PLANT BASED QUORUM SENSING INHIBITORS OF PSEUDOMONAS AERUGINOSA

Phytochemical research has gained a lot of momentum in the medical field for the discovery of new, safe, and effective remedies. In the context of antimicrobial research, various plant sources have been discovered with the potential to disrupt bacterial quorum sensing (QS), which plays a key role in the regulation of virulence in many Gram positive and Gram negative bacteria. Pseudomonas aeruginosa, a Gram negative bacterium, is known to produce multiple QS systems that control the expression of virulence determinants and biofilm development in this pathogen. Hence, the inhibition of QS has been pursued as a promising therapy for treatment of drug resistant Pseudomonas infections. A comprehensive review of the research data available for plant products as QS inhibitors of the organism has been presented here, while further suggesting the future prospects for these inhibitors

ASSESSMENT OF PREVALENCE OF DEPRESSION IN CARDIAC PATIENTS AND ITS ASSOCIATION WITH THE USE OF BETA-BLOCKERS AND STATINS

Objective: The objective of the study is to assess the prevalence of depression among patients with cardiovascular disease and its association with the use of β-blockers and statins.Methods: This is a prospective observational study conducted at a corporate hospital, Hyderabad, Telangana, India, for a period of 6 months. 250 cardiac patients above 16 years are included in the study. The required data are collected from the patients through direct interview using standard questionnaires and also from patients' respective case sheets. The acquired data are evaluated based on the standard questionnaires Patient Health Questionnaire-9 (PHQ-9) and Beck Depression Inventory-II (BDI-II) scales; used to diagnose the severity of depression in cardiac patients.Results: Prevalence of minor to major depressive symptoms according to BDI-II was found to be 17.2%. Prevalence of minor to major depressive symptoms according to PHQ-9 was found to be 19.2%. Among male patients, 13% showed depressive symptoms, whereas among female patients 25% showed depressive symptoms. Among the patients coadministering beta-blockers and statins, 15% were depressed according to BDI-II, and 16% were depressed according to PHQ-9 at visit. After 1 month (first follow-up), the percentage increased by 8% (for BDI-II)-12% (for PHQ-9) and remains almost the same at the second follow-up. As per BDI-II and PHQ-9 scores, the percentage of patients with minor to major depression among the patients using only beta-blockers decreased significantly from the time of visit to the second follow-up. The percentage of patients with minor to major depression among the patients using only statins increased significantly from the time of visit to second follow-up.Conclusion: Prevalence of minor to major depression according to BDI-II was found to be 17%, whereas according to PHQ-9, it was found to be 20% in patients with cardiovascular disease. Cardiovascular diseases have been more prevalent in men than in women, whereas depressive symptoms have been more prevalent in women than in men. Patients using only β-blockers showed a decrease in symptoms of depression. Whereas statins have shown to increase the chances of depression slightly which is often negligible, atorvastatin was associated with a higher level of depression when compared to rosuvastatin. Controversies still exist that statins decrease risk of depression.Â

A retrospective interventional study for evaluation of efficacy and safety of sildenafil citrate in improving intrauterine growth restriction and oligohydramnios using ultrasound Doppler velocimetry

Background: Intrauterine growth restriction (IUGR) is one of the major reasons for neonatal morbidity and mortality. Oligohydramnios is a common finding in IUGR. In majority of these cases diminished utero-placental blood flow is observed. However, in spite of this understanding and identification of high-risk patients, the management options are limited. Sildenafil citrate, a phosphodiesterase type-5 inhibitor improves utero-placental perfusion.Methods: We present a retrospective interventional study involving 50 adult pregnant women diagnosed with early-onset IUGR (n=38) and oligohydramnios (n=12). Vaginal sildenafil citrate 25 mg t.i.d. was started from the day of diagnosis till delivery. Primary efficacy endpoints included changes in Doppler parameters i.e., amniotic fluid index (AFI), uterine artery (UA)- pulsatility index (PI), resistance index (RI) and systolic diastolic ratio (S/D ratio). Secondary endpoints included live birth, birth weight, Apgar score at birth, neonatal survival to hospital discharge and adverse maternal side effects.Results: There was a statistically significant improvement in UA-PI, RI and S/D ratios (p<0.0001) in all cases. In oligohydramnios cases, treatment showed a statistical significant increase in AFI score (2.86±1.33 cm). The mean birth weight on delivery was 2200 gm with good Apgar scores. No major adverse effects were reported by women using sildenafil citrate vaginally.Conclusions: Sildenafil citrate, by increasing utero-placental perfusion, improves uterine artery Doppler patterns, AFI, fetal weight and overall better neonatal survival rates by reducing neonatal morbidity and mortality. Sildenafil citrate may hold a promising treatment strategy for management of IUGR and oligohydramnios

