Effect of immune gene silencing in WSSV infected tiger shrimp Penaeus monodon.

White spot syndrome virus, continues to cause huge economic loss to aquaculture industry. In the absence of effective therapeutics to control WSSV, it is important to understand the host pathogen interaction at the molecular level. Suppression subtractive hybridization (SSH) cDNA library was constructed which led to identification of several differentially expressed genes in response to WSSV infection in Penaeus monodon. The genes expressed in SSH cDNA library of shrimp gill and gut tissues belonged to a wide range of biological functions. The three differentially expressed genes, Single von Willebrand factor type C domain protein (pmSVC), P53 protein gene (pmP53) and ADP ribosylation factor (pmArf) were up-regulated against WSSV infection and were further characterized by gene silencing to study the role of these shrimp immune genes on WSSV multiplication. The sequence-specific knock down of pmSVC, pmP53 and pmArf using the dsRNA revealed that in pmSVC-dsRNA inoculated shrimps WSSV replication was more with increased viral copy numbers when compared with pmP53-dsRNA and pmArf -dsRNA inoculated shrimps. The varied response of immune genes to WSSV infection, indicated that host genes may either inhibit virus replication to some extent or might act as a target to facilitate viral pathogenesis.

Differentially expressed urinary biomarkers in children with idiopathic nephrotic syndrome.

BACKGROUND: We performed a discovery phase of urinary proteomic profile in children with idiopathic nephrotic syndrome and validated selected biomarkers. METHODS: Urinary proteomic profile was performed using isobaric tags for relative and absolute quantitation labeling, coupled with liquid chromatography-matrix assisted laser desorption and ionization analysis. Validation of biomarkers apolipoprotein A1, alpha 2 macroglobulin, orosomucoid 2, retinol binding protein 4 and leucine-rich alpha 2-glycoprotein 1 was done by enzyme-linked immunosorbent assay. RESULTS: Apolipoprotein A1 levels of <0.48 µg/mg of creatinine-differentiated steroid-resistant nephrotic syndrome (SRNS) from first episode nephrotic syndrome, area under curve (AUC) [0.99 (CI 0.9-1.0), 100 % sensitivity and 100 % specificity] and a value of <0.24 µg/mg of creatinine could differentiate SRNS from frequently relapsing nephrotic syndrome/steroid dependent nephrotic syndrome [AUC 0.99 (CI 0.9-1.0), 100 % sensitivity and 100 % specificity]. Alpha 2 macroglobulin could differentiate children with SRNS-focal segmental glomerulosclerosis (FSGS) from SRNS-minimal change disease (MCD) at values >3.3 µg/mg of creatinine [AUC 0.84 (CI 0.62-1.0), 90 % sensitivity and 85 % specificity]. Orosomucoid 2 >1.81 µg/mg of creatinine could distinguish SRNS-FSGS from SRNS-MCD [AUC 0.84 (CI 0.62-1.0), sensitivity 90 % and specificity 85.5 %]. RBP 4 value of >1.54 µg/mg of creatinine differentiated SRNS-FSGS from SRNS-MCD [AUC 0.87 (CI 0.68-1.0), sensitivity 90 % and specificity 85.7 %]. CONCLUSIONS: Lower level of apolipoprotein A1 in urine is suggestive of SRNS. Alpha 2 macroglobulin, retinol binding protein 4 and orosomucoid 2 are markers associated with FSGS, with alpha 2 macroglobulin being most predictive.

A prospective study of haematological changes after switching from stavudine to zidovudine-based antiretroviral treatment in HIV-infected children.

Long-term use of stavudine is associated with a high incidence of lipodystrophy, warranting its substitution with zidovudine in first-line antiretroviral therapy (ART) regimens. In a prospective observational study, we determined the spectrum and severity of haematological changes after switching from stavudine- to zidovudine-based ART in Indian children aged 2-18 years who had received a stavudine-based ART regimen for at least 48 weeks. They were followed for 48 weeks for changes in haematological parameters and CD4 cell counts after switching to zidovudine. Of the 60 children analysed, 45 (75%) showed a significant fall in Hb (>1 g/dl). A majority developed grade 1 anaemia (14 [31%]) while only three (6%) developed grade 4 anaemia. The lowest Hb was recorded between 12 and 16 weeks with spontaneous improvement noticed after 28 weeks. A significant drop in absolute neutrophil count (5067 cells/mm3 to 3625 cells/mm3; p = 0.004) was also observed but none developed severe neutropenia. No significant changes were observed in platelet and CD4 cell counts. Since the incidence of severe drug toxicity was low with zidovudine and the majority of children recovered without intervention, drug toxicity should not preclude its routine use in poor countries.

