A case report and literature review of IgA nephropathy presenting as nephrotic syndrome in polycythemia vera.

A 66-year-old non-smoker presented with a 2-week history of new-onset pedal oedema and gross haematuria. On evaluation, he was found to be hypertensive and oedematous with a haemoglobin of 19.1 g/dl, platelet count of 546,000/mm3, and creatinine of 2.6 mg/dl. Urine examination revealed abundant RBCs with 3+ albumin on three separate occasions. His 24-h urine protein level was 3830 mg/day, with a serum cholesterol level of 303 mg/dl. Secondary erythrocytosis and thrombocytosis tests were negative. Bone marrow examination revealed hypercellularity, erythroid hyperplasia, tight clusters of large megakaryocytes, and megakaryocytic hyperplasia suggestive of polycythemia vera. PCR analysis revealed a JAK2V617 F (exon 14) mutation. In view of nephrotic syndrome, azotemia, and microscopic haematuria, a renal biopsy was performed, which revealed features of IgA nephropathy with advanced interstitial fibrosis and tubular atrophy. He was started on angiotensin receptor blockers with hydroxy urea as a part of treatment. This case report highlights the association of glomerular disease with polycythaemia vera and the need of prompt renal biopsy for diagnosis and management.

Analysing uric acid levels to assess the effectiveness of dapagliflozin.

BACKGROUND: Sodium-glucose cotransporter-2 inhibitors are an innovative diabetes treatment that lowers blood sugar levels without insulin. A growing body of evidence suggests that blood sugar levels are tightly correlated with uric acid levels in their blood and urine. To alleviate type 2 diabetes (T2DM) suffering, we tested dapagliflozin on serum and urinary uric acid levels of patients with T2DM and measured its efficacy in reducing uric acid levels. METHODS: A study was conducted on 60 people with T2DM. Patients were treated with Dapagliflozin doses of 10 mg daily for 3 months. Three months later, we measured body weight, fasting, and postprandial blood glucose levels, Hemoglobin A1C (HbA1c), serum lipids, renal function tests, routine urine, and serum uric acid. RESULTS: A number of clinical parameters of T2DM patients were compared to those of healthy subjects of the same age group. A comprehensive analysis of all parameters was conducted to evaluate dapagliflozin's impact. After 90 days of dapagliflozin treatment, serum uric acid levels dropped significantly from 9.0 to 8 mg/dL in the dapagliflozin group, as well as uric acid percentage in urine changed from 16.1 to 23.6 %. After three months of treatment, HbA1C levels decreased from 9.8 % to 8.5 %. CONCLUSION: Following treatment with dapagliflozin, the patients' Homeostatic Model Assessment for Insulin Resistance decreased to 4.0. Further, multivariate correlation analysis showed a correlation of serum uric acid with glycemic profile positively. In conclusion, dapagliflozin lowers uric acid levels and increases insulin sensitivity in diabetic patients to improve their glycemic control.

Efficiency of solar PV panel by the application of coconut fibres saturated by earthen clay pot water.

Performance of the solar cell is hindered at higher operating temperature. Although higher irradiation increases the yield annually, the higher temperature would do a disservice. As temperature increases, the solar cell undergoes a thermal stress owing a way to carrier recombination. It becomes imperative to ease of the thermal stress, thereby running the PV system effectively and achieving the higher yield. An alternative cooling method is applied by cooling both the sides of the PV panel simultaneously. It includes cooling of the panel top surface by allowing a thin film of water to flow. The rear surface is packed with a biomaterial, which is saturated with the constant supply of water. This makes up a cooling unit. Here, the experimental study is carried out on comparison between a conventional PV panel and PVT panel packed with coir pith at the rear surface. The latter is cooled at two different modes, one among the panel by normal water and the other by earthen pot water. The results show that cooling the coir pith using earthen pot water has increased the overall efficiency up to 64%. It is higher than the other two panels, as the temperature of the pot water is 5°-8°C lower than the ambient air.

Development of L-histidine immobilized CIM(®) monolithic disks for purification of immunoglobulin G.

