Evaluation of the diagnostic potential and DIVA capability of recombinant LigBCon1-5 protein of Leptospira interrogans serovar Pomona in canine leptospirosis.

BACKGROUND: Canine leptospirosis is a serious public health concern. AIMS: This study aims to investigate the feasibility of conserved first to fifth domains of recombinant Leptospira immunoglobulin like protein B antigen (rLigBCon1-5) as a serodiagnostic marker for detecting canine leptospirosis. METHODS: A total of 340 unvaccinated canine serum samples were screened using microscopic agglutination test (MAT) and rLigBCon1-5 based immunoglobulin G (IgG) indirect-enzyme-linked immunosorbent assay (I-ELISA). Further, 60 vaccinated canine sera were screened using MAT and rLigBCon1-5 based latex agglutination test (LAT). RESULTS: Microscopic agglutination test results revealed seropositivity of 28.6%. The relative sensitivity, specificity, and accuracy of IgG I-ELISA in comparison to MAT were 100%, 96.0%, and 97.2%, respectively. Out of 60 vaccinated sera, 46 sera reacted with MAT alone, and eight sera reacted by both tests, while six sera were non-reactive with both tests. Anti-LigB antibodies were detected in eight canine sera by rLigBCon1-5 based LAT. In five LAT reactive sera, agglutinins of locally circulating Leptospira serovars Grippotyphosa (n=4) and Australis (n=1) were detected. In three LAT reactive sera, agglutinins against Icterohaemorrhagiae (n=3) produced due to natural infection were present. CONCLUSION: Immunoglobulin G based indirect ELISA assay (IgG I-ELISA) can be employed as an alternative test instead of MAT. rLigBCon1-5 based LAT detected anti-LigB antibodies in eight vaccinated sera where the vaccine failure occurred partially or totally due to the limited efficacy spectrum of Nobivac® RL and cold chain breakage. This vaccine could not provide cross-protection against locally circulating Leptospira serovars. The recombinant LigBCon1-5 antigen based LAT possesses capability of differentiating infected from vaccinated individuals (DIVA capability) when employed as a pen-side test for detecting canine leptospirosis.

Use of different coloured ovitraps in the surveillance of Aedes mosquitoes in an arid-urban area of western Rajasthan, India.

BACKGROUND & OBJECTIVES: In the absence of any commercially available dengue vaccine or antiviral therapy, controlling the primary mosquito vector, Aedes aegypti, is currently the only means to prevent dengue outbreaks. Ovitraps are being employed as a sensitive method for detecting the presence of Ae. aegypti. Size, colour, material, paddle and use of oviposition attractant have been reported as important factors influencing the efficacy of ovitraps. METHODS: For the study of coloured ovitraps baited with grass infusion, 24 households were randomly selected in a locality and five ovitraps/premises were installed. Weekly ovitrap surveys were conducted during the study for four times. Ovitrap data were analyzed on the basis of its location, day-wise percent positivity, presence of eggs laid and inference of different ovitrap parameters were drawn. RESULTS: Grass infusion, used in the ovitraps has highest attractiveness on Day 6 in comparison to other days. Percent positivity of different coloured ovitraps revealed that red coloured ovitraps have highest positivity (92.7%), followed by black and orange (91.7% each), green (76.3%) and transparent (45.8%). The individual colour-wise ovitrap positivity revealed that the highest ovitrap positivity on Day 1 was recorded for black colour (21.8%), which on D2 and D3, in case of red colour (37.5 and 26%). However, on D4, D5 and D6 highest number of positive ovitraps was recorded in orange coloured (12.5, 21.9 and 31.3%, respectively) and on D7 highest number of positive ovitraps were recorded in transparent (21.9%), followed by green coloured ovitrap (20.8%). The overall location-wise ovitraps positivity revealed that highest positivity was recorded among the ovitraps installed inside bathrooms (92.0%), followed by bedrooms (85.0%), lobby (66.7%) and stores (48.3%). INTERPRETATION & CONCLUSION: The study revealed that for immediate detection of the presence of gravid females in a particular locality during epidemics, the black coloured ovitraps may be useful, however, in normal situations for surveillance of the vector species, the orange coloured ovitraps might be more suitable. Bathrooms were found to be the highest positive location for pitching ovitraps.

