Loss of myoepithelial calponin-1 characterizes high-risk ductal carcinoma in situ cases,which are further stratified by T cell composition

A hallmark of ductal carcinoma in situ (DCIS) progression is a loss of the surrounding ductal myoepithelium. However, whether compromise in myoepithelial differentiation, rather than overt cellular loss, can be used to predict the risk of DCIS progression is unknown. Here we address this question utilizing pure and mixed DCIS cases (N = 30) as surrogates for DCIS at low and high risk for progression, respectively. We used multiplex immunohistochemical staining to evaluate the relationship between myoepithelial cell differentiation and lymphoid immune cell types associated with poor prognostic DCIS. Our results show that myoepithelial calponin-1 discriminates between pure and mixed DCIS lesions better than histological subtype, presence of necrosis, or nuclear grade. Additionally, focal loss of myoepithelial cells associated with increased PD-1+CD8+ T cells, which suggests a link between the myoepithelium and immune surveillance. To identify associations between calponin-1 expression and immune response, we performed unsupervised hierarchical clustering of myoepithelial and immune cell biomarkers on 219 DCIS lesions from 30 cases. Notably, the majority of pure (low-risk) DCIS lesions clustered in a high calponin-1, T cell low group, whereas the majority of mixed (high-risk) DCIS lesions clustered in a low calponin-1, T cell high group, specifically with CD8+ and PD-1+CD8+ T cells. However, a subset of pure DCIS lesions had a similar calponin-1 and immune signature as the majority of mixed DCIS lesions, which have low calponin-1 and T cell enrichment—raising the possibility that these pure DCIS lesions might be at a high risk for progression.     

Prevalence and Associations of Hypertension in Sri Lankan Adults: Estimates from the SLHAS 2018–19 Survey Using JNC7 and ACC/AHA 2017 Guidelines

Background:Sri Lanka lacks robust estimates of hypertension (HTN) prevalence owing to few national studies, hindering optimization of control strategies. Evidence on how the revised 2017 American College of Cardiology/American Heart Association (ACC/AHA) HTN definition affects prevalence in low- and middle-income countries (LMICs) is also limited.Objectives:To make robust estimates of HTN prevalence in the Sri Lankan adult population, and to assess impact of the ACC/AHA 2017 definitions.Methods:Data were sourced from the 2018–2019 first wave of the Sri Lanka Health and Ageing Study (SLHAS), a nationally representative longitudinal study of the noninstitutionalized adult population. After excluding those with missing data and aged <18 years, 6,342 participants (95.1%) were included in the analysis. HTN was defined using either the traditional threshold of systolic BP (SBP) ≥140 mmHg or a diastolic BP (DBP) ≥90 mmHg, or the ACC/AHA 2017 threshold of SBP ≥130 mmHg or DBP ≥80 mmHg, or if taking antihypertensive medication.Results:Estimated prevalence of HTN in all Sri Lankan adults was 28.2% using the traditional definition, and it doubled to 51.3% when applying the ACC/AHA 2017 definition. Of those classified as hypertensive according to the older and ACC/AHA 2017 definitions, 53.4% and 31.2%, respectively, were previously diagnosed. Of the 23.2% of adults reclassified as hypertensive by the ACC/AHA 2017 definition, 16.6% had a history of CVD or diabetes. Increased prevalence was associated with urban residence, socioeconomic status, obesity, and Muslim ethnicity. Prevalence increased with age, but the increase was steeper in women from their 30s.Conclusions:Nearly one in three adult Sri Lankans are hypertensive, requiring antihypertensive treatment. Applying the ACC/AHA 2017 definitions almost doubles numbers, but many of those reclassified would require treatment under recent WHO guidelines. Study findings also suggest that design effects in HTN surveys may be higher than usually assumed.     

Knowledge,attitude, and practice toward mucormycosis among patients presenting to six tertiary eye care hospitals in South India – A multicentric online questionnaire-based survey

