Urinary N-acetyl-beta-D-glucosaminidase (NAG) in lupus nephritis and rheumatoid arthritis

Increased activity of urinary N-acetyl-beta-D-glucosaminidase (NAG) can be used as an early indicator of damage to the tubular epithelium. Systemic lupus erythematosus (SLE) is a multisystem autoimmune rheumatic disease. Nephritis is known as the most serious complication of SLE and the strongest predictor of poor outcome. In this study urinary NAG excretion was investigated in 24 SLE patients with normal renal function (serum creatinine < or =1.2 mg/dL) and the results were compared with those from 26 untreated patients with rheumatoid arthritis (RA) and 27 healthy controls. The SLE patients were divided into two groups according to their urinary total protein levels: group A consisted of 16 patients with < or =3.5 g/day proteinuria, and group B consisted of eight patients with nephrotic-range proteinuria (>3.5 g/day). Serum and urinary creatinine, total urinary protein levels, and urinary NAG excretion were measured in patients with SLE and RA. In addition, serum C3 and C4 levels were determined in the SLE patients. Renal biopsies were performed in all of the SLE patients. Glomerular lesions were classified according to WHO criteria for lupus nephritis (LN) I-V. The Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) was used to assess disease activity. Urinary NAG excretion was significantly higher in the SLE groups than in the healthy controls (P<0.001). In urinary NAG excretion there was also significant difference between SLE groups and RA patients (P<0.001). However, there was no significant difference in NAG excretion between the RA and control groups (P=0.062). Urinary NAG excretion was significantly higher (P<0.05) in group B compared to group A. There were no differences in SLEDAI scores, ages, and serum creatinine levels between study groups (P=0.601, P=0.285, P=0.669, respectively). Elevated SLEDAI values and hypocomplementemia were detected more often in younger patients (P<0.010, r=-0.529 and P<0.010, r=-0.569, respectively). There was a strong positive correlation between proteinuria and urinary NAG activity (P<0.001, r=0.759). These results suggest that the determination of urinary NAG activity may be a useful supplement to the routine biochemical analysis performed on the urine in cases of SLE.     

Comparison of behavioral,neuroprotective, and proinflammatory cytokine modulating effects exercised by (+)‐cis‐EC and (−)‐cis‐EC stereoisomers in a PTZ‐induced kindling test in mice

Epoxy‐carvone (EC) has chiral centers that allow generation of stereoisomers, including (+)‐cis‐EC and (?)‐cis‐EC, whose effects in the kindling tests have never been studied. Accordingly, this study aims to comparatively investigate the effect of stereoisomers (+)‐cis‐epoxy‐carvone and (?)‐cis‐epoxy‐carvone on behavioral changes measured in scores, in the levels of cytokines (IL‐1β, IL‐6, and TNFα) and neuronal protection in the face of continuous treatment with pentylenetetrazol. Swiss mice were divided into five groups (n = 10), receiving vehicle, (+) – cis‐EC, (?) – cis‐EC (both at the dose of 30 mg/kg), and diazepam (4 mg/kg). Thirty minutes after the respective treatment was administered to the animals one subconvulsive dose of PTZ (35 mg/kg). Seven subconvulsives treatments were made on alternate days, in which each treatment several parameters were recorded. In the eighth treatment, the animals receiving the highest dose of PTZ (75 mg/kg) and were sacrificed for quantification of cytokines and histopathologic analysis. All drugs were administered by intraperitoneal route. In the kindling test, (+)‐cis‐EC and (?)‐cis‐EC reduced the average scores. The stereoisomer (+)‐cis‐EC decreased levels of proinflammatory cytokines IL‐1β, IL‐6, and TNFα, whereas comparatively (?)‐cis‐EC did not reduce IL‐1β levels. Histopathological analysis of the mice hippocampi undergoing this methodology showed neural protection for treated with (+)‐cis‐EC. The results suggest that the anticonvulsant effect of (+)‐cis‐EC possibly takes place due to reduction of proinflammatory cytokines involved in the epileptogenic process, besides neuronal protection, yet further investigation of the mechanisms involved is required.     