The pseudobiospecific affinity ligand l-histidine was immobilized on epoxy, carbonyldiimidazole (CDI), and ethylenediamine (EDA) convective interaction media (BIA Separations, Slovenia) monolithic disks to obtain the histidyl affinity column for purification of immunoglobulin G (IgG). The kinetics and the mass transfer properties of the affinity columns were studied to determine the optimum buffer condition, flow rate, and concentration of IgG for maximum IgG adsorption. The binding capacities of all the three affinity columns were higher with zwitterionic buffer morpholinopropanesulfonic acid than with charged buffers such as tris-HCl and phosphate buffers, and the optimum pH was 6.5. The interaction of IgG with histidine immobilized CDI and epoxy disks was found to be predominantly driven by ionic interaction, while the interaction with EDA-histidine disk could be partially governed by multiple non-covalent forces of interaction. The maximum binding capacity (Qm ) of l-histidine immobilized on EDA-, CDI-, and epoxy-activated convective interaction media disks were 19.83 ± 0.25, 15.85 ± 0.18 and 12.11 ± 0.17 mg/ml of support, respectively, and the dissociation constant (Kd ) were calculated to be in the micromolar range for all the three histidyl monolithic columns. Purification of IgG from untreated human serum was also attempted, and the results indicate the high potential of this method for purification of total IgG from complex biological sources and also for separation of IgG1 from other subclasses.

Affinity selection of histidine-containing peptides using metal chelate methacrylate monolithic disk for targeted LC-MS/MS approach in high-throughput proteomics.

In recent years, bottom-up approach has become the popular method of choice for large scale analysis of complex proteome samples. Peptide fractionation determines the efficiency of the bottom-up method and often the resolving power of reverse phase liquid chromatography (RPLC) is insufficient for efficient protein identification in case of complex biological samples. To overcome the inherent limitation of proteomics associated with sample complexity, we evaluated fast flow metal chelate methacrylate monolithic system - CIM (Convective Interaction Media) disk chelated with Cu(II) for targeted affinity selection of histidine-containing peptides. Initially the Cu(II)-IMAC using CIM disk was evaluated using tryptic digest of protein mixtures of 8 model proteins and was found to be highly efficient in capturing His-containing peptides with high degree of specificity and selectivity. Further the efficiency of His-peptide enrichment using CIM-IMAC was also demonstrated using complex biological samples like total Escherichia coli cell lysate. The analysis of the Cu(II)-IMAC retained peptides from tryptic digests of model protein mixture and E. coli not only demonstrated a significant reduction in sample complexity but also subsequently enabled the identification of additional peptides. His-peptide enrichment also enabled the identification of low abundant proteins that were not detected in the analysis of total E. coli digest.

Development of bio/blood compatible polypropylene through low pressure nitrogen plasma surface modification.

Surface modification of polypropylene by nitrogen containing plasma was performed in this work in order to improve the wettability which resulted in enhanced biocompatibility and blood compatibility. Various nitrogen containing functional groups as well as oxygen containing functional groups were found to be incorporated to the polymer surface during plasma treatment and post plasma reaction respectively. Wettability of the polymers was evaluated by static contact angle measurement to show the improvement in hydrophilicity of plasma treated polypropylene. Cross linking and surface modification were reported to be dominating in the case of nitrogen plasma treatment compared to degradation. The effect of various process variables namely power, pressure, flow rate and treatment time on surface energy and weight loss was studied at various levels according to the central composite design of response surface methodology (RSM). Except pressure the other variables resulted in increased weight loss due to etching whereas with increasing pressure weight loss was found to increase and then decrease. The effect of process variables on surface morphology of polymers was evaluated by Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM). Well spread fibroblast cells on nitrogen plasma treated polypropylene due to the presence of CO, NH2+ and NH+ was observed. Reduced platelet adhesion and increased partial thromboplastin time evidenced the increased blood compatibility.

Influence of fillers on NR/SBR blends containing ENR-organoclay nanocomposites: morphology and wear.

In this study the Epoxidized natural rubber (ENR) and Organoclay (Cloisite 20A) were prepared by solution mixing process. The obtained nanocomposites were incorporated in Natural rubber (NR) and Styrene butadiene rubber (SBR) blends in presence of varying types of carbon black as reinforcing fillers. Morphology, curing characteristics, mechanical and thermal properties were characterized and analyzed. Also, the wear characteristics of the nanocomposites against Du-Pont abrader, DIN abrader and different mining rock surfaces were determined and discussed. The morphology of the organoclay incorporated in ENR shows a highly intercalated structure. ISAF type of carbon black shows a significant effect on curing and mechanical properties by reacting at the interface between SBR and NR matrix. All the samples show only one melting peak at same temperature on the DSC curve. Blends containing ISAF N234 type of carbon black shows high abrasion resistant properties against Du-Pont abrader, DIN abrader and different mining rock. The rubber compound containing 70 wt% of NR, 30 wt% of SBR and 10 wt% of ENR/nanoclay with ISAF N231 are found to be the toughest rubber against all types of rock under the present study.

Characterization of metal chelate methacrylate monolithic disk for purification of polyclonal and monoclonal immunoglobulin G.