Purpose:To study the awareness on mucormycosis among outpatients who visited six tertiary eye care hospitals at Madurai, Pondicherry, Coimbatore, Tirunelveli, Chennai, and Tirupati.Methods:This was a telephone-based survey conducted using questionnaires consisting of 38 questions in five sections from July 5 to 25, 2021. Patients visiting the eye hospitals for an examination were contacted over their phones and responses were directly entered onto the Google forms platform.Results:A total of 4573 participants were included in the study. Among all participants, a cumulative 83% of participants had some knowledge of mucormycosis. More than 80% of them reported that their prime source of information was through mass communication like television or radio. Around 34.8% of the respondents were aware that it can occur after treatment for coronavirus disease 2019 (COVID-19) infection, only half of them (54.3%) knew that systemic steroids were the main risk factor. The knowledge scores were higher for participants who were diabetics (n = 1235) or had been affected by COVID-19 earlier (n = 456) or whose friends had mucormycosis earlier (n = 312). Knowledge, attitude, and practice (KAP) scores of nonprofessional health-care workers (n = 103) were much better compared to patients.Conclusion:Such KAP studies give us an idea of the impact of the measures taken for educating the public. In this study, a cumulative 83% of participants had some knowledge of mucormycosis and 86% knew that this was an emergency. More than 50% of the participants were not aware that diabetes is a risk factor for mucormycosis.     

Respiratory symptoms in patients with inflammatory bowel disease and the impact of dietary salicylates

BACKGROUND: Respiratory symptoms are over-represented in inflammatory bowel disease. There are similarities between the epidemiology of inflammatory bowel disease and that of respiratory conditions for which an adverse influence of salicylate has been identified. Natural salicylates exist within our diet. AIMS: To determine whether a lower intake of dietary salicylates is associated with less active inflammatory bowel disease and fewer concurrent respiratory symptoms. PATIENTS AND METHODS: Respiratory status, inflammatory bowel disease activity, quality of life, and dietary habits were established in 73 patients with Crohn's disease and 69 with ulcerative colitis, using a self-administered questionnaire and peak expiratory flow rate readings. Harvey-Bradshaw and Simple Birmingham/Royal Free Colitis indices, an internally validated respiratory score, and estimated weekly dietary salicylate intake, were calculated for each patient. RESULTS: There was at least one respiratory symptom in 63.4% of patients. The commonest underlying respiratory diagnosis was asthma. Respiratory impairment was similar in ulcerative colitis and Crohn's disease; 56.3% of Crohn's disease patients with an active respiratory diagnosis had other extra-intestinal manifestations. The dietary salicylate intake was independent of respiratory status, but inversely correlated with ulcerative colitis activity (dietary salicylate intake 37.0mg versus 21.4mg for low and higher Simple Birmingham/Royal Free Colitis index, respectively; p<0.02). A similar association was not seen in Crohn's disease. CONCLUSIONS: Respiratory impairment is common in inflammatory bowel disease. Higher intake of dietary salicylates is associated with less active colitis and possibly causally so.     

EPCAM mutation update: Variants associated with congenital tufting enteropathy and Lynch syndrome

The epithelial cell adhesion molecule gene (EPCAM, previously known as TACSTD1 or TROP1) encodes a membrane‐bound protein that is localized to the basolateral membrane of epithelial cells and is overexpressed in some tumors. Biallelic mutations in EPCAM cause congenital tufting enteropathy (CTE), which is a rare chronic diarrheal disorder presenting in infancy. Monoallelic deletions of the 3′ end of EPCAM that silence the downstream gene, MSH2, cause a form of Lynch syndrome, which is a cancer predisposition syndrome associated with loss of DNA mismatch repair. Here, we report 13 novel EPCAM mutations from 17 CTE patients from two separate centers, review EPCAM mutations associated with CTE and Lynch syndrome, and structurally model pathogenic missense mutations. Statistical analyses indicate that the c.499dupC (previously reported as c.498insC) frameshift mutation was associated with more severe treatment regimens and greater mortality in CTE, whereas the c.556‐14A>G and c.491+1G>A splice site mutations were not correlated with treatments or outcomes significantly different than random simulation. These findings suggest that genotype–phenotype correlations may be useful in contributing to management decisions of CTE patients. Depending on the type and nature of EPCAM mutation, one of two unrelated diseases may occur, CTE or Lynch syndrome.     

Molecular mechanism of oxalate-induced free radical production and glutathione redox imbalance in renal epithelial cells: effect of antioxidants

