Multistate product traceforward investigation to link imported romaine lettuce to a US cyclosporiasis outbreak - Nebraska, Texas, and Florida, June-August 2013.

During June-August 2013, 25 US states reported 631 cyclosporiasis cases including Nebraska and Iowa where a regional investigation implicated common-source imported salad mix served in two chain restaurants. At least two common-origin growing fields were likely sources of contaminated romaine lettuce. Using producer- and distributor-provided data, we conducted a grower-specific traceforward investigation to reveal exposures of ill US residents elsewhere who reported symptom onset during 11 June-1 July 2013, the time period established in the Nebraska and Iowa investigation. Romaine lettuce shipped on 2-6 June from one of these Mexico-origin growing fields likely caused cyclosporiasis in 78 persons reporting illness onsets from 11 June to 1 July in Nebraska, Texas, and Florida. Nationwide, 97% (314/324) of persons confirmed with cyclosporiasis with symptom onset from 11 June to 1 July 2013 resided in 11 central and eastern US states receiving approximately two-thirds of romaine lettuce from this field. This grower's production practices should be investigated to determine potential sources of contamination and to develop recommendations to prevent future illnesses.

Regional investigation of a cyclosporiasis outbreak linked to imported romaine lettuce - Nebraska and Iowa, June-August 2013.

A regional, multistate investigation into a June-August 2013 cyclosporiasis outbreak was conducted in Nebraska, Iowa, and neighbouring states. Cases were confirmed on the basis of laboratory and clinical findings. Of 227 cases in Iowa (n = 140) and Nebraska (n = 87) residents, 162 (71%) reported dining at chain A/B restaurants - 96% reported house salad consumption. A case-control study identified chain A/B house salad as the most likely vehicle. Traceback was conducted to ascertain production lot codes of bagged salad mix (iceberg and romaine lettuce, red cabbage, and carrots) served as house salad in implicated restaurants. A single production lot code of salad mix supplied by both a common producer and distributor was linked to the majority of confirmed cases in persons reporting regional chain A/B exposure. The salad mix linked to illnesses contained imported romaine lettuce from two separate single-grower fields-of-origin and ⩾1 additional field from another grower.

Temporal and spatial changes in ion homeostasis, antioxidant defense and accumulation of flavonoids and glycolipid in a halophyte Sesuvium portulacastrum (L.) L.

Salinity is an important environmental constraint limiting plant productivity. Understanding adaptive responses of halophytes to high saline environments may offer clues to manage and improve salt stress in crop plants. We have studied physiological, biochemical and metabolic changes in a perennial, fast growing halophyte, Sesuvium portulacastrum under 0 mM (control), 150 mM (low salt, LS) and 500 mM (high salt, HS) NaCl treatments. The changes in growth, relative water content, cation, osmolyte accumulation, H2O2 and antioxidant enzyme activity (SOD, CAT and APX) were observed under different treatment conditions. A positive correlation was revealed for sodium ion accumulation with malondialdehyde (r2 = 0.77), proline (r2 = 0.88) and chlorophyll content (r2 = 0.82) under salt treatment while a negative correlation was observed with relative tissue water content (r2 = -0.73). The roots and leaves showed contrasting accumulation of potassium and sodium ions under LS treatment. Temporal and spatial study of sodium and potassium ion content indicated differential accumulation pattern in roots and leaves, and, high potassium levels in root. Higher H2O2 content was recorded in roots than leaves and the antioxidant enzyme activities also showed significant induction under salt treatment conditions. Gene expression profiling of sodium transporters, Sodium proton exchanger (NHX3), Vacuolar ATPase (vATPase) and Salt overly sensitive1 (SOS1) showed up regulation under salt stress after 6-24 hr of NaCl treatment. Metabolite changes in the salt stressed leaves showed increased accumulation of flavonoids (3,5-dihydroxy-6,4'-dimethoxy-flavone-7-O-[α-L-rhamnopyranosyl-(1→6)-ß-D-glucopyranoside], and3,5-dihydroxy-6,3',4'-trimethoxy-flavone-7-O-[α-L-rhamnopyranosyl-(1→6)-ß-D-glucopyranoside] in both LS and HS treatments, while a glycolipid, 1-O-linolenyl-2-O-(palmitoyl)-3-O-galactopyranosyl glycerol, accumulated more in LS over HS treatments and control. The results suggest that differential spatial and temporal cation levels in roots and leaves, and accumulation of flavanoid and glycolipid could be responsible for salt adaptation of S. portulacastrum.

