Acute pretreatment with chloroquine attenuates renal I/R injury in rats.

BACKGROUND: Acute kidney injury (AKI) still remains an unresolved problem in pharmacotherapy and renal inflammation is a major factor in its development. Chloroquine, a well-known antimalarial drug, posses pleitropic effects as well: antiinflammatory, anticoagulant and vascular actions. The effects of chloroquine on renal function may involve significant increase in urine flow rate, glomerular filtration rate and sodium excretion, as well as stimulation of nitric oxide synthase. However, its role in experimental models of renal I/R injury is unknown. We aimed to analyze the acute effects of a single-dose intravenous chloroquine administered at three different times in the experimental model of I/R injury in rat. METHODS: Rats were subjected to bilateral renal ischemia (45 min) followed by reperfusion with saline lasting 4 hours. Chloroquine was administered in doses of 0.3 mg/kg i.v. and 3 mg/kg i.v. 30 min before ischemia, 30 min before reperfusion and 5 min before reperfusion. Selected a hemodynamic, biochemical and morphological parameters were followed in the Sham-operated animals and rats subjected to I/R injury and pretreated with saline or chloroquine. RESULTS: Chloroquine (0.3 and 3 mg/kg, i.v.) protected the I/R injured kidney in an U-shaped manner. Both doses were protective regarding biochemical and histological markers of the I/R injury (serum urea, creatinine and fractional excretion of sodium, as well as total histological score, tubular necrosis score and KIM-1 staining score) (P<0.05 vs. corresponding controls, i.e. rats subjected to I/R injury and treated with saline only). The protective effects of the lower dose of chloroquine were more profound. Time-related differences between pretreatments were not observed (P>0.05, all). CONCLUSION: Our study shows for the first time that a single dose of chloroquine (0.3 mg/kg i.v.) could afford significant protection of the injured rat kidney

The effects of chloroquine (0.3 mg/kg; 3 mg/kg, i.v. bolus) on histological score of renal I/R injury.

<p>Chloroquine in dose of 0.3/kg, i.v. was injected 30 min before ischemia (IR+H03-30I), 30 min before reperfusion (IR+ H03-30R) and 5 min before reperfusion (IR+H03-5R), and in dose of 3 mg/kg, i.v. was injected 30 min before ischemia (IR+H3-30I). Control groups, Sham+Saline and IR+ Saline received an i.v. bolus of 0.5 ml saline only. The histological parameters evaluated were tubular necrosis, interstitial edema, loss of brush border, and cast formation score (Panels A–D). A minimum of 10 fields for each kidney slide were examined and assigned for severity of changes. The scoring system was as follows: 0, absent; 1, mininal changes; 2, moderate changes and 3, marked changes. Each bar represents mean ± S.E.M. +: P<0.05 <i>vs</i>. IR+Saline, *: P<0.05 <i>vs</i>.Sham+Saline (N = 6–10 rats).</p

Experimental design.

<p>Chloroquine was injected 30(30I-, 30R- and 5R-group, respectively).</p

The effects of chloroquine (0.3 mg/kg; 3 mg/kg, i.v. bolus) on total histological score of renal I/R injury and histological micrographs of renal tissues.

