Association of VDR Gene Variant (rs1544410) with Type 2 Diabetes in a Pakistani Cohort.

The present study was designed to measure the mean values of body mass index (BMI), random blood sugar/ fast ing blood sugar (RBS/FBS) tests, and Hb A1c and to investigate the role of a genetic variant rs1544410 in the VDR gene in a Pakistani cohort. For this purpose, a total of 917 samples including 469 diabetes mellitus type 2 (T2DM), 145 DM type 1 (T1DM), and 303 healthy control were collected. Out of the total sample set, 500 individuals (250 T2DM cases and 250 controls) were genotyped for rs1544410. It was found that 65 (26.0%) cases and 32 (12.8%) controls had homozygous AA, while 69 (27.6%) cases and 139 (55.6%) controls had heterozygous AG, and 116 (46.4%) cases and 79 (31.6%) controls had homozygous GG (χ2 = 41.81, p = 0.0001). In addition, a similar distribution of allele frequency was determined in cases and controls [p value = 0.866; odds ratio (OR) = 0.967; relative risk (RR) = 1.034]. A significant difference was observed in homozygous dominant [OR = 2.394 (1.501-3.816); RR = 1.46 (1.225-1.740); p = 0.003] and homozygous recessive models [OR = 2.970 (2.086-4.227); RR = 1.798 (1.501-2.154); p = <0.0001] analysis of rs1544410 in the VDR gene. These findings suggest that the VDR gene is associated with T2DM and genotype GG of genetic variant rs1544410 is the susceptible genotype in our Pakistani cohort.

Molecular epidemiology of Staphylococcus aureus in food samples based on the protein A gene polymorphic region DNA sequence.

Foodborne disease due to Staphylococcus aureus is a common and important disease worldwide. Molecular typing of S. aureus strains plays a crucial role in epidemiological studies examining the origin and performing surveillance of major infections. In this survey, we collected 913 food samples and detected 93 S. aureus isolates by using culture and biochemical tests. Subsequently, the X region of the protein A gene was amplified by the polymerase chain reaction (PCR) and amplicons were digested with HaeII and HindIII. Seven different patterns, ranging in length between 1200 and 1600 bp, showed gene polymorphism of the spa gene. The most prevalent spa types were D (20%) and C (16%) in dairy products and D (6%) and E (3%) in meat products. Consequently, 16 genotypes were obtained by HaeII digestion. Typeability of PCR - restriction fragment length polymorphism in genotyping of S. aureus strains in our study was perfect. Therefore, this method is a reliable, rapid, and powerful system in epidemiological investigations.

Determination of the efflux pump-mediated resistance prevalence in Pseudomonas aeruginosa, using an efflux pump inhibitor.

In gram negative bacteria, fluoroquinolone resistance is acquired by target mutations in topoisomerase genes or by reducing the permeation of drugs due to the increase in expression of endogenous multidrug efflux pumps that expel structurally unrelated antimicrobial agents. An ongoing challenge is searching for new inhibitory substances in order to block efflux pumps and restore the antibiotic drugs susceptibility. In this research, we sought to investigate the interplay between ciprofloxacin and an efflux pump inhibitor (EPI), phenyl alanine arginyl beta-naphtylamide (PAbetaN), to determine the prevalence of efflux pump overexpression in clinical isolates of Pseudomonas aeruginosa. Ciprofloxacin was tested at different concentrations (256-0.25 microg/ml) with a fixed concentration of PAbetaN (50 microg/ml). The isolates susceptibility profiles were analyzed by disc diffusion and agar dilution methods using 10 antibiotic discs and 4 powders. It was found that in the presence of PAbetaN, resistance to ciprofloxacin was inhibited obviously and MIC values were decreased. The comparison between subgroups of P. aeruginosa isolates with different resistance profiles indicates that efflux pump overexpression (EPO) is present in 35% of ciprofloxacin resistant isolates with no cross resistance and in variable frequencies among isolates showing cross resistance to other tested antibiotics: gentamicin (31%), ceftazidime (29%), and imipenem (18%). Altogether, these results imply that PAbetaN maybe effective to restore the fluoroquinolone drugs susceptibility in clinical treatment procedures. Results also show that increased use of a fluoroquinolone drug such as ciprofloxacin can affect the susceptibility of P. aeruginosa to other different antipseudomonal agents.

