The outcome of proliferative lupus nephritis with pulse cyclophosphamide therapy.

Proliferative lupus nephritis deserves aggressive therapy and cyclophosphamide plays a pivotal role. Thirty nine patients with proliferative lupus nephritis (Class III-7 patients and Class IV- 32 patients) with a median follow up of 38 months were considered for this observational study. All the patients received induction therapy with intravenous methylprednisolone. Cyclophosphamide was given intravenously initially in monthly pulses for six months and later quarterly pulses until remission was achieved or until the target dose (200 mg/kg) was reached. The treatment with intravenous methylprednisolone was repeated in the event of a nephritic flare. Later the corticosteroid was reduced to a minimum effective dose and cyclophosphamide was changed to either azathioprine or mycophenolate mofetil. At the time of the last follow up, 82.05% of the patients were in remission (complete remission 51.28% and partial remission 30.77%). The median interval to achieve remission in responders was 15 months. Early diagnosis (P=0.04), a higher creatinine clearance at presentation (P=0.02), and concurrent use of an ACEI or an ARB (P=007) significantly favored attaining remission. Five patients experienced a doubling of serum creatinine and one of them became dialysis dependent. Risk of doubling of serum creatinine correlated with a low Ccr (P=0.03) at presentation, occurrence of renal flares (P=0.034) and failure to achieve remission (P=0.0001). The parameters like serum creatinine, serum C3, serum C4, activity and chronicity indices on renal biopsy, hypertension were not statistically significant. Therapy with cyclophosphamide, if initiated early, helps in inducing remission and hence can retard the progression to CKD.

Oxidative stress in hemodialysis patients receiving intravenous iron therapy and the role of N-acetylcysteine in preventing oxidative stress.

To determine the contribution of injectable iron administered to hemodialysis (HD) patients in causing oxidative stress and the beneficial effect of N-acetylcysteine (NAC) in reducing it, we studied in a prospective, double blinded, randomized controlled, cross over trial 14 adult HD patients who were randomized into two groups; one group received NAC in a dose of 600 mgs twice daily for 10 days prior to intravenous iron therapy and the other group received placebo. Both the groups were subjected to intravenous iron therapy, 100 mg of iron sucrose in 100 mL of normal saline given over a period of one hour. Blood samples for the markers of oxidative stress were taken before and after iron therapy. After the allowance of a week of wash out period for the effect of N-acetylcysteine we crossed over the patients to the opposite regimen. We measured the lipid peroxidation marker, malondiaaldehyde (MDA), to evaluate the oxidative stress and total anti-oxidant capacity (TAC) for the antioxidant level in addition to the highly sensitive C-reactive protein (HsCRP). Non-invasive assessment of endothelial dysfunction was measured by digital plethysmography before and after intravenous iron therapy. There was an increase of MDA (21.97 + 3.65% vs 7.06 + 3.65%) and highly sensitive C-reactive protein (HsCRP) (11.19 + 24.63% vs 13.19 + 7.7%) after iron administration both in the placebo and the NAC groups. NAC reduced the baseline acute systemic generation of oxidative stress when compared to placebo, which was statistically significant with MDA (12.76 + 4.4% vs 9.37 + 4.40%: P = 0.032) but not with HsCRP though there was a declining trend (2.85 + 22.75 % vs 8.93 + 5.19%: P = 0.112). Pre-treatment with NAC reduced the endothelial dysfunction when compared to placebo, but it was not statistically significant, except for reflection index (RI). We conclude that in our HD patients NAC reduced the oxidative stress before and after the administration of intravenous iron therapy in addition to the endothelial dysfunction induced by this treatment.

Pulse cyclophospamide in severe lupus nephritis: Southern Indian experience.

To evaluate the efficacy and safety of the monthly pulse IV cyclophosphamide (IVC) therapy in patients with severe lupus nephritis, we studied 39 patients of lupus nephritis on IVC therapy between 1998 to 2002. Single monthly cyclophosphamide (0.75-1 g/m(2)) was infused intravenously with oral prednisolone (0.5 mg/kg per day) and appropriate hydration. Of the 39 patients 25 (86.2%) patients were females and 4 (13.8%) were males. Six (2%) cases had irregular follow-up and 3 patients had expired during the initial cycles and were excluded from the study. The mean age was 25.6 + 6.72 years (range 10-40 years). The mean duration of the disease from the onset to renal biopsy was 24.2 + 18.5 months. The clinical presentations included nephrotic syndrome (34.5%), acute glomerulonephritis (31.0%), Pyrexia of unknown origin (PUO) (10.3%), and rapidly progressive renal failure (6.7%). Renal insufficiency was present in 47.2% cases. Twenty-two (75.9%) patients had diffuse proliferative glomerulonephritis (class IV), 6 (20.7%) focal proliferative glomerulonephritis (class III), and one (3.4%) class Vd. After a mean follow-up of 15.8 months, out of 29 patients, 13 (44.8%) had achieved complete remission, 7 (24.1%) partial remission and 9 (31.0%) cases did not respond to the therapy. Side effects of the therapy included vomiting and nausea (100%) and hair loss during the first few doses of IVC. In addition, one case had dysfunctional uterine bleeding and two patients had avascular necrosis of femoral head. We conclude that our data indicate that IVC in severe lupus nephritis is effective in Indian patients though longer follow-up is required.