Larvicidal potential of wild mustard (Cleome viscosa) and gokhru (Tribulus terrestris) against mosquito vectors in the semi-arid region of Western Rajasthan.

Cleome viscosa L. (Family: Capparaceae) commonly known as Tickweed or wild mustard and Tribulus terrestris L. (Family: Zygophyllaceae) commonly known as Gokhru, growing wildly in the desert areas in the monsoon and post monsoon season, are of great medicinal importance. Comparative larvicidal efficacy of the extracts from seeds of C. viscosa and fruits and leaves of T. terrestris was evaluated against 3rd or early 4th stage larvae of Anopheles stephensi (Liston), Aedes aegypti (Linnaeus) and Culex quinquefasciatus (Say) in different organic solvents. 24 and 48 hr LC50 and LC90 values along with their 95% fiducial limits, regression equation, chi-square (chi2)/ heterogeneity of the response was determined by log probit regression analysis. The 24 hr LC50 values as determined for seeds of C. viscosa were 144.1, 99.5 and 127.1 (methanol); 106.3, 138.9 and 118.5 (acetone) and 166.4, 162.5 and 301.9 mg l(-1) (petroleum ether extracts) for all the three mosquito species respectively showing that methanol and acetone extracts were a little bit more effective than the petroleum ether extracts. Experiments were carried out with fruits and leaves of T. terrestris with all the solvents and mosquito species. The 24 hr LC50 values, as determined for fruits of T. terrestris were 70.8, 103.4 and 268.2 (methanol); 74.0,120.5 and 132.0 (acetone) and 73.8,113.5 and 137.4 mg l(-1) (petroleum ether extracts) while the 24 hr LC50 values for leaves were 124.3, 196.8 and 246.5 (methanol); 163.4, 196.9 and 224.3 (acetone) and 135.8, 176.8 and 185.9 mg l(-1) (petroleum ether extracts) for all the three mosquito species respectively. The results clearly indicate that fruit extracts of T. terrestris were more effective as compared to leaves extracts in the three solvents tested. Larvae of An. stephensi were found more sensitive to both fruit and leaves extracts of T. terrestris followed by larvae of Ae. aegypti and Cx. quinquefasciatus. Extracts from the seeds of C. viscosa were found less effective as compared to the fruit extracts of T. terrestris indicating that active larvicidal principle may be present in the fruits of this plant species. The studywould be of great importance while formulating the control strategy, for vectors of malaria, dengue and lymphatic filariasis, based on alternative plant based insecticides in this semi-arid region.

Laboratory observations on the larvicidal efficacy of three plant species against mosquito vectors of malaria, dengue/dengue hemorrhagic fever (DF/DHF) and lymphatic filariasis in the semi-arid desert.

Comparative larvicidal efficacy of aqueous and organic solvent extracts from seeds, leaves and flowers of three desert plants viz. Calotropis procera (Aiton), Tephrosia purpurea (L.) Pers. and Prosopis juliflora (Sw.) DC. was evaluated against Anopheles stephensi (Liston), Aedes aegypti (Linnaeus) and Culex quinquefasciatus (Say). For this purpose larvae of all the three mosquito species were reared in the laboratory and studies carried out on late 3rd or early 4th instars using standard WHO technique. Based on concentration mortality data 24 and 48 hr LC50and LC90 values along with their 95% fiducial limits, regression equation, chi-square (chi2)/ heterogeneity of the response were determined by log probit regression analysis. Experiments were carried out with different solvent extracts of seeds of C. procera which revealed that methanol (24 hr LC50: 127.2, 194.8, 361.0) and acetone (229.9, 368.1,193.0 mg l(-1)) extracts were more effective with the three mosquito species, respectively. Petroleum ether extract was effective only on An. stephensi while aqueous extracts were not effective at all with any of the mosquito species (mortality < 10-30%). Tests carried out with methanol extracts of fresh leaves (24 hr LC50: 89.2, 171.2, 369.7) and flowers (24 hr LC50: 94.7,617.3, 1384.0 mg l-(-1)) of Calotropis showed that preparations from fresh parts were 2-3 times more effective as compared to the stored plant parts. Efficacy was less than 10-30% with both An. aegypti and Cx. quinquefasciatus while An. stephensi was still susceptible to extracts from both leaves and flowers even after two years of storage. The 24 hr LC50 values as observed for methanol extracts of seeds of T. purpurea and leaves of P. juliflora were 74.9, 63.2 and 47.0 and 96.2,128.1 and 118.8 mg l(-1) for the above three mosquito species, respectively. Experiments carried out up to 500 mg l-(1) with leaves (T. purpurea) and seeds (P. juliflora) extracts show only up to 10-30% mortality indicating that active larvicidal principle may be present only in the seeds of Tephrosia and leaves of Prosopis. In general, anophelines were found more susceptible than the culicines to the plant derived derivatives. More studies are being carried outon some other desert plants found in this arid region. The study would be of great importance while formulating vector control strategy based on alternative plant based insecticides in this semi-arid region.

