Risk Factors for Relapse in Childhood Nephrotic Syndrome.

Minimal change nephrotic syndrome is the most common cause of the glomerular disease in children. Despite its high initial response to corticosteroids, relapses are common leading to increased morbidity. Therefore, appraisal of common triggers of relapse becomes an imperative for successful management of childhood nephrotic syndrome. The objective of the study was to identify the risk factors for frequent relapse in childhood nephrotic syndrome. This descriptive cross-sectional study was conducted in the department of Paediatrics, Rangpur Medical College Hospital, Bangladesh from January 2013 to December 2014. A total of 60 patients with relapsed idiopathic childhood nephrotic syndrome fulfilling the selection criteria were enrolled into the study. Serum total protein, serum albumin and urine culture were done. The following potential risk factors for relapse were analyzed using Chi-square test: age at onset of NS, sex, socioeconomic status, atopy, short duration of initial therapy with steriod, serum total protein, serum albumin level and infection. Among 60 patients, short duration of therapy with steriod (<8 weeks) for first attack was significantly higher among frequent relapse (FRNS) cases compared with infrequent relapse (IFRNS) group (63% vs. 27.27%). FRNS group had initial brief period of remission of <6 months, (p=0.04). Difference in mean serum albumin and that of serum total protein between FRNS and IFRNS cases was (1.0±0.66 vs. 2.66±1.6gm/dl) (p<0.0001) and (3.5±1.2 vs. 6.5±2.12gm/dl) (p<0 .0001) respectively. This study concluded that short initial therapy with corticosteroid, brief duration of initial remission; low serum total protein and decreased serum albumin were the risk factors for frequent relapses.

Multiple metals and agricultural use affects oxidative stress biomarkers in freshwater Aegla crabs / Presença de múltiplos metais e agricultura afetam biomarcadores de estresse oxidativo em caranguejos de água doce (Aegla)

Metals and agrochemicals are among the main aquatic contaminants, being able to trigger oxidative stress in exposed organisms. The objective of this work was to evaluate the correlation between the level of oxidative stress biomarkers in Aegla crabs (Crustacea, Anomura) with (i) the set of metals present in the streams sediment and (ii) with land uses of three hydrographic basins. The study was carried out in streams (≤ 2nd order) of hydrographic basins in southern Brazil (Basins of Rio Suzana, Rio Ligeirinho-Leãozinho and Rio Dourado). In these streams were quantified the land uses and Cu, Cr, Cd, Fe, Mn and Zn concentrations in the sediment. The enzymes Catalase (CAT) and Glutathione Reductase (GR), as well as the level of membrane lipid peroxidation (TBARS), were analyzed in adult females. The PCA analysis showed that the distribution of metals was different between the basins. Cd, Cr and Fe were correlated positively with CAT and negatively with TBARS and GR. The Dourado basin had the lowest concentrations of these three metals and the highest levels of TBARS. However, in Dourado basin there is predominance of agriculture land use, and TBARS was positively correlated with agricultural land use. Besides in Dourado basin, GR activity was higher than in the others basins, indicating a compensatory response in relation to CAT inhibition. The basins of Suzana and Ligeirinho-Leãozinho rivers had lower TBARS values, which may be due to the induction of CAT in response to metals accumulated in sediment. In summary, this work indicates that in the basins with a higher concentration of toxic metals there is an adaptive response of CAT induction, which reduces TBARS in Aegla. On the other hand, in the basin with lower metallic contamination, TBARS occurrence was primarily influenced by agricultural land use.