The Protein Extract of Chlorella minutissima Inhibits The Expression of MMP-1, MMP-2 and MMP-9 in Cancer Cells through Upregulation of TIMP-3 and Down Regulation of c-Jun

Objective Considering the bioactivities exhibited by microalgae, the effect of protein extract of Chlorella minutissimma (CP extract) was investigated on the expression of human matrix metalloproteinases-1 (MMP-1) in the breast cancer cell line MDA-MB231, and that of MMP-2 and -9 in hepatocellular cancer cell line HepG2 at different expression levels. The study aimed identification and analysis of inhibitory activity of microalgal components extracted from Chlorella minutissima against human MMPs. Materials and Methods In this experimental study, we analysed the effect of Chlorella extracts on MMP-1, -2, and -9 expression at various levels. Gelatin zymography was performed to study the inhibitory effect of Chlorella exracts on human gelatinases at the activity level, followed by western blotting to analyse the expression of all three MMPs at the protein level. The similar effect at the mRNA level along with the probable mechanism underlying inhibition of MMPs was assessed using real-time polymerase chain reaction (PCR). Results The results reveal that the treatment with CP extract decreased the mRNA expression of MMP-1, MMP-2, and MMP-9 by 0.26-, 0.29-, and 0.40-fold, respectively, at 20 μg/ml concentration as well as inhibited the activity of MMP-2 and MMP-9 by 37.56 and 42.64%, respectively, at 15 μg/ml concentration. Additionally, upregulated mRNA expression of tissue inhibitor of metalloproteinases-3 (TIMP-3) by 1.68-fold was seen in HepG2 cells at 20 μg/ml concentration treatment group. However, CP extract did not induce any change in the mRNA expression of the TIMP-1, -2 and -4 in HepG2 and TIMP-1, -2, -3 and -4 in MDA-MB231 cells. Activator protein-1 (AP-1)-dependent c-Jun-mediated transcriptional regulation of MMP-1, -2, and -9 was also studied to elucidate the appropriate mechanism involved in the inhibition of MMPs. Conclusion The CP extract successfully inhibited MMP-1, -2, and -9 at different expression levels through TIMP-3 upregulation and c-Jun downregulation

Purification and characterisation of alliinase produced by Cupriavidus necator and its application for generation of cytotoxic agent: Allicin

Allicin, an extremely active constituent of freshly crushed garlic, is produced upon reaction of alliin with the enzyme alliinase (EC 4.4.1.4). A bacterium Cupriavidus necator with the ability of alliinase production was isolated from a soil sample and was identified by morphological, biochemical and 16S rRNA sequence. Alliinase production was optimised and it was further purified to apparent homogeneity with 103-fold purification and specific activity of 209 U/mg of protein by using DEAE Cellulose and Sephadex G-100 chromatography. The enzyme is a homodimer of molecular weight 110 kDa with two subunits of molecular weight 55 kDa each. The optimum activity of the purified enzyme was found at pH 7 and the optimum temperature was 35 °C. The enzyme exhibited maximum reaction rate (Vmax) at 74.65 U/mg and Michaelis–Menten constant (Km) was determined to be 0.83 mM when alliin was used as a substrate. The cytotoxic activity of in-situ generated allicin using purified alliinase and alliin was assessed on MIA PaCa-2 cell line using MTT assay and Acridine orange–ethidium bromide staining. This approach of in-situ allicin generation suggests a novel therapeutic strategy wherein alliin and alliinase work together synergistically to produce cytotoxic agent allicin. Keywords: Alliinase, Characterisation, Cupriavidus necator, Purification, MIA PaCa-2 cell

Attenuation of quorum sensing-regulated behaviour by Tinospora cordifolia extract & identification of its active constituents

