Prospective evaluation of outcome of percutaneous nephrolithotomy using the ‘STONE’ nephrolithometry score: a single-centre experience

Abstract OBJECTIVE: To assess the prediction of stone clearance and complications of percutaneous nephrolithotomy (PCNL) using the \u27STONE\u27 nephrolithometry score, assessing stone size, tract length (skin-to-stone distance), degree of obstruction, number of calyces involved and stone essence (density). PATIENTS AND METHODS: This was a prospective study of patients undergoing single-tract PCNL while prone, conducted at a university hospital. All patients had non-contrast-enhanced computed tomography within 3 weeks of the procedure. Only patients with a unilateral procedure and radio-opaque stones were included. The five variables of the STONE nephrolithometry score were calculated before the procedure. The stone-free rates were assessed with a plain abdominal film at 4 weeks and complications were graded using the modified Clavien system. RESULTS: In all, 107 patients were included in the final analysis. Overall, 80% of patients were rendered stone-free. Among the individual variables, a larger stone (P = 0.002) and the involvement of multiple calyces (P = 0.04) were associated with residual stones, while tract length (skin-to-stone distance), stone density and presence of hydronephrosis were not. Patients who were rendered stone-free had a statistically significant lower overall STONE score than those with residual stones, at 7.24 vs. 8.14 (P = 0.02). The score also correlated with operative duration, which was significantly longer with a higher STONE score (P = 0.03). The complication rate was 18% and most complications were Clavien grade 2, with bleeding requiring a blood transfusion (11 patients) being the commonest. There were no deaths within 30 days of surgery, but there was no correlation between the STONE score and complications. CONCLUSION: The STONE nephrolithometry score is a simple and easy to apply system for predicting complexity in stone clearance with PCNL

MAMP-triggered resistance induced by elicitor protein PeBA1 derived from Bacillus amyloliquefaciens NC6 in common bean (Phaseolus vulgaris L.) against green peach aphid (Myzus persicae Sulzer)

Bacterial microbe-associated molecular patterns (MAMPs) play an important role in innate plant immunity. This in vitro study evaluated the putative role of protein elicitor PeBA1 derived from Bacillus amyloliquefaciens NC6 strain in eliciting induced resistance type responses in common bean (Phaseolus vulgaris) plants against green peach aphid Myzus persicae. Nymphal developmental time of aphids was significantly prolonged and the fecundity was significantly reduced by different concentrations of PeBA1 elicitor (i.e. 40.51, 24.91 and 16.38 µg mL-1) applied at three different temperature regimes (i.e. 21, 27 and 30 °C). Moreover, foliar application of PeBA1 elicitor protein strongly up-regulated the expression levels of salicylic acid (SA) pathway-associated genes, while the expression levels of jasmonic acid (JA) pathway-associated genes exhibited a moderate induction. Quantification by LC/MSMS revealed a linear increase of both SA and JA plant defense hormones along with the time of exposure. Our findings suggest that the bacterial elicitor protein PeBA1 could be used as an effective biological pest management tool against phloem-feeding insect pests such as green peach aphids M. persicae

Human Dendritic Cell Subsets Undergo Distinct Metabolic Reprogramming for Immune Response

Toll-like receptor (TLR) agonists induce metabolic reprogramming, which is required for immune activation. We have investigated mechanisms that regulate metabolic adaptation upon TLR-stimulation in human blood DC subsets, CD1c+ myeloid DCs (mDCs) and plasmacytoid DCs (pDCs). We show that TLR-stimulation changes expression of genes regulating oxidative phosphorylation (OXPHOS) and glutamine metabolism in pDC. TLR-stimulation increases mitochondrial content and intracellular glutamine in an autophagy-dependent manner in pDC. TLR-induced glutaminolysis fuels OXPHOS in pDCs. Notably, inhibition of glutaminolysis and OXPHOS prevents pDC activation. Conversely, TLR-stimulation reduces mitochondrial content, OXPHOS activity and induces glycolysis in CD1c+ mDC. Inhibition of mitochondrial fragmentation or promotion of mitochondrial fusion impairs TLR-stimulation induced glycolysis and activation of CD1c+ mDCs. TLR-stimulation triggers BNIP3-dependent mitophagy, which regulates transcriptional activity of AMPKα1. BNIP3-dependent mitophagy is required for induction of glycolysis and activation of CD1c+ mDCs. Our findings reveal that TLR stimulation differentially regulates mitochondrial dynamics in distinct human DC subsets, which contributes to their activation

RIP1 protein-dependent assembly of a cytosolic cell death complex is required for inhibitor of apoptosis (IAP) inhibitor-mediated sensitization to lexatumumab-induced apoptosis

