Molecular Biology Reports最新文献

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.

Background: Triple-negative breast cancer (TNBC) remains a major therapeutic challenge due to the lack of established molecular targets and the presence of highly infiltrating tumor-associated macrophages (TAMs), particularly the immunosuppressive M2 phenotype that drives tumor progression. This study investigated the potential of lutein, a plant-derived carotenoid with anticancer properties, to repolarize human monocyte-derived M2 macrophages into the antitumor M1 phenotype and thereby modulate the TNBC tumor microenvironment.

Methods and results: CD14 + monocytes isolated from peripheral blood mononuclear cells were differentiated into M1 or M2 macrophages using GM-CSF/IFN-γ/lipopolysaccharide or M-CSF/IL-4, respectively. Lutein's effects on macrophage polarization, cell proliferation, cell migration and cytokine secretion were evaluated using flow cytometry, MTT proliferation assays, migration assays and enzyme-linked immunosorbent assays (ELISAs) in MDA-MB-231 cells. Additionally, tumor-related protein expressions from MDA-MB-231 cells were analyzed using protein arrays. Lutein significantly reduced the proliferation and migration of MDA-MB-231 cells when co-cultured with M1/M2 macrophages compared to the untreated group. IL-4 stimulation increased the expression of M2 macrophage marker (CD206) and protumor cytokines (IL-10 and TGF-β1), but lutein effectively reduced these elevated molecules in IL-4-activated macrophages. Proteomic profiling revealed that lutein downregulated several protumor proteins (survivin, VEGF, BCL-X, M-CSF, MMP-9) and upregulated antitumor markers (FKHR and GM-CSF). Furthermore, lutein markedly reduced the secretion of proinflammatory and protumorigenic cytokines and chemokines (IL-6, IL-18BPa, CCL2, CCL8, CCL7, CCL3, CCL20, and CXCL8).

Conclusion: These findings suggest that lutein reprograms M2 macrophages toward an M1-like phenotype and disrupts tumor-promoting signaling within the TNBC microenvironment, highlighting its potential as an adjunct therapeutic agent.

Background: Human sex development is a highly regulated process guiding undifferentiated gonads toward a testicular or ovarian fate. Disruptions in this pathway result in disorders of sexual development (DSD), characterized by atypical chromosomal, gonadal, or anatomical sex. These conditions usually appear as ambiguous genitalia at birth or as atypical pubertal development during adolescence. Different etiologic, phenotypic, and genotypic factors can cause DSD. Advances in next-generation sequencing (NGS) have significantly accelerated the identification of genetic variants through targeted panels, including both known genes involved in sex determination and differentiation, as well as newly discovered genes linked to DSD.

Methods and results: In this study, whole exome sequencing (WES) was performed on a Moroccan patient, born to non-consanguineous parents, who presented with severe hypospadias, micropenis, and cryptorchidism, and exhibited overlapping phenotypic features consistent with congenital disorder of glycosylation (CDG) and primary ciliary dyskinesia (PCD). After variant annotation and prioritization, two heterozygous variants in the MPI (c.305 C > T; p. Ser102Leu) and RSPH1 (c.471 C > G; p. His157Gln) genes were identified and confirmed by Sanger sequencing in family members. Their pathogenic effects on protein structures and functions were subsequently anticipated using bioinformatic tools and molecular dynamics (MD) simulations.

Conclusions: To our knowledge, this is the first report of these specific variants in the context of DSD, shedding light on a unique genotype-phenotype profile associated with the patient's complex clinical presentation. The high genetic variability underlying these disorders has made molecular diagnosis challenging. Yet, genomic approaches could expand our understanding of DSD landscape and improve diagnosis, personalized interventions, and patient management.

Purpose: Inflammatory bowel disease (IBD) is a chronic inflammatory disease characterized by damage to the epithelial barrier and intestinal inflammation; there is currently no standard treatment. Recent evidence suggests that worms and their by-products may have potential to treat IBD, owing to their immunomodulatory effects in humans. We investigated the effects of the recombinant Echinococcus granulosus antigen EgM123 on host immunity and inflammation.

Methods: We produced recombinant EgM123 to test its effects in the dextran sodium sulfate (DSS)-induced colitis mouse model. Mice were immunized with EgM123 three times via intraperitoneal injection and then challenged with 4% DSS in the drinking water for 1 week; they were observed daily for weight, mobility, and stool. After necropsy, fixed tissues/organs were sectioned and stained with hematoxylin and eosin to assess inflammation. Cytokine levels were detected by ELISA and reverse transcription polymerase chain reaction. Colitis severity was assessed by colon length, weight loss, and a semi-quantitative score of morphological changes.

Results: EgM123 had protective effects against colitis in mice. Histopathological analysis of the colon revealed a significant reduction in intestinal inflammation caused by DSS, which was associated with downregulation of the Th1/Th17 response in the colon. Administration of EgM123 before colitis induction limited the severity of intestinal inflammation and the disease index score, inhibited TNF-α secretion, and induced IL-10 expression.

Conclusions: EgM123 may alleviate colitis by inhibiting the Th1 response and inducing Th2 immunity. Immunotherapy with EgM123 may represent a strategy to modulate the inflammatory processes involved in IBD.

Background: Eurasian red squirrels (Sciurus vulgaris) are common in Europe and Asia, but are declining in many regions across their range. As continental European populations are now facing current and future threats from invasive species in addition to existing anthropogenic pressures it will be important to carefully monitor these populations. Non-invasive genetic sampling methods are a useful tool in conservation assessments, but often require techniques such as microsatellite markers that can be used with lower quality DNA. It remains helpful to increase the resolution of these assessments by identifying additional genetic markers. We describe new microsatellite markers developed from European red squirrels from France and use them to assess genetic diversity in populations in southern France.

Methods and results: We used Illumina sequencing to characterize microsatellites from tissue samples of S. vulgaris. Using 7 tissue samples we assessed amplification and polymorphism in 48 microsatellite inserts and further evaluated 16 of these microsatellite loci in hair samples from 120 individuals from four populations. In the 104 samples for which those loci amplified, there was an average of 6.1 alleles amplified per locus, with mean observed and expected heterozygosities of 0.44 and 0.59, respectively. Only one locus showed significant deviation from HWE across all populations. The same locus exhibited a likely presence of null alleles.

Conclusions: We describe 16 new microsatellite loci, with caution required for one locus in analyses sensitive to null alleles. These new loci can help provide increased resolution in population genetic assessments of red squirrels in continental Europe.