Search

Objective To determine the effect of integrated and concurrent delivery of health, nutrition, water, sanitation and hygiene (WaSH), and psychosocial care interventions during the preconception period alone, during pregnancy and early childhood, and throughout preconception, pregnancy, and early childhood on birth outcomes and linear growth at 24 months of age compared with routine care. Design Individually randomised factorial trial. Setting Low and middle income neighbourhoods of Delhi, India. Participants 13 500 women were randomised to receive preconception interventions (n=6722) or routine care (n=6778). 2652 and 2269 pregnant women were randomised again to receive pregnancy and early childhood interventions or routine care. The analysis of birth outcomes included 1290 live births for the preconception, pregnancy, and early childhood interventions (group A), 1276 for the preconception intervention (group B), 1093 for the pregnancy and early childhood interventions (group C), and 1093 for the control (group D). Children aged 24 months by 30 June 2021 were included in the 24 month outcome analysis (453 in group A, 439 in B, 293 in C, and 271 in D). Interventions Health, nutrition, psychosocial care and support, and WaSH interventions were delivered during preconception, pregnancy, and early childhood periods. Main outcome measures The primary outcomes were low birth weight, small for gestational age, preterm, and mean birth weight. At 24 months, the outcomes were mean length-for-age z scores and proportion stunted. Three prespecified comparisons were made: preconception intervention groups (A+B) versus no preconception intervention groups (C+D); pregnancy and early childhood intervention groups (A+C) versus routine care during pregnancy and early childhood (B+D) and preconception, pregnancy, and early childhood interventions groups (A) versus control group (D). Results The proportion with low birth weight was lower in the preconception intervention groups (506/2235) than in the no preconception intervention groups (502/1889; incidence rate ratio 0.85, 98.3% confidence interval 0.75 to 0.97; absolute risk reduction −3.80%, 98.3% confidence interval −6.99% to −0.60%). The proportion with low birth weight was lower in the pregnancy intervention groups (502/2096) than in the no pregnancy intervention groups (506/2028) but the upper limit of the confidence interval crossed null effect (0.87, 0.76 to 1.01; −1.71%, −4.96% to 1.54%). There was a larger effect on proportion with low birth weight in the group that received interventions in the preconception and pregnancy periods (267/1141) compared with the control group (267/934; 0.76, 0.62 to 0.91; −5.59%, −10.32% to −0.85%). The proportion stunted at 24 months of age was substantially lower in the pregnancy and early childhood intervention groups (79/746) compared with the groups that did not receive these interventions (136/710; 0.51, 0.38 to 0.70; −8.32%, −12.31% to −4.32%), and in the group that received preconception, pregnancy, and early childhood interventions (47/453) compared with the control group (51/271; 0.49, 0.32 to 0.75; −7.98%, −14.24% to −1.71%). No effect on stunting at 24 months was observed in the preconception intervention groups (132/892) compared with the no preconception intervention groups (83/564). Conclusions An intervention package delivered during preconception, pregnancy, and early childhood substantially reduced low birth weight and stunting at 24 months. Pregnancy and early childhood interventions alone had lower but important effects on birth outcomes and 24 month outcomes. Preconception interventions alone had an important effect on birth outcomes but not on 24 month outcomes. Trial registration Clinical Trial Registry—India CTRI/2017/06/008908.

Variability studies were carried out among different accessions of Argyrolobium roseum for 12 metric traits under natural and cultivated conditions. Almost all characters showed higher values in cultivated than natural population. Leaf breadth recorded highest CV (30.59%). Highest percentage of vitexin and D-pinitol (0.208 and 0.773% dwb) was observed in RAR-7 and RAR-6, respectively, under cultivated conditions. Phenotypic coefficient was higher than genotypic coefficient of variation. The association analysis revealed that root yield had positive significant correlation with 100 seed weight (r=0.839) biomass yield (r=0.601), where biomass yield had a positive significant correlation with leaf length, single pod weight. High heritability (97.4–99.8%) coupled with moderate genetic advance ranged between (49–67%) as a percent of mean was observed for leaf breadth, pod length, total foliage biomass yield and total root yield/plant. This suggests that direct selection for these traits is suitable for the improvement of this crop.