Comparative larvicidal potential of different plant parts of Withania somnifera against vector mosquitoes in the semi-arid region of Rajasthan.

Larvicidal potential of the extracts from different parts viz. green and red fruits, seeds, fruit without seeds, leaves and roots of Withania somnifera in different solvents was evaluated against larvae of Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus, the important disease vectors prevalent in the semi-arid region. Experiments were carried out on late 3rd or early 4th instar larvae of these mosquitoes using standard WHO technique. 24 and 48 hr LC50 values along with their 95% confidence limits, regression equation, chi-square (chi2)/heterogeneity of the response have been determined by log probit regression analysis. The 24 hr LC50 values as observed for whole green fruits in water, methanol and petroleum ether were 350.9, 372.4, 576.9; 115.0, 197.1, 554.6; 154.9, 312.0, 1085.0 while corresponding values for red fruits were 473.5, 406.4,445.2; 94.7, 94.5, 1013.0; 241.8, 535.0, 893.3 mg l(-1) for An. stephensi, Ae. aegypti and Cx. quinquefasciatus respectively showing that methanol extracts were more effective against anophelines as compared to culicines when whole fruits were taken. The 24 hr LC50 values as observed for seeds in acetone, methanol and petroleum ether were 188.1, 777.5, 822.5; 245.5, 769.0, 1169.0; 140.3, 822.9, 778.4 and for fruit without seeds were 80.2, 97.6, 146.6; 88.4, 404.4, 1030.0; 30.0, 44.5, 54.2 mg l(-1) for the above mosquito species respectively showing that extract of fruit without seeds were most effective in petroleum ether followed by acetone and methanol extracts. However, experiments conducted with methanol extracts of leaves and roots of this plant species did not show any appreciable larvicidal activity and a 20-40% mortality was observed up to 500 mg l(-1) of the extracts. Overall larvae of anophelines were found more susceptible as compared to culicines to all the extracts tested. Petroleum ether extract of fruit without seeds was found most effective against all the mosquito species showing that active ingredient might be present in this part of the plant species. The study would be of great importance while planning vector control strategy based on alternative plant derived insecticides.

Revisiting the CNS tuberculosis with emphasis on giant tuberculomas and introducing the "outer rim excrescence sign".

Central nervous system tuberculosis is a leading cause of morbidity and mortality in developing countries with Mycobacterium tuberculosis as the main etiological agent. Classical imaging findings pose no problem for diagnosis of CNS tuberculosis. Cases of giant or solitary ring enhancing lesions are increasing which mimic other infectious and noninfectious pathologies. This is a retrospective study of 40 patients (28 male and 12 female) of either surgically or clinically proven CNS tuberculosis. Solitary tuberculomas (ring or solid enhancing) were seen in 19 patients. Four patients showed presence of two to three lesions and the lesion more than one centimeter size was categorized as 'giant tuberculoma'. Multiple lesions with other associated findings were noted in 21 patients. Total 33 patients showed solitary or multiple ring enhancing lesions. Seven patients showed solid enhancing lesions (giant tuberculoma). The "Outer rim excrescence sign" was noted in a total of 16 patients. Both brain and spinal tuberculosis was noted in six patients. Seven patients presented with meningitis and hydrocephalous which is considered as the most common manifestation in the literature. Spinal leptomeningeal involvement was seen in five patients, whereas intramedullary cord tuberculomas were noted in four patients. Profound T2W hypointensity is a reliable predictor for giant tuberculomas with intense contrast enhancement, especially in deep locations. 'Outer rim excrescence' sign may contribute in cases of ring and solid enhancing lesions, however it requires further comprehensive study with MT Imaging and MR Spectroscopy to be recorded as a differentiating feature.

