Development of probe-based real-time loop-mediated isothermal amplification for detection of Brucella.

AIM: To develop a rapid, sensitive and specific diagnostic assay for the detection of Brucella. METHODS AND RESULTS: The probe-based RT-LAMP was carried out by using a set of four or six primers and different LAMP chemicals to compare its results with real-time PCR. Detection of gene amplification is done within 40 min and can be seen by amplification curve, turbidity and addition of DNA-binding dye at the end of the reaction results in colour difference under normal day light and in UV. The sensitivity of probe-based real-time LAMP assay was found 10-fold higher than Taqman-based qPCR. The specificity of the developed assay was validated by the absence of any cross-reaction with other pathogenic bacteria. CONCLUSION: The developed probe-based RT-LAMP assay is extremely rapid, cost effective, highly specific and sensitive, and has potential usefulness for rapid Brucella surveillance. SIGNIFICANCE AND IMPACT OF THE STUDY: The developed probe-based RT-LAMP is a powerful gene amplification technique which is a specific, fast diagnostic tool for early detection and identification of Brucella.

Molecular characterization of kisspeptin gene and effect of nano-encapsulted kisspeptin-10 on reproductive maturation in Catla catla.

Kisspeptin, a member of the RF-amide-related peptide family, has emerged recently as an essential gatekeeper of various reproductive processes via its ability to activate kisspeptin receptors at puberty. In this study, the kiss1 gene and its receptor kiss1rb were cloned and characterized from the brain of Catla catla. Further, the effects of kissppetin-10 (K-10) and chitosan-encapsulated K-10 nanoparticles (CK-10) on gene expression were assessed. The full-length complementary DNA sequence of kiss1 is 754 bp with an open reading frame of 351 bp that encodes a putative protein of 116 amino acids. The kiss1rb complementary DNA is 1,280 bp long and contains a 5'-untranslated region of 30 bp, 3'-untranslated region of 149 bp, and an open reading frame (open reading frame) of 1,101 bp. The expression patterns of kiss1 and kiss1rb messenger RNA (mRNA) in basal tissues revealed that they are mainly expressed in the brain, pituitary gland, and gonads. CK-10 nanoparticles with a particle size of 125 nm and a zeta potential of 36.45 mV were synthesized and compared with K-10. Chitosan nanoparticles showed 60% entrapment efficiency for K-10. The mRNA expression of reproductive genes (GnRH, LH, and FSH) in fish injected with K-10 declined after 6 h, whereas those injected with CK-10 showed controlled and a sustained surge of mRNA expression of these genes with a peak at 12 h. Histologic examination of ovaries indicated a pronounced effect of CK-10 on maturation and gonadal development. The study reports that this sustained release delivery system will help in increasing the half-life of K-10 and other therapeutic protein drugs in the biological system. Besides, the nanoformulation developed in the present study may be useful for developing therapies against various reproductive dysfunctions in vertebrates.

Relationship between Serum Soluble Interleukin-2 Receptor and Renal Allograft Rejection: A Hospital-Based Study in KashmirValley.

BACKGROUND: Even after adequate immunosuppression therapy, acute rejection continues to be the single most important cause of graft dysfunction after renal transplantation. Renal allograft biopsy continues to be the reference standard, though certain clinical and biochemical parameters are helpful in assessment of these patients. Renal allograft rejection is mediated by T lymphocytes, expressing cell surface interleukin-2 receptors (IL-2R) which has been suggested as a marker of acute rejection episodes after organ transplantation. OBJECTIVE: To determine the pre- and post-transplantation serum soluble IL-2R levels in live related kidney transplant patients to predict acute rejection episodes. METHODS: Serial serum samples from 75 recipients and 41 healthy controls were assessed for soluble IL-2R levels by ELISA. The outcome of the graft was also determined for each recipient. RESULTS: The mean±SD serum soluble IL-2R levels in renal allograft recipients with rejection were significantly (p<0.001) higher than those without rejection (329.85±59.22 vs 18.12±11.22 pg/mL). The elevation of serum soluble IL-2R was evident in acute rejection episodes and found before elevation of serum creatinine. The higher values of serum soluble IL-2R in the rejection group were significantly reduced after recovery of allograft function by adequate anti-rejection therapy. 36.4% of patients in the rejection group had proven positive biopsies for the rejection and higher creatinine values, which was found to be statistically significant (p<0.001). A cohort of 41 healthy controls showed significantly (p<0.05) lower serum soluble IL-2R concentrations (15.27±7.79 pg/mL) when compared with the rejection group. CONCLUSION: Serum soluble IL-2R concentrations showed significant correlation with the acute rejection episodes in the renal allograft recipients. Prediction of soluble IL-2R levels might help the early detection of rejection episodes, which may pave way for the management of immunosuppression regimes and better graft functioning.

