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Cystatin C and beta-2-microglobulin (B2M) are filtration markers associated with adverse outcomes in nontransplant populations, sometimes with stronger associations than for creatinine. We evaluated associations of estimated glomerular filtration rate from cystatin C (eGFR cys ), B2M (eGFR B 2M ), and creatinine (eGFR cr ) with cardiovascular outcomes, mortality, and kidney failure in stable kidney transplant recipients using a case-cohort study nested within the Folic Acid for Vascular Outcome Reduction in Transplantation (FAVORIT) Trial. A random subcohort was selected (N = 508; mean age 51.6 years, median transplant vintage 4 years, 38% women, 23.6% nonwhite race) with enrichment for cardiovascular events (N = 306; 54 within the subcohort), mortality (N = 208; 68 within the subcohort), and kidney failure (N = 208; 52 within the subcohort). Mean eGFR cr , eGFR cys , and eGFR B 2M were 46.0, 43.8, and 48.8 mL/min/1.73m 2 , respectively. After multivariable adjustment, hazard ratios for eGFR cys and eGFR B 2M <30 versus 60+ were 2.02 (95% confidence interval [CI] 1.09-3.76; p = 0.03) and 2.56 (1.35-4.88; p = 0.004) for cardiovascular events; 3.92 (2.11-7.31) and 4.09 (2.21-7.54; both p < 0.001) for mortality; and 9.49 (4.28-21.00) and 15.53 (6.99-34.51; both p < 0.001) for kidney failure. Associations persisted with additional adjustment for baseline eGFR cr . We conclude that cystatin C and B2M are strongly associated with cardiovascular events, mortality, and kidney failure in stable kidney transplant recipients., (© 2017 The American Society of Transplantation and the American Society of Transplant Surgeons.)

Background: High intra-hepatic fat (IHF) content is associated with insulin resistance, visceral adiposity, and increased morbidity and mortality following liver resection. However, in clinical practice, IHF is assessed indirectly by pre-operative imaging [for example, chemical-shift magnetic resonance (CS-MR)]. We used the opportunity in patients undergoing liver resection to quantify IHF by digital histology (D-IHF) and relate this to CT-derived anthropometrics, insulin-related serum biomarkers, and IHF estimated by CS-MR., Methods: A reproducible method for quantification of D-IHF using 7 histology slides (inter- and intra-rater concordance: 0.97 and 0.98) was developed. In 35 patients undergoing resection for colorectal cancer metastases, we measured: CT-derived subcutaneous and visceral adipose tissue volumes, Homeostasis Model Assessment of Insulin Resistance (HOMA-IR), fasting serum adiponectin, leptin and fetuin-A. We estimated relative IHF using CS-MR and developed prediction models for IHF using a factor-clustered approach., Results: The multivariate linear regression models showed that D-IHF was best predicted by HOMA-IR (Beta coefficient(per doubling): 2.410, 95% CI: 1.093, 5.313) and adiponectin (β(per doubling): 0.197, 95% CI: 0.058, 0.667), but not by anthropometrics. MR-derived IHF correlated with D-IHF (rho: 0.626; p = 0.0001), but levels of agreement deviated in upper range values (CS-MR over-estimated IHF: regression versus zero, p = 0.009); this could be adjusted for by a correction factor (CF: 0.7816)., Conclusions: Our findings show IHF is associated with measures of insulin resistance, but not measures of visceral adiposity. CS-MR over-estimated IHF in the upper range. Larger studies are indicated to test whether a correction of imaging-derived IHF estimates is valid., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Introduction: Pre-eclampsia (PE) is characterized by failed remodeling of maternal vessels perfusing the placenta. Blood vessels and lymphatic system are involved in vessel remodeling and flow homeostasis in the uterus during pregnancy. This study aims to investigate the involvement of angiogenesis and lymphangiogenesis in PE., Methods: Placental and decidual tissues were obtained from pregnancies with PE (n = 90), including PE cases with decidual vasculopathy (DV) (n = 52) and without DV (n = 38), and healthy pregnancies (control, n = 20). The clinical characteristics of these groups were analyzed. The expression levels of VEGF1, CD34, PROX-1, VEGFR3, and CD31 in the placenta and decidua were detected through immunohistochemistry, reverse-transcription polymerase chain reaction, and Western blot., Results: The lymphangiogenic markers PROX-1 and VEGFR3 were negatively expressed in the placenta but positively expressed in the decidua. The expression levels of the angiogenic markers VEGF1 and CD34 and the panendothelial marker CD31 were significantly lower in the placenta and decidua of the PE group than in those of the control group. The expression levels of VEGF1, CD34, and CD31 were significantly lower in the placenta and decidua with DV than in those without DV. Furthermore, the expression trends of PROX-1 and VEGFR3 was similar to those of VEGF1, CD34, and CD31 among the groups., Conclusions: Lymphangiogenesis occurred in the decidua but not in the placenta. Impaired angiogenesis and lymphangiogenesis were associated with PE, particularly in the presence of DV., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In planning, designing, procuring and ensuring delivery of improved services ('commissioning') for the school age population, the outcomes should be students who are healthy to learn and who learn to be healthy. Intuitively, linking education and health development together within the wider learning environment seems a good start to planning school health. However there has been a shortage of either theoretical models that can span different settings or experimental research that demonstrates improved community health. Is there evidence that the wider learning environment provided in a school is valuable in improving health? An initial scoping exercise identified domains of health where there was a promise of health gain. International literature on school health outcomes using the framework of Asset-Based Community Development (ABCD) has been reviewed. It was found that research on a variety of interventions was relevant to schools as an asset for public health. Effective areas for health gain were identified for local planning and evaluation using this community model. However, none of the studies reviewed was originally designed to test schools as assets and most of the research lacked methodological rigour, especially regarding children in low income countries. The ABCD model could help national governments develop resources for both education and health, but there is a global need to generate better quality evidence. Then people who commission for their local communities can make more effective use of these multifaceted assets to improve health and education outcomes for children., (Copyright © 2014 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.)