Larvicidal activity of the extracts from different parts of the plant Solanum xanthocarpum against important mosquito vectors in the arid region.

Larvicidal efficacy of the aqueous and methanol extracts from green unripe and yellow ripe fruits, seeds, leaves and roots of the plant Solanum xanthocarpum was evaluated against Anopheles culicifacies, Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus, the important mosquito vectors prevalent in the arid region. Studies were carried out on late 3rd orearly 4th instarlarvae of these species using standard WHO technique. Based on concentration mortality data LC50 and LC90 values along with their fiducial limits, regression equation, chi-square (chi2)/heterogeneity of the response have been determined bylog probit regression analysis. The 24 hr LC50 values as observed for aqueous extracts for green unripe and yellow ripe fruits were 112.7, 498.2 and 846.3 mg l(-1) and 104.7, 267.7and 832.2 mg l(-1) for Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus respectively. However, the 24 hr LC50 values formethanol extracts of yellow ripe fruits and seeds were 51.6, 52.2, 118.3 and 157.1 mg l(-1) and 66.9, 73.7, 123.8 and 154.9 mg l(-1) for the above four vector species respectively. The results showed that larvae of anophelines were much more susceptible as compared to culicines to both the aqueous and methanol extracts from fruits and seeds of the plant tested. The methanol extracts were found 2-5 times more effective as compared to the aqueous extracts. The chi-square values calculated during the analysis did not show any heterogeneity of the response. Experiments conducted with aqueous and methanol extracts from leaves and roots of this plant species didn't show any larvicidal activity against any of the mosquito species tested. The study would be of great importance while formulating vector control strategy based on alternative plant based insecticides.

Evaluation of larvicidal efficacy of Solanum xanthocarpum storage against vector mosquitoes in north - western Rajasthan.

Efficacy of the methanol extracts from fruits without seeds, whole yellow ripe fruits and seeds of the plant Solanum xanthocarpum was evaluated against larvae of Anopheles culicifacies, Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus, the important vector mosquitoes prevalent in the arid region. Experiments were carried out on late 3rd or early 4th instar larvae of these mosquitoes using standard WHO technique. 24 and 48 hr LC50 and LC90 values along with their fiducial limits, regression equation, chi-square (chi2)/heterogeneity of the response have been determined by log probit regression analysis. The 24 hr LC50 values as observed for fruit without seeds, whole fruits and seeds after six months were 79.6, 91.7 and 131.7; 131.4, 186.9 and 195.6; 273.4, 290.9 and 377.6 and 384.9, 450.6 and 520.0 mg l(-1) for An. culicifacies, An. stephensi, Ae. aegypti and Cx. quinquefasciatus respectively. However, the 24 hr LC50 values for fresh yellow ripe fruits and seeds determined previously were 51.6 and 66.9; 52.2 and 73.7; 118.3 and 123.8 and 157.1 and 154.9 mg l(-1) for the above four vector species respectively showing that the efficacy was decreased two to three times more after six months of storage. The 24 hr LC50 values were also determined after one year of storage for fruit without seeds, whole fruits and seeds. The values were 103.3, 120.1 and 195.7; 146.3, 224.3 and 251.2; 316.9, 336.5 and 426.2 and 393.0, 500.3 and 656.7 mg l(-1) as compared to the 24 hr LC50 values of 47.4, 51.6 and 66.9; 52.3, 52.2 and 73.7; 108.0, 118.3 and 123.8 and 141.1, 157.1 and 154.9 mg l(-1) of all the three fresh preparations for the above four vector species respectively showing that the efficacy was further decreased after one year of storage. Further the extracts should be used when they have been prepared from the fresh plant parts instead of the old stored parts. Larvae of anophelines were found more susceptible as compared to culicine. HPLC and IR studies showed that active component might be present in the fruits and seeds of this plant species. The study would be of great importance while planning vector control strategy based on alternative plant derived insecticides.

Relative susceptibility of some common mosquito vector larvae to synthetic insecticidal compounds in north-western Rajasthan.