Detection of chandipura virus from sand flies in the genus Sergentomyia (Diptera: Phlebotomidae) at Karimnagar District, Andhra Pradesh, India.

A total of 191 adult sand flies belonging to the genus Sergentomyia were collected from seven villages in Karimnagar and Warangal districts of Andhra Pradesh State, India, after an outbreak of encephalitis due to Chandipura virus (CHPV). Fifteen pools, each containing two specimens, were tested by reverse transcriptase-polymerase chain reaction and sequencing. One pool of Sergentomyia from Kolanur village in Karimnagar District was positive for CHPV.

Isolation of Ganjam virus from ticks collected off domestic animals around Pune, Maharashtra, India.

Studies on viruses of zoonotic importance in certain villages around Pune were undertaken between December 2000 and January 2002. A total of 1,138 adult ticks belonging to six different species were collected off domestic animals and processed for virus isolation. Six virus isolates were obtained. All six isolates were identified as Ganjam virus by Quick Complement Fixation test and reverse transcriptase-polymerase chain reaction using RNA nucleocapsid gene amplification. Five isolates were from the pools of adult Hemaphysalis intermedia ticks, and one isolate was from a pool of adult Rhipecephalus hemaphysaloides. This is the first report of isolation of Ganjam virus from Maharashtra state of India.

Incidence of neutralizing antibodies to Chandipura virus in domestic animals from Karimnagar and Warangal Districts of Andhra Pradesh, India.

In order to determine the possible role of domestic animals in the outbreak of acute encephalitis associated with Chandipura virus (CPV) among children in Andhra Pradesh in 2003, a serological survey of domestic animals was carried out during the epidemic in July 2003. Out of 180 animal sera from highly affected areas of the Karimnagar and Warangal districts of Andhra Pradesh 33 (18.3%) had virus neutralizing (VN) antibodies to CPV. The positive animals consisted of pigs (30.6%), buffalos (17.9%), cattle (14.3%), goats (9.3%) and sheep (7.7%). Isolation of CPV and detection of CPV antibodies in patients with encephalitis reported earlier and the evidence of antibodies to CPV in domestic animals shown here suggest that CPV circulates in this region and should be considered an emerging virus of public health importance.

Determination of trace impurities in uranium, thorium and plutonium matrices by solvent extraction and inductively coupled plasma atomic emission spectrometry.

Studies on the determination of trace metallic impurities in nuclear materials such as uranium, thorium and plutonium are described. The bulk of the matrix is separated by batch extraction from their nitric acid solutions using 2-ethylhexyl hydrogen 2-ethylhexyl phosphonate (KSM-17, equivalent to PC88-A). The final aqueous phase containing the metallic impurities is fed to a high-temperature source inductively coupled plasma and the analysis is carried out employing a computer-controlled multichannel direct-reading spectrometer. The studies also included the recovery of impurities at various acidities and spectral interferences of the above matrices over the analyte elements. Based on the above studies, methods were standardized for the determination of 19 elements, viz. Al, B, Be, Ca, Cd, Cr, Cu, Fe, Mg, Mn, Ni, Pb, Si, Zn, Ce, Dy, Eu, Gd and Sm, in U/Th/Pu solutions. The relative standard deviation for various elements is in the range 1-5%.

Investigation of buffalopox outbreaks in Maharashtra State during 1992-1996.

During 1992-96, outbreaks of buffalopox zoonosis were reported from different villages in Jalgaon, Dhule and Beed districts of Maharashtra State. In humans, pox lesions were observed on the hands whereas in affected buffaloes and cows the lesions were noticed mainly on the teats and udder. Twenty two virus strains were isolated from the skin scabs collected from infected humans and milch animals. Neutralizing antibodies were detected not only in the sera of affected humans but also in their contacts. Detection of antibodies in young individuals from endemic area, who were neither vaccinated for smallpox nor had any contact with buffaloes or history of any poxvirus disease, is suggestive of occurrence of subclinical infection. A few children who had no contact with infected animals also showed clinical manifestations with disseminated lesions on the face, arm and buttocks, and thus suspected to have acquired infection through their infected parents or other family members indicating a possible man to man transmission. Therefore, in the light of discontinuation of smallpox vaccination, buffalopox outbreaks need to be monitored carefully as this may emerge as a serious zoonotic disease in India.

