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.

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.

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.

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.

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.

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.

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.

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.

In vitro and in vivo antifungal activity of essential oils of Cymbopogon martini and Chenopodium ambrosioides and their synergism against dermatophytes.

The present study was carried out to evaluate the antifungal efficacy of essential oils (EO) of Cymbopogon martini, Chenopodium ambrosioides and of their combination against dermatophytes and some filamentous fungi in vitro as well as in vivo using a guinea pig model. The minimum inhibitory concentrations of EOs and of their combination were found between 150 and 500 ppm, while those of known antifungal drugs ranged from 1000 to 5500 ppm. EO ointments were prepared and applied against induced ringworm in guinea pig model and disease removal was observed in 7-21 days, and the hair samples showed negative results for fungal culture in a time-dependent manner after the application of EO ointments. Chemical constituents of EOs were determined by GC-MS. Both the EOs and their combination displayed strong antifungal effects. The results provide a scientific validation for the use of these EOs in the treatment of dermatophyte infections and may be recommended as an alternative to synthetic drug for topical application.

Swine-origin influenza A (H1N1) in Indian children.

BACKGROUND: Swine-origin influenza A H1N1 (S-OIV) has not been systematically studied in Indian children. OBJECTIVES: To study the clinical characteristics, morbidity and mortality pattern in children with S-OIV infection. METHODS: This prospective study was conducted during the 'containment phase' of the pandemic in New Delhi from 10 June to 5 August 2009. All children suspected of being infected by S-OIV were admitted to the isolation wards and clinically evaluated according to WHO guidelines. Nasal and throat swabs were collected immediately for real-time reverse transcriptase polymerase chain reaction (RT-PCR). Haemoglobin, total leucocyte and platelet counts and chest radiography were undertaken in all patients. Those who tested positive for S-OIV infection were treated with oseltamivir for 5 days in isolation wards. RESULTS: Thirty-seven children fulfilled the inclusion criteria. Twenty-one tested positive for S-OIV by RT-PCR and 16 tested negative. Comparison of the clinical characteristics of the two groups showed that duration of cough was longer in children with S-OIV (p<0.03). Total leucocyte and lymphocyte counts were significantly less in the S-OIV group (p<0.001 and , 0.02, respectively). Oseltamivir-related gastritis was seen in 38% of children. All improved and were discharged. CONCLUSION: S-OIV infection in Indian children had features similar to those of seasonal influenza. Lymphopenia is an important feature of S-OIV.

Supplemental ultraviolet-B induced changes in essential oil composition and total phenolics of Acorus calamus L. (sweet flag).

The effect of supplemental UV-B radiation (sUV-B) was evaluated on the essential oil contents of sweet flag (Acorus calamus L.), a medicinal plant grown under natural field conditions. After the emergence of two leaves, plants were exposed to sUV-B radiation of 1.8 kJ m(-2) above the ambient level of UV-B. The level of essential oil and phenol contents increased with exposure to sUV-B. Exposure of sUV-B resulted in significant increase in p-cymene and carvacrol contents of essential oil. Decrease in the level of major component beta-asarone due to sUV-B treatment is of prime importance, because of its toxicological concern to human health.

Evaluation of Chenopodium ambrosioides oil as a potential source of antifungal, antiaflatoxigenic and antioxidant activity.

Essential oil extracted from the leaves of Chenopodium ambrosioides Linn. (Chenopodiaceae) was tested against the aflatoxigenic strain of test fungus Aspergillus flavus Link. The oil completely inhibited the mycelial growth at 100 microg/ml. The oil exhibited broad fungitoxic spectrum against Aspergillus niger, Aspergillus fumigatus, Botryodiplodia theobromae, Fusarium oxysporum, Sclerotium rolfsii, Macrophomina phaseolina, Cladosporium cladosporioides, Helminthosporium oryzae and Pythium debaryanum at 100 microg/ml. The oil showed significant efficacy in inhibiting the aflatoxin B1 production by the aflatoxigenic strain of A. flavus. During in vivo investigation it protected stored wheat from different storage fungi for one year. Chenopodium oil also exhibited potent antioxidant activity when tested by ABTS method. All these observations suggest the possible exploitation of the Chenopodium oil as potential botanical fungitoxicant in ecofriendly control of post harvest biodeterioration of food commodities from storage fungi.

Evaluation of Cymbopogon martinii oil extract for control of postharvest insect deterioration in cereals and legumes.

The essential oil of Cymbopogon martinii was tested for its potency as a botanical pesticide to protect stored wheat (Triticum aestivum) and gram (garbanzo bean, Cicer arietinum) from insect infestation. The C. martinii oil was potent as a fumigant in stored gram. The oil was an effective repellent against the beetles Callosobruchus chinensis and Tribolium castaneum. Geraniol, the major component of the oil, was not as effective as the oil itself. C. martinii oil significantly affected oviposition, adult development, and mortality of C. chinensis in cow peas (Vigna unguiculata). The C. martinii oil when used as fumigant did not affect viability, germination, and seedling growth of gram. Because of its insecticidal and semiochemical nature, the oil could be used as an alternative to synthetic pesticides in an integrated pest management program to protect stored food commodities.

Townes-Brocks syndrome with hypothyroidism.

Townes-Brocks syndrome (TBS) is an autosomal dominant disorder with multiple malformations and variable expression. Major findings include external ear anomalies, hearing loss, limb deformity, imperforate anus, and renal malformations. Hypothyroidism is not a recognized feature of TBS. We are reporting a case of TBS with hypothyroidism, a rare association.

Life threatening exacerbation in idiopathic pulmonary hemosiderosis salvaged by cyclophosphamide infusion.

A seven-year-old girl presented with frequent fever, cough and shortness of breath of three months duration. On the basis of her clinical features, peripheral blood and sputum findings, she was diagnosed as a case of idiopathic pulmonary hemosiderosis. After initial stabilisation with steroids and chloroquine, she presented four years later with massive pulmonary hemorrhage and respiratory failure, which responded dramatically to cyclophosphamide infusion. The rare occurrence of pulmonary hemosiderosis and different treatment regimens is discussed.