Meningitis and intracranial bleed in a child with steroid-resistant nephrotic syndrome.

Meningitis and associated intracranial bleeding have been rarely reported in patients with steroid-resistant nephrotic syndrome. We present such a case with raised intracranial tension in a 13-year-old child and discuss the management issues. Prompt recognition and appropriate treatment of these complications can be life saving in a child with nephrotic syndrome.

Efficacy of chemically characterized Foeniculum vulgare Mill seed essential oil in protection of raw tobacco leaves during storage against fungal and aflatoxin contamination.

AIMS: To report fungal and aflatoxin contamination in stored tobacco leaves and the potential of Foeniculum vulgare (fennel) seed essential oil (EO) as a plant-based preservative in protection of tobacco during storage. METHODS AND RESULTS: Mycological analysis of tobacco samples was done by surface sterilization and serial dilution tests. The Aspergillus flavus isolates were screened for their toxigenicity. Both in vivo and in vitro tests were done to evaluate antifungal and antiaflatoxigenic efficacy of chemically characterized EO. The mycoflora analysis revealed 108 fungal colonies belonging to five genera and nine species. All A. flavus isolates were found aflatoxigenic during screening. Gas chromatography and mass spectrometry analysis of EO identified 19 components (99·66%); estragole being the major component (47·49%). The EO showed broad fungitoxicity at 1·25 µl ml(-1) and 100% inhibition to AFB1 production as well as ergosterol synthesis at 1·0 µl ml(-1) concentration. EO showed 100% protection of stored tobacco samples from aflatoxin B1 contamination. CONCLUSIONS: The fennel EO can thus be formulated as a plant-based preservative for food items. SIGNIFICANCE AND IMPACT OF THE STUDY: The present investigation comprises the first report on antiaflatoxin efficacy of fennel oil and its potency in the protection of tobacco leaves from fungal and aflatoxin contamination during storage.

Efficacy of Angelica archangelica essential oil, phenyl ethyl alcohol and α- terpineol against isolated molds from walnut and their antiaflatoxigenic and antioxidant activity.

Mold association, aflatoxin B1 contamination as well as oxidative deterioration of agri-food items during storage and processing are some global task for food industries. In view of the adverse effects of some synthetic preservatives on treated food items and subsequently on consumers health, recently plant based chemicals are encouraged by food industries as better alternatives of synthetics. The present study recommends the combination (1:1:1) of Angelica archangelica essential oil: Phenyl ethyl alcohol (PEA): α- terpineol as botanical preservative against molds, aflatoxin contamination and oxidative deterioration of walnut samples. Eight mold species were procured from stored walnut samples, including some aflatoxigenic Aspergillus flavus strains. The combination inhibited growth of aflatoxigenic strain Aspergillus flavus NKDW-7 and aflatoxin B1 production at 2.25 and 2.0 µL mL(-1) respectively. The IC50 value of the combination was recorded as 3.89 µL mL(-1), showing strong antioxidant potential. The antifungal action of the combination showed > 90 % decrease in ergosterol content in plasma membrane of A. flavus at 2.0 µL mL(-1). The LD50 of the combination, through oral administration on mice, was 9562.9 µL kg(-1) body weight, indication favourable safety profile as a plant based preservative. The combination may be recommended as safe preservative against molds, aflatoxin contamination and oxidative deterioration of walnut samples.

Insecticidal activity of Ageratum conyzoides L., Coleus aromaticus Benth. and Hyptis suaveolens (L.) Poit essential oils as fumigant against storage grain insect Tribolium castaneum Herbst.

Essential oils (EOs) from Ageratum conyzoides L., Coleus aromaticus Benth. and Hyptis suaveolens (L.) Poit were extracted and tested against Tribolium castaneum Herbst, the storage grain insect. The EOs were found effective against Tribolium castaneum during in vitro as well as in vivo fumigant testing. The EOs of H. suaveolens and A. conyzoides showed 100 % mortality of test insect at 250 ppm while C. aromaticus at 350 ppm. During in vivo fumigant testing of wheat samples against Tribolium castaneum, the essential oils of A. conyzoides and C. aromaticus completely checked the damage of wheat grains by the insect at 1000 ppm while essential oil of H. suaveolens checked the grain damage completely even at 500 ppm concentration. There was no adverse effect on seed germination as well as on seedling growth of EOs treated seeds showing non-phytotoxic nature of the oils. Hence, these EOs may be recommended as botanical insecticide against insect invasion of stored food commodities, thereby enhancing their shelf life.

