Exploring the Therapeutic Efficacy of Zingerone Nanoparticles in Treating Biofilm-Associated Pyelonephritis Caused by Pseudomonas aeruginosa in the Murine Model.

Biofilms of Pseudomonas aeruginosa can cause complicated urinary tract infections especially in people with indwelling catheters which may result in pyelonephritis. Microorganisms in biofilm demonstrate high resistance to both antibiotics and host protection mechanisms, often resulting in chronic and difficult-to-treat infections. This study is aimed to assess in vivo and ex vivo efficacy of Zingerone nanoparticles (Z-NPs) against P. aeruginosa biofilm-associated murine acute pyelonephritis. In the present study, Zingerone and chitosan acted synergistically in the form of Z-NPs and found to be nontoxic to the kidney cell lines as depicted in MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay demonstrating their cytocompatibility. In vivo experiments indicated that Z-NPs (100 mg/kg) treatment reduced P. aeruginosa pathogenicity and enhanced the clearance of bacterial count from the renal and bladder tissue. Z-NPs improved the disease outcome by lowering the levels of various inflammatory markers, and histopathological examination revealed better recovery in renal and bladder tissue. Besides, ex vivo efficacy also confirmed that Z-NPs enhanced serum bactericidal effect along with increased phagocytic uptake and intracellular killing of P. aeruginosa as confirmed by fluorescent microscopy. To the best of our knowledge, this is the first study to provide evidence that Z-NPs are effective therapeutic agents for combating P. aeruginosa associated pyelonephritis.

Nanolipoidal α-terpineol modulates quorum sensing regulated virulence and biofilm formation in Pseudomonas aeruginosa.

Aim:Pseudomonas aeruginosa has emerged as a major opportunistic pathogen meaning there is an immediate need to develop efficient antivirulence agents which offer a new class of superior therapeutics. Methods: Nanostructured lipid carriers (NLCs) containing α-terpineol (αT) were developed and characterized to determine expression profiles of quorum sensing regulated genes, antivirulence activity and antibiofilm effects against P. aeruginosa. Results: The αT-NLCs had a size of 145.4 nm, polydispersity index of 0.242 and ζ-potential of -31.4 mV. They exhibited pronounced effects on the inhibition of quorum sensing mediated virulence and biofilm formation which were confirmed by molecular docking analysis and gene expression profiles. Conclusion: αT-NLCs show promise as effective antivirulence agents against P. aeruginosa in the postantibiotic era.

Screening for paroxysmal nocturnal hemoglobinuria (PNH) in patients presenting with cerebral sinovenous thrombosis (CSVT): Results of a FLAER based flowcytometry study in Indian patients.

Patients with paroxysmal nocturnal hemoglobinuria (PNH) may present with thrombosis at unusual sites, of which cerebral sinovenous thrombosis (CSVT) is one and screening for PNH is recommended in this condition. Though many patients diagnosed with PNH develop CSVT, it is unclear how many patients with PNH would present for the first time with thrombosis. We analysed the results of screening for PNH by flowcytometry in our patients with CSVT. The laboratory data of patients referred for thrombophilia and PNH testing in CSVT was examined to assess the frequency of PNH at presentation in these patients. FLAER and CD24 on granulocytes and FLAER and CD14 on monocytes respectively were used to screen the leucocytes for PNH by flowcytometry. The data for Protein C, S and Antithrombin deficiency, antiphospholipid antibodies and the Factor V Leiden mutation was examined and circumstantial risk factors were also assessed. Of the 180 cases of CSVT screened by flowcytometry for PNH, not a single case tested positive. Positivity for anti-phospholipid antibodies was the most common thrombophilic risk factor (5%). Pregnancy was the most common circumstantial risk factor. Our data on FLAER based flowcytometry in the North Indian population with CSVT suggests that PNH is not a common risk factor in our patients with thrombosis at this unusual site.

Coagulation factor VIII, IX and XI levels in north Indian patients with venous thromboembolism: first study from India.