Searching for new strategies to trigger apoptosis in rhabdomyosarcoma (RMS), we investigated the effect of two novel classes of apoptosis-targeting agents, i.e. monoclonal antibodies against TNF-related apoptosis-inducing ligand (TRAIL) receptor 1 (mapatumumab) and TRAIL receptor 2 (lexatumumab) and small-molecule inhibitors of inhibitor of apoptosis (IAP) proteins. Here, we report that IAP inhibitors synergized with lexatumumab, but not with mapatumumab, to reduce cell viability and to induce apoptosis in several RMS cell lines in a highly synergistic manner (combination index <0.1). Cotreatment-induced apoptosis was accompanied by enhanced activation of caspase-8, -9, and -3; loss of mitochondrial membrane potential; and caspase-dependent apoptosis. In addition, IAP inhibitor and lexatumumab cooperated to stimulate the assembly of a cytosolic complex containing RIP1, FADD, and caspase-8. Importantly, knockdown of RIP1 by RNA interference prevented the formation of the RIP1·FADD·caspase-8 complex and inhibited subsequent activation of caspase-8, -9, and -3; loss of mitochondrial membrane potential; and apoptosis upon treatment with IAP inhibitor and lexatumumab. In addition, RIP1 silencing rescued clonogenic survival of cells treated with the combination of lexatumumab and IAP inhibitor, thus underscoring the critical role of RIP1 in cotreatment-induced apoptosis. By comparison, the TNFα-blocking antibody Enbrel had no effect on IAP inhibitor/lexatumumab-induced apoptosis, indicating that an autocrine TNFα loop is dispensable. By demonstrating that IAP inhibitors and lexatumumab synergistically trigger apoptosis in a RIP1-dependent but TNFα-independent manner in RMS cells, our findings substantially advance our understanding of IAP inhibitor-mediated regulation of TRAIL-induced cell death

National hepatitis registry in Pakistan: a dire need for hepatitis surveillance and control

Abstract Hepatitis is a major public health issue in Pakistan, with an estimated 11.55% prevalence of HCV infection in the adult population. The country ranks second globally in terms of hepatitis C virus (HCV) infections, with approximately one in every 20 Pakistanis already infected. The mortality rates due to HBV and HCV stand at 563,000 and 366,000 annually, respectively. However, the absence of a national registry or database system and the lack of coordination among provinces pose significant obstacles in combating this disease effectively. To address this issue, the establishment of a centralized national database registry is crucial, allowing comprehensive analysis, tracking of hepatitis prevalence, and identification of high-risk areas for targeted interventions. By fostering collaboration among provinces, the government, and non-governmental organizations, the registry would facilitate joint decision-making, minimize duplication of efforts, and address inconsistencies in diagnosis and treatment. Collaborating with student-run organizations and leveraging enhanced laboratory capacities post-COVID era can strengthen the hepatitis control program. The centralized approach and unified efforts are necessary to achieve the goal of a hepatitis-free Pakistan, where a healthier future can be realized

Association of demographics, lumbar active range of motion and disability in chronic low back: a baseline data analysis of a randomized controlled trial from Pakistan

Abstract Background The disability and significant economic costs accredited to Low back pain (LBP) are likely to rise which is an essential problem in low and middle-income countries like Pakistan. The associated factors of LBP are age, sex, and race including physical activity, high spinal load, lifting, bending, and twisting occupations. The literature highlighted there is substantial differences in associated factors of LBP within available studies in developing countries. The objective is to investigate the association of demographic factors and lumbar range of motion with disability in patients with chronic low back. Methods A baseline data analysis was performed as an analytical cross-sectional study among 150 patients with chronic low back in a randomized controlled trial with a duration from March 2020 and January 2021. After recording demographics, Modified-Modified Schober’s test was used to measure lumbar flexion and extension and Oswestry disability index for disability. After the descriptive analysis the continuous variables, age and pain were analyzed with Spearman’s correlation. Variables that were significant in bivariate analysis were then fitted in a multivariable linear regression. The Kruskal–Wallis test was used to analyze variations of disability in gender, marital status, work status, education level, and duration of pain. The p-value of 0.05 was significant. Results The results showed a significant correlation between age and pain in sitting (rh=-0.189, p = 0.021 and rh = 0.788, p  0.05). Conclusions The female gender and unmarried marital status are associated with functional disability. Decreased lumbar range of motion accompanies more disability, while age, education level, and work status do not effect on disability

Exploring relationships between Lipid parameters and Serum Vitamin-D deficiency.