This paper examines the development and achievements of the Teaching Public Health Networks (TPHNs) in England; an initiative that aimed to catalyse collaborative working between the public health workforce and further and higher education, to enhance public health knowledge in the wider workforce with a view to enhancing capacity to tackle inequalities and meeting public health targets. This paper highlights activities under three outcomes: mobilizing resources, people, money and materials; building capacity through training and infrastructure development; and raising public and political awareness. The TPHN approach is shown to have led to innovative developments in public health education and training, including engagement with professionals that have not previously had exposure to public health. This paper aims to disseminate the learning from this complex public health initiative, now in its third year of development, and to share examples of good practice. It is hoped that other countries can use the TPHN approach as a model to address the various common and country-specific challenges in public health workforce development.

Arrest of proliferation is one of the prerequisites for differentiation of cytotrophoblasts into syncytiotrophoblasts, and thus during differentiation telomerase activity, as well as human telomerase reverse transcriptase (hTERT) expression, is down-regulated. Considering this, it is of interest to investigate whether syncytium formation can be delayed by prolonging the expression of telomerase in cytotrophoblasts. BeWo cells were transfected with pLPC-hTERT retroviral vector and the reverse transcription-polymerase chain reaction analysis for hTERT mRNA concentrations in the transfected cells revealed a several-fold increase in hTERT mRNA compared with the cells transfected with empty vector, and this confirmed that the transfection was successful. An increase in the proliferation, as assessed by bromodeoxyuridine incorporation assay, as well as an increase in mRNA and protein concentration of various cyclins and proliferating cell nuclear antigen, was noticed. The effect of hTERT transfection was also assessed after the addition of forskolin to induce differentiation and it was observed that cell-cell fusion was delayed and differentiation did not occur in hTERT-transfected cells. However, the effects seen were only transient as stable transfection was not possible and the cells were undergoing apoptosis after 72 h, which suggested that apart from hTERT other factors might be important for immortalization of BeWo cells.

Cytokines are important modulators of inflammation. The balance between pro- and anti-inflammatory cytokines determines whether the intensity of inflammatory response is within physiological limits or in the pathological range. The cytokine network is highly complex, containing interactive cascades of gene activation and suppression. Both chronic kidney disease (CKD) and end-stage renal disease (ESRD) are characterized by elevated levels of proinflammatory cytokines and markers of inflammation. Cytokines may modulate the risk for progression of renal disease and the susceptibility to cardiovascular disease (CVD). Polymorphisms of cytokine genes may influence gene transcription and cytokine secretion and thereby modulate the risk of progression of renal and CVDs. The observed inconsistencies in the data regarding associations between single-nucleotide gene polymorphisms (SNPs) and their presumed phenotypic expression emphasize the need to recognize several conceptual and methodological aspects such as haplotypic rather than single SNP variations and the influence of pathway genes with synergistic or antagonistic effects that ultimately determine the phenotype. It is conceivable that when a patient with a high-risk cytokine genotype develops CKD, the risk for CVD is increased. Early interventions in CKD patients with high-risk genotypes may slow the progression of renal disease and also decrease CV mortality and morbidity.

Polyamidoamine dendrimers are protein-like nanomaterials that have many potential biological applications. Research studies using dendrimers often require their detection and quantitation. This article describes a simple, inexpensive, and rapid fluorometric assay based on the ability of dendrimers to bind the fluorescent probe, 8-anilino-1-naphthalene-sulfonic acid. This interaction leads to an enhanced fluorescence intensity of the dye that shows a linear relationship with dendrimer concentration. The procedure is applicable to dendrimers that have various surface functional groups and internal cores. Amine- and hydroxyl-terminated dendrimers gave the best calibration curves under alkaline conditions (pH 7.4). Carboxyl-terminated dendrimers required acidic conditions for their measurement. The assay for dendrimers with an amine surface and an ethylene diamine core was accurate between 0 and 100 microM and produced a within-run coefficient of less than 5%. Under the assay conditions used, the fluorescence signal strength was dependent on type of surface group and core length of the dendrimer.

