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Background: This study investigates the relative expression of the Na+, HCO3- cotransport gene NBCn1, and caspase-3 within the tumor microenvironment of human breast cancer, considering the in vivo microenvironment. Method: In this experimental study, breast cancer MDA-MB-231 cells were cultured under normoxia/hypoxia conditions for 24, 48, and 72 hours with varying glucose concentrations (5.5, 11, and 25 mM). The mRNA expression of NBCn1 and caspase-3 was evaluated using real-time polymerase chain reaction. The stability and binding pocket of NBCn1 were assessed using DispHred and the Computed Atlas of Surface Topography of proteins (CASTp) servers, respectively. The location prediction of the protein was determined using the Transmembrane Helices; Hidden Markov Model (TMHMM) server. Results: Normoxia led to an increase in NBCn1 expression during all three time periods, displaying heterogeneity. The expression was particularly elevated at glucose concentrations of 25 and 5.5 mM. In hypoxic conditions, gene expression was reduced; however, an increase in glucose concentration enhanced SLC4A7 expression. Specifically, a glucose concentration of 25 mM led to decreased caspase-3 expression under hypoxic conditions. In silico studies revealed that SLC4A7 becomes disordered when the pH falls below 7, with most amino acids in the binding pocket being nonpolar. Conclusion: The heightened risk of breast cancer metastasis may be linked to the upregulation of SLC4A7 and downregulation of caspase-3 expression, underscoring their fundamental roles in cancer treatment and prevention. SLC4A7 is a transmembrane protein, and its folding is pH-dependent. [ABSTRACT FROM AUTHOR]

Brucellosis is a momentous zoonotic disease. Unpasteurized milk and milk products are the leading sources of Brucella transmission to humans. The present research was conducted to investigate the contamination of local crumbled Kope cheeses distributed in Urmia city with Brucella species and evaluate the antibiotic -resistance pattern of the isolates. Fifty samples of local cow’s Kope cheese were randomly collected from traditional dairy retailers in different regions of Urmia by sterile conditions in 2022. The samples were cultured in Brucella enrichment broth and then in Brucella selective agar with a supplement. Molecular identification of Brucella spp. was done using specific primers by polymerase chain reaction (PCR). Antimicrobial susceptibility testing on the isolates was performed by the Kirby -Bauer disc diffusion method. Among the tested samples, two samples were contaminated with Brucella (4%). One was contaminated with B. abortus bv. 1, 2, and 4 (2%), and the other with B. melitensis (2%). The isolates were sensitive to azithromycin, imipenem, doxycycline, rifampin, and co -trimoxazole antibiotics and were resistant to ampicillin, amoxicillin -clavulanic acid, tetracycline, gentamicin, and ceftriaxone antibiotics. Also, the B. abortus strain was sensitive to ciprofloxacin, but the B. melitensis strain was resistant. The isolates showed multi -drug resistance (MDR) characteristics. From the findings, it can be concluded that Brucella contamination in local Kope cheeses distributed in Urmia city is low; However, due to the high pathogenicity of B. melitensis for humans, it is recommended to screen infected cows, vaccinate sheep and goat herds, and prevent the supply of unpasteurized milk and milk products. It is also recommended to evaluate azithromycin and imipenem antibiotics along with other common antibiotics in brucellosis. [ABSTRACT FROM AUTHOR]

Introduction: Cancer is an intricate disorder/dysfunction of cells that can be defined as a genetic heterogeneity in human disease. Therefore, it is characterized by several adaptive complex hallmarks. Among them, the pH dysregulation appears as a symbol of aberrant functions within the tumor microenvironment (TME). In comparison with normal tissues, in the solid tumors, we face with an irregular acidification and alkalinization of the extracellular and intracellular fluids. Methods: In this study, we comprehensively discussed the most recent reports on the hallmarks of solid tumors to provide deep insights upon the molecular machineries involved in the pH dysregulation of solid tumors and their impacts on the initiation and progression of cancer. Results: The dysregulation of pH in solid tumors is fundamentally related to the Warburg effect and hypoxia, leading to expression of a number of molecular machineries, including: NHE1, H+ pump V-ATPase, CA-9, CA-12, MCT-1, GLUT-1. Activation of proton exchangers and transporters (PETs) gives rise to formation of TME. This condition favors the cancer cells to evade from the anoikis and apoptosis, granting them aggressive and metastasis phenotype, as well as resistance to chemotherapy and radiation therapy. This review aimed to discuss the key molecular changes of tumor cells in terms of bio-energetics and cancer metabolism in relation with pH dysregulation. During this phenomenon, the intra- and extracellular metabolites are altered and/or disrupted. Such molecular alterations provide molecular hallmarks for direct targeting of the PETs by potent relevant inhibitors in combination with conventional cancer therapies as ultimate therapy against solid tumors. Conclusion: Taken all, along with other treatment strategies, targeting the key molecular machineries related to intra- and extracellular metabolisms within the TME is proposed as a novel strategy to inhibit or block PETs that are involved in the pH dysregulation of solid tumors. [ABSTRACT FROM AUTHOR]

Introduction: Testis-specific gene antigen 10 ( TSGA10 ) is a less-known gene, which is involved in the vague biological paths of different cancers. Here, we investigated the TSGA10 expression using different concentrations of glucose under hypoxia and also its interaction with the hypoxia-inducible factor 1 (HIF-1). Methods: The breast cancer MDA-MB-231 and MCF-7 cells were cultured with different concentrations of glucose (5.5, 11.0 and 25.0 mM) under normoxia/hypoxia for 24, 48, and 72 hours and examined for the HIF-1α expression and cell migration by Western blotting and scratch assays. The qPCR was employed to analyze the expression of TSGA10 . Three-dimensional (3D) structure and the energy minimization of the interacting domain of TSGA10 were performed by MODELLER v 9.17 and Swiss-PDB viewer v4.1.0/UCSF Chimera v1.11. The UCSF Chimera v 1.13.1 and Hex 6.0 were used for the molecular docking simulation. The Cytoscape v 3.7.1 and STRING v11.0 were used for protein-protein interaction (PPI) network analysis. The HIF-1a related hypoxia pathways were obtained from BioModels database and reconstructed in CellDesigner v4.4.2. Results: The increased expression of TSGA10 was found to be significantly associated with the reduced metastasis in the MDA-MB-231 cells, while an inverse relationship was seen between the TSGA10 mRNA level and cellular migration but not in the MCF-7 cells. The C-terminal domain of TSGA10 interacted with HIF-1α with high affinity, resulting in PPI network with 10 key nodes ( HIF-1α, VEGFA, HSP90AA1, AKT1, ARNT, TP53, TSGA10, VHL, JUN, and EGFR ). Conclusions: Collectively, TSGA10 functional expression alters under the hyper-/hypo-glycemia and hypoxia, which indicates its importance as a candidate bio-target for the cancer therapy.