Os metais e agroquímicos estão entre os principais contaminantes aquáticos, podendo desencadear estresse oxidativo em organismos expostos. O objetivo deste trabalho foi avaliar uma possível correlação entre o nível de biomarcadores de estresse oxidativo em Aegla (Crustacea, Anomura) com (i) o conjunto de metais presentes no sedimento e (ii) com os usos da terra, em três bacias hidrográficas distintas. O estudo foi realizado em riachos(≤ 2ª ordem) de bacias hidrográficas do Sul do Brasil (Bacias do Rio Suzana, do Rio Ligeirinho-Leãozinho e do Rio Dourado), as quais foram caracterizadas em função do percentual de usos da terra e do nível de Cu, Cr, Cd, Fe, Mn e Zn no sedimento. As enzimas Catalase (CAT) e Glutationa Redutase (GR), bem como o nível de peroxidação lipídicadas membranas (TBARS), foram analisadas em fêmeas adultas. Uma análise de PCA mostrou que a distribuição de metais foi distinta entre as bacias. Cd, Cr e Fe no sedimento correlacionaram positivamente com a CAT e negativamente com TBARS e GR. Entretanto, a bacia do Dourado apresentou os menores níveis destes três metais e os maiores níveis de TBARS, o que pode ser justificado pelo predomínio da agricultura nesta bacia, já que o TBARS correlacionou positivamente com o percentual de uso agrícola. Nesta bacia, a atividade da GR foi mais alta do que nas outras, indicando uma resposta compensatória em relação a inibição da CAT. As bacias do rio Suzana e rio Ligeirinho-Leãozinho apresentaram valores menores de TBARS, o que pode decorrer da indução da CAT em função dos metais acumulados no sedimento. Em síntese, este trabalho indica que nas bacias com maior concentração de metais tóxicos ocorre uma resposta adaptativa de indução da CAT, o que reduz os níveis de TBARS em Aegla. Por outro lado, na bacia com menor contaminação metálica os níveis de TBARS foram primariamente influenciados pelo uso agrícola.

Multiple metals and agricultural use affects oxidative stress biomarkers in freshwater Aegla crabs

Abstract Metals and agrochemicals are among the main aquatic contaminants, being able to trigger oxidative stress in exposed organisms. The objective of this work was to evaluate the correlation between the level of oxidative stress biomarkers in Aegla crabs (Crustacea, Anomura) with (i) the set of metals present in the streams sediment and (ii) with land uses of three hydrographic basins. The study was carried out in streams ( 2nd order) of hydrographic basins in southern Brazil (Basins of Rio Suzana, Rio Ligeirinho-Leãozinho and Rio Dourado). In these streams were quantified the land uses and Cu, Cr, Cd, Fe, Mn and Zn concentrations in the sediment. The enzymes Catalase (CAT) and Glutathione Reductase (GR), as well as the level of membrane lipid peroxidation (TBARS), were analyzed in adult females. The PCA analysis showed that the distribution of metals was different between the basins. Cd, Cr and Fe were correlated positively with CAT and negatively with TBARS and GR. The Dourado basin had the lowest concentrations of these three metals and the highest levels of TBARS. However, in Dourado basin there is predominance of agriculture land use, and TBARS was positively correlated with agricultural land use. Besides in Dourado basin, GR activity was higher than in the others basins, indicating a compensatory response in relation to CAT inhibition. The basins of Suzana and Ligeirinho-Leãozinho rivers had lower TBARS values, which may be due to the induction of CAT in response to metals accumulated in sediment. In summary, this work indicates that in the basins with a higher concentration of toxic metals there is an adaptive response of CAT induction, which reduces TBARS in Aegla. On the other hand, in the basin with lower metallic contamination, TBARS occurrence was primarily influenced by agricultural land use.

Resumo Os metais e agroquímicos estão entre os principais contaminantes aquáticos, podendo desencadear estresse oxidativo em organismos expostos. O objetivo deste trabalho foi avaliar uma possível correlação entre o nível de biomarcadores de estresse oxidativo em Aegla (Crustacea, Anomura) com (i) o conjunto de metais presentes no sedimento e (ii) com os usos da terra, em três bacias hidrográficas distintas. O estudo foi realizado em riachos ( 2ª ordem) de bacias hidrográficas do Sul do Brasil (Bacias do Rio Suzana, do Rio Ligeirinho-Leãozinho e do Rio Dourado), as quais foram caracterizadas em função do percentual de usos da terra e do nível de Cu, Cr, Cd, Fe, Mn e Zn no sedimento. As enzimas Catalase (CAT) e Glutationa Redutase (GR), bem como o nível de peroxidação lipídica das membranas (TBARS), foram analisadas em fêmeas adultas. Uma análise de PCA mostrou que a distribuição de metais foi distinta entre as bacias. Cd, Cr e Fe no sedimento correlacionaram positivamente com a CAT e negativamente com TBARS e GR. Entretanto, a bacia do Dourado apresentou os menores níveis destes três metais e os maiores níveis de TBARS, o que pode ser justificado pelo predomínio da agricultura nesta bacia, já que o TBARS correlacionou positivamente com o percentual de uso agrícola. Nesta bacia, a atividade da GR foi mais alta do que nas outras, indicando uma resposta compensatória em relação a inibição da CAT. As bacias do rio Suzana e rio Ligeirinho-Leãozinho apresentaram valores menores de TBARS, o que pode decorrer da indução da CAT em função dos metais acumulados no sedimento. Em síntese, este trabalho indica que nas bacias com maior concentração de metais tóxicos ocorre uma resposta adaptativa de indução da CAT, o que reduz os níveis de TBARS em Aegla. Por outro lado, na bacia com menor contaminação metálica os níveis de TBARS foram primariamente influenciados pelo uso agrícola.