Diffusion tensor imaging can provide the fundamental information required for viewing structural connectivity. However, robust and accurate acquisition and processing algorithms are needed to accurately map the nerve connectivity. In this paper, we present a novel algorithm for extracting and visualizing the fiber tracts in the CNS, specifically in the brain. The automatic fiber tract mapping problem will be solved in two phases, namely a data smoothing phase and a fiber tract mapping phase. In the former, smoothing of the diffusion-weighted data (prior to tensor calculation) is achieved via a weighted TV-norm minimization, which strives to smooth while retaining all relevant detail. For the fiber tract mapping, a smooth 3D vector field indicating the dominant anisotropic direction at each spatial location is computed from the smoothed data. Neuronal fibers are then traced by calculating the integral curves of this vector field. Results are expressed using three modes of visualization: (1) Line integral convolution produces an oriented texture which shows fiber pathways in a planar slice of the data. (2) A streamtube map is generated to present a 3D view of fiber tracts. Additional information, such as degree of anisotropy, can be encoded in the tube radius, or by using color. (3) A particle system form of visualization is also presented. This mode of display allows for interactive exploration of fiber connectivity with no additional preprocessing.

This study assessed the effects of ambient UV light on the development of two native species of anurans, Rana blairi and Hyla chrysoscelis, during their normal breeding season in Oklahoma. Additionally, the effects of ambient UV light and water contaminated with landfill leachate in Rana blairi were examined. Embryos were collected from the field and distributed equally among replicates of four filter treatments of ambient UV light in experimental tubs filled with either FETAX solution or landfill leachate diluted to 25, 10, and 5% concentrations. Three endpoints (mortality, teratogenesis, and growth) were compared between filter treatments. By itself, UV-B caused no significant effects. Leachate at 10 and 25% concentrations caused 100% mortality across all filter treatments. There was a significant interaction between filter treatment and water toxicity at leachate concentrations of 5% for both malformation and growth. Increased UV-B exposure decreased the malformation rate and increased growth in the leachate treatments.

We studied the uptake, transport and metabolism of cocaine in human intestine using the colonic T-84 monolayers as a model. T-84 cells were grown in DMEM/Ham's F-12 medium containing 6% newborn calf serum at 37 degrees C on 1.0 microm collagen inserts. The cells develop into a polarized monolayer with the apical surface facing the upper chamber and the basolateral surface facing the lower. The monolayers develop a transepithelial resistance of > or = 600 ohms cm2 in 7 days. Varying concentrations of cocaine HCI was added in a serum free medium to the luminal side only, and after 30 min and 60 min samples from the luminal and serosal aspect were removed for analysis. Cocaine and its metabolites were measured by GC/MS. Cocaine transport across T-84 monolayers increased linearly with increasing concentration of cocaine, with no significant difference between 30 min and 60 min of exposure. Transepithelial resistance did not change even at 800 ng of cocaine suggesting no effect on monolayer viability. The metabolites, benzoylecgonine (BE) and ecgonine methyl ester (EME) but not norcocaine were detected in both luminal and serosal side. The concentrations of BE and EME in the luminal side were significantly higher than in the serosal. Combined recovery of cocaine, BE and EME from luminal and serosal sides were 52 - 80% of total added cocaine. While fresh medium did not metabolize cocaine, media previously exposed to the monolayer (cell-free medium) caused a significant breakdown into BE and EME, suggesting that esterases may be released into the medium. These results indicate transfer of cocaine across this model of intestinal epithelial cell line is by simple diffusion and is concentration dependent. These studies imply that cocaine in swallowed amniotic fluid can be absorbed by the fetal gastrointestinal tract.

Nonnative protein structures having a compact secondary, but not rigid tertiary structure, have been increasingly observed as intermediate states in protein folding. We have shown for the first time during acid-induced unfolding of xylanase (Xyl II) the presence of a partially structured intermediate form resembling a molten globule state. The conformation and stability of Xyl II at acidic pH was investigated by equilibrium unfolding methods. Using intrinsic fluorescence and CD spectroscopic studies, we have established that Xyl II at pH 1.8 (A-state) retains the helical secondary structure of the native protein at pH 7.0, while the tertiary interactions are much weaker. At variance, from the native species (N-state), Xyl II in the A-state binds 1-anilino-8-sulfonic acid (ANS) indicating a considerable exposure of aromatic side chains. Lower concentration of Gdn HCl are required to unfold the A-state. For denaturation by Gdn HCl, the midpoint of the cooperative unfolding transition measured by fluorescence for the N-state is 3.5 +/- 0.1 M, which is higher than the value (2.2 +/- 0.1 M) observed for the A-state at pH 1.8. This alternatively folded state exhibits certain characteristics of the molten globule but differs distinctly from it by its structural stability that is characteristic for native proteins., (Copyright 2001 Academic Press.)

The high incidence of oral submucous fibrosis (OSF), a potentially malignant condition of the oral cavity, in the Indian subcontinent is causally associated with commonly prevailing habit of chewing areca nut and tobacco. Knowledge of molecular alterations in OSF is meagre. OSF is characterised by progressive accumulation of collagen fibres in lamina propria and oral submucosa. Colligin/HSP47 is a 47KDa stress protein which acts as a chaperone for collagen. We hypothesized that since colligin plays a vital role in folding and assembling collagen it may be involved in the pathogenesis of OSF. The present study was undertaken in tobacco and areca nut chewing Indian OSF patients to investigate the correlation, if any, between the expression of colligin and collagen type I proteins in OSF lesions. Immunohistochemical analysis showed overexpression of colligin and collagen type I proteins in 16/23 (70%) and 15/23 (65%) of OSF cases, respectively. The hallmark of the study was the significant association between the increased expression of type I collagen and its chaperone, colligin, in OSF lesions (P=0.0494). The data suggest that the increased levels of colligin in OSF may contribute to the deposition of collagen and consequent increased fibrosis in the oral submucosa in OSF lesions.