Six phenotypic characters and three withanolide markers were assessed in 25 accessions of Withania somnifera collected from different states of India for studying genetic variability. The variability ranges observed at phenotypic and chemotypic levels were polymorphic. Based on D2 values and PCA (Principal Component Analysis) of phenotypic traits like plant height, no. of branches/plant, no. of seeds/berry, root length, root diameter and root yield, these 25 accessions were grouped in five clusters. The relative contribution of each character towards genetic divergence was worked out. Five accessions–AGB002 (Rajasthan), AGB003 (J&K), AGB004 (Madhya Pradesh), AGB006 (J&K) and AGB009 (Punjab) representing clusters 2 and 4 exhibited maximum intra and inter-cluster divergence. Cluster 5 representing accession AGB053 (Andhra Pradesh) was having mixed traits. Chemically most of the accessions in cluster 3 showed uniformity in presence of three marker withanolides Withaferin A, Withanone and Withanolide A in the leaves.

Andrographis paniculata (Burm.f.) Nees (Acanthaceae), commonly known as Kalmegh is used both in Ayurvedic and Unani system of medicines for a number of ailments related to digestion, hepatoprotection, hypoglycemic, and as anti-bacterial, analgesic, anti-inflammatory, vermicidal and antiacene. Ten accessions of A. paniculata from different sources were screened to assess the yield and its components besides andrographolides on three harvesting dates viz. 70, 100, 130 days after plantation corresponding to calendar months of September, October and November. The results revealed its successful growth in the subtropical region of Jammu, with last week of October ideal for obtaining maximum dry herbage (931.3 kg/ha) and total andrographolide yield (61.83 kg/ha), respectively. On the basis of leaf/stem ratio the relative estimates of total andrographolide is higher in October (46%) than September (15%) or November (39%). Accession Acc.1 and Acc.9 have been identified as the best sources for obtaining higher drug yield.

Six accessions of geranium (Pelargonium sp. ‘hybrid’) were introduced in temperate and subtropical regions of Jammu and Kashmir. The herbaceous parts in the commercial distillation gave 0.09–0.13% oil on fresh weight basis. The calculated herbage yield (tonnes/ha) and oil yield (litre/ha) varied from 31.5 ± 1.87 to 49.1 ± 2.73 and 32.40 to 57.12, respectively. The geranium oil produced in different regions of Jammu and Kashmir was comparable to that produced in other geranium-growing areas in India in terms of its major constituents, citronellol (22.5–34.5%) and geraniol (13.9–24.7%), respectively. This suggests that geranium can be successfully grown in the state of Jammu and Kashmir. Copyright © 2006 John Wiley & Sons, Ltd.

Six varieties/strains belonging to three species of Cymbopogon, viz., C. flexuosus, C. khasianus and C. pendulus, or their hybrids, were evaluated for oil yield across four years (1997-98-2000-01) as very limited information is available on variety-by-environment interaction from multi-environment trials. The main effects, varieties and years, were significant, but variety-by-year interaction was not significant. Log-transformation of data reduced coefficients of variation from 45.52% to 16.7% and increased R-square for the linear model from 70.8% to 81.9%. A traditional analysis of variance indicated that CKP25 and F2-38 were not different in oil yield and were superior to other varieties. Principal component analysis of GGE biplot methodology also revealed CKP25 to be the highest yielding genotype in all four years, followed by F2-38. Variety CKP25 could be used in a breeding program to develop new high yielding, consistent-performing varieties. Both untransformed and log-transformed data analy...

The effect of seasonal changes on oil content of a new clone of Jamrosa ‘RL‐931’ (Cymbopogon nardus var. confertiflorus × C. jwarancusa) was investigated during 1998–99. The pre‐monsoon and onset of the monsoon period (May‐July) was characterised by a higher oil content, whereas the post‐monsoon and winter period (August‐December) showed a comparatively lower oil content. The quality of the oil is best in May‐July. High temperature and low humidity favours the accumulation of geraniol, geranyl acetate, and a low citral content in the oil. Maximum oil content was recorded when both maximum and minimum temperatures were high.

