Chronic High-Salt Diet Activates Tumor-Initiating Stem Cells Leading to Breast Cancer Proliferation.

Several chronic inflammatory diseases have been linked to high-salt (HS) diets. Chronic inflammation is an established causative hallmark of cancer. However, a direct role of HS diets in tumorigenesis is yet to be defined. Previous orthotopic murine breast tumor studies have shown that short-term HS diets caused inhibition of tumor growth through the activation of cytotoxic adaptive immune responses. However, there have been experimental challenges in developing a viable chronic HS-diet-based murine tumor model. To address this, we have developed a novel chronic HS diet tumor model through the sequential passaging of tumor cells in mice under HS dietary conditions. Two orthotopic murine triple-negative breast cancer models, 4T1 tumor cells injected into BALB/c mice and Py230 tumor cells injected into C57Bl/6 mice, were utilized in our study. For the HS diet cohort, prior to orthotopic injection with tumor cells, the mice were kept on a 4% NaCl diet for 2 weeks. For the regular salt (RS) diet cohort, the mice were kept on a 1% NaCl diet. Following syngeneic cancer cell injection, tumors were allowed to grow for 28 days, following which they were collected to isolate immune cell-depleted cancer cells (passage 1, P1). The tumor cells from P1 were reinjected into the next set of non-tumor-bearing mice. This procedure was repeated for three cycles (P2-P4). In P1, compared to the RS diet cohort, we observed reduced tumor kinetics in both murine tumor models on the HS diet. In contrast, by P4, there was significantly higher tumor progression in the HS diet cohort over the RS diet cohort. Flow cytometry analysis demonstrated an 8-fold increase in tumor-initiating stem cells (TISCs) from P1 to P4 of the HS diet cohort, while there were no significant change in TISC frequency with sequential passaging in the RS diet cohort. Molecular studies showed enhanced expression of TGFßR2 and CD80 on TISCs isolated from the P4 HS diet cohort. In vitro studies demonstrated that TGFß stimulation of these TISCs increased the cellular expression of CD80 molecules. Further, the chronic HS diet selectively induced the glycolytic metabolic phenotype over the mitochondrial oxidative phosphorylation phenotype in TISCs, which is needed for the production of metabolites during tumor cell differentiation and proliferation. The infiltrating CD8 and CD4 T-lymphocytes in P4 tumors demonstrated increased expression of the immune checkpoint inhibitor (ICI) CTLA4, a known binding partner of CD80, to cause immune exhaustion and pro-tumorigenic effects. Interestingly, anti-TGFß monoclonal antibodies (mAbs) played a synergistic role in further enhancing the anti-tumor effect of anti-CTLA4 mAb. In summary, our findings demonstrated that chronic HS diet increased the frequency of TISCs in tumors leading to blunting of cytotoxic adaptive immune responses causing tumor proliferation. Furthermore, a combination of anti-TGFß with current ICI-based immunotherapies could exert more favorable anti-cancer clinical outcomes.

Genomic Features of Homologous Recombination Deficiency in Breast Cancer: Impact on Testing and Immunotherapy.

Genomic instability is one of the well-established hallmarks of cancer. The homologous recombination repair (HRR) pathway plays a critical role in correcting the double-stranded breaks (DSB) due to DNA damage in human cells. Traditionally, the BRCA1/2 genes in the HRR pathway have been tested for their association with breast cancer. However, defects in the HRR pathway (HRD, also termed 'BRCAness'), which has up to 50 genes, have been shown to be involved in tumorigenesis and treatment susceptibility to poly-ADP ribose polymerase inhibitors (PARPis), platinum-based chemotherapy, and immune checkpoint inhibitors (ICIs). A reliable consensus on HRD scores is yet to be established. Emerging evidence suggests that only a subset of breast cancer patients benefit from ICI-based immunotherapy. Currently, albeit with limitations, the expression of programmed death-ligand 1 (PDL1) and tumor mutational burden (TMB) are utilized as biomarkers to predict the favorable outcomes of ICI therapy in breast cancer patients. Preclinical studies demonstrate an interplay between the HRR pathway and PDL1 expression. In this review, we outline the current understanding of the role of HRD in genomic instability leading to breast tumorigenesis and delineate outcomes from various clinical trials. Furthermore, we discuss potential strategies for combining HRD-targeted therapy with immunotherapy to achieve the best healthcare outcomes in breast cancer patients.

Efficacy of Autoinoculation in Treatment of Multiple Viral Warts - A Single Arm Study.