microRNA146a Expression in Lupus Patients With and Without Renal Complications

microRNA146a (miRNA) expression profiles are likely to become important diagnostic and prognostic tools in many disease aspects. This work aimed to study the expression of miRNA146a in lupus patients with and without renal complications and to assess its association with disease activity. Patients enrolled in the study included 52 females affected by systemic lupus erythematosus (SLE) and another 60 females with lupus nephritis (LN). Forty‐eight age‐matched healthy females were enrolled as a control group. miRNA146a expression was assessed using real‐time PCR. In SLE patients, miRNA146a was underexpressed as compared to healthy controls and lower levels were detected among lupus patients with renal affection. In addition, miRNA146a expression was low and serum Interferon‐α (IFN‐α) level was high in patients with active lupus as compared to those with inactive disease. miRNA146a expression was inversely correlated to serum IFN‐α level and to anti ds‐DNA titer in the three studied groups. In conclusion, miRNA146a might be implicated in lupus pathogenesis. Moreover, miRNA146a expression correlates negatively to lupus activity and LN, whereas serum IFN‐α has a direct correlation to both disease activity and nephritis; hence, both miRNA146a expression and serum IFN‐α could be potentially important diagnostic biomarkers and potential novel strategies for therapeutic interventions, which may possibly be implied to enhance the sensitivity and specificity for the prediction of flares and prognosis in SLE patients.     

Improved method for analysis of cysteinyldopa in human serum

5‐S‐l‐Cysteinyl‐l‐dopa is a well‐known pigment intermediate and analysis of its serum concentration is well suited for evaluation of treatment and follow‐up of stage III and IV malignant melanoma. A simplified analytical method is described using organic extraction followed by clean‐up on a boronate gel to capture the compound containing vicinal hydroxyls. Weak acid solution elutes the 5‐S‐cysteinyldopa suitable for high‐performance liquid chromatography (HPLC). The absolute recoveries of cysteinyldopa and its diastereomer 5‐S‐D‐cysteinyl‐l‐dopa (used as an internal standard) were 81.5±2.8% and 81.3±2.7%, respectively, and use of the internal standard for the whole procedure gave an analytical recovery of 101±0.8%. The limit of quantitation was 1.5?nmol/L and the imprecision of the method was <5.0% over the analytical range 1.5–500?nmol/L. The method is cheap and easy to perform and compares well with other described techniques. The use of the method is illustrated by results obtained during treatment of a patient with metastatic malignant melanoma.     

Behavioral and sensory changes after direct ischemia-reperfusion injury in rats.

Complex regional pain syndromes (CRPS) are disabling pain syndromes that can develop after trauma or minor tissue injury affecting a limb. Characteristics of CRPS are sensory signs and symptoms, autonomic abnormalities, trophic changes and an impaired motor function. Pathophysiological mechanisms for the development of CRPS are still a matter of investigation. Based on clinical data and investigations of CRPS patients it is hypothesized that tissue hypoxia and inflammation are important for the development of CRPS. The aim of the current study was therefore to examine if direct ischemia-reperfusion injury can induce behavior in rats with symptoms present in patients with CRPS. After baseline behavior measurements the femoral artery of Wistar rats was ligated for 3h with consecutive reperfusion. Sham-operated rats underwent the same preparation except ligation of the artery. Subsequent behavioral testing (observations of spontaneous pain behavior, paw withdrawal to mechanical, noxious mechanical, cold and heat stimuli) was performed up to two months after surgery. Both in rats that underwent ischemia and in sham-operated rats no obvious changes of hindpaw tissue were observed after ischemia-reperfusion injury (trophic changes, edema, differences in skin color or temperature). In behavioral tests only minor changes were observed, these being not different between postischemic rats and sham-operated rats. Using Wistar rats, our data do not support the idea that an ischemia-reperfusion injury can play a major role in the development of CRPS.     

Effects of lipids and lipid-lowering therapy on hemostatic factors in patients with myocardial infarction.

BACKGROUND: The risk of cardiovascular disease (CVD) is associated with specific hemostatic markers and lipid profiles, and evidence indicates that there are associations between lipid profiles and the levels of certain hemostatic factors. The disturbances in hemostasis and the risk of CVD can be ameliorated by lipid-lowering therapy. OBJECTIVE: We investigated the associations of lipid profiles with factor (F)VIIa, von Willebrand factor (VWF), D-dimer and plasminogen activator inhibitor-1 (PAI-1), and examined whether lipid-lowering statin therapy would affect the levels of these hemostatic markers. PATIENTS AND METHODS: This cross-sectional study analyzed 1045 postmyocardial infarction patients. RESULTS: In multivariate regression analyses (without adjusting for clinical covariates) HDL-cholesterol (HDL-C) and HDL size were independent and significant predictors of FVIIa; HDL size was a predictor of VWF; HDL size, HDL-C and LDL size were predictors of D-dimer; and triglyceride and HDL size were predictors of PAI-1. After adjusting for clinical covariates, HDL-C, lipoprotein (Lp)(a), apolipoprotein B (apoB) and warfarin were independent and significant predictors of FVIIa; HDL size, age, diabetes mellitus, insulin, race and warfarin were predictors of VWF; HDL-C, HDL size, LDL size, age, warfarin, hypertension and gender were predictors of D-dimer; and triglyceride, HDL size, body mass index, insulin and hypertension were predictors of PAI-1. Patients on statin therapy had significantly lower levels of D-dimer than those who were not on this therapy. CONCLUSION: There are significant associations of lipid profiles with hemostatic factors, the directions of which suggest novel pathways by which dyslipidemia may contribute to coronary heart disease.     