We prospectively examined the incidence of bacteriuria in malnourished patients between 6 months and 5 years of age. For each patient, a normally nourished control matched for age, sex, and presence of fever and diarrhea was included. Of 112 patients (65 boys), 55 had moderate and 57 had severe malnutrition; 43 had diarrhea and 35 had fever. Clean-catch and suprapubic urine specimens were examined microscopically and cultured. Significant bacteriuria was found in 17 (15.2%) malnourished and 2 (1.8%) control subjects ( P

SummaryWe have prepared human recombinant antibody molecules against the glycoprotein antigen of the rabies virus (GPRV) based on the single chain variable fragment (scFv) format. Anti-GPRV scFvs were selected from a human synthetic scFv phage display library with a repertoire of approximately 109 specificities. After three rounds of selection against the PV11 strain of the virus, 40% of the clones tested recognized the rabies antigen. Of the 20 positive clones that were sequenced, five distinct sequences were identified. These distinct scFvs were cloned into a mammalian expression vector carrying the human IgG1 Fc region. The specificity of the resulting scFv-Fc molecules for GPRV was established by ELISA, dot blot and western blot analyses and membrane immunofluorescence. Two of the scFv-Fc fusion proteins neutralized the PV11 strain in a standard in vivo neutralization assay where the virus was incubated with the scFv-Fc molecules before intracranial inoculation in mice. These anti-GPRV scFv-Fc molecules have the potential to be used as an alternative to the presently available HRIG, for use in post-exposure preventive treatment.

Gamma irradiation treatment (30 Gy) of stem cuttings with single leaf and an axillary bud of sexually sterile patchouli [Pogostemon cablin (Blanco) Benth.] has resulted in isolation of an erect plant mutant ‘PM-12’ with thick, dark green, illuminated and shiny leaves. This single plant with distinctly different morphology on vegetative propagation produced all erect plants with dark green shiny leaves. The radio mutant phenotype persisted over generations by stem cutting propagation. The essential oil quality profile of radio mutant revealed a significant higher percentage of patchouli alcohol, the main component of patchouli oil compared to its parental clone (60.3% against 47.8% in control). Creation of variability in patchouli alcohol content of the oil has more commercial viability because it imparts the patchouli oil a characteristic earthy piquant tenacious aroma.

Tanacetum gracile, an alpine desert aromatic herb collected from Ladakh Himalaya, on steam distillation yielded 0.46% (DWB) essential oil having a characteristic blue colour. The essential oil contained lavendulol (21.5%) as a major chemical constituent. The other compounds include lavandulyl acetate (1.7%), 1,8-cineole (15.2%), cis-β-ocimene (6.4%) borneol (6.1%) and chamazulene (3.7%). The major components were identified by 1 HNMR, 13 CNMR, GC-MS and

Enteroaggregative Escherichia coli (EAggEC) is a recently discovered diarrhoeal pathogen implicated as a cause of persistent diarrhoea in children. EAggEC strains exhibit a characteristic pattern of adherence when incubated with HEp-2 cells. Because of the difficulty in identifying this group of bacteria, the epidemiological significance of this pathogen as a diarrhoeal agent has not been fully realised. A gene probe was developed from the 60-MDa plasmid associated with EAggEC strains that encodes the genes for adherence and fimbriae. The sensitivity of the gene probe was 93% and the specificity 98% for detecting EAggEC isolates and is potentially useful for diagnostic and epidemiological studies.

Dormant vegetative slips of Jamrosa were subjected to various doses of gamma-rays. Plants raised from them were screened with a view to isolate improved clones of the crop. Five mutant clones isolated, exhibited variation in quality/quantity of essential oil. These changes in oil characters were attributed to microlevel mutations induced by gamma-rays.

Fibromatosis, arare non-neoplastic spindle cell proliferation of unknown aetiology, can occur anywhere in the body. Though extra-abdominal sites are commonly involved, intra-abdominal fibromatosis has also been described. Described herein is an unusual case of diffuse intra-abdominal fibromatosis in a 9-year-old boy, who could not be salvaged despite extensive medical management.

Phenotypic and genotypic coefficient of variability, heritability in broad sense and genetic advance were determined investigating the characters of 15 accessions of Silybum marianum. Seed yield/plant and number of capsules/plant had highest estimates of genotypic variation, heritability and genetic advance which suggest that direct selection for these traits is suitable for the improvement of this crop. Number of capsules/plant had a significant positive correlation with the number of branches/plant and leaf length (r=0.3398, 0.7547), whereas seed yield/plant had a positive significant correlation with leaf length, stem diameter, capsule diameter and silymarin content (r=0.6830, 0.3140, 0.3484, 0.2925).