OBJECTIVE: To test the effectiveness of autoinoculation of a wart in patients with multiple viral warts. STUDY DESIGN: An experimental study. PLACE AND DURATION OF STUDY: Department of Dermatology, Pak Emirates Military Hospital (PEMH), Rawalpindi, from February 2021 to February 2022. METHODOLOGY: Patients above 12 years of age, having more than 5 warts were included. Those getting any other treatment for warts were excluded. The procedure was performed on a total of 70 patients with warts on 3 sites i.e. face/neck, palmoplantar and multiple sites. The outcome was categorised into cleared (no wart), partially cleared (reduced number of warts than baseline), and no response (same or increased number of warts than baseline). RESULTS: Out of 70 patients, 54 (77.1%) patients showed complete response, 6 (8.6%) showed partial response and 10 (14.3%) showed no response at all. Out of 54 patients who showed complete response, 36 (66.6%) were cleared after only one inoculation, fifteen (27.7%) were cleared after two inoculations, and 3 (5.5%) were cleared after three inoculations. The results were statistically significant with p-value <0.05. CONCLUSION: Autoinoculation is a simple and minimally invasive procedure which proved to be very effective against all types of warts, with very promising results against palmoplantar warts. KEY WORDS: Viral warts, Multiple viral warts, Autoinoculation, Delayed hypersensitivity, Human papilloma virus (HPV), Palmoplantar warts, Immunity.

Analysis of aerosol liquid water content and its role in visibility reduction in Delhi.

Aerosols undergo significant changes due to water uptake under high RH conditions, leading to changes in physical, optical, and chemical properties. Detailed assessment and investigation are needed to understand better aerosol liquid water content (ALWC) characteristics in highly polluted regions like Delhi. Therefore, in this study, we examined the mass concentration and the factors governing the ALWC associated with PM2.5 in Delhi for two winters (Dec 2019 to Jan 2020 and Dec 2020 to Feb 2021) using the real-time measurements of NR-PM2.5 from Aerodyne aerosol chemical speciation monitor (ACSM) and the application of thermodynamic modeling (ISORROPIA II). The average NR-PM2.5 mass concentration in the 2020-2021 winter was 152 µg/m3, about 50 % higher than the average mass concentration of 102 µg/m3 in 2019-2020. Consequently, the ALWC was also 60 % higher during 2020-2021, with an average mass concentration of 150 µg/m3. ALWC increased exponentially with RH and is significant when RH > 80 %. Further, all the inorganic components of NR-PM2.5 were found to contribute significantly to ALWC uptake; however, the relative contribution varied in different RH conditions. Ammonium sulphate dominated the ALWC uptake among the inorganic components at low RH, but ammonium nitrate was the dominant contributor at high RH. The decreased chloride mass fraction in inorganics in the recent winters reduced its relative contribution to ALWC. High ALWC mass concentration during high PM2.5 and high RH leads to a significant reduction in visibility. We further validated this visibility reduction by estimating the enhanced light scattering coefficient (f(RH)) and found that the hygroscopic growth is responsible for the enhanced visibility reduction during high RH conditions (> 85 %) when light scattering efficiency increased by a factor of >3.5. Sensitivity tests of f(RH) on mass concentration of inorganic salts showed that all the salts contributed almost equally. As revealed in our study, variations in PM2.5 mass concentration and composition despite similar meteorological conditions between different winters indicate changing regional aerosol emissions. Therefore, long-term observations of ALWC and PM2.5 chemical composition are required to arrive at actionable measures and mitigation strategies. Further, the focus should be on reducing the overall inorganic mass concentrations of PM2.5 in general, decreasing the absolute ALWC, and improving visibility.

Association of hypothyroidism with acute COVID-19: a systematic review.

OBJECTIVE: There is an increasing body of literature on the impact of COVID-19 on the pituitary-thyroid axis. Therefore, we conducted a systematic review to assess the prevalence of hypothyroidism in patients with COVID-19. METHODS: A literature review was conducted using LitCOVID for study selection in PubMed and MEDLINE till May 2021. All relevant original articles evaluating thyroid dysfunction were included and information regarding the prevalence of hypothyroid disease in COVID-19 was retrieved from the eligible articles. RESULTS: Out of 32 articles, six articles qualified for the final analysis which included 1160 patients. There was significant heterogeneity among the included articles. Most of the patients had lower mean triiodothyronine (T3) and normal or low thyroid-stimulating hormone (TSH). Increased TSH ranged from 5.1% to 8% while low T3 was present in up to 28% of the patients. In these studies, the prevalence of altered thyroid hormones was significantly more in COVID-19 patients as compared to control groups. A positive correlation between low mean T3 and clinical severity of COVID-19 was reported. CONCLUSION: This systematic review reveals a significant proportion of hypothyroidism associated with COVID-19. Therefore, routine assessment of thyroid function is warranted in hospitalized COVID-19 patients.

Post-acute COVID-19 syndrome and its prolonged effects: An updated systematic review

ObjectiveThis systematic review aimed at estimating the demographics, clinical characteristics, and prevalence of post-acute COVID-19 symptoms in view of published literature that studied prolonged clinical manifestations after recovery from acute COVID-19 infection. MethodsAfter protocol setting, relevant articles were searched on various databases including PubMed, Medline, the Cumulative Index to Nursing and Allied Health Literature (CINAHL), Embase, and Web of Sciences using MeSH keywords. The data regarding demographic, clinical characteristics, and prevalence of each persisting symptom were carefully studied and tabulated. Data analysis was carried out using Statistical Package for Social Sciences (SPSS) version 26. ResultsOut of the 153 articles reviewed, 21 articles qualified for the final analysis. The most common persistent clinical manifestations were fatigue (54.11%), dyspnea (24.38%), alopecia (23.21%), hyperhidrosis (23.6%), insomnia (25.98%), anxiety (17.29%), and arthralgia (16.35%). In addition to these symptoms, new onset hypertension, diabetes, neuropsychiatric disorders, and bladder incontinence were also reported. ConclusionClinical features of post-acute COVID-19 infection can manifest even after 60 days of initial infection. Constitutional symptoms include fatigue, dyspnea, anosmia, and insomnia were the most commonly reported in literature. Multidisciplinary care along with regular follow-up must be a provided to such patients. Curation and modification of guidelines is required for the assessment of discharged hospitalized patients for better management of their post-acute COVID-19 syndrome.