BACKGROUND: Peroxidation of renal cells is a critical event in the nucleation and formation of calcium oxalate crystals under hyperoxaluric conditions. We previously demonstrated that oxalate-induced peroxidative injury is one of the major mechanisms in promoting crystal attachment to renal epithelial cells. METHODS: In this study we have demonstrated that the mechanism of oxalate-induced peroxidative injury is through the induction of TGF-beta1 and glutathione (GSH) redox imbalance in LLC-PK1 cells. RESULTS: LLC-PK1, renal epithelial cells exposed to oxalate had significantly higher reactive oxygen species (ROS) production; higher TGF-beta1 levels, as measured by ELISA (1.89 +/- 0.035 fold increase) or Western blot (1.65 +/- 0.01 fold increase); increased malondialdehyde formation; increased LDH release, and loss of cell viability. In addition, oxalate exposure significantly decreased GSH content, glutathione reductase, glucose-6-phosphate dehydrogenase activities, and increased oxidized GSH content. Treatment with vitamin E, neutralizing anti-TGF-beta antibody, or diphenylene iodium, an inhibitor of NAD(P)H oxidase, significantly inhibited oxalate-induced ROS production and prevented peroxidative injury and cytolysis. Vitamin E, catalase, or desferoxamine treatment also significantly restored the oxalate-induced cellular GSH redox status toward the control level, and vitamin E treatment significantly attenuated the oxalate-mediated increase in TGF-beta1 protein in cultured LLC-PK1 cells. CONCLUSIONS: This is the first study to demonstrate that the mechanism of oxalate-induced free radical production in renal tubular epithelial cells is through the activation of NAD(P)H oxidase via cytokine TGF-beta1 induction. These results also provide direct evidence that antioxidant therapy might prevent calcium oxalate nucleation and kidney stone formation by preventing oxalate-mediated peroxidative injury and GSH redox imbalance.     

Rapid Detection of Vancomycin-Intermediate Staphylococcus aureus by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

Vancomycin is the standard of care for the treatment of invasive methicillin-resistant Staphylococcus aureus (MRSA) infections. Infections with vancomycin-nonsusceptible MRSA, including vancomycin-intermediate S. aureus (VISA) and heterogeneous VISA (hVISA), are clinically challenging and are associated with poor patient outcomes. The identification of VISA in the clinical laboratory depends on standard susceptibility testing, which takes at least 24 h to complete after isolate subculture, whereas hVISA is not routinely detected in clinical labs. We therefore sought to determine whether VISA and hVISA can be differentiated from vancomycin-susceptible S. aureus (VSSA) using the spectra produced by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS). Strains of MRSA were characterized for vancomycin susceptibility phenotype by broth microdilution and modified population analysis. We tested 21 VISA, 21 hVISA, and 38 VSSA isolates by MALDI-TOF MS. Susceptibility phenotypes were separated by using a support vector machine (SVM) machine learning algorithm. The resulting model was validated by leave-one-out cross validation. Models were developed and validated by using spectral profiles generated under various subculture conditions, as well as with and without hVISA strains. Using SVM, we correctly identified 100% of the VISA and 97% of the VSSA isolates with an overall classification accuracy of 98%. Addition of hVISA to the model resulted in 76% hVISA identification, 100% VISA identification, and 89% VSSA identification, for an overall classification accuracy of 89%. We conclude that VISA/hVISA and VSSA isolates are separable by MALDI-TOF MS with SVM analysis.     

Electrochemical behaviour of certain biomimetic copper(II) complexes in aqueous and aqueous micellar solutions

Voltammetric studies of biomimetic Cu(II) complexes of bis(pyrid-2-yl)dithia (N₂S₂) and -trithia (N₂S₃) and bis(benzimidazol-2-yl)dithia ligands in aqueous and sodium dodecylsulphate (SDS, anionic), cetyltrimethylammonium bromide (CTAB, cationic) and Triton X-100 [α-[p-(1,1,3,3-tetramethylbutyl)phenyl]-w-hydroxypolyoxyethylene(9.5), nonionic] micellar solutions have been carried out on a glassy carbon electrode. The electronic spectral properties of Cu(II) complexes in micellar solutions are consistent with their incorporation into hydrophobic microenvironments. For most of the complexes, the adsorption of Cu(I) species observed in aqueous solution is absent and the reversibility of the Cu(II)/Cu(I) couple is increased in SDS and Triton X-100 micellar solutions. From the dependence of I_pc values on SDS concentration, the micellar binding constant K_M has been estimated. Ligand enlargement and the structure of the Cu(II) complexes and Cu(I) forms of complexes enhance the binding with micelles. Compared with aqueous isotropic solutions, the E_1/2 values in micellar solutions either decrease or increase depending upon the nature of the surfactant and the hydrophobicity and structure of the complexes. The ratio of equilibrium binding constants K₊/K₂₊ of Cu(I) and Cu(II) species does not follow the same trend in SDS, CTAB and Triton X-100 solutions, indicating that the interaction of the cationic complexes depends on a delicate balance between hydrophobic and electrostatic interactions. Interestingly, for complexes with the CuN₂S₂ chromophore the Cu(I)/Cu(0) couple is also observed in Triton X-100 micelles.