In developing countries like India, unlike in the West, diagnosis of celiac disease is a dilemma because the histological changes consistent with celiac disease may not be pathognomonic of the disease. We need clarity on how to use one or more of the serological antibodies, namely IgG and IgA anti-gliadin and IgA anti-endomysial antibodies as independent disease markers or as adjuncts to clinico-histological diagnosis. In this review, various possible algorithms have been discussed. The serological antibodies can be used as reliable screening tests to decide whom to biopsy and to interpret histological changes in our settings where malnutrition and intercurrent enteric infection are common.

Chlamydia trachomatis infection in pregnant women is suspected to result in low birth-weight and premature infants. We conducted studies to ascertain the prevalence of this infection among pregnant women in our setting and whether its presence is a risk factor for low birth-weight or prematurity.In the first study, 94 pregnant women between 26 and 30 weeks of gestation were screened for infection with Chlamydia trachomatis. The second investigated a cohort of 172 pregnant women presenting in spontaneous labour. The infection status was related to perinatal outcome in terms of birth-weight and gestation. In both the studies, Chlamydia trachomatis infection was diagnosed using the Chlamydiazyme test performed on endocervical swabs.The prevalence of Chlamydia trachomatis infection in mid-pregnancy and at labour was 17% (16/94) and 18.6% (32/172), respectively. Women with infection were relatively older than those without it [mean (SD) age: 26.6 (4.5) years v. 24.8 (3.6) years, p = 0.01]. The mean (SD) birth-weight [2869 (611) g v. 2814 (496) g], gestation [38.5 (2.6) weeks v. 38.3 (2.0) weeks], and incidence of low birth-weight [18.7% v. 20.7%] as well as prematurity [9.4% v. 10.7%] were similar among neonates born to women with or without infection. Neonates born to infected mothers experienced purulent conjunctivitis more frequently than those born to non-infected mothers [12.5% v. 2.8%, p = 0.04].Chlamydia trachomatis is a relatively common infection in pregnant women. However, it was not associated with either low birth-weight or prematurity.

This prospective study was undertaken to study the occurrence of histologic chorioamnionitis and determine its association with prematurity; and to assess whether colonization of the genital tract of pregnant women by genital mycoplasmas or Chlamydia trachomatis is a risk factor for histologic chorioamnionitis. A total of 268 women with singleton pregnancies of over 26 weeks gestation were subjected to high vaginal cultures of genital mycoplasmas and endocervical specimens for chlamydia antigen. Placental histopathology was performed on multiple sections. Histologic chorioamnionitis was documented in 22.4 per cent (60/208) placentae. Genital tract colonization with Ureaplasma urealyticum or C. trachomatis was not a risk factor for histologic chorioamnionitis. Neonates born in association with histologic chorioamnionitis had a mean birth weight 111 g lower than those born without this lesion (2626.9 +/- 702 g vs 2737.8 +/- 500 g, NS). The relative risk (95% confidence interval) of prematurity in the presence of histologic chorioamnionitis was 1.49 (0.87-2.53). Analysis of linear trend in proportions for prevalence of histologic chorioamnionitis with decreasing gestation showed a significant association (P = 0.047, 1-tail). These results taken together suggest that histologic chorioamnionitis may be a risk factor of prematurity, but of only a modest magnitude.

A randomized, controlled trial was undertaken to compare nutrient absorption from low fat (22.2% energy as fat) and high fat (48% energy as fat)-containing mixed diets in 39 (26M, 13F) malnourished children aged2 y. Subjects of both dietary groups, standard (SDG, n = 18) and experimental (EDG, n = 21), were offered 184.5 ml of feeds/kg body weight/24 h that provided 146 kcal with the low fat diet and 216 kcal with the high fat diet, respectively. The baseline clinical and biochemical parameters and the volume of feeds consumed by the patients in the two groups were comparable. Thus, the subjects in the EDG were ingesting a mean (95% CI) of 50% (23.8%, 76.2%) extra energy than that eaten by the SDG. Coefficients of absorption of energy, nitrogen and carbohydrate were similar in the two dietary groups (p = 0.08-0.98). Median (binomial exact 95% CI) fat absorption among subjects receiving the high fat diet (96.3; 92.9, 98.6) was 7% more than in those consuming the low fat diet (89.3; 86.7, 94.1) (p = 0.01). Subjects of the EDG thus, retained almost 50% additional energy as compared to that by SDG.