Multiple metals and agricultural use affects oxidative stress biomarkers in freshwater Aegla crabs / Presença de múltiplos metais e agricultura afetam biomarcadores de estresse oxidativo em caranguejos de água doce (Aegla)

Metals and agrochemicals are among the main aquatic contaminants, being able to trigger oxidative stress in exposed organisms. The objective of this work was to evaluate the correlation between the level of oxidative stress biomarkers in Aegla crabs (Crustacea, Anomura) with (i) the set of metals present in the streams sediment and (ii) with land uses of three hydrographic basins. The study was carried out in streams (≤ 2nd order) of hydrographic basins in southern Brazil (Basins of Rio Suzana, Rio Ligeirinho-Leãozinho and Rio Dourado). In these streams were quantified the land uses and Cu, Cr, Cd, Fe, Mn and Zn concentrations in the sediment. The enzymes Catalase (CAT) and Glutathione Reductase (GR), as well as the level of membrane lipid peroxidation (TBARS), were analyzed in adult females. The PCA analysis showed that the distribution of metals was different between the basins. Cd, Cr and Fe were correlated positively with CAT and negatively with TBARS and GR. The Dourado basin had the lowest concentrations of these three metals and the highest levels of TBARS. However, in Dourado basin there is predominance of agriculture land use, and TBARS was positively correlated with agricultural land use. Besides in Dourado basin, GR activity was higher than in the others basins, indicating a compensatory response in relation to CAT inhibition. The basins of Suzana and Ligeirinho-Leãozinho rivers had lower TBARS values, which may be due to the induction of CAT in response to metals accumulated in sediment. In summary, this work indicates that in the basins with a higher concentration of toxic metals there is an adaptive response of CAT induction, which reduces TBARS in Aegla. On the other hand, in the basin with lower metallic contamination, TBARS occurrence was primarily influenced by agricultural land use.

Os metais e agroquímicos estão entre os principais contaminantes aquáticos, podendo desencadear estresse oxidativo em organismos expostos. O objetivo deste trabalho foi avaliar uma possível correlação entre o nível de biomarcadores de estresse oxidativo em Aegla (Crustacea, Anomura) com (i) o conjunto de metais presentes no sedimento e (ii) com os usos da terra, em três bacias hidrográficas distintas. O estudo foi realizado em riachos (≤ 2ª ordem) de bacias hidrográficas do Sul do Brasil (Bacias do Rio Suzana, do Rio Ligeirinho-Leãozinho e do Rio Dourado), as quais foram caracterizadas em função do percentual de usos da terra e do nível de Cu, Cr, Cd, Fe, Mn e Zn no sedimento. As enzimas Catalase (CAT) e Glutationa Redutase (GR), bem como o nível de peroxidação lipídica das membranas (TBARS), foram analisadas em fêmeas adultas. Uma análise de PCA mostrou que a distribuição de metais foi distinta entre as bacias. Cd, Cr e Fe no sedimento correlacionaram positivamente com a CAT e negativamente com TBARS e GR. Entretanto, a bacia do Dourado apresentou os menores níveis destes três metais e os maiores níveis de TBARS, o que pode ser justificado pelo predomínio da agricultura nesta bacia, já que o TBARS correlacionou positivamente com o percentual de uso agrícola. Nesta bacia, a atividade da GR foi mais alta do que nas outras, indicando uma resposta compensatória em relação a inibição da CAT. As bacias do rio Suzana e rio Ligeirinho-Leãozinho apresentaram valores menores de TBARS, o que pode decorrer da indução da CAT em função dos metais acumulados no sedimento. Em síntese, este trabalho indica que nas bacias com maior concentração de metais tóxicos ocorre uma resposta adaptativa de indução da CAT, o que reduz os níveis de TBARS em Aegla. Por outro lado, na bacia com menor contaminação metálica os níveis de TBARS foram primariamente influenciados pelo uso agrícola.