Ethnic differences in the association between body mass index and impedance index (Ht2/Z) in adult women and men using a leg-to-leg bioimpedance method.

BACKGROUND: Ethnic differences in the association between body mass index (BMI) and body fat suggest that body composition varies across ethnic groups. OBJECTIVE: To investigate the association between impedance index - a measure of tissue resistivity - and BMI in adults of different ethnic groups (Asian Indians, West Africans and White Caucasians) living in their native countries. METHODS: Male (n=329) and female (n=277) adult subjects (18-50 years) living in urban areas in the UK, The Gambia and Pakistan were studied. Body weight and height were measured and BMI calculated. The same leg-to-leg bioimpedance instrument was used in each study and impedance index (height(2) (cm)/impedance (Omega)) used as measure of tissue resistivity. RESULTS: In women, Asian Indians and West Africans had a significantly greater increase in impedance index per unit increase in BMI compared with white Caucasians (P<0.001). In men, Asian Indians had a significantly lower impedance index compared with West Africans and white Caucasians (P<0.001). CONCLUSION: Different ethnic groups may have different tissue resistivity for the same BMI indicative of systematic differences in body composition.

Mucosal immunity.

Mucosal defense is provided by a number of host factors countering the specific virulence factors of the many microorganisms infecting the mucous membranes. Secretory IgA antibodies presumably play an important role. Increase of the sIgA antibodies may most advantageously be attained by parenteral immunization, following mucosal priming. This was demonstrated in a rat model, where it was also noted that antigen injection into PP induced high milk IgA antibody levels. In man, parenteral vaccination against polio increased the sIgA antibody levels in the milk of mothers previously exposed naturally to the poliovirus. The response was relatively short-lived. In the previously unexposed, there was little or no response. By contrast peroral immunization with live poliovirus vaccine did not increase, or even decrease, the milk sIgA poliovirus antibody levels. Although salivary sIgA antibodies against antigens of colonizing E. coli appear during the first days of life, they are slow to increase. This deficiency is richly compensated for by all the sIgA antibodies that are provided the baby through the milk. No transfer of dimeric IgA into the milk could be shown in lactating rats, in contrast to what has been reported in mice. There is no evidence for a contribution to milk sIgA from serum in man. Close to parturition, human milk often contains some 7S IgA and various sizes of free SC, in addition to the dominating 11S sIgA. A few days later there is almost exclusively monomeric SC and 11S sIgA. IgG antibodies also play a role at the mucosal level. IgG2 antibodies against the bacterial polysaccharide capsule are as slow to appear as sIgA in ontogeny, possibly explaining the prevalence of infections with encapsulated bacteria and the poor response to polysaccharide vaccines in early childhood. Other defense factors preventing infections by way of mucous membranes may be important. Thus, oligosaccharides present in human milk seem to specifically prevent pneumococcal attachment to retropharyngeal cells. This anti-attachment capacity, in addition to that provided by milk and salivary IgA antibodies, may explain why breast-fed babies have less otitis media than formula-fed ones.

Interaction of P-fimbriated Escherichia coli with human meconium.

The ability of Escherichia coli with different receptor specificities to interact with meconium was studied. E. coli strains expressing P-fimbriae, specific for Gal alpha 1-4Gal beta-containing receptors, were agglutinated by meconium at high titres. This reaction was inhibited by globotetraosylceramide. The attachment of P-fimbriated E. coli to human colonic epithelial cells of the HT-29 cell line was inhibited by meconium. Some type 1 fimbriated strains were agglutinated by meconium, but the agglutination was rarely blocked by methyl alpha-D-mannoside. The attachment by type 1 fimbriated strains to HT-29 cells was reduced by meconium only in some cases. These results suggest that meconium interacts with the P-fimbriae of E. coli, in a way that may influence bacterial colonization of the neonatal intestine.

Immunogenicity and reactogenicity of rhesus rotavirus vaccine given in combination with oral or inactivated poliovirus vaccines and diphtheria-tetanus-pertussis vaccine.