The rotavirus nonstructural glycoprotein NSP4 may represent the first identified viral enterotoxin. We have sequenced reverse transcription-polymerase chain reaction (RT-PCR)-generated fragments of 16 NSP4 genes of human rotavirus (HRV) strains from six different countries, representing seven different G and P type combinations. Based on the amount of sequence divergence between these and 11 previously sequenced NSP4 genes of human and animal rotaviruses, three distinct genetic groups could be recognized. Most strains within a group were closely related to each other at the nucleotide (nt) and amino acid (aa) levels (usually10% divergence) but more distantly related (maximum 30.0% nt divergence and 24.7% aa divergence) to members of the other groups. Intergroup variation occurred in two highly variable regions of NSP4 (aa 16-34 and aa 131-148). The NSP4 "toxic peptide" (aa 114-135) exhibited aa variation at its carboxy terminus both within and between genetic groups. The largest group (genetic group II) contained HRV strains of subgroup II specificity (including genotypes P[8]G1, P[8]G3, P[6]G3, and P[8]G5 and serotype P8[11]G9), and the smaller group (genetic group I) contained HRV strains of subgroup I specificity (genotype P[4]G2). The NSP4 sequence of the rhesus rotavirus vaccine strain was distinct from all other strains and formed the third group (genetic group III). The NSP4 genes of animal rotaviruses UK, NCDV, and SA11 (genetic group I) and YM (genetic group II) and two possible human-animal rotavirus reassortant strains, Brazilian P[8]G5 and Indian P[11]G9 (genetic group II), could also be classified into one of these groups, suggesting a close evolutionary relationship between human and animal NSP4 genes. These results will facilitate studies of the host immune response to NSP4, which may be relevant to future HRV vaccine design.

In a zinc supplementation trial (with a significant impact on diarrheal morbidity), to evaluate effect of zinc supplementation on cellular immune status before and after 120 days of supplementation.A double blind, randomized controlled trial with immune assessment at baseline and after 120 days on supplement.Community based study in an urban slum population.Randomly selected children (zinc 38, control 48), had a Multitest CMI skin test at both times. In 66 children (zinc 22, control 34), proportions of CD3, CD4, CD8, CD16, CD20 cells and the CD/CD8 ratio were also estimated using a whole blood lysis method and flowcytometry.Zinc gluconate to provide elemental zinc 10 mg daily and 20 mg during diarrhea.Regarding CMI, the percentage of anergic or hypoergic children (using induration score) decreased from 67% to 47% in the zinc group, while in the control group it remained unchanged (73% vs 71%) (p = 0.05). The percentage of children deteriorating between first and second tests was significantly lower in the zinc group (13% vs 33%, p = 0.03). Regarding lymphocyte subsets, the zinc group had a significantly higher rise in the geometric means of CD3 (25%, p = 0.02), CD4 (64% p = 0.001), and CD4/CD8 ratio (73% p = 0.004) with no difference in CD8 and CD20. The rise in CD4 was significantly higher in the zinc as compared to the control group; the ratio of geometric means was 1.45 (95% CI, 1.03-2.01).Zinc supplementation improves cellular immune status, which may have been one of the mechanisms for observed impact of zinc supplementation on diarrheal morbidity.

We have sequenced the genes encoding the inner capsid protein VP6 and the nonstructural proteins NSP1 and NSP4 of the Indian neonatal serotype P8[11]G9 human/bovine reassortant candidate vaccine rotavirus strain 116E. These three genes share a high degree of sequence and deduced amino acid homology with human prototype strain Wa. Our results confirm and extend those of previous RNA-RNA hybridization studies which suggested that these genes are of human origin, and will facilitate examination of the host immune response to 116E induced by natural infection and vaccination.

To investigate whether supplementation of zinc in preschool children is associated with improvement in observed activity levels.On 2 consecutive days, we performed 5-hour observations with momentary time sampling (instant activity every 10 minutes) in children selected from an ongoing double-blind, randomized trial of zinc supplementation. The study was conducted in Kalkaji, a low-socioeconomic urban population of New Delhi with high diarrheal incidence and rates of malnutrition. A total of 93 children (48 zinc and 45 control) 12 to 23 months of age from an ongoing community-based, randomized, controlled trial received supplements for at least 1 month before study; 71% had received supplementation for more than 120 days. Zinc gluconate (10 mg of elemental zinc) was given daily, with both zinc and control groups receiving vitamins A, B1, B2, B6, D3, and E and niacinamide in addition.Outcomes were percentages of time spent in each of five activity levels and two groups representing high and low movement and overall rating by two activity scores. Children in the zinc group spent 72% more time performing activities in the high-movement group. Among the zinc-supplemented children, the activity rating by the children's activity rating score was 12% higher and by the energy expenditure score was 8.3% higher than in the control group.In conclusion, zinc supplementation, given along with selected vitamins, was associated with significantly greater activity levels in children. The relationship between the activity increase and locomotor development needs to be investigated, as do the long-term implications of zinc supplementation in terms of developmental status and school performance.