Analysis of a clinically important interaction between phenytoin and Shankhapushpi, an Ayurvedic preparation.

During the course of routine plasma drug level monitoring an unexpected loss of seizure control and reduction in plasma phenytoin levels was noticed in two patients who were also taking 'Shankhapushpi' (SRC), an Ayurvedic preparation. Therefore, the present study was undertaken in rats to investigate any SRC-phenytoin interaction from both pharmacokinetic (serum levels) and pharmacodynamic (electroshock seizure prevention) aspects. Single dose SRC and phenytoin (oral/i.p.) coadministration did not have any effect on plasma phenytoin levels but decreased the antiepileptic activity of phenytoin significantly. On multiple-dose coadministration, SRC reduced not only the antiepileptic activity of phenytoin but also lowered plasma phenytoin levels. SRC itself showed significant antiepileptic activity compared to placebo and is worth further investigation. However, the clinical combination of SRC with phenytoin is not advised.

Serological evidence for Japanese encephalitis virus and West Nile virus infections in water frequenting and terrestrial wild birds in Kolar District, Karnataka State, India. A retrospective study.

In a serological survey of birds in a Japanese encephalitis (JE) endemic area of Kolar District, Karnataka State, India, 859 bird sera were tested by hemagglutination-inhibition test (HIT) for JE encephalitis and West Nile encephalitis (WNE) viruses. Only 2 (0.002%) and 178 (20.72%) sera were positive for JE virus (JEV) and WNE virus (WNV), respectively. Only 160 (18.63%) of 859 sera could be subjected to neutralizing test (NT). Of these, 20 (12.50%) and 62 (38.75%) were positive for JEV and WNV antibodies, respectively. These findings indicate that bird species such as Pond Herons and Little Egrets among ardeid birds and Grey Partridges and Quails among terrestrial birds are infected with JEV and WNV and play probably a role in the maintenance of these viruses in the abovementioned part of India.

Isolation of bovine viral diarrhea virus 1, a pestivirus from autopsied lamb specimen from Tamil Nadu, India.

An epizootic of febrile illness among the Madras red breed of sheep had occurred in 1994 in Verrapuram, Chennai, India. The epizootic was suspected as Rift Valley fever (RVF)-like sickness based on clinical features. However, its etiological agent could neither be isolated nor implicated conclusively. During the post-epizootic period a male lamb died of similar clinical features and the spleen was immediately collected. Inoculation of spleen suspension in infant mouse brain yielded a virus that was serially passaged in infant mice and rhabdomyosarcoma (RD) cells. Electron microscopic observations revealed virus particles resembling flaviviruses. RT-PCR performed on extracted total RNA from infected cells and mouse brains with flavivirus-specific or RVF-specific primers gave negative results. However, an amplicon of 280 bp was obtained with pestivirus-specific primers from the 5'-UTR. Further, a nested PCR yielded a product of 157 bp. Nucleotide sequencing of the 157 bp product showed 100% homology to BVDV-1. Western blot analysis with a flavivirus envelope protein-specific MAb revealed three proteins of 33 K, 45 K and 55 K. Further studies suggested that the 33 K and 55 K proteins were glycosylated. This is the first report of isolation of BVDV-1 from a lamb in India.

Isonicotinic acid hydrazide inhibits cell population growth during teratogenesis of chick embryo.

In chick embryos treated with a 4 hr pulse of 7.2 X 10(-5) M isonicotinic acid hydrazide (INH) the cell population growth is inhibited with an increased population doubling time. Teratogenised blastoderm cells complete their ongoing cell cycle and arrest in G1 phase. A chase with an equimolar concentration of pyridoxal-5-phosphate restores the growth rate after a lag of 4 hr equivalent to the duration of treatment with INH. Presumptive mesoblast cells invaginated through the primitive streak and neuroectoblast cells induced prior to the application of INH differentiate, while the teratogen inhibits morphogenesis and organization of organ primordia.