Efficacy of moxifloxacin for treatment of penicillin-, macrolide- and multidrug-resistant Streptococcus pneumoniae in community-acquired pneumonia

This pooled analysis of six prospective, multicentre trials aimed to determine the efficacy of moxifloxacin in community-acquired pneumonia (CAP) due to penicillin-, macrolide- and multidrug-resistant Streptococcus pneumoniae (MDRSP). At a central laboratory, isolates were identified and antimicrobial susceptibility determined (microbroth dilution). MDRSP was defined as resistance > or =3 drug classes. Patients received oral or sequential intravenous/oral 400 mg moxifloxacin once daily for 7-14 days. The primary endpoint was clinical success at test-of-cure for efficacy-valid patients with proven pretherapy S. pneumoniae infection. Of 140 S. pneumoniae isolated (112 respiratory, 28 blood), 23 (16.4%) were penicillin resistant, 26 (18.6%) macrolide resistant and 31 (22.1%) MDRSP. The moxifloxacin MIC90 was 0.25 microg/ml. Clinical cure with moxifloxacin was 95.4% (125/131) overall, and 100% (21/21) for penicillin-, 95.7% (22/23) for macrolide- and 96.4% (27/28) for multidrug-resistant strains. Moxifloxacin provided excellent clinical and bacteriological cure rates in CAP due to drug-resistant pneumococci.     

IGF-1, IGFBP-3, VEGF and MMP-9 levels and their potential relationship with renal functions in patients with compensatory renal growth

BACKGROUND: Mechanisms of compensatory renal growth (CRG) still remain a mystery. Various growth factors, including growth hormone, insulin-like growth factor-1 (IGF-1) have been implicated in different forms of CRG. AIMS: To investigate the serum levels of IGF-1, vascular endothelial growth factor (VEGF - role in vascular remodelling), matrix metalloproteinase-9 (MMP-9 - essential for normal nephrogenesis) and correlation of renal function in patients with unilateral nephrectomized, agenesis and hypoplasic kidney. METHODS: Thirty patients were included in this study. In group I, there were 10 patients with unilateral nephrectomy, while in group II, there were 10 patients with unilateral agenesis. As for group III, there were 10 patients with unilateral hypoplastic kidney. The serum levels of IGF-1, IGF-binding protein-3 (IGFBP-3), VEGF and MMP-9 were studied in all the cases. Clearance of creatinin (Ccr) and protein excretion were examined in the 24 h urine. CRG was determined with ultrasonography and scintigraphy. Twenty-six control subjects were also studied. RESULTS: The levels of IGF-1, IGFBP-3, VEGF and MMP-9 were significantly higher in patients than in the control subjects (P < 0.001). Ccr and protein excretion levels were different in study groups than in those of the control group (P < 0.01). There were positive correlations between the serum levels of IGF-1 with IGFBP-3; IGF-1 with MMP-9; IGFBP-3 with MMP-9 (r = 0.825, P = 0.0001; P < 0.001 r = 0.611; P < 0.001 r = 0.585, respectively). There were negative correlations between GFR and the serum levels of IGF-1, IGFBP-3 and MMP-9 (P < 0.01 r = -0.708; P = 0.002 r = -0.803; P < 0.05 r = -0.442, respectively). Furthermore, there were positive correlations between proteinuria and the serum levels of IGF-1, IGFBP-3 and MMP-9 (P = 0.039 r = 0.600; P < 0.05 r = 0.456; P < 0.05 r = 0.424). CONCLUSIONS: Increased IGF-1, IGFBP-3, VEGF and MMP-9 were observed in CRG in the follow-up period. IGF-1 and MMP-9 seemed to have increased in patients with CRG in defiance of the development of fibrosis. Moreover, IGF-1 and MMP-9 seem to be associated with reduced renal function and proteinuria.     

Role of oxidative stress in pathophysiology of peripheral neuropathy and modulation by N-acetyl-L-cysteine in rats.

