Background: Carbonic anhydrase (CA) catalyzes the formation of bicarbonate ions and protons from carbon dioxide and water, and also the reverse reaction, thus regulating acid-base balance and processes requiring HCO₃^- in the cell. Previous data from us and others have suggested that CAs participate in the regulation of insulin secretion. The aim of this study was to examine sublines of insulin-secreting MIN6 cells in terms of CA isoform transcript levels, and to analyze their roles in insulin secretion.

Methods: Differential expression of CA isoforms in MIN6 cells differing in glucose responsiveness were analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Next, we established stable cell lines with a doxycycline-inducible system overexpressing several of the CA isoforms. We also generated MIN6 sublines with shRNA-mediated suppression and studied glucose-stimulated insulin secretion, as well as glucose metabolism, using radiolabeled glucose in these cells.

Results: We confirmed expression of a mitochondrial CA isoform, CA VB, and a cytosolic isoform, CA XIII, was significantly greater (4.18-fold, p < 0.01 for CA VB; 48.3-fold, p < 0.01 for CA XIII), while expression of CA XI, a cytosolic isoform that lacks catalytic activity, was lower (0.50-fold, p < 0.01), in highly glucose-responsive sublines. CA VB overexpression increased (1.32-fold, p < 0.05), while its knockdown suppressed (0.77-fold, p < 0.05), glucose-stimulated insulin secretion. In contrast, CA XIII overexpression and CA XIII knockdown failed to modulate insulin secretion evoked by glucose. CA XI overexpression produced no changes in insulin secretion. Changes in insulin secretion due to CA VB overexpression or knockdown were not accompanied by changes in [5-³H]glucose and [U-¹⁴C]glucose metabolism.

Conclusions: CA VB serves as a positive regulator of glucose-stimulated insulin secretion in MIN6 cells.

Background: Tuberculosis is a bacterial infection caused by Mycobacterium tuberculosis (MTB) that affects both humans and animals. It is one of the world's most widespread serious public health challenges. The present study aimed to check the prevalence of tuberculosis through GeneXpert and association with possible risk factors in suspected tuberculosis positive patients in district Narowal, Pakistan.

Methods: A designed questionnaire was filled out to collect data from suspected patients including age, gender, area, marital status, diet, economic status, smoking status, home condition, education, and close contact with tuberculosis (TB) patients. A Lung function test was performed. Statistical analysis was done by applying a chi-square test in SPSS software (version 22) to evaluate the association between GeneXpert and acid-fast staining technique.

Results: A total of 500 samples were collected and they were analyzed through auramine rhodamine staining to get efficient results samples were further analyzed to GeneXpert. Out of 500 samples, 195 samples were positive for staining and 282 were positive for GeneXpert from both male and female. The prevalence rate was recorded higher in age groups >50 while a lower prevalence rate was recorded in less than 40 age groups. Rifampicin (RIF) resistance was detected in 63 patients in which males showed high resistance to rifampicin with (52.4%) as compared to females with (47.6%) resistance.

Conclusion: The actual prevalence of suspected tuberculosis cases is relatively high, and the GeneXpert MTB/RIF assay is superior to traditional acid fast staining in detecting the disease and identifying rifampicin-resistant patients at the same time, which is an important guideline for the diagnosis and treatment of the disease.

Background: Understanding the mechanisms underlying chemoresistance is crucial for improving the outcomes of chemotherapy. Nuclear factor κB (NF-κB) signaling can be triggered by a variety of chemotherapeutic drugs and may be involved in the development of tumour resistance. However, its role in oral squamous cell carcinoma (OSCC) resistance to cisplatin (DDP) remains unclear. Here, we sought to investigate it.

Methods: OSCC cells were divided into DDP and DDP+pyrrolidine dithiocarbamate (PDTC, an NF-κB inhibitor) groups, and their survival rates as well as the IC50 of DDP were examined after the 5-diphenyltetrazolium bromide solution (MTT) assay. The messenger RNA (mRNA) and protein levels of NF-κB, P-glycoprotein (P-gp), cleaved caspase-3, and B-cell lymphoma/leukaemia-2 (Bcl2) were analysed by reverse transcriptase–polymerase chain reaction (RT-PCR) and western blotting, respectively.