Postpartum thyroiditis in India: prevalence of postpartum thyroiditis in Kashmir Valley of Indian sub-continent.

Various studies have reported a spectrum of thyroid dysfunction in the postpartum period. Postpartum thyroiditis is a syndrome of thyroid dysfunction that occurs in the first year after parturition. Prevalence of postpartum thyroiditis has been reported to vary from 3 to 6 percent in different regions of the world. Kashmir Valley is inhabited by a relatively homogeneous racial group and the Valley has been documented to have significant iodine deficiency. We studied the prevalence and pattern of postpartum thyroiditis in an urban region of this Valley. 120 women were registered within first month of postpartum period for the study along with one hundred controls. Of these 120 women, 104 reported for follow-up at 3 months postpartum and 106 reported for follow-up at 6 months postpartum. Initial and subsequent clinical details at follow-up were recorded on a pre-determined questionnaire. Overall, postpartum thyroiditis (PPT) was seen in 8 (7%) study subjects. Of these 8 patients with PPT, 4 had biochemical evidence of thyrotoxicosis at first month, 3 developed biochemical thyrotoxicosis at 3-month follow-up while as one study subject developed thyrotoxicosis at 6 months. Most of these subjects were antithyroid antibodies (anti-microsomal and anti-thyroglobulin) positive. We conclude that iodine deficient status of the community doesn't seem to influence the incidence of PPT.

Impact of sociomedical factors on pre-school malnutrition -- an appraisal in an urban setting.

PIP: The nutritional status of 584 preschool children in an urban slum of Srinagar district was assessed through interviews of mothers conducted in a house-to-house survey, the clinical assessment of child nutritional status, and the anthropometric measurement of the children using accepted standard techniques. The anthropometric measurements of the children were compared to the 50th percentile of the Harvard Standard, while the classification recommended by the Indian Academy of Pediatrics was adopted for the categorization and grading of protein energy malnutrition. 60.45% of the children were malnourished, comprised of 33.22%, 20.89%, and 6.34% of children with grade I, II, and III degrees of malnutrition, respectively. Higher prevalence of malnutrition is associated with higher age, female sex, lower per capita income, higher birth order, and lower parental literacy. Parental literacy status followed by birth order and per capita income were the most important factors associated with preschool child malnutrition.^ieng

Efficiency of various anthropometric measurements in determining low birth weight babies.

PIP: Anthropometric measurements were applied to 119 healthy, singleton, full-term infants born during June 1993-January 1994 in the obstetric ward of the community medicine department of SKIMS in Srinagar, Kashmir, India. Researchers aimed to identify an alternative simple and accurate measurement to birth weight for field conditions. The anthropometric measurements included weight, crown-heel length, mid-arm circumference, head circumference, thigh circumference, calf circumference, and chest circumference. The researchers applied their findings to a separate cohort of 250 newborns at the child health clinic of the same department to determine specificity, sensitivity, and predictive values. The mean birth weight was 2.8 kg. Mean mid-arm, calf, thigh, chest, and head circumferences were 10.45, 10.15, 015.73, 32.18, and 34.96 cm, respectively. Mean crown-heel length was 49.78 cm. Birth weight had a very significant correlation with calf circumference (r = 0.87), thigh circumference (r = 0.7), mid-arm circumference (r = 0.7), and chest circumference (r = 0.40). Calf circumference accounted for 75.69% of the total variance. Thigh circumference and mid-arm circumference accounted for 49% and 46.24%, respectively. The critical level value of birth weight to 2.5 kg was 9.78 cm for calf circumference, 14.8 cm for thigh circumference, 9.94 cm for mid-arm circumference, 32.23 cm for chest circumference, 34.72 cm for head circumference, and 49.58 cm for chest circumference. Calf circumference had the highest sensitivity and specificity (93.55% and 98.92%, respectively), compared to 77.42% and 92.63%, respectively, for thigh circumference and 64.52% and 93.55%, respectively, for mid-arm circumference. The likelihood of missing low birth weight infants was lowest for calf circumference (7-8% vs. 22-23% for thigh circumference and 30-35% for mid-arm circumference). These findings show that calf circumference is the most effective alternative anthropometric measurement to birth weight for identifying low birth weight newborns.^ieng

Parental awareness and practices in acute diarrhea.