Immunogenicity and reactogenicity of the oral rhesus rotavirus vaccine (RRV) were assessed among 72 infants (6 weeks old) in Lahore, Pakistan, from August to December 1985. Special emphasis was placed on the possible interaction or interference caused by giving RRV at the time infants received their first polio immunization. RRV was given to the infants at the same time as diphtheria-tetanus-pertussis (DTP), oral poliovirus vaccine (OPV), or inactivated poliovirus vaccine (IPV). The immune response to RRV was assessed by plaque-reduction neutralization 3 weeks after immunization and serum immunoglobulin (Ig) G and IgA antibody levels to poliovirus type 1 were tested by enzyme-linked immunosorbent assay (ELISA) after polio immunizations. Of the infants in the group given RRV with OPV, 50% had a two- to four-fold rise in neutralization titre against rotavirus, compared with 22% in the group given RRV with DTP and 20% in the group given RRV and IPV (P less than 0.05). Interference by live oral polio vaccination in the response to RRV seems unlikely. We observed no significant difference in rates of seroconversion of IgG antibodies to poliovirus type 1 among infants aged 18 and 21 weeks who received RRV and OPV (81%), RRV with delayed OPV (67%), or RRV and IPV (59%). Administration of RRV was safe and was not associated with adverse reactions in the 6 weeks old infants. The low rate of seroconversion to rotavirus suggests that a more antigen-rich vaccine or multiple doses of the same vaccine might produce a better immune response.

Growth stunting in early life in relation to the onset of the childhood component of growth.

AIMS: Growth stunting is prevalent in developing countries. The prevalence of stunting ranges from 10-80% or more. The critical period for growth faltering in length is between six and 18 months. The aims of this study were 1) to investigate the association between the age at onset of the childhood component and growth stunting in early life; 2) to develop a screening method for identifying any individual infant subject to stunting in early life. MATERIALS & METHODS: This community-based longitudinal study was carried out in Lahore, Pakistan. Three studied areas and one local control group were included in the study. There were 425 subjects for whom data were available for determining the age at onset of the childhood component. The onset was defined as an abrupt increase in length velocity. It was determined individually using the infancy-childhood-puberty growth model. RESULTS: The medians of the age at onset of childhood component were 15, 13, 10 and 9 months in the periurban, village, urban and control groups, respectively. The onset was significantly related to both length and length velocity during the critical period of stunting. Based on the distribution of individual length increment from 12 to 15 months, 2.0 and 3.0 cm were chosen as the cut-off points to identify a stunting infant. In the normal onset group, 84% of the children were identified as growing normally; in the delayed onset group, 61% were identified as stunting. Using this screening method, 97% of children have a delayed onset if their length increase is below 2 cm from 12 to 15 months; 99% of them have a normal onset if the increase is above 3 cm over the same period. CONCLUSIONS: The age at onset of the childhood component can significantly explain the variations in both length and length velocity in early life. The stunting screening method reported here is simple. It can be used in almost any situation, such as a pediatric clinic or community-based survey.

The mammary gland-infant intestine immunologic dyad.

The human infant has a very small immune system and needs the support of the mother with the transplacentally arrived IgG antibodies to protect tissues with inflammatogenic and energy-consuming defense. The mucous membranes, where most infections occur, need support via the specialized secretory IgA antibodies and the many other mucosal defense mechanisms provided via the mother's milk. This defense is not inflammatogenic and energy-consuming. We learn about additional defense factors in the milk, like the anti-secretory factor, which seems to protect against diarrhoea. The milk contains numerous growth factors and cytokines, like leptin, which may promote the development of the intestine as well as the immune system. Results are appearing giving interesting evidence for enhanced protection against infection also after the termination of breastfeeding. This may occur via the priming of the infant's immune system after uptake of anti-idiotypic antibodies and lymphocytes from the milk. A breastfeeding motivation study in a large Pakistani village resulted in a 50% decrease of diarrhoea and infant mortality. Deep interviews with the mothers and the traditional birth attendants suggested that even better results may be obtained.

Hypercoagulation in diabetes mellitus.