The pH induced conformational and structural changes of Xyl II have been investigated from the alkalophilic thermophilic Bacillus sp. using kinetic, circular dichroism and fluorescence spectroscopy studies. The systematic studies on the folding and stability of cellulase-free xylanases are important, since their biotechnological applications require them to function under extremes of pH and temperature. The Trp fluorescence and the kinetic constants were found dependent on the pH. Above pH 8, the enzyme exhibited unfolding transitions as revealed by a red shift in the emission maximum as well as decreases in the fluorescence intensity. Circular dichroism studies revealed a decrease in the CD ellipticity at 222 nm at pH 9 and 10. The reduced catalytic activity of Xyl II at alkaline pH is correlated to the pH induced unfolding and ionization or protonation of key protein residues. The pH profile of Xyl II showed apparent pK values of 5.5 and 7 for the free enzyme and 5.6 and 6.7 for the enzyme-substrate complex. The abnormally high pK of 6.7 indicated the participation of a carboxyl group present in a non-polar environment. The pH dependence of inactivation kinetics of Xyl II with Woodward's reagent K corroborates evidence for the presence of a catalytically important carboxyl residue. The sequence alignment studies of Xyl II, in combination with kinetic and chemical modification data provide strong evidence for the participation of Asp94 in the catalytic function. The Xyl II produced from an alkalophilic source, was stable at pH 10 with a t(1/2) of 24 h. However, the enzyme exhibited pH optimum at near neutral values, which can be explained by the ionization and microenvironment of the active site residues.

A key component in the evaluation of efficacy of a vaccine to protect against disease caused by an antigenically diverse infectious pathogen in a preventative vaccine trial is assessing how vaccine-induced protection depends on genotypic and phenotypic variations of the exposing pathogen. This assessment is made by comparing pathogen isolates between infected vaccinated subjects and infected unvaccinated subjects. A survey of efficacy trial reports reveals a lack of systematic, quantitative investigation in this question. Analysis tools for testing if vaccine protection against disease is superior against some pathogen strains, and for estimating the magnitude of this differential vaccine protection, are described. The broad applicability of the methods is illustrated through analysis of isolates taken from persons infected while participating in vaccine trails for cholera, HIV-1, hepatitis B, rotavirus, and pneumococcus. These analyses reveal intriguing trends for Genentech's monovalent rgp120 HIV-1 vaccine, for two whole-killed-cell oral cholera vaccines, and for other vaccines.

An enzyme-linked immunosorbent assay (ELISA) utilizing antigen coated on hydrophobic polyvinyldiene fluoride (PVDF) membranes is described for detecting antibodies that bind to squalene (SQE). Because of the prior lack of availability of validated antibodies to SQE, positive controls for the assay were made by immunization with formulations containing SQE to create monoclonal antibodies (mAbs) that reacted with SQE. Among eight immunogens tested, only two induced detectable murine antibodies to SQE: liposomes containing dimyristoyl phosphatidylcholine, dimyristoyl phosphatidylglycerol, 71% SQE, and lipid A [L(71% SQE+LA)], and, to a much lesser extent, an oil-in-water emulsion containing SQE, Tween 80, Span 85, and lipid A. In each case, lipid A served as an adjuvant, but neither SQE alone, SQE mixed with lipid A, liposomes containing 43% SQE and lipid A, nor several other emulsions containing both SQE and lipid A, induced antibodies that reacted with SQE. Monoclonal antibodies produced after immunizing mice with [L(71% SQE+LA)] served as positive controls for developing the ELISA. Monoclonal antibodies were produced that either recognized SQE alone but did not recognize squalane (SQA, the hydrogenated form of SQE), or that recognized both SQE and SQA. As found previously with other liposomal lipid antigens, liposomes containing lipid A also induced antibodies that reacted with the liposomal phospholipids. However, mAbs were also identified that reacted with SQE on PVDF membranes, but did not recognize either SQA or liposomal phospholipid. The polyclonal antiserum produced by immunizing mice with [L(71% SQE+LA)] therefore contained a mixed population of antibody specificities and, as expected, the ELISA of polyclonal antiserum with PVDF membranes detected antibodies both to SQE and SQA. We conclude that SQE is a weak antigen, but that antibodies that specifically bind to SQE can be readily induced by immunization with [L(71% SQE+LA)] and detected by ELISA with PVDF membranes coated with SQE.