Endothelial dysfuntion in children with idiopathic nephrotic syndrome.

BACKGROUND: Impaired endothelial function is the initial step in atherogenesis, which is largely responsible for ischaemic heart disease and thrombotic strokes decades later. METHODS: Fourty two children with first episode nephrotic syndrome (FENS) aged 1-16 years and 40 controls were enrolled. Soluble thrombomodulin (sTM), tissue plasminogen activator (t-PA), plasminogen activator inhibitor -1 (PAI-1) and von-willebrand factor (vWF) levels were measured in plasma in FENS, at 12 weeks of drug induced remission and in steroid resistant nephrotic syndrome (SRNS) patients at diagnosis. RESULTS: PAI-1, sTM, vWF and t-PA were significantly raised at the onset of nephrotic syndrome (p<0.0001). All the markers had a fall after 12 weeks of steroid treatment, but were still raised. Children with SRNS had higher levels of sTM, tPA, vWF as compared to infrequent relapsers, at onset and at 4 weeks of steroid treatment. CONCLUSION: Children with idiopathic nephrotic syndrome have endothelial dysfunction which is largely dependent upon disease activity.

Homocysteine metabolism in children with idiopathic nephrotic syndrome.

BACKGROUND: Homocysteine metabolism is altered in children with idiopathic nephrotic syndrome. Hyperhomocysteinemia is a risk factor of early atherosclerosis and glomerulosclerosis and may occur at time of first occurrence of idiopathic nephrotic syndrome. METHODS: Thirty children with first episode of idiopathic nephrotic syndrome (FENS) aged 1-16 years along with 30 age- and sex-matched healthy controls were enrolled in this study. Homocysteine and cysteine were measured with HPLC; vitamin B12 and folic acid were measured with electro-chemilumiscence immunoassay. Primary outcome measure was plasma homocysteine level in children with FENS and in controls. Secondary outcome measures were (1) plasma and urine homocysteine and cysteine levels in children with FENS at 12 weeks and 1 year (remission) and (2) plasma and urine levels of vitamin B12 and folic acid in children with FENS, at 12 weeks and 1 year (remission). RESULTS: Plasma homocysteine and cysteine levels were comparable to controls in children with FENS, at 12 weeks and 1-year remission. Plasma levels of vitamin B12 and folic acid were significantly decreased compared to controls in FENS due to increased urinary excretion, which normalize during remission at 12 weeks and 1 year. Urinary homocysteine and cysteine levels were significantly raised in FENS compared to controls and continued to be raised even at 12-week and 1-year remission. CONCLUSION: Homocysteine metabolism is deranged in children with FENS. Renal effects of long-term raised urinary homocysteine levels need to be studied.

Comparison of oral ibuprofen with oral indomethacin for PDA closure in Indian preterm neonates: a randomized controlled trial.

Oral ibuprofen is being used as an alternative to indomethacin in medical management of patent ductus arteriosus (PDA), but limited data exist on oral efficacy of these drugs for PDA closure in India. To assess and compare the efficacy of oral ibuprofen and oral indomethacin for PDA closure in preterm Indian neonates, we designed a randomized controlled study on clinically diagnosed and echocardiographically confirmed hemodynamically significant PDA in preterm neonates. Patients were assigned to receive either oral ibuprofen at a dosage of 10, 5, 5 mg/kg every 24 h or three doses of oral indomethacin (0.20-0.25 mg/kg every 24 h) starting on the third day of life or when diagnosed. A second course of ibuprofen/indomethacin was given, if PDA failed to close within 48 h after the first course. Patients were monitored for complications like oliguria, bleeding, necrotizing enterocolitis, intraventricular hemorrhage, oxygen dependency, and gastrointestinal side effects. The baseline characteristics were comparable in both groups. Of the 83 children enrolled, 57.8 % received oral ibuprofen and 42.1 % received oral indomethacin. The overall closure rate of PDA was 60 and 65.7 % in the ibuprofen and indomethacin groups, respectively. Closure rate was significantly higher when the drugs were administered at an early postnatal age (<8 days) (83.3 % [p = 0.02] in the indomethacin group and 75 % [p = 0.03] in the ibuprofen group) in neonates >28 weeks (ibuprofen group 66.7 % [p = 0.02]; indomethacin group 65.5 % [p = 0.04]) and in babies with birth weight >1,000 g (ibuprofen group 62.2 %; indomethacin group 70 % [p = 0.04 in both groups]). Complications were similar in both groups. The efficacy of both drugs was similar. Poor closure in our study could be because of genetic differences in pharmacokinetics of drug metabolism in the Indian population. Regimens with higher doses or increased duration of treatment may increase the frequency of closure. Studies with larger numbers of subjects with evaluation of pharmacokinetic parameters are therefore required.