<p>Chloroquine in dose of 0.3/kg, i.v. was injected 30 min before ischemia (IR+H03-30I), 30 min before reperfusion (IR+ H03-30R) and 5 min before reperfusion (IR+H03-5R), and in dose of 3 mg/kg, i.v. was injected 30 min before ischemia (IR+H3-30I). Control groups, Sham+Saline and IR+Saline received an i.v. bolus of 0.5 ml saline only. A minimum of 10 fields for each kidney slide were examined and assigned for severity of changes. The scoring system was as follows: 0, absent; 1, mininal changes; 2, moderate changes and 3, marked changes. Total histological score was calculated by addition of all scores (Panel A). Each bar represants mean ± S.E.M. +: P<0.05 <i>vs</i>. IR+Saline, *: P<0.05 <i>vs</i>.Sham+Saline (N = 6–10 rats). Histological micrographs of renal tissues: kidney sections taken from Sham-operated rats or rats subjected to renal I/R injury. Periodic acid–Schiff (PAS) stain coloring. Original magnification ×20. Figures were randomly chosen from the series of at least 6 experiments (Panels B–G). Panel B: Sham-operated animals treated with saline only (Sham+Saline group) - normal renal parenchyma (PAS staining). Panel C: Rats subjected to renal I /R injury, pretreated with chloroquine at 0.3 mg/kg, i.v. 30 min before ischemia (IR+H03-30I group) - moderate kidney damage, about half of proximal tubules show loss of brush border, dilatation of lumen and loss of nuclei in some epithelial cells. Panel D: Rats subjected to renal I /R injury, pretreated with chloroquine at 0.3 mg/kg, i.v. 30 min before reperfusion (IR+ H03-30R group) - moderate kidney damage, loss of brush border was observed in half of proximal tubules, in addition to dilatation of lumen and loss of nuclei in some epithelial cells. Panel E: Rats subjected to renal. I /R injury, pretreated with chloroquine at 0.3 mg/kg, i.v. 5 min before reperfusion (IR+H03-5R group) - moderate kidney damage, two thirds of proximal tubules show loss of brush border, dilatation of lumen and loss of nuclei in numerous epithelial cells. Panel F: Rats subjected to renal. I /R injury, pretreated with chloroquine at 3 mg/kg, i.v. 30 min before ischemia (IR+H3-30I group) - moderate kidney damage, two thirds of proximal tubules show loss of brush border, dilatation of lumen and loss of nuclei in majority of epithelial cells (marked necrosis). Panel G: Rats subjected to renal I / R injury, pretreated with saline only (IR+Saline-group) - marked kidney damage, interstitial edema diffusely present, proximal tubules show loss of brush border and lumen dilatation and loss of nuclei in some epithelial cells.</p

The effects of chloroquine (0.3 mg/kg, i.v; 3 mg/kg, i.v.) on KIM-1 staining score of renal I/R injury and histological micrographs of renal tissues.

<p>Chloroquine in dose of 0.3/kg, i.v. was injected 30 min before ischemia (IR+H03-30I), 30 min before reperfusion (IR+ H03-30R) and 5 min before reperfusion (IR+H03-5R), and in dose of 3 mg/kg, i.v. was injected 30 min before ischemia (IR+H3-30I). Control groups, Sham+Saline and IR+ Saline received instead of drug i.v. bolus of 0.5 ml saline only. The following scoring system was used: 0, no staining; 0.5+, focal weak fine granular staining; 1+, weak fine granular staining; 2+, moderate granular staining; and 3+, strong large granular staining. The KIM-1 staining intensity score for each sample was graded by two experts in the field of pathology who was blinded to the group assessments. Finally, KIM-1 staining density score for each sample calculated using mean score of these two experts (mean score ± S.E.M.) (Panel A). Each bar represants mean ± S.E.M. +: P<0.05 <i>vs</i>. IR+Saline, *: P<0.05 <i>vs</i>.Sham+Saline (N = 6–10 rats). Histological micrographs of renal tissues: kidney sections taken from Sham-operated rats or rats subjected to renal I/R injury. Kidney injury molecul-1 (KIM-1) staining. Original magnification ×20. Figures were randomly chosen from the series of at least 6 experiments (Panels B–G). Panel B: Sham-operated animals treated with saline only (Sham+Saline group) - absence of immunoreactivity for KIM-1. Panel C: Rats subjected to renal I /R injury, pretreated with chloroquine at 0.3 mg/kg, i.v. 30 min before ischemia (IR+H03-30I group) – most of proximal and some distal tubules show mild staining for KIM-1. Panel D: Rats subjected to renal I /R injury, pretreated with chloroquine at 0.3 mg/kg, i.v. 30 min before reperfusion (IR+ H03-30R group) - nearly half of proximal tubules and rare distal tubules show mild to moderate KIM staining.Panel E: Rats subjected to renal I /R injury, pretreated with chloroquine at 0.3 mg/kg, i.v. 5 min before reperfusion (IR+H03-5R group) - nearly half of proximal tubules and rare distal tubules show moderate KIM staining. Panel F: Rats subjected to renal I /R injury, pretreated with chloroquine at 3 mg/kg, i.v. 30 min before ischemia (IR+H3-30I group) - most of proximal and some distal tubules show moderate staining for KIM-1. Panel G: Rats subjected to renal I / R injury, pretreated with saline only (IR+Saline-group) - proximal and distal tubules show moderate to intensive positive KIM staining.</p

The effects of chloroquine (0.3 mg/kg, i.v; 3 mg/kg, i.v.) on renal dysfunction and injury caused by I/R.