PIP: In 1992, in Srinagar, India, the Pediatric Outpatient Department of the Institute of Medical Sciences interviewed the parents of 1600 infants with acute diarrhea to learn their awareness and practices in infantile diarrhea. The parents were divided into two groups: Group A (physicians, engineers, teachers, professors, lawyers, clerks, and educated business professionals; N = 660) and Group B (all others and uneducated parents; N = 940). Most infants with acute diarrhea were boys (66.2%) and aged 6-9 months (58.7%). Only 15% of all parents knew the definition of diarrhea (3 loose stools/day) with Group B parents being more likely to know it than Group A parents (18.5% vs. 10%; p .001). 81.9% of all parents incorrectly considered frequent stools as constituting diarrhea. The parents considered weaning, maternal diet, and teething to be the major causes of diarrhea (58.1%, 42.5%, and 34.4%, respectively). Group A parents were more likely than Group B parents to report infections (27.4% vs. 9.5%) and formula and dirty feeds (7.6% vs. 2%) to be causes of diarrhea (p .001). Group B parents were more likely have no knowledge about the danger signs of diarrhea than Group A parents (30.2% vs. 9.1%; p .001). Group B parents were more likely to treat infantile diarrhea with antidiarrheal and antispasmodic drugs than Group A parents (77.7% vs. 22.7%; p .001). Group A parents were more likely to use both these drugs and oral rehydration therapy (ORT) than Group B parents (65.2% vs. 7.9%; p .001). Few parents (.6%) administered only ORT. Group A parents were much more likely than Group B parents to know the composition of various ORT brands, reconstitution of the solution, and their utility in diarrhea (65.6% vs. 7.7%; p .001). Group B parents preferred complete dietary restrictions during a diarrheal episode than Group A parents (28.7% vs. 6.2%; p .001). These findings reflect limited parental knowledge about diarrhea and its treatment.^ieng

Who is responsible for artificial feeding? An evaluation.

PIP: Artificial feeding has been found to be on the increases in India. In this research study, 320 infants who were artificially fed before 6 months of age and attended the child health clinic of the Department of Community Medicine, SKIMS Srinagar, between March and November, 1991, were surveyed. Information was obtained from mothers on personnel who advised artificial feeding by type of medical care unit. The sample of mothers included 59.3% who were illiterate. 52.25% of all mothers were housewives and 43.75% had a middle socioeconomic status, while 31.5% had a low socioeconomic status. 53.75% of the artificially fed babies were born at a hospital, of which 2.5% received intensive care. 46.25% were born at home. 76.87% were first or second births. 3.55% of babies were completely artificially fed from birth; at 3 months, 18.76% were exclusively artificially fed. Mixed feeding of breast and artificial milk was 10.76% at birth and 25.97% by 3 months and 42.10% by 6 months. 51.25% of mothers were advised by pediatricians to use artificial feeding; 24.65% of mothers were advised by general practitioners and 10.31% by paramedical. 40% were advised at private clinics and 26.25% in hospital wards at the time of discharge. 18.12% were advised in outpatient departments, and 15.62% at other health centers, such as immunization clinics, health clinics, or nutrition education centers. 66.25% received recommendations to use tinned milk (Lactogen/Milk-care) and 30%, for cows milk. 45.31% of mothers received advice from pediatricians about tinned milk and 5.93% about cow's milk. 5.93% of infants received advice from general practitioners about tinned milk and 5.31 about cow's milk. Over 80% of mothers were given instructions on the amount of dilution and frequency of feeding. 60.93% of mothers received information on feeding equipment and 56.15% on sterilization of feeding equipment. Only 59.37% and 65.37% were able to understand instructions on equipment and sterilization, respectively. The reasons given by mothers for using artificial feeding were low milk output (49.68%), nonacceptance of breast milk by the baby (22.50%), working mothers (12.18%), and baby illness (8.12%). Mothers attributed infections, colic, and regurgitation to artificial feedings. Clearly, the professional community is not complying with the professional code promoted by the World Health Assembly and Indian legislation in 1986 on not recommending milk formula.^ieng

A study on the impact of maternal knowledge and practice on the nutritional status of infants.