Dynamic binding capacity (DBC) of commercial metal-chelate methacrylate monolith-convective interaction media (CIM) was performed with commercial human immunoglobulin G (IgG) (Cohn fraction II, III). Monoliths are an attractive stationary phase for purification of large biomolecules because they exhibit very low back pressure even at high flow rates and flow-unaffected binding properties. Adsorption of IgG onto CIM-IDA disk immobilized with Cu(2+), Ni(2+) and Zn(2+) were studied with Tris-acetate (TA), phosphate-acetate (PA) and MMA (MES, MOPS and acetate) buffer systems at different flow rates. Adsorption and elution of IgG varied with different buffers and adsorption of IgG was maximum with MMA buffer. Adsorption of human IgG from Cohn fractions (II, III) was high when Cu(2+) was used as ligand. CIM-IDA disk showed dynamic binding capacity in the range of 14-16 mg/ml with Cu(2+) and 7-9 mg/ml with Ni(2+) for human IgG with MMA buffer. In the case of CIM-IDA-Zn(2+) column, the binding capacity was only about 0.5mg/ml of support. Different desorption strategies like lowering of pH and increasing of competitive agent were also studied to achieve maximum recovery. Chromatographic runs with human serum and mouse ascites fluid were also carried out with metal chelate methacrylate monolithic disk and the results indicate the potential of this technique for polyclonal human IgG and monoclonal IgG purification from complex biological samples.

Antigen-dependent selection of T cells that are able to efficiently regulate free cytoplasmic Ca2+ levels.

In T helper cells, the process of memory acquisition is reflected in the expression of phenotypic markers. However, little is known regarding the functional changes that occurred in T helper cells selected after a primary Ag-specific response. We now present data that indicate that such T helper cells acquire the ability to down-regulate high concentrations of free cytoplasmic Ca2+. This property renders them resistant to intense inductive stimuli, such as high concentrations of ionomycin. The accumulation of cells that display this ability parallels the progressive proliferative enrichment of Ag-specific T cells and correlates with the surface expression of the CD45RB(low) isoform. As previously shown, persistent high levels of cytoplasmic Ca2+ are responsible for death via apoptosis in virgin T cells. In contrast, the ability to regulate cytosolic Ca2+ allows survival and clonal expansion. We suggest that memory T cells behave differently from virgin T cells when interacting with Ag-presenting B cells (as reported by others) because they have acquired this new physiologic property. Ag-presenting B cells deliver an intense stimulatory signal to T cells because of a high multiplicity of cognate interactions. Thus, Ag-driven T cell proliferation results in the selection of "resistant" T helper cells that can be successfully stimulated by memory B cells. In contrast, naive T cells, which cannot modulate high levels of cytosolic Ca2+, are deleted as a consequence of Ag presentation by B cells.

Selection of pulmonary CD4-, CD8-, alpha beta+ T cells expressing V beta 8 T cell receptors.

The double negative (CD4-CD8-) alpha beta+ T cells constitute about 20% of all alpha beta+ T cells in murine lungs. We find that in BALB/c mice 60% of the double negative alpha beta+ pulmonary T cells express receptors of the V beta 8 family whereas only 33% of single positive (CD4+/CD8+) pulmonary T cells express V beta 8. However, in C57BL/6 mice, equal frequencies (25%) of double negative and single positive alpha beta+ pulmonary T cells express V beta 8. The high frequency of double negative V beta 8+ pulmonary T cells is dominantly inherited in (C57BL/6 X BALB/c) F1 offsprings. Further studies exclude the involvement of classic MHC region genes in determining the level of V beta 8 usage among double negative pulmonary T cells. Upon exposure to mycobacterial antigens, double negative alpha beta+ pulmonary T cells are co-enriched in vitro in parallel with gamma delta+ T cells.

In Mls-1a mice, fetal-type beta-gene rearrangements are frequent among self-anergic V beta 6 T cells.

T lymphocytes generated in the fetal and neonatal period are characterized by T cell receptor (TCR) gene rearrangements that lack N region nucleotides (fetal-type TCR). Using fetal-type TCR as a lineage marker, we show that such T cells are long-lived and persist in the periphery of adult mice. Moreover, in both neonatal and adult environments, upon encounter with self-antigens, they are less likely to be deleted. Inefficient clonal deletion could be due to the intrinsic properties of the T cells generated during this period, or to yet unknown properties of the perinatal thymus. Such anergic T cells constitute a subset that can further expand in vivo in an antigen-independent fashion, leaving open the possibility for self-aggression under the appropriate triggering conditions.

Selective proliferation of gamma delta T lymphocytes exposed to high doses of ionomycin.

