax rufa.

Methods and results: Markers were tested on 246 specimens from Reunion and Mayotte islands in the southwestern Indian Ocean. Allelic diversity ranged from 1 to 21 for the 5 species, and 9 loci were estimated to have null allele frequencies ranging from 10 to 37%. Observed and expected heterozygosities ranged from 0.03 to 1 and from 0.03 to 0.93, respectively. 12 loci showed data significantly out of Hardy-Weinberg equilibrium. Cross-amplification was performed in 8 species, of which 3 showed high successful amplification rates (53 to 93%).

Conclusion: The estimated genetic metrics were similar to those reported for other hydroid and cnidarian marker sets. Cross-amplification showed a contrasting transferability between species, often related to the hydroid phylogeny. These newly developed markers will be relevant to the study of hydroid population genetics and coral reef conservation.

This study conducts an in-depth review of the correlation between testis tissue changes and circulating microRNAs (miRNA) in diabetes-induced male reproductive complications, drawing upon both animal and clinical studies. The original articles published in English that specifically investigate miRNAs linked to male infertility in humans or animals with either type I or ΙΙ diabetes mellitus were included. The relevant articles were gathered from the PubMed, Google Scholar, Cochrane Library, and ScienceDirect databases. The quality of study was assessed utilizing the Joanna Briggs Institute (JBI) Critical Appraisal Checklist for Prevalence Studies. We collected an overall number of 1989 citations relating to our research subject. Following the elimination of articles based on the criteria, a total of 20 papers were included in the study. Aberrant expression profiles of 25 miRNAs were identified in diabetes associated with male reproductive issues, with 15 miRNAs exhibiting increased expression and 10 miRNAs showing decreased expression. Among the chosen publications, eighteen were identified as low-risk and two were classed as moderate quality. The dysregulated miRNAs were linked to testicular injury, disrupted steroid production, decreased sperm development and quality, and erectile dysfunction. The results demonstrate that the miRNA-mRNA network is linked to the pathological progression of diabetic testicular damage or erectile dysfunction. From a therapeutic perspective, the identification of circulating miRNAs could be beneficial in the timely identification and prevention of diabetes problems, such as diabetes-induced male infertility. Among all signaling pathways influenced by modified miRNAs, the Bax-caspase-3, MAPK, PI3K-Akt, and eNOS-cGMP-PKC were the main deregulated pathways.

Introduction: Cataranthine is an alkaloid used in the development of anti-cancer drugs. In this study, the effect of cataranthine is assessed by measuring the levels of miR-34 and miRNA-29, which are effective regulators of BCL-2 and NRF-2 gene expression, and their relation to the survival of HCC cells.

Methods: This study used cataranthine, and the HepG2 cell line. The MTT test was used to determine the appropriate concentration of cataranthine for treatment (IC50). Oxidative stress status was assessed by evaluating TAC (total antioxidant capacity), TOS (total oxidant status), and MAD (malondialdehyde) levels. Flow cytometry was used to investigate apoptosis. The expression levels of Nrf2, Bcl2, miRNA34, and miRNA29 genes in HepG2 were evaluated by RT-PCR.

Results: We observed that cataranthine significantly reduced the levels of oxidative markers (MAD, and TOS) and, conversely, increased the level of antioxidant markers in HepG2 cells. Treatment of HepG2 cells with different doses of cataranthine significantly increased the expression of Nrf2 and Bcl-2 genes, while significantly decreasing the expression of miR29 and miR34 genes.

Conclusion: These findings suggest that cataranthine may exert its anticancer effects by reducing oxidative stress and promoting apoptosis, while decrease in miR34 and miR29 as well as increase in Nrf2 and Bcl2 may act as resistance mechanisms in cancer cells. The results highlight the dual potential of cataranthine in regulating cellular responses to oxidative stress and cell death in liver cancer, with dose-dependent modulatory effects.