PIP: In India, health workers interviewed 123 mothers of infants attending the child health clinic of the S.K. Institute of Medical Sciences in Srinagar to determine whether maternal knowledge and practice were associated with the nutritional status of the infants. 28 children were considered to be well nourished, while the remaining 95 children were determined to be in various degrees of malnutrition. Mothers whose infants were well nourished had a higher level of breast feeding knowledge than did those whose infants were moderate to severely malnourished (score, 27.13 vs. 16.01-18.75; p .0001). None of the mothers of malnourished infants had an excellent score on breast feeding practices. Differences in the mean score values for breast feeding practices between all consecutive grades of nutrition were significant (20.22 for excellent score, 16.85, 15.53, and 14.18 for grades I, II, and III, respectively; p .001). The only mothers who had an excellent score for infant weaning awareness were 3 mothers whose infants had an excellent nutritional status. A significant difference in mean score values for knowledge of infant weaning between mothers of well nourished infants and grade I malnourished infants (26 vs. 17.19; p .001) as well as between those of grade II malnourished infants and grade III malnourished infants (17.06 vs. 13.64; p .01) were significant. Little difference in infant nutritional status existed between mothers who scored fair and those who scored poor, but, among mothers of well nourished infants, those who scored well were more likely to be have infants of good nutritional status than those who did not score well (47.05% v. 19.56% and 18.33%; p .001). These findings show a decreasing trend between awareness and practice of breast feeding/infant weaning, suggesting that further improvement of health education is needed to reduce the lag between breast feeding awareness and practice.^ieng

Low birth weight: an outcome of poor socio-obstetric interaction.

PIP: Factors contributing to low birth weight (LBW) include poverty, ignorance, and inability to use health care services. Early marriage and low family income lead to poor maternal nutrition reserves, which lead to reduced fetal nutrition. Poor maternal nutrition is also the result of ignorance, short birth intervals, multi-parity, and lack of prenatal care. Both heavy manual labor and smoking contribute to placental ischemia, which, along with reduced fetal nutrition, leads to intrauterine growth retardation (IUGR). In developing countries, IUGR accounts for over 66% of all LBW neonates. About 7 million Indian babies annually are LBW. This study examined the incidence of LBW among 178 mothers delivering single births at the maternity hospital associated with the Department of Community Medicine of SKIMS, Srinagar, Kashmir, India, during 1989-90. 26.40% (47) of the 178 births were LBW (2500 g). Among 71 first-borns, marriage age was found to be statistically significantly associated with LBW. 31.82% of mothers younger than 20 years had LBW babies, compared to only 6.12% of mothers older than 20 years. The impact ratio, which measured excess LBW, was 4.20. Birth interval was statistically significantly associated with LBW outcome. 55.81% of women with a birth interval of less than 18 months had LBW babies, compared to 20.31% of mothers with longer birth intervals. The impact ratio was 1.75. Gravidity was also statistically associated with LBW babies. 34.58% of multigravida mothers had LBW babies, compared to 14.08% of primigravidae. The impact ratio was 1.46. Presence of prenatal care was statistically associated; the impact ratio was 1.42. 31.30% of illiterate women had LBW babies, compared to only 17.46% among literate women, which indicated significant associations with LBW. Other significant factors were manual labor, maternal smoking, and monthly family income. Reduction of LBW by 10-30% nationally by the year 2000 will be difficult and best accomplished by a high risk approach supplemented by health and nutrition education.^ieng

Maximum thigh circumference as an indicator of birth weight.