Comparison of WHO Clinical Criteria and Radiological Findings of Pneumonia in Children Aged 02-59 Months.

Despite recent advances, pneumonia contributes substantially to childhood mortality in low and middle-income countries. To reduce case fatality, World Health Organization (WHO) adopted a working formula to identify pneumonia clinically in resource constrained setting. Chest X-ray is the gold standard test to diagnose pneumonia but this tool is not readily available or affordable in primary health facility in developing countries even when it is indicated. Objective of the study was to compare WHO clinical criteria and radiological findings of pneumonia in children aged 02-59 months. This cross-sectional observational study was conducted at inpatient department of Paediatrics of Rangpur Medical College Hospital from July 2015 to June 2017. Total 112 patients aged 2 to 59 months fulfilling selection criteria were enrolled into this study. Data were collected and chest X-ray was done for each patient. Data were analyzed through SPSS software (version 16.0). Minimum level of significance was predetermined as p<0.05. Among 112 patients, mean age of chest radiograph positive cases of pneumonia was 7.64±7.08 months and that of negative cases 10.75±10.95 months. There was no statistically significant difference of age (p=0.083) between chest radiography positive and negative cases. Radiological findings were positive in 52(46.43%) cases and negative in 60(53.57%). Fever, cough and fast breathing were present in all patients with sensitivity 100%. Chest indrawing was significantly more (p=0.003) among 52(46.43%) cases who demonstrated positive radiological findings with sensitivity 63.46% (95% CI = 48.96% to 76.37%). There was no statistically significant difference of fever, cough and fast breathing between radiograph positive and negative cases. Chest indrawing was significantly more in radiograph positive cases. The results highlighted the importance of adoption of positive radiograph of WHO guideline to identify pneumonia.

Multiple metals and agricultural use affects oxidative stress biomarkers in freshwater Aegla crabs.

Metals and agrochemicals are among the main aquatic contaminants, being able to trigger oxidative stress in exposed organisms. The objective of this work was to evaluate the correlation between the level of oxidative stress biomarkers in Aegla crabs (Crustacea, Anomura) with (i) the set of metals present in the streams sediment and (ii) with land uses of three hydrographic basins. The study was carried out in streams (≤ 2nd order) of hydrographic basins in southern Brazil (Basins of Rio Suzana, Rio Ligeirinho-Leãozinho and Rio Dourado). In these streams were quantified the land uses and Cu, Cr, Cd, Fe, Mn and Zn concentrations in the sediment. The enzymes Catalase (CAT) and Glutathione Reductase (GR), as well as the level of membrane lipid peroxidation (TBARS), were analyzed in adult females. The PCA analysis showed that the distribution of metals was different between the basins. Cd, Cr and Fe were correlated positively with CAT and negatively with TBARS and GR. The Dourado basin had the lowest concentrations of these three metals and the highest levels of TBARS. However, in Dourado basin there is predominance of agriculture land use, and TBARS was positively correlated with agricultural land use. Besides in Dourado basin, GR activity was higher than in the others basins, indicating a compensatory response in relation to CAT inhibition. The basins of Suzana and Ligeirinho-Leãozinho rivers had lower TBARS values, which may be due to the induction of CAT in response to metals accumulated in sediment. In summary, this work indicates that in the basins with a higher concentration of toxic metals there is an adaptive response of CAT induction, which reduces TBARS in Aegla. On the other hand, in the basin with lower metallic contamination, TBARS occurrence was primarily influenced by agricultural land use.

Efficacy of F-75 & F-100 Recipes in theTreatment of Severe Acute Malnutrition: A Randomized Controlled Trial.