The alpha beta T cell repertoire is primarily shaped in the thymus. However, extrathymic positive selection has been demonstrated for many gamma delta T cell clonotypes. This latter type of selection is the result of a peripheral clonal expansion which could be facilitated by special physiological properties of gamma delta T cells, distinguishing them from most alpha beta T cells. In studying the behavior of T cells under conditions of polyclonal activation, we noticed a differential sensitivity between alpha beta and gamma delta T cells to strong stimulatory signals. When induced with high doses of ionomycin, a large fraction of peripheral gamma delta T cells and a small fraction of alpha beta T cells are able to proliferate exponentially while most alpha beta T cells die. This phenomenon appears to be related to intracellular regulation of high concentrations of cytosolic Ca2+. The ability to proliferate under strong stimulatory conditions is a striking feature of many peripheral gamma delta T cells but not of gamma delta thymocytes. In general, T cells selected in the periphery by clonal expansion might be characterized by resistance to strong stimuli and typically, by their ability to "handle" higher concentrations of free cytoplasmic calcium.

Self heat shock and gamma delta T-cell reactivity.

We have investigated the effects of heat shock on T-cell induction and selection in vitro. We find that when cell preparations containing T lymphocytes are incubated for 30 min at 42 degrees C, a selective proliferation of gamma delta + T cells bearing the gamma delta T-cell antigen receptor follows. A greater enrichment of gamma delta + T cells is observed, upon preexposure to mycobacterial antigens in vivo. By comparing the effects of heat shock with that of mitogen or specific T-cell triggering by conventional antigens and by analyzing the gamma delta T-cell receptor genes expressed in cells that proliferate as a result of heat shock induction, we conclude that a subset of murine gamma delta T cells react to antigens on self cells in which a heat shock response was induced.

Regulation of growth and differentiation of pre-activated B lymphocytes.

The differential effects of H-2 IAk-specific T helper cells, their soluble factors and Sepharose-coupled anti-mu antibodies on the growth and differentiation of pre-activated B cells were studied. It was found that the prolonged growth of pre-activated B cells required activation signals from major histocompatibility complex (MHC)-restricted T helper cells or Sepharose-coupled anti-mu antibodies. The MHC-restricted T helper cells induced both prolonged growth and differentiation of activated B cells. Anti-mu antibodies together with T helper cell-derived soluble factors induced prolonged growth of activated B cells, but no differentiation into Ig secretion was detected. The inhibition of Ig secretion in anti-mu cultures could be overcome to some extent by either MHC-restricted T helper cells or lipopolysaccharide together with soluble factors. It was also observed that T helper cell interactions were needed for long-term in vitro culture of pre-activated B lymphocytes.

Heterogeneity in the specificities of an alloreactive T helper cell line.

Heterogeneity in antigen recognition of an alloreactive helper T-cell line was studied by repeated limiting dilution cloning. Analysis of the fine specificities of these clones showed that the gain of reactivity to new antigens by the T-cell line included the generation of non-cross-reactive T cells specific to new major histocompatibility (MHC) antigens. Analysis of function revealed that all the T-cell clones studied had similar cell surface antigens and secreted interleukin 4 (IL-4) and IL-5/IL-3 but not IL-2 or gamma interferon (IFN-gamma). The T-cell clones served as helper cells for B cells expressing the appropriate MHC antigens to both clonal proliferation and differentiation into antibody-producing cells.

Prolonged growth of activated B lymphocytes requires interaction with helper T cells.

Resting B lymphocytes acquire upon activation responsiveness to soluble factors that support their growth and differentiation. We have studied the growth requirements of anti-mu pre-activated B lymphocytes in vitro. We found that activated B lymphocytes did respond to soluble factors from helper T cells by entering the cell cycle but cell-cell contact with specific helper T cells was required for prolonged growth of activated B lymphocytes. Thus, activated B lymphocytes must receive activation signals from helper T cells in order to stay in the proliferative phase of the cell cycle.

HLA antigens in South India: II. Selected caste groups of Tamil Nadu.

HLA-A, B antigen and haplotype frequencies were studied in four different caste groups of Tamil Nadu living in Madurai. A total number of 101 Nadars, 36 Kallars, 54 Iyers and 57 Telugu-speaking Naidus were studied. HLA A3 and B15 were significantly higher in Nadars; A10 & B8 in Kallars and Aw19, B12 & B35 in Iyers. HLA A-B haplotypes A10-B7, A28-B17 & A24-B- were characteristic of Nadars; A10-B8 & A1-B-, Kallars; Aw19-B12 & A1-B15, Iyers and A2-B-, Naidus. Negative linkage disequilibria for Aw19-B7, A28-B15 & A9-B51 were significant in Nadars; A1-B5, A1-B12 & Aw19-B- in Iyers and A2-B17 in Naidus. Heterogeneity chi-square based on antigen frequency and genetic distance also suggest the heterogeneous nature of the population of South India. Will these caste groups with such diverse haplotypic combinations differ from one another in their immune response and susceptibility to a given epidemic or infection?