Background: Optimal nutrient intake is important in the maintenance of a positive nitrogen balance in hemodialysis (HD) patients. The objectives of this study were (1) to assess the influence of two levels of protein intakes on nitrogen balance in stable adult HD patients, and (2) to identify a minimum level of protein intake that would result in a negative nitrogen balance, so that preliminary recommendations may be made in Indian patients on maintenance HD (MHD)., Methods: Stable, adult, nondiabetic MHD patients were recruited after informed consent into a cross over trial with a high-protein (HP) diet [1.2 g/kg ideal body weight (IBW)/day), followed by a low-protein (LP) diet (0.6 g/kg IBW/day] after appropriate periods of equilibration; for both diets, 50% of protein was of high biological value, and calorie intake was 35 kCal/kg IBW/day. Duplicate meals and residues were weighed, homogenized, and stored at -20 degrees C for analysis of dietary N by the Kjeldahl method, used to check the consistency of the N content of the diet supplied. Pre- and post- (30-minute equilibrated) blood urea samples were drawn, and details of weights and other HD parameters were recorded. Interdialytic urine collections for urea were obtained. N input came from dietary protein calculated as 16% of the weight of biological protein; N output was calculated using blood-side urea measurements and urinary urea excretion and was the sum of urea N (UN) and nonurea N (NUN) losses (assumed to be equal to 0.031 g N/kg/day)., Results: Fifteen patients were recruited. Twelve patients completed both limbs of the study. The mean age was 30.3 +/- 12.7 years. The body mass index was 18.9 +/- 2.4. Serum albumin was 3.8 +/- 0.35 g/dL, and Kt/V (equilibrated) was 1.17 +/- 0.3 g/dL. Protein consumed was 1.06 +/- 0.18 g/kg IBW/day in the HP limb versus 0.61 +/- 0.1 g/kg IBW/day in the LP limb (P = 0.000). Energy intake was 33 +/- 6.5 vs. 32.8 +/- 6. 7 kCal/kg IBW/day, respectively (P = 0.8). The normalized protein N appearance (nPNA) was 0.88 +/- 0.2 g/kg/day in the HP limb versus 0. 78 +/- 0.2 g/kg/day in the LP limb (P = 0.02). Dietary N was 73.5 +/- 15.3 g in the HP week and 42.5 +/- 7.5 g in the LP week (P = 0. 000). The difference between this and the sum of (UN + NUN) losses over the week was 29 +/- 13.2 g versus 1.2 +/- 8.1 g, respectively (P = 0.001), showing a strong, uniformly positive nitrogen balance with HP diet and neutral to negative nitrogen balance with LP diet. The ratio of dietary protein intake (DPI) to nPNA was significantly lower (anabolic) in the HP limb (0.7 +/- 0.2 vs. 1.12 +/- 0.3, P = 0. 000). On a scatter plot of nPNA to DPI, a catabolic relationship was demonstrated below a DPI of 0.75 g/kg/day (95% CI, 0.65 to 0.85 g/kg/day)., Conclusion: A DPI of approximately 1.1 g/kg/day produces a positive nitrogen balance and 0.6 g/kg/day a neutral to negative nitrogen balance, demonstrating protein anabolism as a function of protein intake. It is suggested that a protein intake of 0.85 g/kg/day should be considered unsafe. These conclusions apply in stable nondiabetic adult HD patients in the setting of adequate dialysis and adequate calorie intake.

A 1.0 kilobase gene fragment from the genomic DNA of an alkaliphilic thermophilic Bacillus was found to code for a functional xylanase (XynII). The complete nucleotide sequence including the structural gene and the 5' and 3' flanking sequences of the xylanase gene have been determined. An open reading frame starting from ATG initiator codon comprising 402 nucleotides gave a preprotein of 133 amino acids of calculated molecular mass 14.090 kDa. The occurrence of three potential N-glycosylation sites in XynII gene is a unique feature for a gene of bacterial origin. The stop codon was followed by hairpin loop structures indicating the presence of transcription termination signals. The secondary structure analysis of XynII predicted that the polypeptide was primarily formed of beta-sheets. XynII appeared to be a member of family G/11 of xylanases based on its molecular weight and basic pI (8.0). However, sequence homology revealed similar identity with families 10 and 11 of xylanases. The conserved triad (Val-Val-Xaa, where Xaa is Asn or Asp) was identified only in the xylanases from alkaliphilic organisms. Our results implicate for the first time the concept of convergent evolution for XynII and provide a basis for research in evolutionary relationship among the xylanases from alkaliphilic and neutrophilic organisms., (Copyright 1999 Academic Press.)

Background: Calculation of the incidence of typhoid fever during preschool years is important to define the optimum age of immunisation and the choice of vaccines for public-health programmes in developing countries. Hospital-based studies have suggested that children younger than 5 years do not need vaccination against typhoid fever, but this view needs to be re-examined in community-based longitudinal studies. We undertook a prospective follow-up study of residents of a low-income urban area of Delhi, India, with active surveillance for case detection., Methods: A baseline census was undertaken in 1995. Between Nov 1, 1995, and Oct 31, 1996, we visited 8172 residents of 1820 households in Kalkaji, Delhi, twice weekly to detect febrile cases. Blood samples were obtained from febrile patients, and those who tested positive for Salmonella typhi were treated with ciprofloxacin., Findings: 63 culture-positive typhoid fever cases were detected. Of these, 28 (44%) were in children aged under 5 years. The incidence rate of typhoid per 1000 person-years was 27.3 at age under 5 years, 11.7 at 5-19 years, and 1.1 between 19 and 40 years. The difference in the incidence of typhoid fever between those under 5 years and those aged 5-19 years (15.6 per 1000 person-years [95% CI 4.7-26.5]), and those aged 19-40 years (26.2 [16.0-36.3]) was significant (p<0.001 for both). The difference between the incidence of typhoid at 5-19 years and the incidence at 19-40 years was also significant (10.6 [6.3-14.8], p<0.001). Morbidity in those under 5 and in older people was similar in terms of duration of fever, signs and symptoms, and need for hospital admission., Interpretation: Our findings challenge the common view that typhoid fever is a disorder of school-age children and of adults. Typhoid is a common and significant cause of morbidity between 1 and 5 years of age. The optimum age of typhoid immunisation and the choice of vaccines needs to be reassessed.