Severe malnutrition is an important cause of death in children. According to new WHO-growth chart 2006, the proportion of children with severe wasting is 3.1% thus the total number being 4, 65000 (BDHS 2014). Overall risk of death among children with severe acute malnutrition (SAM) is 9 times more than well nourished children. The death rate among hospitalized children of SAM was as high as 15%. Once properly treated, severely malnourished children would grow up leading a normal life. Severe malnutrition in children can be successfully treated by using WHO guidelines that have been shown to be feasible and sustainable even in small district hospital with limited resources. A randomized controlled trial was conducted at department of Pediatrics, Dhaka Medical College Hospital, Dhaka, Bangladesh from July 2014 to September 2015. The trial enrolled 92 SAM patients (46 cases + 46 controls) aged 06 months to 59 months of either sex who meet the inclusion criteria consecutively. Enrolled children were randomized by lottery method into two groups, Group I and Group II. Patients in Group I was treated with F-75 and F-100 recipes and managed in two phases, initial stabilization phase with F-75 recipes then subsequently rehabilitation phase by F-100 recipes. Patients in Group II was treated with prepackaged F-75 & F-100 formulae and feeding was given in two phases i.e. initial stabilization phase and subsequently rehabilitation phase according to national guidelines for the management of severe malnourished children. Then play therapy was given for half an hour daily with red colored toy in Ashic play centre Dhaka Medical College Hospital for patients of both groups. The time (days) taken to return of appetite (mean±SD) was 5.1±1.16 and 4.8±1.34 in Group I and Group II respectively, disappearance of edema (mean±SD) 4.8±1.53 in and 4.9±1.05 for Group I Group II respectively, to gain target weight (mean±SD) 13.8±2.20 days and 13.5±1.74 days in Group I and Group II respectively, rate of weight gain (mean±SD) 17.70±7.07gm/kg/day for Group I and 16.20±4.63gm/kg/day for Group II. The side effects, diarrhea was equal in both group, vomiting was more in Group II, combined diarrhea and vomiting was more in Group I but the differences were not statistically significant. The treatment cost (mean±SD) was higher in Group II (97.2±78.24 BDT/child/day) than in Group I (58.5±54.36 BDT/child/day). Return of appetite and disappearance of oedema and target weight gain were similar in both groups but treatment cost was higher in Group II than Group I, which was statistically significant.

Diagnostic Performance of RBC Indices in Beta Thalassaemia Trait.

Beta thalassaemia trait is one of the most common forms of thalassaemia in Bangladesh. This clinical entity usually remains undiagnosed because they present with mild anaemia or sometimes no anaemia. Determination of trait is the mainstay of prevention of thalassaemia. RBC indices can provide valuable diagnostic tool for beta thalassaemia trait. This was a hospital based cross-sectional, analytical study, done at Department of Paediatrics, Rangpur Medical College Hospital, Rangpur, Bangladesh from July 2014 to June 2016. A total of 150 anaemic children, aged 1-15 years from inpatient and outpatient department were included by purposive sampling. Blood samples were collected and sent to the Department of Biochemistry, Rangpur Medical College to determine the value of RBC indices. Out of 150 anaemic children 37(24.66%) children were diagnosed as E-trait, 9(9%) E-disease 6(4%) as E-Beta thalassaemia, 1(0.66%) had error in the report, a total of 53 children were excluded from the study. Finally, 32(21.3%) children, diagnosed as beta thalassaemia trait and 65(43.33%) children, diagnosed as normal Hb variant, so a total of 97 children were enrolled in this study. Age of the children was 1-15 years. Their mean age was 6.4 years. Male female ratio was 1.15:1. There was significant difference of HbA2 between BTT and non BTT group of participants (p value <0.001). Value of MCV and MCH were significantly greater in BTT than non BTT group (p value <0.05). Sensitivity, Specificity, Positive predictive value, Negative predictive value and Youden index of MCV were 81.25%, 90.76%, 81%, 90% and 71 respectively. Sensitivity, Specificity, Positive predictive value, Negative predictive value and Youden index of MCH were 84.37%, 92.30%, 84%, 92% and 76 respectively.

Shape and size constraints on dust optical properties from the Dome C ice core, Antarctica.