<p>Chloroquine in dose of 0.3/kg, i.v. was injected 30 min before ischemia (IR+H03-30I), 30 min before reperfusion (IR+ H03-30R) and 5 min before reperfusion (IR+H03-5R), and in dose of 3 mg/kg, i.v. was injected 30 min before ischemia (IR+H3-30I). Control groups, Sham+Saline and IR+ Saline received an i.v. bolus of 0.5 ml saline only. Panel A, B, and C: serum creatinine and urea concentrations and fractional excretion of Na⁺. Each bar represants mean ± S.E.M. +: P<0.05 <i>vs</i>. IR+Saline, *: P<0.05 <i>vs</i>.Sham+Saline (N = 6–10 rats).</p

Treatment of peripartum depression with antidepressant and other psychotropic medications: a synthesis of clinical practice guidelines in Europe

This study examined (1) the availability and content of national CPGs for treatment of peripartum depression, including comorbid anxiety, with antidepressants and other psychotropics across Europe and (2) antidepressant and other psychotropic utilization data as an indicator of prescribers’ compliance to the guidelines. We conducted a search using Medline and the Guidelines International Network database, combined with direct e-mail contact with national Riseup-PPD COST ACTION members and researchers within psychiatry. Of the 48 European countries examined, we screened 41 records and included 14 of them for full-text evaluation. After exclusion of ineligible and duplicate records, we included 12 CPGs. Multiple CPGs recommend antidepressant initiation or continuation based on maternal disease severity, non-response to first-line non-pharmacological interventions, and after risk-benefit assessment. Advice on treatment of comorbid anxiety is largely missing or unspecific. Antidepressant dispensing data suggest general prescribers’ compliance with the preferred substances of the CPG, although country-specific differences were noted. To conclude, there is an urgent need for harmonized, up-to-date CPGs for pharmacological management of peripartum depression and comorbid anxiety in Europe. The recommendations need to be informed by the latest available evidence so that healthcare providers and women can make informed, evidence-based decisions about treatment choices

Toxicity evaluation of two polyoxotungstates with anti-acetylcholinesterase activity

A toxicity evaluation of two Keggin-type heteropolytungstates, K-7[Ti2PW10O40].6H(2)O and K6H [SiV3W9O40].3H(2)O, with different inhibitory potencies toward acetylcholinesterase activity (IC50 values of 1.04 x 10(-6) and 4.80 x 10(-4) mol/L, respectively) was performed. Wistar albino rats were orally treated with single doses (5 and 50 mg/kg) of both investigated compounds. The biochemical parameters of renal (serum urea and creatinine) and liver function (direct and total bilirubin, alanine transaminase, and aspartate aminotransferase) were determined after 24 h and 14 days. A histopathological analysis of liver tissue was carried out 14 days after the polyoxotungstate administration. Both applied doses of the investigated compounds did not induce statistically significant alterations of the renal function markers. However, the polyoxotungstate treatment caused an increase in the activities of serum alanine transaminase and aspartate aminotransferase in a time- and concentration -dependent manner, although statistically significant changes in bilirubin concentrations were not observed. Furthermore, the detected hepatotoxic effect was confirmed by histhopathological analysis that suggested some reversible liver tissue damage two weeks after the treatment, especially in the case of K6H [SiV3W9O40]-3H(2)O. Accordingly, the toxicity of these two polyoxotungstates with anti-acetylcholinesterase effect cannot be considered as a severe one, but their potential clinical application would require a more complex toxicological study

Immune Cell and Biochemical Biomarkers in Advanced Laryngeal Cancer

Objective The aim of this study was to evaluate cell and biochemical biomarkers and establish their prognostic value in patients with advanced laryngeal cancer. Material and Methods A prospective study included 52 patients with advanced laryngeal carcinoma surgically treated at the tertiary referral center. Tumor tissue was immunohistochemically stained for T-cell markers (CD4 and CD8), and levels of cytokines (IL-6 and IL-8) and C-reactive protein were analyzed from blood samples. Results Overall 3-year survival (OS) of patients included in the study was 69.2% and the disease specific survival (DSS) 72.5%. Higher expression of CD4 + and CD8 + were significant prognostic factors with positive impact on both OS and DSS in univariate analysis, but not in multivariate analysis. Levels of IL-8 were a significant predictor of 3-year OS and DSS survival in patients with advanced laryngeal cancer but not levels of IL-6 and CRP values. Conclusion Though high expression of CD4 and CD8 were demonstrated in the tumor tissue, but their prognostic role was not established. Higher values of IL-8 proved to be significant negative predictor of DSS. This could further collaborate the inclusion of combination of biomarkers in assessment of favorable treatment choice in patients with advanced laryngeal carcinoma