Recombinant Escherichia coli (pATBX 1.8) secreting extracellular xylanase was used as a model system to study the application of an aqueous two phase system for extractive cultivation. An increase in the polymer concentrations from 6 to 20% in the polyethylene glycol phosphate aqueous two phase system resulted in an increase in the phase volume ratio with a concomitant decrease in the partition coefficient (K) and recovery of xylanase in the top phase. However, varying phosphate concentrations from 8 to 16% decreased both the phase volume ratio and the partition coefficient of xylanase. The polyethylene glycol (6%) and phosphate (12%) system was found to be optimum for extracellular cultivation of E. coli, where extracellular xylanase was selectively partitioned to the top phase giving a purification ratio of above 1.0. The process was extended to a semicontinuous operating mode at the optimal condition, wherein the top phase containing xylanase was recovered and the surviving cells were recycled together with the new top phase. The maximum recovery of xylanase was obtained after 12 h in the top phase with a twofold increase in the specific activity as compared to the one obtained in the reference fermentation. In the present work, we report for the first time the use of the two phase system for the extractive cultivation of recombinant E. coli (pATBX 1.8) with the purpose of obtaining a simple and inexpensive separation procedure and achieving the maximal extraction of xylanase to one phase., (Copyright 1999 Academic Press.)

Microenvironment and conformation of the active site of xylanase from an extremophilic Bacillus was deciphered for the first time using fluorescence spectroscopy. NBS modified enzyme showed complete inactivation and the kinetic analysis implicated the presence of an essential tryptophan at the active site of xylanase. Xylan (0.5%) protected the enzyme completely from inactivation with NBS, whereas it afforded 35% protection against the loss of fluorescence, suggesting that not all the tryptophans are involved at the substrate binding site. Quenching studies revealed that acrylamide was more efficient than KI and CsCl as indicated by the higher Stern-Volmer quenching constants (Ksv). The steric factor represented by the percentage accessibility of the tryptophan residues of XylII was higher with the positively charged Cs+ (80) than with the negatively charged I- (10), suggesting that the tryptophan residues are located in a relatively electronegative environment. In the presence of 6 M Gdn HCl the fluorescence shifted to 350 nm with increased accessibility of the fluorophore to the quenchers. The proximity of the essential carboxyl groups with a high pKa value of 6.9 [Chauthaiwale and Rao (1994) Biochim. Biophys. Acta] probably contributes to the electronegative environment of the tryptophan residue. Our results on sequence analysis of the gene encoding for XylII (Accession Number U83602 in the GenBank database) have shown that Trp 61 is highly conserved and may play a role in the structure-function relationship of the enzyme.

Contaminated groundwater poses a significant health hazard and may also impact wildlife such as amphibians when it surfaces. Using FETAX (Frog Embryo Teratogenesis Assay-Xenopus), the developmental toxicity of ground and surface water samples near a closed municipal landfill at Norman, OK, were evaluated. The groundwater samples were taken from a network of wells in a shallow, unconfined aquifer downgradient from the landfill. Surface water samples were obtained from a pond and small stream adjacent to the landfill. Surface water samples from a reference site in similar habitat were also analyzed. Groundwater samples were highly toxic in the area near the landfill, indicating a plume of toxicants. Surface water samples from the landfill site demonstrated elevated developmental toxicity. This toxicity was temporally variable and was significantly correlated with weather conditions during the 3 days prior to sampling. Mortality was negatively correlated with cumulative rain and relative humidity. Mortality was positively correlated with solar radiation and net radiation. No significant correlations were observed between mortality and weather parameters for days 4-7 preceding sampling.

Objectives: There are several reports of the pulmonary findings in children with HIV disease; however, the occurrence of bronchiectasis rarely has been noted. We evaluated occurrence of bronchiectasis in a large group of children referred to us with AIDS pneumopathy., Methods: From January 1984 to April 1996, 203 children with AIDS and respiratory problems were referred to the pediatric pulmonary division at Children's Medical Center of Brooklyn. Medical records for 164 of these children were available and retrospectively reviewed., Results: Uncomplicated pneumonia was present in 75, 24 had recurrent pneumonia, and 18 had unresolved pneumonia; lymphocytic interstitial pneumonitis (LIP) was diagnosed in 47 patients, worsening with time in all patients. Bronchiectasis was observed in 26 patients (26/164, 15.8%), diagnosed by chest radiograph in 26 (26/26, 100%), confirmed by CT scan of chest in 10 (10/26, 38.4%), and by histology in three (3/26, 11.5%). Median age at time of diagnosis of bronchiectasis was 7.5 years (range, 1 to 16 years). Sixteen children with LIP developed bronchiectasis (16/47, 34.0%). Three patients with recurrent pneumonia (3/24, 12.5%) developed bronchiectasis. Five patients with unresolved pneumonia (5/18, 27.7%) developed bronchiectasis. One infant developed bronchiectasis after Pneumocystis carinii pneumonia; another child developed bronchiectasis after P. carinii and Mycobacterium tuberculosis pneumonia. The CD4+ T-cell counts measured within 6 months of diagnosis of bronchiectasis were available in 23/26 patients and, all were < 100 cells per cubic millimeter., Conclusion: We conclude, from our experience, that there is a significant occurrence of bronchiectasis in children with AIDS and pulmonary disease, especially in children developing LIP, recurrent pneumonia and unresolved pneumonia, and CD4+ T-cell counts < 100 cells per cubic millimeter.