The pathogenesis of diabetic vascular disease is not completely known. Many authors have suggested that hemorheological disturbance is an important factor in the development of vascular complication. The hypercoagulable state also suggested to cause this complication. In order to assess the role of blood coagulation in diabetic patients, we investigated hypercoagulation in diabetic patients and its relation with duration of the disease, smoking habits, blood pressure, blood glucose, diabetic retinopathy, diabetic ulcers and patient treatment. A case control study was done to evaluate these factors in 60 diabetic patients in out-and in-patient clinics of the Department of Internal Medicine, General Hospital Dr M Jamil Padang, during December 1990-March 1991. The average age was 51.55 years, average duration of the disease 5, 8 years. Fibrinogen levels is higher in diabetic patient (mean +/- SD = 442.42 +/- 86.79 mg/dl), compared to normal person (mean +/- SD = 349.2 +/- 35.26 mg/dl), PT is shorter (mean +/- SD = 10.1 +/- 1.31 sec), compared to normal person (mean +/- SD = 11.04 +/- 0.93 sec), APTT is also shorter (mean +/- SD = 29.2 +/- 3.69 sec), compared to normal person (mean +/- SD = 32.16 +/- 3.77 sec). There was a significantly high fibrinogen and short PT and APTT in diabetic patients, especially those who suffered from diabetes for a long time and followed by chronic complications.

Early child health in Lahore, Pakistan: IV. Child care practices.

Child care practices and hygiene measures were studied at 6 months of age in a longitudinally followed cohort of 1476 infants born between September 1984 to March 1987 in four socio-economically different areas in and around Lahore, Pakistan. Although, 76-98% of the mothers looked after their infants during health and 96-98% during a diarrhoeal illness, child care practices and hygiene measures differed significantly between the four areas. During a diarrhoeal episode, the mothers from the upper middle class took timely medical help, fed ample food and Oral Rehydration Salts (ORS) to the sick infants and provided uncontaminated food to them in clean surroundings. The mothers from the village and the periurban slum took their sick child, mostly after the second day of illness, to a doctor, but preferred home remedies. Fourteen percent of the mothers in the village and 6% in the periurban slum did not seek any medical help at all. One-third of the families, from these two areas, fed food to children 12 hours after cooking; the surroundings of the child were dirty with large numbers of flies present throughout the year, though the food was commonly kept covered with a lid. We constructed a simple measure of the surroundings of the child, rated as dirty, medium or clean; it was found to be associated to both parental illiteracy and child growth, but not with housing standard. The main conclusion is that any attempt to improve child-care practices and the hygienic environment for the child, should focus on maternal literacy and simple health messages.

Early child health in Lahore, Pakistan: XIII. Primary teeth emergence.

The aim of this study was to investigate the number of emerged primary teeth at various ages from 6 to 24 months in 1347 longitudinally followed Pakistan infants sampled from four socio-economically different areas in Lahore, Pakistan; from a very poor periurban slum to a privileged upper middle class group. The emergence of the primary teeth was found to be little, or not all related to sex or to the area of living. However, in comparison with studies conducted in other continents, the Indo-Pak subcontinent population lags behind in primary teeth emergence, especially in early life. This genetic difference makes it necessary to create specific standards of primary teeth emergence for this population.

Early child health in Lahore, Pakistan: III. Maternal and family situation.

The family situation for mothers, in three areas differing in degree of urbanization and an upper middle class control group, in Lahore, Pakistan was described. Area differences in socio-economic, family composition, and housing and sanitary conditions were investigated. Data from a longitudinal sample (n = 1476 newborns) were compared with data from a cross-sectional population survey (n = 2998 families). Risk factors for child mortality and morbidity were common in the village and periurban slum area; conditions were somewhat better in the urban slum community. The sample was concluded to be representative of the population in the three areas and also for Pakistan in general. Two indices for cross-study comparisons were proposed, one for socio-economic background and the other for housing standard. The two indices were shown to be related to maternal weight for height at 9 months of pregnancy in the urban slum area; the socio-economic level was also functionally related to the weight for height measure in the village. The lower socio-economic and housing standard level, the lower was the mothers' relative weight just before childbirth. The findings were discussed in terms of risk factors for infant mortality, morbidity and psychological development.

Early child health in Lahore, Pakistan: XI. Growth.