Previous studies have shown increased stool output when children with persistent diarrhea (PD) received milk as the predominant source of nutrition.We evaluated the efficacy of milk given in modest amounts as a part of a mixed diet in children with PD. One hundred sixteen children 3 to 24 months of age with diarrhea for between 14 days and 12 weeks were allocated to milk-based (n = 60) or milk-free (n = 56) cereal dietary regimens. The two diets were isocaloric (86.9 calories/100 g foror = 9 months; 95.6 cal/100 g for9 months) consisting of puffed rice cereal, sugar, and oil differing in only their source of protein, which was either milk or egg white, respectively. An average of 30% of the calories were constituted by milk in the milk-cereal diet. Both diets were offered at the rate of 150 kcal/kg per day. Children receiving milk-cereal consumed an average of 1.9 g/kg lactose per day.The baseline characteristics in the two groups were similar. Comparable amounts of diet were consumed in both groups. The milk-cereal group did not have higher median (range) stool output (g/kg/h) compared with the milk-free group during a 0- to 48-hour (milk-cereal, 1.7 [0.2 to 8.7]; milk-free, 1.5 [0.1 to 6.6]) or 0- to 120-hour (milk-cereal, 1.6 [0.4 to 7.2]; milk-free, 1.3 [0.1 to 7.6]) period. The percentage of weight gain was similar in the two groups, and there were no significant differences in the duration of diarrhea. Overall, 23 children had treatment failures, 10 (17%) in the milk-cereal and 13 (23.6%) in the milk-free groups.Our findings suggest that modest intakes of milk are well tolerated as a part of mixed diet during PD.

Diarrhoea that begins acutely but lasts longer than two weeks is defined to be persistent. Revised estimates in developing countries including India showed that acute diarrhoea accounts for 35 per cent, dysentery 20 per cent and non-dysenteric persistent diarrhoea (PD) for 45 per cent of total diarrhoeal deaths. PD also often changes marginal malnutrition to more severe forms. Factors that increase the risk of acute diarrhoea becoming persistent have been identified in India and other developing countries. These include antecedent malnutrition, micronutrient deficiency particularly for zinc and vitamin A, transient impairment in cell mediated immunity, infection with entero aggregative Escherichia coli and cryptosporidium, sequential infection with different pathogens and lack of exclusive breast feeding during the initial four months of life particularly use of bovine milk. Several issues regarding the management of persistent diarrhoea in hospitalized children in India have been resolved. Diets providing modest amounts of milk mixed with cereals are well tolerated. In those who fail on such diets providing carbohydrate as a mixture of cereals and glucose or sucrose hasten recovery. The role of antimicrobial agents and individual micronutrients in PD is currently being investigated. A management algorithm appropriate for India and other developing countries has been developed and found to substantially reduce case fatality in hospital settings to about 2-3 per cent. Recent epidemiological and clinical research related to persistent diarrhoea is also reviewed.

Enteroaggregative Escherichia coli (EAggEC) is a distinct category of diarrheal pathogen implicated as the cause of persistent diarrhea. The pathogen exhibits a characteristic "stacked-brick" pattern of aggregation when incubated with HEp-2 cells. The outer membrane protein (OMP) profile of a prototype EAggEC strain (F03) reflected the presence of one major 30-kDa protein. The OMP is expressed in the presence of the 60-MDa plasmid that the strain harbors. Antibodies were raised against the OMP by injecting the protein into a rabbit. The manifestation of an adherence phenotype on HEp-2 cells was observed for F03 and other strains that express OMP in the presence and absence of anti-OMP serum. Clumps of bacteria forming an aggregative pattern were observed in the HEp-2 cell assay in the absence of OMP antibodies, whereas a few bacteria attached to the cells in the presence of OMP antibodies. Mannose-resistant hemagglutination of human erythrocytes observed in the presence of EAggEC strains was inhibited in the presence of anti-OMP serum. Sequence analysis of a peptide generated by protease digestion of OMP exhibited 90% homology to a peptide of flagellin protein encoded by the hag gene of Serratia marcescens. Immunolabeling of the outer membrane by colloidal gold confirmed the protein to be an OMP. Our results suggest that the OMP of EAggEC have common antigenic properties. Antibodies raised against the protein can prevent adherence in vitro and could potentially interrupt the natural disease.