Inactivation of xylose reductase (XR) by p-hydroxy-mercury benzoate (PHMB) was found to be biphasic with second-order rate constants of 80 and 6 M-1s-1 for the fast (kf) and slow (ks) phase respectively. Spectroscopic studies indicated that the inactivation was due to modification of one Cys residue per molecule of XR and not due to subsequent disruption of the quaternary structure. The binding of NADPH to XR (Kd 0.9 microM) was depressed on modification of the enzyme by PHMB (Kd 2.3 microM). The dependence of PHMB induced inactivation of XR in the presence of alcohols and varying temperature revealed that the Cys residue is situated in a hydrophobic microenvironment and is not involved in hydrogen bonding. Our present investigation using o-phthalaldehyde (OPTA) as the chemical initiator for fluorescent chemo-affinity labeling and double inhibition studies indicates that Cys residues involved in the reaction with PHMB (SHI) and OPTA (SHII) are distinctly different. Experimental evidence presented here serves to implicate that SHI located in a hydrophobic microenvironment at the high affinity NADPH binding site of XR plays a role in the binding of the coenzyme to XR, whereas SHII serves to maintain the conformation of the active site essential for catalysis by interacting with the NH2 group of an essential lysine residue.

Army families in the UK have been observed to have a higher infant mortality compared with the civilian population. Little is known about the reasons for the higher mortality. We report a comparative study of the health of army and civilian babies and their families in a UK garrison town, the aim being to investigate any differences in risk factors which may have explained the differences in mortality. A total of 250 first born army and civilian babies fulfilling the selection criteria were recruited to the study. Army babies had a lower birth weight distribution and both maternal and paternal smoking rates were higher in army families. In addition, social support and in particular, contact with close family members appeared to be available to fewer army mothers. The study highlighted the needs of this population and prompted a number of local health services to work together to address the issues it raised.

To determine the energy savings caused by sucrose taste, we measured heat loss through direct calorimetry for 23 premature and normal term infants who were studied a total of 31 times. Following stabilization in the calorimeter, crying was induced by applying 1 ml cold water to the infant's foot. After 4 min, 0.1 or 0.2 ml sucrose were delivered intraorally through a remote syringe to arrest crying. Crying was accompanied by a 13.2% increase in metabolic rate that was quickly and completely reversed during crying cessation caused by sucrose taste. Heat loss was inversely and linearly related to infant body weight. The implications of these findings for minimizing crying and energy expenditure in normal newborns and especially in ill or small newborns are discussed.

The genomic library of alkalophilic thermophilic Bacillus was constructed in E.coli. A single recombinant [pATB 507 I] out of 10,000 colonies showed growth at 50 degrees C, which produced large, opaque and mucilaginous colonies on LB. The IMViC biochemical tests suggested that the strain was E.coli, and scanning electron microscopy revealed increased cell length. The plasmid DNA from the thermophilic recombinant showed multiple bands. The cured strain showed ampicillin resistance (10 micrograms/ml) and presence of insert DNA in the genome, as revealed by southern blot analysis. The thermophilic trait was transformable with 10(-4) frequency of expression amongst the pATB 507I transformants. The hybridization of E.coli genomic DNA with labeled genomic DNA from alkalophilic thermophilic Bacillus suggested that the genetic recombination(s) of the insert DNA was an essential prerequisite for expression of thermophilic trait. The conjugative plasmid (RP4) mediated mobilization of thermophilic trait resulted in thermophilic conjugants after 2 h incubation. Southern blot analysis revealed that a single locus [5.6 kb Bam HI fragment] was involved in the transfer of thermophilic trait.

Chromosomal integration and xylanase gene amplification were demonstrated for the first time in an alkalophilic thermophilic Bacillus sp. (NCIM 59). The integrants were characterized by larger zone of xylan clearance than the parent culture and hybridization with vector (pUC8) DNA. Repeated transformation strategy was used for further amplification of the xylanase gene. The results of Southern blot analysis indicated the occurrence of homologous recombination in the 6.5 kb xylanase gene region of the genomic DNA and suggested a non campbell mode of recombination. The integrants were checked for xylanase production up to ten subcultures and consistently showed two fold higher xylanase activity than the parent strain with the maximum xylanase productivity (U/ml/h) at 16 h.