Need for 25 mg tablets of phenytoin.

Phenytoin is widely used for the treatment of generalized tonic clonic and partial seizures. Monitoring of serum phenytoin levels is essential to optimize therapy. Of 320 patients monitored, 190 patients whose seizures were uncontrolled were followed up before and after dosage adjustment was carried out. Plasma phenytoin estimation was done by HPLC method. Of all the patients receiving the drug, 20% and 8% of patients were finally on dosages requiring 50 and 25 mg fraction administration respectively. Administration of 100 mg fractions resulted in either loss of seizure control or toxicity. This emphasizes the need for providing tablets of 25 mg strength, presently not available in this country.

Food reduces isoniazid bioavailability in normal volunteers.

Food and drug interaction is a major variable in bioavailability of drugs. Isoniazid is the most common antitubercular drug used in India. We studied the effect of standard Indian breakfast and lunch on the bioavailability of isoniazid in a single dose crossover study in normal male volunteers. The standard breakfast and lunch significantly reduced plasma AUC, Cmax and Kabs values of isoniazid. Isoniazid, thus, should not be administered with food.

"An analysis of epileptic patients nonresponsive to drugs".

Fifty-four consecutively referred patients with uncontrolled epilepsy were subjected to Therapeutic Drug Monitoring on an out patient basis. Regular 2 weekly follow up for a minimum period of 2 months was done, after altering the drug dosage and bringing plasma level(s) within therapeutic range. Plasma levels of Phenobarbitone, Phenytoin and Carbamezepine were done by High Pressure Liquid Chromatography. Eventually, 24 patients were controlled and 30 remained uncontrolled. Significant differences between these 2 groups were found, as regards, duration of epilepsy (p < 0.01), associated mental retardation (p < 0.02), initial carbamazepine dosage and plasma levels in patients on carbamazepine montherapy (p < 0.02 and P < 0.01, respectively) and final phenytoin plasma levels in patients on combined therapy with phenobarbitone and phenytoin (p < 0.05). This study emphasizes the importance of early diagnosis and treatment of epilepsy with the help of plasma level monitoring of anti-epileptic drugs.

Multiple neural inductions in area opaca by grafts of the Hensen's node do not retard host chick embryo development.

Four hundred and forty four full primitive streak stage chick embryos were cultured in vitro and 261 were transplanted with 1, 3 or 5 Hensen's nodes in the area opaca. Irrespective of the number of grafts, neural induction was observed in 90% cases. The development of control and grafted embryos and the size of blastoderm area were monitored at the time of grafting and after 20 hr. We find that the induced neural tissue and differentiated tissue of graft-origin neither fuse with the host embryonic axis, nor retard its development.

A study of phenobarbital and dilantin in neonatal seizures.

A total of 40 cases of neonatal convulsions of different nonmetabolic aetiological factors were studied. Patients with kernicterus were included in the study. Peak plasma phenobarbital concentrations after incremental loading doses of phenobarbital i.e. 10 mg/kg, 15 mg/kg, and 20 mg/kg were determined. Diphenylhydantoin was added if phenobarbital alone was unable to control seizures. In three patients, a combination of phenobarbital and diphenylhydantoin was used as the initial loading therapy. Increase in the loading dose of phenobarbital was associated with an increase in its peak plasma concentration. Despite increase in the plasma phenobarbital concentration beyond the 'therapeutic' levels suggested by the Western studies, doses of 15 mg/kg and 20 mg/kg of phenobarbital were unable to score over the traditional regimen of 10 mg/kg. Convulsions were controlled in 50% of the patients with any of these three regimens, irrespective of the aetiology. Convulsions were controlled in 7 out of the 9 cases where diphenylhydantoin was added, because of the failure of phenobarbital in controlling the convulsions as a single drug. Convulsions of all the three patients, in whom a combination of phenobarbital and diphenylhydantoin was used by random selection as the initial bolus, were controlled. Seizure effects were difficult to distinguish from drug effects but major side effects were not encountered despite the fluctuating drug levels in the sick neonate.