OBJECTIVES: The objectives of this study were to examine the role of reactive oxygen species and oxidative stress in peripheral neuropathy and behavioural pain responses in experimentally induced chronic constriction injury (CCI) of sciatic nerve of rat. Effect of N-acetyl-L-cysteine (NAC) administered intraperitoneally, was also investigated on CCI-induced neuropathic pain in rats. METHODS: Neuropathy was induced by CCI of the right sciatic nerve in ketamine anaesthetized rats. Effect of intraperitoneally administered NAC in rats was also investigated using nociceptive behavioural tests. Malondialdehyde, an index of oxidative stress and antioxidant enzymes was also estimated in ligated sciatic nerve. RESULTS: Behavioural tests, mechanical, thermal and cold stimuli confirmed the development of neuropathic pain after the CCI. The malondialdehyde levels of ligated sciatic nerves were significantly increased compared to non-ligated sciatic nerves (sham operated). The antioxidant enzyme reduced, glutathione was inhibited, while superoxide dismutase increased. However, catalase remained unaffected in the injured sciatic nerves. Intraperitoneal administration of NAC resulted in significant reduction of hyperalgesia in CCI-induced neuropathic rats. CONCLUSIONS: This study identifies antioxidants superoxide dismutase and reduced glutathione, and oxidative stress as important determinants of neuropathological and behavioural consequences of CCI-induced neuropathy, and NAC may be a potential candidate for alleviation of neuropathic pain.     

Serum levels of IGF-I and IGFBP-III and their relation with carcinoembryonic antigen and carbohydrate antigen 19-9 in cases of esophageal cancer

Tumour markers are used for diagnosis, staging, evaluation of response to treatment, prognosis and detection of recurrences in clinical oncology. In this study, we aim to investigate the levels of insulin-like growth factor (IGF)-I and IGF-binding protein (IGFBP)-III in cases with oesophageal carcinoma. We investigated their possible use as tumour markers and their relation to other tumour markers. Forty patients who were diagnosed as having oesophageal carcinoma by histopathological evaluation of endoscopic biopsies between January 2003 and July 2004 and 40 healthy people as the control group were included in the study. The serum levels of tumour markers including IGF-I, IGFBP-III, carcinoembryonic antigen (CEA) and carbohydrate antigen (CA) 19-9 were measured in both groups. Data were compared statistically, and the importance of IGF-I and IGFBP-III levels were investigated in cases with oesophageal carcinoma. IGF-I levels were significantly higher in patients with oesophageal carcinoma when compared with the control group (p < 0.05), whereas IGFBP-III levels were significantly lower (p < 0.05). The increase in CEA levels was not statistically significant when compared with the control group. The increase in CA 19-9 levels was statistically significant when compared with the control group (p < 0.05). No correlation was detected between levels of IGF-I and IGFBP-III and levels of CEA and CA 19-9. We suggest that the serum IGF-I level may be used as a tumour marker in oesophageal carcinoma. A low level of serum IGFBP-III is also significant in cases with oesophageal carcinoma. We believe that drugs which inhibit IGF-I function or which stimulate the function of IGFBP-III may open new horizons in extra-surgical modalities for the treatment of oesophageal cancer.     

Outcome of patients hospitalized for COVID‐19 and exposure to angiotensin‐converting enzyme inhibitors and angiotensin‐receptor blockers in France: results of the ACE‐CoV study

Data are lacking on the impact of ACEI/ARB exposure on unfavorable outcome in the population of patients hospitalized for COVID‐19 with hypertension/cardiovascular disease, particularly in Europe. The ACE‐CoV study was designed to assess this question. The study was conducted in the Covid‐Clinic‐Toul cohort, which contains data about all patients hospitalized at Toulouse University hospital, France with a SARS‐CoV‐2 infection since March, 2020. We selected the patients with a history of cardiovascular disease (heart failure or coronary disease) and/or arterial hypertension. We conducted a subgroup analysis in patients with arterial hypertension. ACEI/ARB exposures at admission were assessed. The outcome was composite: admission to intensive care unit, need of mechanical ventilation or death during the 14 days after admission to hospital. We used logistic regression models with propensity scores (PS) weighted by overlap weighting (OW) and inverse probability of treatment weighting (IPTW). Between March 2020 and April 20, 2020, the Covid‐Clinic‐Toul included 263 patients. Among them, 111 were included in the ACE‐CoV study population. In OW‐PS‐adjusted analyses, the association of exposure to ACEIs or ARBs with outcome occurrence was OR: 1.56 (95% CI: 0.73–3.33). It was 0.99 (95% CI: 0.68–1.45) for ACEIs and 1.64 (95% CI: 0.77–3.50) for ARBs. Analyses with weighting by the IPTW‐PS method gave similar results. Results were similar when considering the subgroup of patients with arterial hypertension. The ACE‐CoV study found no association between exposure to ACEIs or ARBs and unfavorable outcome in hospitalized patients for COVID‐19 with a history of cardiovascular disease/arterial hypertension.