From May, 1990, a new schedule of immunisation against diphtheria, tetanus, and pertussis (at 2, 3, and 4 months) replaced the previous more widely spaced schedule. A report that children had lower concentrations of diphtheria and tetanus antibodies a month after an accelerated schedule led us to undertake a controlled study to assess antibody response and the persistence of antibodies a year after immunisation in children receiving vaccine according to widely spaced and accelerated schedules. Concentrations of antibodies to diphtheria and tetanus toxoids and to Bordetella pertussis filamentous haemagglutinin (FHA) were measured by solid-phase radioimmunoassay (SP-RIA). We studied 57 children who received accelerated immunisation at median ages of 11, 16, and 21 weeks and two control cohorts (total n = 82) who received vaccine at median ages of 15, 21, and 45 weeks. 6-8 weeks after the third dose the accelerated-schedule group had lower (p < 0.0001) geometric mean concentrations of antibody to tetanus (0.522 [95% CI 0.383-0.710] vs 3.43 [2.45-4.81] IU/mL), diphtheria (0.266 [0.179-0.396] vs 2.39 [0.616-3.53] IU/mL), and FHA (0.044 [0.030-0.063] vs 0.270 [0.196-0.374] units/mL) than the longer-schedule group. 12 months after the third dose the differences between the groups had narrowed (tetanus 0.197 vs 0.341 IU/mL, p = 0.29; diphtheria 0.100 vs 0.131 IU/mL, p = 0.64; FHA 0.014 vs 0.016 units/mL, p = 0.72). At that time all children had tetanus antibody concentrations above protective levels (0.01 IU/mL); only 2 of 31 in the accelerated-schedule group and 3 of 31 in the longer-schedule group had diphtheria antibody concentrations below the protective level. The use of an accelerated schedule of diphtheria, tetanus, and pertussis vaccination is unlikely to lead to an increase in the proportion of children unprotected against these diseases before the preschool booster.

Experimental evidence for the specific interaction of guanidine hydrochloride with the carboxyl group of xylanase has been presented for the first time. Guanidine hydrochloride (0.1 M) inactivated the xylanase from alkalothermophilic Bacillus sp. to 50% without affecting the conformation of the protein as determined by the fluorometric profile. The kinetic analysis indicated a competitive type of inhibition and a requirement of 1.4 molecules of guanidine hydrochloride per molecule of the enzyme for inhibition. Maximum inhibition occurred at the pH which is optimum for the enzyme activity. The reaction of guanidine hydrochloride with the enzyme prior to modification by Woodward's Reagent K, a specific inhibitor of the carboxyl group, made it inaccessible for modification as indicated by absorbance data at 340 nm. Urea, sodium dodecyl sulphate, LiCl, KCl and NaCl at 0.1 M concentration each had negligible effect on the enzyme activity.

Membrane-associated neurotransmitter stimulating factor (MANS) can modulate sympathetic neurotransmitter expression and promote ciliary neuron survival in cell culture. Previous studies have shown that its biological effects and biochemical properties are similar to those of ciliary neurotrophic factor (CNTF). In addition, CNTF is present in spinal cord, the source of MANS. These observations raised the possibility that MANS preparations contain CNTF. We find that partially purified MANS fractions contain a 24-kD protein that is recognized in Western blots by an antiserum generated against recombinant rat CNTF (rCNTF). This antiserum immunoprecipitates virtually all the cholinergic-inducing and the ciliary neurotrophic activities present in MANS preparations. When iodinated rCNTF is incubated with spinal cord membranes, a significant proportion of the labeled CNTF segregates with the membrane pellet. The membrane-associated exogenous CNTF can be eluted from the membrane fraction by treatment with high-salt solutions, similar to that used to solubilize MANS from spinal cord membranes. Our data suggest that a substantial portion of the cholinergic differentiation and ciliary neurotrophic activities present in MANS preparations can be attributed to CNTF or a CNTF-like molecule.

Although the literature well describes chronic eosinophilic pneumonia in the adult, there are no reports of this entity in the pediatric age group. We describe a child with chronic eosinophilic pneumonia, emphasizing the specific radiologic features, common conditions in the differential diagnosis, and dramatic response in corticosteroid administration, which is a unique feature of this entity.

A simple, inexpensive acrylate polymer which has a capacity to absorb 170 ml of water per g has been developed. It can be used to concentrate dilute solutions of macromolecules such as proteins, nucleic acids, and carbohydrates. The polymer absorbs only low-molecular-weight substances such as glucose, sucrose, and inorganic salts. It can replace the various conventional concentration methods. No special device or electricity is needed for the concentration. The inexpensive polymer, molded in the form of rods, can be very conveniently used as "disposable concentration sticks."

Sixty-one cases of acute cervical lymphadenitis in Kings County Hospital Center were reviewed. Staphylococcus aureus accounted for 50 percent of the cases and Group A β-hemolytic streptococcus accounted for 15 percent; 35 percent had no growth on culture. Of the variables considered (age, sex, temperature, leukocytosis, previous antibiotic treatment, duration of symptoms, lymph node consistency) only age appeared to be a predictor of the causative organisms. Children under 1 year of age had a higher incidence of S aureus (65 percent) than children above 3 years of age (25 percent); the difference was statistically significant (P <.01). Therapeutic antimicrobial regimens should include coverage for S aureus in this age group.

Ten children who had developed pneumatoceles during the course of pneumonia are described. Seven of the ten patients had persistence of pneumatoceles for 12 months or more. There was no evidence of an underlying disorder in any of the patients, and all recovered completely without complications. The etiologic agent was identified in two of the ten cases as Staphylococcus aureus. In our experience, a delay in the resolution of pneumatoceles for a period of one or more years after an episode of pneumonia is not an unusual occurrence in normal healthy children.