Mineral dust aerosol (dust) is widely recognized as a fundamental component of the climate system and is closely coupled with glacial-interglacial climate oscillations of the Quaternary period. However, the direct impact of dust on the energy balance of the Earth system remains poorly quantified, mainly because of uncertainties in dust radiative properties, which vary greatly over space and time. Here we provide the first direct measurements of the aerosol optical thickness of dust particles windblown to central East Antarctica (Dome C) during the last glacial maximum (LGM) and the Holocene. By applying the Single Particle Extinction and Scattering (SPES) technique and imposing preferential orientation to particles, we derive information on shape from samples of a few thousands of particles. These results highlight that clear shape variations occurring within a few years are hidden to routine measurement techniques. With this novel measurement method the optical properties of airborne dust can be directly measured from ice core samples, and can be used as input into climate model simulations. Based on simulations with an Earth System Model we suggest an effect of particle non-sphericity on dust aerosol optical depth (AOD) of about 30% compared to spheres, and differences in the order of ~10% when considering different combinations of particles shapes.

Heat shock protein bystander antigens for peptide immunotherapy in autoimmune disease.

Mucosal administration of an antigen eliciting bystander suppression at the site of inflammation results in effective antigen-specific immunotherapy for autoimmune diseases. Heat shock proteins are bystander antigens that are effective in peptide-specific immunotherapy in both experimental and human autoimmune disease. The efficacy of preventive peptide immunotherapy is increased by enhancing peptide-specific immune responses with proinflammatory agents. Combining peptide-specific immunotherapy with general suppression of inflammation may improve its therapeutic effect.

Translational medicine 2.0: from clinical diagnosis-based to molecular-targeted therapies in the era of globalization.

Translational medicine encompasses the itinerary from the conception of an idea to its implementation in the diagnosis, prognosis, and treatment of human diseases. It cannot be identified as a stand-alone field; rather, it comprises a fragmented patchwork of activities, competencies, and knowledge distributed along diverse and often disparate disciplines. In standard practice, the success of a translational project is often less dependent on the validity of the original idea than on the outcome of the combination of several stochastic events.

Heat shock proteins (HSP) for immunotherapy of rheumatoid arthritis (RA).

Rheumatoid arthritis (RA) is an inflammatory disease that primarily involves the joints and has a worldwide prevalence of about one percent, with a female to male ratio of 3:1. This chapter summarizes some of the recent progress in molecular immunology, and discusses the application of this new knowledge for therapeutic purposes. We focus on our recent experiences and that of others in modulation of antigen specific responses as a tool for manipulating autoimmune inflammation. Particular emphasis is given to the concept of exploiting for therapeutic purposes a natural mechanism of immune regulation. This mechanism is based on sequential cross recognition of bacterial and human derived heat shock protein peptides.

A genomewide screen in multiplex rheumatoid arthritis families suggests genetic overlap with other autoimmune diseases.

Rheumatoid arthritis (RA) is an autoimmune/inflammatory disorder with a complex genetic component. We report the first major genomewide screen of multiplex families with RA gathered in the United States. The North American Rheumatoid Arthritis Consortium, using well-defined clinical criteria, has collected 257 families containing 301 affected sibling pairs with RA. A genome screen for allele sharing was performed, using 379 microsatellite markers. A nonparametric analysis using SIBPAL confirmed linkage of the HLA locus to RA (P < .00005), with lambdaHLA = 1.79. However, the analysis also revealed a number of non-HLA loci on chromosomes 1 (D1S235), 4 (D4S1647), 12 (D12S373), 16 (D16S403), and 17 (D17S1301), with evidence for linkage at a significance level of P<.005. Analysis of X-linked markers using the MLOD method from ASPEX also suggests linkage to the telomeric marker DXS6807. Stratifying the families into white or seropositive subgroups revealed some additional markers that showed improvement in significance over the full data set. Several of the regions that showed evidence for nominal significance (P < .05) in our data set had previously been implicated in RA (D16S516 and D17S1301) or in other diseases of an autoimmune nature, including systemic lupus erythematosus (D1S235), inflammatory bowel disease (D4S1647, D5S1462, and D16S516), multiple sclerosis (D12S1052), and ankylosing spondylitis (D16S516). Therefore, genes in the HLA complex play a major role in RA susceptibility, but several other regions also contribute significantly to overall genetic risk.