PIP: This study had the objective of estimating the strength of association between birth weight and maximum thigh circumference (MTC), sole length, and mid arm circumference (MAC), and on the basis of positive correlation, to identify the ideal anthropometric measurement that can give a reasonable estimate of the birth weight, especially where weight measurement facilities are nonexistent. A random sample of 75 singleton normal neonates was studied who had normal birth in the maternity hospital of the S.K. Institute of Medical Sciences, Srinagar, Kashmir, India, in the first half of 1990. The anthropometric measurements recorded within 24 hours of birth were birth weight, MTC, MAC, and sole length using the left arm and foot. The mean +or- SEM values derived were 3.20 +or- 0.05 kg for weight, 14.68 +or- 0.14 cm for MTC, 7.71 +or- 0.05 cm for sole length, and 9.55 +or- 0.09 cm for MAC. The correlation coefficients of weight were +0.50 for MTC, +0.16 for sole length, and +0.21 for MAC. Only MTC showed a correlation coefficient value of high significance (p .001). MTC was significantly interrelated and interdependent on birth weight inasmuch as MTC can safely be used as an indicator of birth weight in all situations. On the basis of this observation, a regression equation was developed exhibiting the form: Y = (1.48) x +9.4944, where Y represented the MTC. The corresponding values of MTC for birth weight of 2.5 kg and 2.0 kg obtained were 13.644 cm and 12.904 cm, respectively. The lower values constitute the dividing line for low birth weight babies by Indian standards. The comparable values reported by Sharma (1989) were 14.5 cm and 13.5 cm, respectively, presumably due to higher altitude in Kashmir associated with low birth weight. A graphic representation indicating the critical zone was developed for ready reference of paramedical staff in rural areas where weighing facilities for the newborns are not available. MTC, sole length, and MAC are all positively correlated with birth weight.^ieng

Ill effects with use of pacifiers in children.

PIP: Various ill effects among children who used pacifiers in an urban area of Kashmir, India, are described. 170 children exposed to pacifiers were compared with a control group of 145 children. Both groups were selected from the Well Baby Clinic of the Sher-i-Kashmir Institute of Medical Sciences from September 1990 to March 1991 by a sampling procedure comparing the two cohorts in relation to socioeconomic correlates. Information related to sucking habits and feeding practices was collected. Mothers whose children had completed one year after exposure to pacifiers were questioned about morbidity profile and beliefs about pacifiers. Among children using pacifiers, 64.29% had illiterate mothers, 55.39% had working mothers, and 53.5% had a family income of 1500 rupees or more per month. The use of pacifiers in middle class socioeconomic groups amounted to 56.70%. About 20% of mothers had given pacifiers to siblings beforehand. 77% of children used honey-based pacifiers and 5.8% of children used empty bottle teats. 70% of children had pacifiers readily available for use. 67.06% of children started pacifier use between 3 and 6 months of age. The mean age of weaning among children using pacifiers was 5.9 months vs. 6.8 months among nonusers. About 85% of children discontinued pacifier use 1.5-2 years later. The main reason for introducing pacifiers was to keep the baby busy (75.29%), to calm the child (70.O%), to facilitate dentation (11.76%), and for nutritive purposes (7.06%). Children using pacifiers also had the habit of thumb (64.71%), knuckle (16.43%), toe (3.53%) and cloth (5.29%) sucking. The morbidity pattern in the pacifier group revealed that 70.5% had diarrhea, 55.29% had respiratory tract infections, and 31.19% had both diarrhea and respiratory tract infections. The frequencies were significantly higher in comparison to children not using pacifiers. Only about 28% of mothers felt that pacifiers might have bad effects on the health of the baby.^ieng

Persistent limb pain and raised serum alkaline phosphatase the earliest markers of subclinical hypovitaminosis D in Kashmir.

The present study was an attempt to assess the cause of persistent pain in lower limbs among children from Kashmir. The study was conducted on one hundred children attending Paediatric out-patient department of Sher-i-Kashmir Institute of Medical Sciences, Srinagar. All the children were in the age group of 5 to 14 years. They showed markedly raised levels of serum alkaline phosphatase, whereas serum phosphorus, serum calcium levels and antistreptolycin O-titres were normal in 93% cases. None of them had any rheumatic or rheumatoid pathology. Among 15 suspected clinical rickets only three were established radiologically. Dietary and socio-economic history revealed deficient vitamin D intake and less exposure to sun. It was hypothesized that sub-clinical vitamin D deficiency could be a major cause of persistent pain in lower limbs and raised serum alkaline phosphatase could be the earliest marker of vitamin D deficiency. It was confirmed by injecting single dose of vitamin D (3 lac I. U.) which relieved bone pain and lowered the levels of serum alkaline phosphatase to normal within 14 weeks of initiation of therapy.