Background & objectives: The pathogenicity of the nosocomial pathogens, Pseudomonas aeruginosa and Acinetobacter baumannii is regulated by their quorum sensing (QS) systems. The objective of the present study was to examine the effect of the cold ethyl acetate extract of Tinospora cordifolia stem on virulence and biofilm development in the wild type and clinical strains of P. aeruginosa and A. baumannii. The study was further aimed to identify the probable active constituents in the plant extract. Methods: P. aeruginosa virulence factors viz., LasA protease, LasB elastase and pyocyanin production were analyzed spectrophotometrically. Biofilm formation was studied using crystal violet staining-microtitre plate assay. The plant extract was fractionated using silica gel column chromatography and the most active fraction was derivatized using silylation and analyzed by gas chromatography-mass spectrometry (GC-MS). In silico testing of the molecules identified in GC-MS was performed, for binding to the P. aeruginosa LasI and LasR proteins, to predict the QS inhibitory molecules. Results: The plant extract inhibited three major virulence factors in P. aeruginosa; it exhibited enhanced biofilm formation in P. aeruginosa while decreased biofilm development in A. baumannii. The most active fraction obtained from column chromatography, exhibited suppression of virulence as well as biofilm in both the organisms. Docking scores were calculated for all the molecules identified in GC-MS, and high docking scores were obtained for 2,3,4-triacetyloxybutyl acetate, methyl 16-methyl heptadecanoate, 2-(5-ethenyl-5-methyloxolan-2-yl)propan-2-ol, methyl hexadecanoate and 2-methoxy-4-vinyl phenol. Interpretation & conclusions: The compounds showing high docking scores could probably be the QS inhibitors. These molecules can be screened further for the development of new anti-infective drugs

Impact of acute kidney injury in patients with acute decompensated heart failure: Cardiorenal syndrome

Cardiorenal syndrome (CRS) is a complex interdependent relationship between the heart and kidneys, prevalent in hospitalized patients with acute decompensated heart failure (ADHF). The main aim of this study is to evaluation of cardiac and renal function, treatment factors, and outcomes in view of mortality and persistent renal dysfunction in acute decompensated heart failure (cardio renal syndrome type 1) patients. We studied 100 patients hospitalised with ADHF and acute kidney injury (AKI). Patients were evaluated clinically, biochemically, ultrasonographically, and echocardiographically to assess demographics, etiologic and risk factors, cardiac and renal function, and outcomes in view of mortality and persistent renal dysfunction. The study monitored the patients until discharge and follow up with three months to one year. Record information about functional improvement, worsening symptoms, and mortality. The majority of the patients were males (72%), with dyspnea being the most common symptom (92%) followed by decreased urinary output (82%). The mean age of the patients was 62.60 years. Low level of Mean arterial pressure (MAP) 18.97 (95% CI 4.59 to 78.37, P 0.0001), estimated glomerular filtration rate (eGFR) 0.92(95% CI 0.87 to 0.99; P 0.02), maximum creatinine 3.08 (95% CI 1.67 to 5.67, P 0.0001), maximum level of urea 1.02(95% CI, P 0.001), lower Left ventricular ejection fraction (LVEF) 1.05 (95% CI 0.15 to 0.84, P 0.04) were independently predictors of in-hospital mortality. CRS-1 is associated with increased risk of mortality (25%), residual renal dysfunction (16%) at one year follow up. Persistent renal dysfunction, renal replacement therapy possibly improves for the treating persistent renal dysfunction, and recurrent HHF (more than 2 admissions) post hospitalisation index within twelve months were predictors of mortality (25%) at one-year

Epidemiological and clinical characteristics of pulmonary arterial hypertension in Indian patients: A hospital-based observational study

Context: Pulmonary hypertension (PH) is a condition characterized by an increase in pulmonary artery pressure, leading to morbidity and poor survival. In India, the high prevalence of certain diseases contributes to the burden of PH. To address this, this study provides epidemiological data and clinical characteristics of pulmonary artery hypertension. Materials and Methods: A cross-sectional study was conducted at a tertiary cardiac care hospital, recruiting 964 patients with severe pulmonary arterial hypertension (PAH). Patient data were obtained through a detailed history and physical examination, including 12-lead electrocardiogram, two-dimensional echocardiography, coronary angiography, right heart catheterization, and hematological and serological investigations. Results: The majority of patients (28.94%) were in the age group of 31–40 years, with a female preponderance in PAH disease. Idiopathic PAH (IPAH) was the most common type, accounting for 69.78% of all cases. The most common comorbid conditions were hypertension (48.55%) and Type-2 diabetes mellitus (44.81%). Dyspnea on exertion (98.34%) and edema of extremities (91.18%) were the most common symptoms. The mean survival rate was 50.98 months, with Type 1 PAH having the worst survival rate of 47.56 months. The statistical analysis showed a significant association between smoking and PAH. Conclusion: This study provides epidemiological data and clinical characteristics of PAH patients in India, emphasizing the need for early diagnosis, prompt management, and smoking cessation programs. The high prevalence of IPAH and female preponderance was also observed in this study. These findings can help in the development of targeted interventions and management strategies for PH patients in India