Artificial antigen-presenting cells as a tool to exploit the immune 'synapse'.

Recent progress in molecular medicine has provided important tools to identify antigen-specific T cells. In most cases, the approach is based on oligomeric combinations of recombinant major histocompatibility complex-peptide complexes fixed to various rigid supports available for binding by the T-cell receptor. These tools have greatly increased our insight into mechanisms of immune responses mediated by CD8+ T cells. Examples of the diverse fields of application for this technology include immunization, viral infections and oral tolerance induction.

Genetic immunization for the recovery and purification of recombinant proteins.

A system that uses genetic immunization for recombinant protein recovery and purification is described. The genetic sequence encoding a target protein is subcloned into both a eukaryotic and a prokaryotic vector. With the eukaryotic construct, a rabbit is genetically immunized and specific polyclonal antibodies to the encoded protein raised. The prokaryotic construct is used for bacterial transformation and expression of recombinant protein. Recovery and purification of target recombinant protein are obtained by passing the lysate of expressing bacteria through an immunoaffinity column prepared with the polyclonal antibodies raised in the genetically immunized animal. This method allows purification of recombinant protein without fusion tails and can be applied to purify any protein whose encoding genetic sequence is known.

Cell-mediated DNA transport between distant inflammatory sites following intradermal DNA immunization in the presence of adjuvant.

After intradermal genetic immunization, naked DNA is transported from the site of injection to regional lymph nodes. Little is known on how inflammation influences this process and whether DNA is transported beyond local lymph nodes. In the experiments herein reported, we injected naked DNA in the presence of adjuvant to address questions related to 1) the fate of naked DNA in the presence of inflammation; 2) the generation of immune responses to the encoded protein during inflammation; and, more in general, 3) the fate of ingested molecules beyond regional lymph nodes during inflammation. Two sites of inflammation were induced in vivo in mice. Naked DNA was injected in the nape together with adjuvant, and adjuvant only was injected at a distant peritoneal site. Injected DNA, uptaken at the primary dermal site of inflammation, was transported beyond regional lymph nodes to distant organs such as the spleen and to the distant peritoneal site of inflammation. This transport, mediated by CD11b+ cells, was cumulative during chronic inflammation. These results indicate a novel route of transport of DNA beyond regional lymph nodes and may have specific implications for DNA-based immune modulation.

Ontogeny of synonymous T cell populations with specificity for a self MHC epitope mimicked by a bacterial homologoue: an antigen-specific T cell analysis in a non-transgenic system.

By means of a novel technique for identification and isolation of MHC class II-restricted antigen-specific T cells, we describe here in non-transgenic BALB / c mice physiological positive selection of an oligoclonal population of T cells which recognizes both a self MHC-derived peptide (Ialpha52) and a bacterial homologoue (Hi15). The results support a model for self peptide-mediated generation of T cells which have specificity for microbial antigens through molecular mimicry. This mechanism may be a model for the ontogeny of a physiological T cell response to infectious agents. Loss of control of these circuits may be part of the inciting factors of autoimmunity.

Isolation of an IgG monoclonal anti-dnaJ antibody from an immunoglobulin combinatorial library from a patient with rheumatoid arthritis.

OBJECTIVE: Previously, we showed that rheumatoid arthritis (RA) had both antibodies and T cells specific for the QKRAA-encompassing Escherichia coli dnaJ protein. These findings suggest that the bacteria induced anti-dnaJ responses may cross react with the human homolog of bacterial dnaJ in the joint, resulting in tissue damage. METHODS: We used the combinatorial library technique to isolate and characterize an IgG monoclonal anti-dnaJ antibody (designated CG1) from the blood of a patient with RA. RESULTS: Sequence analysis of CG1 revealed that its heavy and light chain V regions were respectively most homologous to the 3d279d VH4 and the O18 Vk1 genes. Interestingly, 3d279d is frequently expressed by B cells stimulated with staphylococcal enterotoxin; and O18 is the main gene employed by the Vk1 IgG antibodies against Haemophilus influenzae. CONCLUSION: The combinatorial immunoglobulin library method represents an interesting model of how to approach the isolation and characterization of antibody-like reagents in the elucidation of autoantigens in RA.