The ability of HEp-2 cell adherent Esch. coli of aggregative phenotype (EA-Agg EC) to cause diarrhoea and to colonize the bowel of rabbits was studied. Thirty six rabbits were challenged with one of three EA-Agg EC strains (F23A; H766C and F17A-15, 3 and 3 rabbits respectively) or a control strain (K12-15 rabbits) in reversible ileal-tie in adult rabbit diarrhoea (RITARD) model. The animals were sacrificed 72 h post challenge. Severe diarrhoea occurred in greater number of F23A challenged rabbits than the controls (P0.05). Mucosal cultures from proximal and distal small intestine and colon yielded about 1000 times more Esch. coli in the test than control rabbits (P0.001 in each case). EA-Agg EC were consistently grown from mucosa in the test rabbits who commonly showed mild to moderate villous stunting and grade + to nuclear fragmentation (karyorrhexis) in the small and large bowel epithelium. The control animals had either normal villi or very mild villous stunting. Results comparable to F23A were obtained with the other two EA-Agg EC strains tested in a smaller number of animals.

A collection of 44 enteroaggregative Escherichia coli (EAggEC) strains isolated from infants with diarrhea in India and the United Kingdom were examined for their ability to adhere in vitro to human intestinal mucosa and by electron microscopy for production of putative adherence factors. None of the strains adhered to human duodenal mucosa, and six strains tested did not adhere to ileal mucosa; all 44 strains, however, adhered to human colonic mucosa in localized aggregates. Electron microscopy of infected colonic mucosa indicated fimbrially mediated adhesion of the EAggEC strains. Four morphologically distinct kinds of fimbriae, including a new morphological type of E. coli fimbriae consisting of bundles of fine filaments, were identified among the EAggEC strains; this new type of fimbria was observed in 43 of the 44 EAggEC strains. Forty-three of the 44 EAggEC strains were positive with a DNA probe developed to identify EAggEC, and most of the strains belonged to serotypes unrelated to the other major classes of diarrheic E. coli. These results suggest that EAggEC may be a large-bowel pathogen and colonize the colon by a fimbrially mediated adhesion mechanism.

The present study describes a nonisotopic DNA-DNA hybridization assay for the detection of enterotoxigenic Escherichia coli (ETEC) in which the probes were labelled with the hapten molecule digoxigenin and after hybridization, DNA hybrids were detected by antidigoxigenin alkaline phosphatase conjugate. A blinded study carried out on a battery of enterotoxigenic and nonenterotoxigenic Esch. coli by dig-probe hybridization assay were compared with the results of a radiolabelled toxin gene probe hybridization assay (performed earlier). The three digoxigenin labelled probes gave a 100 per cent specificity and sensitivities of 95.45, 100 and 100 per cent for LT, STh and STp respectively. These results were comparable to those with the radioactive probes.

This randomized, double-blind trial determined whether adding 90 mmol/L of alanine with a reduction in glucose to 90 mmol/L (alanine ORS) improves the efficacy of the standard oral rehydration solution (WHO-ORS). One hundred twenty-nine males aged 3-48 months with weight for length greater than or equal to 70% of NCHS, diarrheal duration less than or equal to 96 h, and clinical signs of mild to moderate dehydration were randomly allocated to either treatment group. During 0-6 h of treatment, ORS was offered at 120 ml/kg for rehydration without food or water. Beyond 6 h, ORS was offered as a volume-to-volume replacement for stool losses and a mixed diet of uniform composition was offered in amounts standardized for body weight. The most frequently isolated pathogens in alanine ORS and WHO-ORS groups were rotavirus (42 and 48%, respectively) and enterotoxigenic Escherichia coli (15 and 12%, respectively). In the 0-6 h period when food was withheld, median urine output in ml/kg (8;5, p less than 0.05) and percentage decrease in total serum solids (9:7%, p = 0.06) was significantly greater in alanine ORS than in WHO-ORS; median ORS intake and stool output were marginally lower in the alanine group but the differences were statistically not significant. Between 0 h and recovery, although the median values for duration of diarrhea (56.5 and 65.0 h), ORS consumption (260 and 323 ml/kg), and stool output (188.4 and 216.3 g/kg) were lower in the alanine ORS group, these differences with the WHO-ORS group were not statistically significant.(ABSTRACT TRUNCATED AT 250 WORDS)