Studies have shown elevated levels of certain coagulation factors as risk factors for venous thromboembolism (VTE). In this study, we investigated the levels of coagulation factor VIII (FVIII), FIX and FXI in north Indian patients with VTE. A total of 123 patients with VTE were screened prospectively for FVIII, FIX and FXI levels and the conventional risk factors - deficiencies of protein C, S and antithrombin, positivity for antiphospholipid antibodies and the factor V Leiden mutation. Age-matched and sex-matched controls were included. VTE was secondary to known circumstantial and thrombophilic risk factors in 66 (53.7%) patients. In 46.3% (idiopathic VTE) patients, no cause was identified. The mean FVIII levels in idiopathic (187 IU/dl) and secondary VTE patients (185.4 IU/dl) were significantly higher compared with controls (129.6 IU/dl; P < 0.001). However, there was no statistically significant difference in the levels of FIX and FXI between patients and controls (P = 0.214 and 0.198, respectively). Patients with elevated FVIII levels had increased risk of VTE compared with controls (odds ratio: 9.4, 95% confidence interval: 4.7-18.79). On logistic regression analysis after adjusting for surgery and presence of antiphospholipid antibodies, this risk remained unchanged (odds ratio: 9.54, 95% confidence interval: 4.68-19.44). A dose-response relationship was observed with progressive increase in FVIII levels. Elevated FVIII levels constitute an independent risk factor for VTE in the north Indian population. Elevated levels of FIX and FXI were not associated with increased risk of VTE.

Thrombotic Primary Antiphospholipid Syndrome: the profile of antibody positivity in patients from North India.

AIM: We evaluated the frequency of antiphospholipid antibody syndrome (APS) in patients presenting with thrombosis of various vascular beds from North India and report the antibody profiles encountered. PATIENTS AND METHODS: A retrospective analysis was performed on the laboratory results of aCL (anticardiolipin), aß2 Gp1 (anti-ßeta-2 glycoprotein 1) antibody and LAC (lupus anticoagulant) of 1222 consecutive patients referred to the coagulation laboratory work-up for a hypercoagulable/thrombophilic state over a period of 4 years between 2009 and 2013. LAC was screened with dRVVT (diluted Russel Viper Venom Test) and KCT (Kaolin clotting time), and aCL and aß2 Gp1 antibodies with commercial enzyme-linked immunosorbent assy kits. RESULTS: The current APS criteria was satisfied in 3.85% of all patients and 4.2% of pediatric patients with thrombosis. The venous circulation was more frequently affected (59.6%). Cerebral arterial and intra-abdominal vein involvement was common. Transient antibody positivity was seen in 44 (3.6%) cases. aß2 Gp1, aCL and LAC were positive in 95%, 54.5% and 23% of patients with APS, respectively, during the initial visit and 93.6%, 23% and 17%, respectively, during the follow-up visit. Persistent triple positivity was seen in only three cases. At initial testing, positivity for both aCL and aß2 Gp1 was the most frequent pattern (38% of cases). CONCLUSIONS: aß2 Gp1 antibody was the commonest antibody that was persistently positive in patients with thrombosis. Triple positivity for all antibodies had the highest specificity and positive predictive value to diagnose APS in the first visit, whereas aß2 Gp1 antibody had the highest sensitivity and negative predictive value.

Paroxysmal Nocturnal Hemoglobinuria is rare cause for thrombosis of the intra-abdominal veins in the ethnic Indian population - results from FLAER-based flowcytometry screening.

BACKGROUND: Paroxysmal nocturnal hemoglobinuria (PNH) may present as cytopenia, hemolysis, or thrombosis at unusual sites including splanchnic vessels. Thrombosis of the portal veins and hepatic veins are associated with thrombophilic risk factors: deficiencies of protein C, protein S, and antithrombin, positivity for antiphospholipid antibodies, and factor V Leiden mutation. There is limited information regarding PNH presenting primarily as a thrombotic event. We prospectively screened 142 consecutive patients with intrabdominal thrombosis and 106 controls with fluorescently labeled inactive toxin aerolysin (FLAER)-based flowcytometry to assess the frequency of PNH as a thrombophilic risk factor in patients with intra-abdominal thrombosis. METHODS: Granulocytes of patients and controls were screened with CD 24 and FLAER and monocytes with CD 14 and FLAER. Dual negativity of >1% events in both lineages was interpreted as a positive PNH clone. Screening for thrombophilia risk factors was carried out. RESULTS: Two (1.4%) cases had large PNH clones. RBC also demonstrated the PNH defect. Thrombophilia risk factors were as follows: deficiency of protein S, protein C, and antithrombin in 13.4%, 4.9%, and 2.1%, respectively, and positivity for anti-beta-2 glycoprotein 1, anticardiolipin antibodies, and lupus anticoagulant in 9.2%, 1.4%, and 0.7%, respectively. Factor V Leiden mutation was seen in 1.4% patients. CONCLUSION: PNH was uncommon in patients with intra-abdominal thrombosis in the ethnic Indian population. Despite low positivity, screening by flowcytometry for PNH is of value in this group of patients because it provides an opportunity to rapidly establish the diagnosis of this treatable disorder, which might otherwise be missed if the initial presentation is only thrombotic.