Fungal peritonitis in patients on continuous ambulatory peritoneal dialysis: a single-centre experience in India.

AIMS: The clinical features, treatment and outcome of fungal peritonitis (FP) in continuous ambulatory peritoneal dialysis (CAPD) patients were examined. METHODS: Dialysis records of all 303 end-stage renal disease (ESRD) patients initiated on CAPD treatment between January 1998 and February 2008 were reviewed retrospectively. RESULTS: In the 303 patients dialysed between January 1998 and February 2008, a total of 137 bacterial peritonitis and 43 FP episodes were recorded. The incidence rate of FP was 0.67/100 patient months or 1/148.67 months. It accounted for 23.88% of all peritonitis episodes. Three factors appeared to predict mortality: the presence of non-Candida species, the catheter being left in situ and a serum albumin level <3 g/dl. Multivariate analysis yielded only the latter 2 as predictors of mortality. The use of intraperitoneal antibiotics in the 3 months before infection and low serum albumin have been identified as risk factors for contracting FP. CONCLUSION: Risk factors for contracting FP and for mortality due to FP have been identified.

Activated pericyte attenuates endothelial functions: nitric oxide-cGMP rescues activated pericyte-associated endothelial dysfunctions.

Hepatic stellate cells are liver-specific pericytes and exist in close proximity with endothelial cells. The activation of liver pericytes is intrinsic to liver pathogenesis, and leads to endothelial dysfunction, including the low bioavailability of nitric oxide (NO). However, the role of nitric oxide in pericyte-endothelium cross-talk has not yet been elucidated. This work examines the cellular mechanism of action of NO in pericyte-mediated endothelial dysfunction. We used in vitro coculture and conditioned medium systems to study the effects of activated liver pericytes on endothelial function, and an egg yolk vascular bed model was used to study the effects of activated pericytes on angiogenesis. This study also demonstrates that activated pericytes attenuate the migration, proliferation, permeability, and NO production of endothelial cells. Our results demonstrate that activated pericytes restrict angiogenesis in egg yolk vascular bed models, and NO supplementation recovers 70% of the inhibition. Our results also demonstrate that supplementation with NO, sildenafil citrate (phosphodiesterase inhibitor), and 8-bromo-cGMP (cGMP analog) partially recovers activated-pericyte-mediated endothelium dysfunction. We conclude that NO-cGMP alleviates activated-pericyte-associated endothelial dysfunction, including angiogenesis, in a cGMP-dependent manner.

Antibacterial nanosized silver substituted hydroxyapatite: synthesis and characterization.

The silver (0.5-3 at %) substituted nanosize hydroxyapatites (AgHAs) were synthesized by microwave processing. The X-ray diffraction (XRD) peaks are very broad, indicating that the AgHAs were of nanosize (30 nm). Transmission electron microscopy analysis shows needle-like morphology of AgHA, having length 60-70 nm and width 15-20 nm. The AgHA phase was stable up to 700 degrees C without any secondary phases. The antibacterial effect of AgHA against Escherichia coli and Staphylococcus aureus was observed by spread plate method, even for low concentration of silver ions (0.5%) with 1 x 10(5) cells/mL of respective bacterial culture, after a 48 h incubation period. However, some colonies of E. coli were seen with a high dose of 1 x 10(8) cells/mL after 24 h. The zone of inhibition by disc diffusion test method was found to vary with the amount of silver in the sintered AgHA pellets, for both the bacteria, after 24 h of inoculation. Osteoblast cell attachment in varying density was noticed on AgHA samples with 0.5, 1.0, and 1.5% silver substitution. However, osteoblast spreading was significantly greater on 0.5% AgHA compared to 1.0 or 1.5% substituted AgHA samples. Thus, the low amount of AgHA has a potential of minimizing the risk of bacterial contamination, without compromising the bioactivity, and is expected to display greater biological efficacy in terms of osseointegration.