This work gives growth reference values at birth to 24 months of age for Pakistan based on upper middle class infants. Growth rate reference values are also included and they are given for various interval lengths. The growth was differently affected in infants living in three poorer areas; the stunting incidence at 24 months of age was 63% in periurban slum, 54% in the village and 26% in the urban slum. Less differences could be seen between the areas in weight for length. There was an age dependency in the incidence of reduced growth; a normal length gain was seen at birth to about six months of age, but they were highly reduced at 6 to 18 months of age. The weight gain was to some degree reduced during the first 12 months of life, followed by a catch-up growth period. The seasonal influence was also age dependent; weight was highly affected during the summer at birth to 24 months of age, but not in the winter. The seasonal effect in length was marginal at birth to 6 months, little at 6 to 12 months (although, constant below the normal) and large at 12 to 24 months of age. We did not see any seasonality of growth in the reference group. The incidence of reduced growth reflects the socio-economic differences in one restricted geographic area, i.e., in the city of Lahore, Pakistan.

Early child health in Lahore, Pakistan: XII. Milestones.

Psychomotor development was assessed in 1476 infants from four different levels of urbanization in and around Lahore, Pakistan. Development was assessed monthly at birth to 24 months of age by using a set of 10 milestones selected from the Denver Developmental Screening Test and the Developmental Screening Inventory. The psychomotor development of children in the upper middle (Um) class was consistent with reference population groups in Europe and North America, hence this group was used as control. There was significant delay in the psychomotor development of infants belonging to the poorer areas compared to the Um class. There was no sex difference in the development of the milestones in any study area, except for girls in the Um class who were earlier for about one month than boys in talking. Infants in the poorer areas were, on average about 3 months, delayed in their walking and fine motor activity (building a tower of 3 cubes) in comparison with the Um class. Observed disturbance in psychomotor development at an early age may affect the abilities and achievements in later life.

Early child health in Lahore, Pakistan: II. Inbreeding.

The prevalence of consanguineous marriages was studied in 940 families belonging to four different socio-economic groups in and around Lahore, Pakistan. The births occurring in these families from September 1984 to March 1987 were also investigated for birth defects. The overall prevalence of consanguineous marriages was 46%. The first cousin marriages were most common (67%), followed by the marriages between second cousins, 19%. The prevalence of consanguineous marriages was clearly associated with the socio-economic status of the study groups; 50% of the marriages was related in the periurban slum, 49% in the village, 44% in the urban slum and 31% in the upper middle class. The birth defects were also more prevalent in the poorer areas, being highest in the periurban slum (17.7%) followed by the urban slum (15.6%) and then the village (14.8%) and lowest in the upper middle class (12.3%). Although, the frequency of both consanguinity and birth defects were related with the socio-economic levels of the study groups, there was no association between inbreeding and birth defects. Perhaps, deleterious recessive genes for birth defects have been "bred out", because of continuous inbreeding over generations in this population. There was a significant predilection of major birth defects in boys without clear sex linkage. The conclusion is that the rate of consanguineous marriages was high, especially in the poorer areas, but the relationship between consanguineous marriages and birth defects was little or none. However, other child health indicators may be affected by consanguineous marriages.

Early child health in Lahore, Pakistan: VI. Morbidity.

Morbidity patterns were studied according to the age, area of living, sex and season among 1476 monthly followed infants born during 1984-1987 in four socio-economically different areas in Lahore, Pakistan. Infections were responsible for 87.0% of the morbidity during the first two years of life. The overall monthly based morbidity was 77.0% between birth and 24 months of age; diarrhoeal diseases 30.3%, upper and lower acute respiratory tract infections (ARI) 22.4%, skin and eye infections 6.7% and skin rash 6.2%. The vaccine-preventable diseases were only 0.5% of the total. Anaemia and rickets were rare (2.0%), but commonly seen among the nutritional deficiencies. Diarrhoea, tetanus, septicaemia, ARI and infections of the skin and eyes were reported more during earlier ages and from the three poorer areas of living. Diarrhoeal diseases, respiratory tract, skin and eye infections, in particular, followed clear seasonal patterns, while scabies prevailed throughout the year. The presence of these many preventable infections and illnesses implies that proper planning of interventions must be forthcoming.