Antifungal and antiaflatoxigenic properties of Cuminum cyminum (L.) seed essential oil and its efficacy as a preservative in stored commodities.

The study reports potential of Cuminum cyminum (cumin) seed essential oil (EO) as a plant based shelf life enhancer against fungal and aflatoxin contamination and lipid peroxidation. The EO showed efficacy as a preservative in food systems (stored wheat and chickpeas). A total of 1230 fungal isolates were obtained from food samples, with Aspergillus flavus LHP(C)-D6 identified as the highest aflatoxin producer. Cumin seed EO was chemically characterized through GC-MS where cymene (47.08%) was found as the major component. The minimum inhibitory concentration and minimum aflatoxin inhibitory concentration of EO were 0.6 and 0.5 µl/ml respectively. The EO showed toxicity against a broad spectrum of food borne fungi. The antifungal action of EO on ergosterol content in the plasma membrane of A. flavus was determined. The EO showed strong antioxidant potential having IC50 0.092 µl/ml. As a fumigant in food systems, the EO provided sufficient protection of food samples against fungal association without affecting seed germination. In view of the antifungal and antiaflatoxigenic nature, free radical scavenging potential and efficacy in food system, cumin seed EO may be able to provide protection of food commodities against quantitative and qualitative losses, thereby enhancing their shelf life. The present investigation comprises the first report on antifungal mode of action of cumin seed EO and its efficacy as fumigant in food systems.

Subclinical non-autoimmune hypothyroidism in children with steroid resistant nephrotic syndrome.

BACKGROUND: Thyroid status has not been studied well in children with steroid resistant nephrotic syndrome (SRNS). METHODS: In this cross sectional study we recruited 20 children aged 1-16 years with SRNS and similar number of controls. Serum levels of FT3, FT4 and TSH were measured in all the subjects. Overt hypothyroidism was defined as low FT4 (normal values: 0.7-2.0 ng/mL) and elevated serum TSH above reference values (0.45-4.5 mIU/L). Subclinical hypothyroidism (SH) was defined as an elevation in serum TSH with a normal serum FT4 concentration. The primary outcome measure was serum levels of FT3, FT4 and TSH in children with SRNS. RESULTS: Thirty per cent of the children (n = 6) with SRNS had non-autoimmune subclinical hypothyroidism (2 children each with grade I, II and III). Children with SRNS had a median TSH value [3.9 mIU/L (0.5-13)] within normal range, but levels were high as compared to controls. Out of 6 children with SH, 3 were in partial remission, 3 were in complete remission. The TSH levels normalized on thyroxine supplementation in grades II and III subclinical hypothyroidism. CONCLUSION: Subclinical non-autoimmune hypothyroidism is present in a significant proportion of children with SRNS despite partial or complete remission. Thyroid profile should be evaluated routinely in this subset of patients.

Survival with 98% methemoglobin levels in a school-aged child during the "festival of colors".

Methemoglobin levels more than 70% have almost always been reported to have been fatal. The case of a 4-year-old boy who survived with methemoglobin levels of 98% is presented here. He was brought to the emergency department with complaints of vomiting, pain abdomen, and altered sensorium following accidental ingestion of paint thinner mixed with "Holi" colors. On examination, the child was in altered sensorium, cyanosed with saturations of 55%, who did not respond despite positive pressure ventilation with 100% oxygen. A possibility of toxic methemoglobinemia was considered and confirmed by finding of elevated methemoglobin levels of 98%. The child survived with definitive therapy with methylene blue and aggressive goal-directed approach.

The Tei index and asymptomatic myocarditis in children with severe dengue.