Pharmacy services during COVID-19 pandemic: experience from a tertiary care teaching hospital in Pakistan.

The coronavirus disease 19 (COVID-19) is rapidly spreading across the world. Pharmacy services play a vital role in public health in preventing and containing the COVID-19 pandemic. All over the world, especially in the developed countries pharmacists have responded smartly and speedily for public health, such as establishing professional protective and service guidance for pharmacy staff and services, creating and updating drug formularies, addressing the issues of drug shortages, providing public education for prevention and management of infection, contributing in drug evaluation and clinical trials. In this commentary, we review the exclusive demands from pharmacy services in Pakistan during coronavirus disease 2019 pandemic and sharing the responses of our hospital pharmacy to these demands and needs with the international pharmacy community, especially of the low and middle-income countries like Pakistan.

Neonatal fluid and electrolytes profile effect on amphotericin B associated nephrotoxicity in neonatal tertiary care unit of Karachi-Pakistan.

OBJECTIVES: Amphotericin-B (d-AmB) has a broader anti-fungal spectrum and is used for neonatal invasive-fungal-infections especially invasive-candidiasis (IC). To prevent d-AmB-induced nephrotoxicity, renal protective effect of fluid and electrolyte management has been established among adults; in this study, the authors determined this effect among neonates. METHODS: In this retrospective cohort study, the authors reviewed neonatal medical records, admitted to neonatal intensive care unit and received d-AmB therapy. Patients were divided into, renal-insufficiency-group (RIG) and the non-renal-insufficiency-group (NIG). RESULTS: A total of 90 cases were analyzed, 41 composed RIG and 49 NIG. Renal insufficiency (RI) was developed on 1.7 (0.84) and 7.8 (1.21) days of d-AmB therapy in 26 (63%) and 15 (37%) cases respectively. Bivariate and multivariate analysis demonstrate that >4 m Eq/kg/d sodium intake across all-time points was significantly (p < 0.0001) associated with reduced risk of RI [(phase-I: AOR: 0.96; 95% CI: 0.91-0.99), (phase-II: AOR: 0.84; 95% CI: 0.68-0.92) and (phase-III: AOR: 0.90; 95% CI: 0.86-0.95)]. While adequate fluid intake reduced the likelihood of RI if started before and initial 2 days of d-AmB therapy. CONCLUSIONS: Adequate hydration before and 48 hours after d-AmB therapy and >4 mEq/kg/day sodium intake before and through d-AmB therapy may protect neonatal RI.

Efficacy of colistin in multidrug-resistant neonatal sepsis: experience from a tertiary care center in Karachi, Pakistan.

OBJECTIVE: Infections with multidrug-resistant organisms (MDROs) such as Gram-negative bacteria have high morbidity and mortality with limited treatment options. Colistin, an antibiotic active against MDRO, was rarely used due to frequent adverse effects, but its use has now been recommended among adults. In this study, we determined the efficacy of colistin for the treatment of sepsis in neonates. DESIGN/SETTING/PATIENTS/OUTCOMES: We conducted a retrospective record review of all neonates admitted to the neonatal intensive care unit of Aga Khan University Hospital, Karachi, Pakistan, between June 2015 and June 2018, who had sepsis and received colistin by intravenous, inhalation and/or intrathecal routes. Predictors of colistin efficacy, for neonatal survival and microbial clearance, were assessed using multiple logistic regression. RESULTS: 153 neonates received colistin; 120 had culture-proven sepsis; and 93 had MDR-GNB (84 colistin-sensitive). 111 (72.5%) neonates survived and were discharged from hospital; 82.6% had microbial clearance. Neonates with colistin-sensitive bacteria (adjusted OR (AOR)=3.2, 95% CI 2.8 to 4.0), and those in which colistin therapy started early (AOR=7.2, 95% CI 3.5 to 13.6) were more likely to survive. Neonates with increased gestational age (AOR=1.9, 95% CI 1.5 to 3.0), higher weight (AOR=5.4, 95% CI 3.3 to 11.8) and later onset of sepsis (AOR=4.3, 95% CI 2.0 to 9.0) had higher survival. Adverse events included nephrotoxicity in 5.2%; 13.7% developed seizures and 18.3% had electrolyte imbalance. CONCLUSIONS: Colistin therapy was associated with survival among neonates suffering from MDR-GNB sepsis. The frequency of side effects was moderate.