Abstract Background: Vitamin D deficiency is a prevalent global health concern. To investigate the relationship between serum vitamin D levels and lipid parameters. Methodology: A prospective observational cohort study was carried out at Sindh Government Hospital Liaquatabad from October 2021 to February 2022. The study included a total of 171 participants, comprising both males and females aged 18 years or older, with vitamin D levels falling below 30 ng/ml. Participant selection employed a consecutive non-probability sampling method. Serum vitamin D levels were quantified using the COBAS method. Dyslipidemia and hypertriglyceridemia were defined based on established clinical guidelines. Exclusion criteria encompassed individuals with specific medical conditions and those currently using medications. Statistical analysis was conducted using SPSS version 22.0. Results: Out of the 171 participants, the majority were female, constituting 129 (75.43%), while 42 (24.56%) were male. Interestingly, a higher prevalence of vitamin D deficiency was observed among younger patients, with the age group of 30-39 years displaying the highest rate of vitamin D deficiency. Regarding lipid profiles, dyslipidemia was significantly more common in female patients (50.87%) compared to their male counterparts (5.26%). However, it's noteworthy that male patients exhibited higher triglyceride levels (167.21 ± 89.64 mg/dl) than females. Conclusion: This study reveals a strong relationship between vitamin D deficiency and lipid parameters

Extracellular acidification induces ROS- and mPTP-mediated death in HEK293 cells

The extracellular pH (pHe) is a key determinant of the cellular (micro)environment and needs to be maintained within strict boundaries to allow normal cell function. Here we used HEK293 cells to study the effects of pHe acidification (24 h), induced by mitochondrial inhibitors (rotenone, antimycin A) and/or extracellular HCl addition. Lowering pHe from 7.2 to 5.8 reduced cell viability by 70% and was paralleled by a decrease in cytosolic pH (pHc), hyperpolarization of the mitochondrial membrane potential (Δψ), increased levels of hydroethidine-oxidizing ROS and stimulation of protein carbonylation. Co-treatment with the antioxidant α-tocopherol, the mitochondrial permeability transition pore (mPTP) desensitizer cyclosporin A and Necrostatin-1, a combined inhibitor of Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) and Indoleamine 2,3-dioxygenase (IDO), prevented acidification-induced cell death. In contrast, the caspase inhibitor zVAD.fmk and the ferroptosis inhibitor Ferrostatin-1 were ineffective. We conclude that extracellular acidification induces necroptotic cell death in HEK293 cells and that the latter involves intracellular acidification, mitochondrial functional impairment, increased ROS levels, mPTP opening and protein carbonylation. These findings suggest that acidosis of the extracellular environment (as observed in mitochondrial disorders, ischemia, acute inflammation and cancer) can induce cell death via a ROS- and mPTP opening-mediated pathogenic mechanism

Mitochondrial complex I inhibition triggers a mitophagy-dependent ROS increase leading to necroptosis and ferroptosis in melanoma cells

Inhibition of complex I (CI) of the mitochondrial respiratory chain by BAY 87-2243 (‘BAY’) triggers death of BRAFV600E melanoma cell lines and inhibits in vivo tumor growth. Here we studied the mechanism by which this inhibition induces melanoma cell death. BAY treatment depolarized the mitochondrial membrane potential (Δψ), increased cellular ROS levels, stimulated lipid peroxidation and reduced glutathione levels. These effects were paralleled by increased opening of the mitochondrial permeability transition pore (mPTP) and stimulation of autophagosome formation and mitophagy. BAY-induced cell death was not due to glucose shortage and inhibited by the antioxidant α-tocopherol and the mPTP inhibitor cyclosporin A. Tumor necrosis factor receptor-associated protein 1 (TRAP1) overexpression in BAY-treated cells lowered ROS levels and inhibited mPTP opening and cell death, whereas the latter was potentiated by TRAP1 knockdown. Knockdown of autophagy-related 5 (ATG5) inhibited the BAY-stimulated autophagosome formation, cellular ROS increase and cell death. Knockdown of phosphatase and tensin homolog-induced putative kinase 1 (PINK1) inhibited the BAY-induced Δψ depolarization, mitophagy stimulation, ROS increase and cell death. Dynamin-related protein 1 (Drp1) knockdown induced mitochondrial filamentation and inhibited BAY-induced cell death. The latter was insensitive to the pancaspase inhibitor z-VAD-FMK, but reduced by necroptosis inhibitors (necrostatin-1, necrostatin-1s)) and knockdown of key necroptosis proteins (receptor-interacting serine/threonine-protein kinase 1 (RIPK1) and mixed lineage kinase domain-like (MLKL)). BAY-induced cell death was also reduced by the ferroptosis inhibitor ferrostatin-1 and overexpression of the ferroptosis-inhibiting protein glutathione peroxidase 4 (GPX4). This overexpression also inhibited the BAY-induced ROS increase and lipid peroxidation. Conversely, GPX4 knockdown potentiated BAY-induced cell death. We propose a chain of events in which: (i) CI inhibition induces mPTP opening and Δψ depolarization, that (ii) stimulate autophagosome formation, mitophagy and an associated ROS increase, leading to (iii) activation of combined necroptotic/ferroptotic cell death