Results: The survival rate of OSCC cells and the IC50 of DDP in the DDP+PDTC group were lower than those in the DDP group. Low-dose DDP activated the NF-κB pathway and increased Bcl2 expression in a time-dependent manner, whereas P-gp expression showed no significant changes. PDTC inhibited DDP-induced activation of the NF-κB pathway by upregulating recombinant inhibitory subunit of I Kappa B Alpha (IκBα) expression, which suppressed the expression of the target gene P-gp, resulting in reduced Bcl2 levels and increased caspase-3 activation, ultimately enhancing the sensitivity of OSCC cells to DDP.

Conclusion: Low-dose DDP activates NF-κB signalling in OSCC cells, upregulating Bcl2 expression and leading to drug resistance. In contrast, PDTC inhibits NF-κB, which downregulates Bcl2 and P-gp expression and activates caspase-3, enhancing OSCC cell sensitivity to DDP. Our findings suggest that NF-κB signalling plays an important role in the resistance to chemotherapeutic drugs.

Background: Preeclampsia remains a leading cause of maternal and neonatal morbidity and mortality. It is characterized by altered local and systemic immune regulation and a rise in proinflammatory cytokines. The inflammasome is a cytosolic protein complex that mediates innate immune responses through promoting the secretion of interleukin-1beta (IL-1β). This study aimed to investigate the nucleotide oligomerization domain (NOD)-like receptor family, pyrin domain-containing protein 3, inflammasome activation in preeclampsia (PE), its relation to IL-1β and their association with PE outcomes.

Methods: Placenta and blood were collected from 25 control pregnant women and 50 preeclamptic women. Pyrin domain-containing protein 3 (NLRP3) and IL-1β gene expression were quantified by real-time polymerase chain reaction (PCR).

Results: Placental and blood relative gene expression levels of NLRP3 and IL-1β were significantly higher in mild and severe PE than in controls. A significantly higher blood expression of NLRP3 was noticed in the low birth weight subgroup compared to the normal birth weight subgroup (p = 0.03) in the severe PE group. Both biomarkers levels in placenta and blood showed significant negative correlations with the weight of newborn. The strong positive correlation (p < 0.0001, r = 0.9) between NLRP3 and IL-1β suggested that IL-1β response mostly depend on NLRP3 inflammasome.

Conclusions: These results suggest the presence of excessive activation of NLRP3 and subsequently increased production of active IL-1β that may predispose placental inflammation in severe PE and subsequently, low neonatal birth weight and shortened gestational age.

Objectives: Octylphenol (OP) is a member of the category of substances known as endocrine-disrupting chemicals and can interfere with the actions of steroid hormones, having negative implications on reproductive health. Alpha lipoic acid (αLA), a naturally occurring nutraceutical, has been shown to provide therapeutic benefits due to its antioxidant activity and capacity to reverse oxidative damage. Consequently, the current study is attending to estimate the protective effects of αLA against OP-induced male infertility.

Methods: Forty male albino rats were randomly divided equally into four groups. Group I (control group) didn't received any materials, Group II (αLA-group) received alpha lipoic acid (40 mg/Kg/day), group III (OP-group) received octylphenol (50 mg/Kg/day), and group IV (protective group) received octylphenol (50 mg/Kg/day) plus alpha lipoic acid (40 mg/Kg/day) for 30 days. Serum reproductive hormones (gonadotropin releasing hormone (GnRH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone, total oxidant status (TOS), and total antioxidant capacity (TAC) levels were measured. Histological and histochemical examinations of the hypothalamus, pituitary gland and testis, as well as immunohistochemical detection for tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the pituitary gland and testis were performed.

Results: Rats of OP-group showed significant reduction of serum GnRH, FSH, LH, testosterone and TAC levels and significant rise of TOS levels. In parallel with these outcomes, several histopathological and histochemical abnormalities were observed in their hypothalamus, pituitary and testicular tissues. Additionally these structural abnormalities were associated with increased TNF-α and IL-6 expressions in both pituitary and testicular tissues. On other hand, rats of protective group showed enormous improvements in biochemical, histopathological, and immunohistochemical derangements induced by OP. Insignificant differences in the biochemical, histopathological, and immunohistochemical examinations were observed between control group and αLA-group.

Conclusions: Short-term exposure of OP at low doses disturbs the hypothalamic-pituitary-gonadal (HPG) axis associated with increased oxidative stress, inflammation and structural abnormalities with successful reversion of theses derangements by αLA via its antioxidant and anti-inflammatory properties.