The associated factors in 80 children (less than 2 yrs) with protracted diarrhea (greater than 21 days duration) and weight loss were: secondary carbohydrate intolerance (36): enteric pathogens (non typhoidal salmonella (11), enteropathogenic E. coli 'EPEC' (6), giardia (4), and shigella (3); cow's milk protein intolerance (3), gluten intolerance (3); miscellaneous (5); and undiagnosed enteropathy (9). Three of the EPEC showed localised pattern of adherence in vitro with HEP-2 cells. Most patients with salmonella and EPEC had severe secretory diarrhea with large fecal sodium losses. All 6 patients who died had secretory diarrhea and very high fecal sodium. All but 4 patients could be effectively managed with a chicken puree-glucose-coconut oil based diet.

Brush border lactase, sucrase and glucoamylase activities were assessed in jejunal mucosal biopsy specimens from 34 children (median age 11 months; range 1.5-38) having protracted diarrhoea with failure to thrive and 8 well nourished children with normal jejunal mucosal histology (median age 10.2 months; range 2-37). All enzymes showed progressive decrease in activity which was directly in relation to increasing degree of mucosal injury (P less than 0.002). Lactase was significantly reduced even in patients with protracted diarrhoea and normal mucosa (P less than 0.05). Glucoamylase and sucrase were significantly reduced only in the presence of mucosal injury (P less than 0.01). Our data suggest that most children with protracted diarrhoea may not tolerate lactose containing feeds and may need lactose-free diets preferably based on starch. A small number of children with protracted diarrhoea, who have severe mucosal injury may not be able to handle even starch and may require diets based on short chain glucose polymers. The findings of this study, need to be corroborated with well-controlled metabolic balance studies.

ADHERENCE PROPERTIES OF ENTEROAGGREGATIVE ESCHERICHIA COLI: A NEWLY DESCRIBED CLASS OF DIARRHOEAGENIC E.COLI

An oral rehydration therapy program was introduced in a rural community in India. During the initial 20 months (phase 1), packeted oral rehydration solution (ORS) was promoted and made available through the dispensary and also through 5 village health workers for treatment of all diarrheal illness, regardless of the severity. In the subsequent 11 months (phase 2), the supply of ORS packets was restricted only to cases of obvious dehydration when treatment was sought at the local dispensary. Preparation and correct use of the sugar-salt solution for early treatment of diarrhea were explained to all of the mothers in the villages. The diarrheal episodes and use of ORS were monitored through household visits by data collectors throughout the 31-month period. In phase 1, ORS packets were used in 68.6% of the diarrheal episodes whereas in phase 2, sugar-salt solution was used for 79.3% of the episodes. The mean intake (in liters)/diarrheal episodes was higher (p0.05) for sugar-salt (1.9 +or- 1.6) as compared to the ORS solutions prepared from packets (1.2 +or- 0.7). The percentage of samples with sodium concentration 120mE q/1 was 4.9% with ORS packets and 20% with sugar and salt (0.05). The diarrheal deaths/100 episodes were 0.73 (ORS packets phase) and 0.50 (sugar-salt phase), respectively (p0.05).

The safety of 2 oral rehydration solutions (ORS) containing 60 and 90 mEq/1 of sodium respectively was evaluated in 50 children with mild to moderate dehydration secondary to noncholera diarrhea. Hypernatremia developed in 1 patient (3.7%) on high sodium formula. The risk of hypernatremia and hyponatremia in the 2 groups did not differ significantly. 3 patients (6.1%) showed hypokalemia 24 hours after oral rehydration was initiated. While these data confirm that ORS containing 90 mEq/1 of sodium is safe, an increase in potassium content should be considered.

The significance of IgM and IgG class antibodies to hepatitis B virus (HBV) core component (anti-HBc) was investigated in a study of maternal-fetal HBV transmission. An IgM anti-HBc response was lacking in the majority (49/53) of HBV-infected infants. This antibody thus cannot be used as an indicator of transplacental infection. However, most infants who became HBsAg positive during the first 6 months of life acquire infection in the perinatal period rather than transplacentally. Passively transferred maternal IgG anti-HBc in the infant and additional IgM anti-HBc positively in the carrier mother have no modulating influence on HBV infection of infants born to HBV carrier women.