In the last decades the survival of metastatic gastrointestinal (GI) cancer patients could have been significantly extended due to the introduction of targeted- and immunotherapy. However, only the minority of patients will experience long-lasting survival. Hence, novel therapeutics are clearly necessary for GI cancer patients. Molecular high-throughput profiling techniques have revealed potential novel targetable molecular alterations, emphasizing the necessity for tailored therapeutic approaches. Nuclear export proteins, particularly Exportin-1 (XPO1), have emerged as promising targets in cancer therapy due to their crucial role in cellular homeostasis and regulation of key cellular functions. Dysregulation of XPO1-mediated nuclear export leads to the functional loss of tumor suppressors and pro-apoptotic factors, facilitating cancer progression. Selinexor, a XPO1 inhibitor, has shown promising activity in preclinical and clinical studies, particularly in hematological malignancies. However, its efficacy in GI cancers remains underexplored. This review aims to elucidate the functional and pathophysiological role of XPO1 in GI cancers. Despite the potential of XPO1 inhibitors in suppressing cell proliferation and inducing apoptosis, comprehensive molecular landscape data and validation of selective inhibitors in GI cancers are lacking. Targeting XPO1 presents a significant therapeutic potential for the treatment of GI cancer patients. Further research is necessary to fully elucidate the molecular landscape according to XPO1 expression in GI tumors and to validate the efficacy of selective XPO1 inhibitors. These efforts are expected to contribute to the development of more effective and personalized therapeutic strategies for GI cancer patients.

Background: Molecular cytogenetics, utilizing DNA probes, serves as a critical tool for mapping genes to the physical structures of chromosomes.

Methods: In this study, we examined three Allium species: A. cepa L., A. sativum L., and A. fistulosum L., using in situ hybridization to localize 45S rDNA and 5S rDNA genes.

Results: We observed variation in both the chromosomal localization and signal intensity of the 45S and 5S rDNA probes across the species. Notably, in A. sativum, additional 5S rDNA signals were detected on chromosome 8, in a heterozygous condition. Additionally, we aimed to explore the feasibility of localizing genes associated with pigment biosynthesis in A. cepa, specifically the PAL and FLS genes. For this, we employed TSA-FISH on both meiotic and mitotic chromosomes. Preliminary results suggested that the PAL gene was localized to meiotic metaphase chromosomes, while the single-copy FLS gene was detected on mitotic chromosomes.

Conclusion: The TSA-FISH technique proved neither routine nor robust for consistent localization of these specific probes in plant chromosomes. The findings based on rDNA analysis also offer insights into potential evolutionary implications among the different Allium species studied.

Background: Premature ovarian insufficiency (POI) is a refractory disease that severely affects female fertility. The PERK/eIF-2α/ATF4/CHOP pathway is one of the classical pathways involved in the unfolded protein response to endoplasmic reticulum stress by regulating protein synthesis and promoting apoptosis. This study aimed to investigate the functional role and mechanism of human umbilical cord mesenchymal stem cells (hUCMSCs) in the POI animal model through the PERK/eIF-2α/ATF4/CHOP pathway.

Methods and results: Forty-five sexually mature female C57 mice were divided into a blank control group, POI model group, and hUCMSCs intervention group. To establish the POI model, mice received intraperitoneal injections of cyclophosphamide (CTX) (70 mg/kg) daily for 14 consecutive days, while the control group received saline only. In the hUCMSC intervention group, mice were given hUCMSCs on days 14 and 28, based on CTX modeling in the POI model group. The hUCMSCs were isolated, labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindotricarbocyanine iodide (DiR) fluorescent dye, and tail vein-injected, and the distribution of the DiR signal was monitored in the mice using a fluorescence imaging detection method. The ovarian tissues were hematoxylin and eosin stained to observe the ovarian structure, and the number of primordial follicles were counted. An enzyme-linked immunosorbent assay was used to detect the serum levels of estradiol, anti-mullerian hormone, and follicle-stimulating hormone. Terminal deoxynucleotidyl transferase dUTP nick end labeling was used to detect the apoptosis of granulosa cells (GCs). The reactive oxygen species (ROS) content of ovarian tissue was detected by flow cytometry assay. The RNA expression of PERK, eIF-2α, ATF4, and CHOP was determined by quantitative real-time polymerase chain reaction, and protein levels of the targets were determined by western blot and immunohistochemistry. We identified hUCMSCs using surface antigenic markers (CD90, CD44, CD105, and CD73), and osteoblasts and chondroplast differentiation assays. Our studies demonstrated that hUCMSC intervention significantly restored ovarian function by improving the irregular estrous cycle, increasing the number of follicles, decreasing ROS, and inhibiting GC apoptosis in POI mice. Moreover, hUCMSCs suppressed CTX-induced PERK/eIF-2a/ATF4/CHOP pathway activation.

Conclusions: HUCMSCs can migrate to the damaged ovaries of POI mice, and improve the ovarian function of POI mice by inhibiting oxidative stress, down-regulating the expression of the PERK/eIF-2α/ATF4/CHOP pathway, and reducing the apoptosis of GCs.