Early child health in Lahore, Pakistan: VII. Diarrhoea.

This study was carried out to obtain reliable prospective data on the incidence, home management and duration of infantile diarrhoeal disease in a developing and rapidly urbanizing community. Fourteen hundred and seventy six infants from 4 different communities of Lahore, representing different stages of urbanization, were followed prospectively from birth to 2 years of age. Each child had an average of 3.6 episodes of diarrhoea during one year; 4.3 episodes per child per year in the village, 4.4 in the periurban slum, 3.4 in the urban slum and 1.4 in the upper middle (Um) class control group. The maximum incidence of diarrhoeal episodes occurred in children between 9-10 months of age. No sex difference was found. The seasonal variation showed a peak incidence during April-June. Bloody diarrhoea, in 3-5% of the episodes, was predominantly seen in winter months and in older infants. The maximum use of ORS was seen in the Um class (53%). Antibiotics were used in 75% of all diarrhoeal episodes maximally in the Um class, and morphine derivatives in 1% of all cases. The overall incidence of persistent diarrhoea was 14% of all acute cases, being highest in the village (18%), followed by the periurban slum (14%), Um class (10%) and urban slum (8%) with peak incidence occurring in June and July. Our results confirm the magnitude of the problem of diarrhoea.(ABSTRACT TRUNCATED AT 250 WORDS)

Early child health in Lahore, Pakistan: X. Mortality.

Mortality during the first two years of life among 1476 Pakistani infants born between September 1984 and March 1987 is described according to age, causes of death, area of living, season of the year and sex. The mortality rate (deaths under two years/1000 live births) projected over the whole population of Pakistan was 127 and under one year 114. During the first two years of life, the mortality rate was 133/1000 in the village, 159 in the periurban slum, 107 in the urban slum and 17 in the upper middle class group. In the latter group all deaths had occurred within 72 hours after birth. The overall major causes of death were acute and prolonged diarrhoea (36%), asphyxia neonatorum (13%), respiratory infections (13%), septicaemia (11%) and tetanus (9%). A clear age dependency was noted with 14% of deaths occurring during the first 24 hours of life (asphyxia neonatorum in 86%), and 57% dying within the first 28 days of life. In the later age groups, infections were mainly responsible for 82% of total deaths. Early mortality was therefore extremely high in the poorer areas studied and the cause of death was highly age dependent. Any interventions for reducing mortality must therefore be directed towards better antenatal care and safe delivery and postnatally, towards preventing infections, especially during the first six months of life.

Early child health in Lahore, Pakistan: IX. Perinatal events.

In Pakistan there are a number of acute problems related to maternal and infant health in the perinatal period. There is also lack of reliable data needed for the formulation of action strategies. To provide a database 1490 women have been followed from the 5th month of pregnancy in four different areas at various levels of urbanization and socio-economic development. After adjusting for gestational age, the proportion of newborns with weight for length < -2SDS in relation to the Swedish National Standard was 12-31% for boys and 12-25% for girls, the figure being highest in the most deprived area. Preterm birth was infrequent compared with IUGR. The overall prevalence rate of birth defects was 21% out of which 8% were severe defects. The overall perinatal mortality rate was 56/1000 births, with rates of 60, 75, 36 and 33/1000 births for the village, periurban slum, urban slum and the upper middle class. Two thirds of the deaths were related to either a continuation of intrauterine disturbances or severe congenital defects incompatible with life. One third of the deaths were due to infection; mostly diarrhoea, clinical sepsis and ARI. Neonatal mortality was significantly related to birth length (< -2SDS, odds ratio 5.5) and length of gestation (< 37 weeks, odds ratio 5.6) and was to a lesser extent related to weight (< -2SDS, odds ratio 2.0) and weight for length (< -2SDS, odds ratio 1.3). Forty percent of the mothers had weight for height below -2SDS, 23-35% had height < -2SDS. Forty percent of mothers from a subset within the cohort had a hemoglobin < 10 gm/dl and 20% showed signs of pre-eclampsia. This presentation raises the issue of expanding the current Child Survival Programs into the perinatal period as well.