Permeability of R6G across Cx43 hemichannels through a novel combination of patch clamp and surface enhanced Raman spectroscopy.

We have measured the permeability of rhodamine-6G across Cx43 hemichannels reconstituted on a pipette tip. Cx43 hemichannels were overexpressed in Sf9 cells, and affinity-purified. The hemichannels were reconstituted in a lipid bilayer on a pipette tip by the tip-dip method. R6G in the pipette permeated across the channels into the bath. The permeability of R6G was quantified by measuring R6G concentration in the bath after several hours by surface enhanced Raman spectroscopy (SERS) with 100 nm silver colloid particles. The ratio of the permeability of dye to salt, as extracted by this combined electrical-SERS technique, is compatible with similar ratios for other dyes across whole gap junction channels. The results for the permeability ratio were further compared to fluorescence measurements. The novel combination of patch and SERS techniques can be extended to quantifying the transport of biologically significant non-fluorescent molecules, such as cAMP and IP3, across 1 nm sized pores, such as the gap junction channel.

Large-area topography analysis and near-field Raman spectroscopy using bent fibre probes.

We present a method for combined far-field Raman imaging, topography analysis and near-field spectroscopy. Surface-enhanced Raman spectra of Rhodamine 6G (R6G) deposited on silver nanoparticles were recorded using a bent fibre aperture-type near-field scanning optical microscope (NSOM) operated in illumination mode. Special measures were taken to enable optical normal-force detection for control of the tip-sample distance. Comparisons between far-field Raman images of R6G-covered Ag particle aggregates with topographic images recorded using atomic force microscopy (AFM) indicate saturation effects due to resonance excitation.

Influence of colon degradation of polysaccharide on the oral bioavailability of theophylline from controlled release hydrophilic matrices.

Hydrophilic matrices of gum karaya (GK) and guar gum (GG) using theophylline (TH) as a model drug were prepared for oral controlled release. In vitro release studies were performed for these matrix systems to find out the suitable drug-carrier ratio, which extend the drug release up to 24 h. Promising matrix systems were subjected for in vitro degradation studies in the presence of rat caecal contents. These matrices were also evaluated for their in vivo performance in healthy human volunteers. Matrix systems containing 40% w/w of polysaccharide (GK or GG) have shown uniform and similar in vitro drug release profile for 24 h in the Sorenson's phosphate buffer (pH 7.4). However, TH release from GG-TH matrix system in the presence of rat caecal contents was significantly higher than that from GK-TH matrix system. This is because of the susceptibility of GG for degradation by microorganisms present in the rat caecal content. Though there was no significant difference between the peak plasma concentration (Cmax) and time of its occurrence (Tmax) for TH from GG-TH and GK-TH matrix systems, it was found that oral bioavailability of TH from former matrix was significantly higher than that of later. Therefore, the present study disclosed that the usage of colon degradable polymer offers an advantage in the design of controlled release dosage forms of drugs, which has good absorption properties throughout the gastrointestinal tract.

Multivariate evaluation of doxorubicin surface-enhanced Raman spectra.

Multivariate evaluation of surface-enhanced Raman spectra of doxorubicin in plasma was performed. In a principal component analysis (PCA) all spectral features were modelled into three principal components. The major variation of the data was shown to be the variation of doxorubicin Raman signal together with the doxorubicin fluorescence, whereas the variation due to plasma was of minor importance. It was also shown that the surface-enhanced Raman scattering (SERS) measurements were independent on such factors as measurement occasion and silver colloids. The presented results show that with some improvements, quantification of doxorubicin directly in plasma could be possible.

Method evaluation of an enzymatic method for serum creatinine.

Selecting the correct method for routine analysis by 'method evaluation' is an important component of quality assurance. It is a step-wise procedure that evaluates various analytical parameters like accuracy, precision etc of the given method. Finally reference intervals are established for selected population. We evaluated an enzymatic method for serum creatinine. The results show that it is an acceptable method based on the above mentioned criteria.

Dietary patterns and selected anthropometric indices in reproductive age women of a slum in urban: Kurnool.

Dietary intake in pregnant, lactating and non-pregnant nonlactating women of an urban slum in Kurnool, A.P. are about 30% less than the ICMR recommended daily allowances. Heights and weights of the subjects are similar to the National Nutrition Monitoring Bureau's values for slum women and some inter-religious difference is observed in the anthropometry of the subjects.