This study aimed to assess myocardial involvement in infants and children with severe dengue (as per the new World Health Organization [WHO] classification 2009) using the Tei index. This prospective observational study was conducted in the Department of Pediatrics, PGIMER and the associated Dr. RML Hospital, New Delhi from August to December 2010. The study included 67 children (ages 3 months-14 years) who satisfied the WHO criteria for the diagnosis of probable dengue fever with warning signs or severe dengue and tested positive for dengue via immunoglobulin-M (IgM) capture enzyme-linked immunoassay (MAC-ELISA). The patients were subjected to a complete blood count, liver function tests, renal profile, electrocardiography, myocardial band enzymes of creatine phosphokinase (CPK-MB), chest x-ray, abdomen ultrasonography, and two-dimensional echocardiography with color-flow Doppler mapping. Ejection fraction and Tei index measurements were performed. Significantly fewer patients with severe dengue were found to have myocardial involvement at admission by ejection fraction (48 %) and E/E' (37 %), than by the Tei index (70 %). Of the 67 patients with severe dengue, one died, giving a case fatality rate of 1.5 %. At discharge, the Tei index persisted on the high side for patients with myocardial involvement, whereas the ejection fraction improved for the majority of them. Most of the patients with severe dengue had asymptomatic myocarditis, as evident by a deranged Tei index, which improved but did not normalize by the time of discharge, necessitating a longer follow-up period. For the majority of the patients, inotropic support was not required to maintain hemodynamic stability.

Hearing status in children with frequently relapsing and steroid resistant nephrotic syndrome.

BACKGROUND: Children with idiopathic nephrotic syndrome (INS) are at risk of hearing impairment due to nephrotoxic drugs and biochemical impairments. METHODS: Forty children with INS aged 5-16 years [20 patients with frequently relapsing nephrotic syndrome (FRNS)/steroid dependent nephrotic syndrome (SDNS) and 20 with steroid resistant nephrotic syndrome (SRNS)] and 20 normal healthy controls were enrolled in this study. Pure tone audiometry was done using the ALPS AD 2000 audiometer. Sensorineural hearing loss was diagnosed when the bone conduction level was >20 dB and the difference in air to bone gap was ≤15 dB. Based on the air conduction (AC) threshold, deafness was graded into the following categories: mild (26-40 dB), moderate (41-55 dB), moderately severe (56-70 dB), severe (71-91 dB) and profound (>91 dB). RESULTS: Children with FRNS/SDNS had a higher threshold for hearing at frequencies of 250 and 500 Hz, respectively, than the controls. Of the children in the FRNS/SDNS category, three (15 %) had mild sensorineural hearing impairment. These children had a low serum calcium level (P < 0.03) and received higher cumulative doses of furosemide (P < 0.04). Children with SRNS had a higher threshold for hearing at frequencies of 250, 500, 1,000, and 2,000 Hz, respectively, than the controls. Of the 20 children with SRNS, ten (50 %) had sensoineural hearing impairment (8 mild, 2 moderate). Children with SRNS with a hearing defect had received a higher cumulative dose of furosemide (P < 0.03). CONCLUSIONS: Children with FRNS/SDNS and SRNS are at risk of sensorineural hearing impairment. The risk factors associated with this impairment were higher cumulative doses of furosemide and hypocalcemia. Larger prospective cohort studies are required to evaluate this association.

Safety profile assessment and efficacy of chemically characterized Cinnamomum glaucescens essential oil against storage fungi, insect, aflatoxin secretion and as antioxidant.

The study explores the efficacy of Cinnamomum glaucescens essential oil (EO) as insecticidal, antifungal, antiaflatoxin and antioxidant agent so as to recommend its application as plant based preservatives for food commodities. The study reports the chemical characterization of C. glaucescens oil and its 100% insecticidal activity against insect pest Callosobruchus chinensis on 12 h exposure and 98.74% oviposition deterrency at 0.15 µl/ml. The EO significantly inhibited growth and aflatoxin production by toxigenic strain of Aspergillus flavus LHP-10 at 4.5 and 3.5 µl/ml respectively. EO also showed appreciable antioxidant activity (IC(50) value=15.1 µl/ml), non phytotoxic nature on chickpea seed germination and in vivo potential as fumigant in food system providing 71.07% protection of chickpea samples from fungal contamination and 100% antifeedant activity against the insect invasion. The EO exhibited non-mammalian toxicity showing high LD(50) (3971.34 µl/kg) during oral toxicity on mice.