Relative susceptibility of three important mosquito vector larvae viz., Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus, prevalent in the arid region was determined against four organophosphates (malathion, fenitrothion, fenthion, temephos) and three synthetic pyrethroid (alphamethrin, deltamethrin and fanvalerate) compounds. Studies were carried out on late 3rd or early 4th instar larvae of these species using standard WHO technique. Based on concentration mortality data LC50 and LC90 values along with their fiducial limits, regression equation, chi-square (chi2)/heterogeneity of the response have been determined by log probit regression analysis. LC50 values as observed for the above seven insecticides were 0.8097, 0.0398. 0.0432, 0.0035, 0.0025, 0.0092, 0.1006; 1.2370, 0.0531, 0.0655, 0.0076, 0.00004, 0.00004, 0.0046 and 1.4980, 0.0719, 0.0817, 0.0056, 0.00021, 0.00073, 0.0112 mg/l for the above three mosquito species respectively Among the four organophosphates tested temephos was the most effective followed by fenitrothion, fenthion and malathion. In general, Anopheles was found more susceptible as compared to the other two culicines to the above four compounds. The results also showed that larvae of Ae. aegypti were most susceptible followed by Cx. quinquefasciatus and An. stephensi to all the three pyrethroids tested. Among the three compounds tested alphamethrin was found to be the most toxic followed by deltamethrin while fanvalerate was the least toxic. The study would be of great importance while planning use of these insecticides for the control of different vector species in this area.

Bioanalytical method development and validation for a large peptide HIV fusion inhibitor (Enfuvirtide, T-20) and its metabolite in human plasma using LC-MS/MS.

A method for measuring a human immunodeficiency virus (HIV) cell membrane fusion inhibitor (T-20/Ro 29-9800) and its metabolite (M-20/Ro 50-6343) in human plasma by liquid chromatography tandem mass spectrometry (LC-MS/MS) was developed. The relatively large peptide analytes and their corresponding deuterated (d(10)) peptides used as internal standard were isolated from plasma by protein precipitation with two volumes of acetonitrile to plasma. A large pore size reversed-phase C(18) column was employed to elute the peptides. A triple quadrupole mass spectrometer with electrospray interface operating in positive ion and multiple reaction monitoring modes with transitions m/z 1124-->1343 for both T-20 and M-20 was utilized for peak detection. The advantages of the method were a simple sample preparation, specific and sensitive MS/MS detection, and a wide dynamic range of 10-2000 ng/ml for T-20. The method was validated and used for analyzing samples from clinical studies to provide pharmacokinetic profiles of the HIV fusion inhibitor peptide drug and its metabolite.

Study on serum and urinary cortisol levels of asthmatic patients after treatment with high dose inhaled beclomethasone dipropionate or budesonide.

BACKGROUND: The potential for long-term adverse effects from inhaled corticosteroids relates to their systemic absorption. With increasing use of high dose inhaled corticosteroids, there is need to establish whether similar doses of beclomethasone dipropionate (BDP) and budesonide (BUD) produce clinically important differences in untoward side effects specially hypothalamo-pituitary-adrenal (HPA) axis suppression. METHODS: Fifteen asthmatic patients were started on BDP or BUD (2000 microg/day) through spacer for six weeks. Serum cortisol (9 AM and 4 PM), 24-hour urinary steroid and pulmonary function testing parameters were performed. RESULTS: The serum cortisol levels were not found to be suppressed with either BDP or BUD. Similarly no significant changes were found in 24 hours urinary excretion of steroids with either of the drugs. Significant improvement was found in values of forced expiratory volume in the first second (FEV1) with BDP. With BUD the changes in forced vital capacity (FVC) and FEV1 were found to be significant. CONCLUSION: BDP or BUD in high doses of 2000 microg/day given upto six weeks through spacer are equally effective for treatment of bronchial asthma and do not cause any significant change in serum and urinary cortisol levels, and adrenal function/HPA axis.

Selective beta(1)-adrenoreceptor blocking activity of newly synthesized acyl amino-substituted aryloxypropanolamine derivatives, DPJ 955 and DPJ 890, in rats.

The in-vivo beta-adrenoreceptor antagonistic activity of test compounds DPJ 955 and DPJ 890 was assessed against beta-adrenoreceptor agonist (isoprenaline) induced tachycardia in anaesthetized rats. The selectivity to block isoprenaline responses on different &beta-adrenoreceptor subtypes (beta(1), beta(2) and beta(3)) of the test compounds was carried out on isolated rat right atria, isolated rat uterus and isolated rat colon preparations, respectively. Intravenous injection of isoprenaline alone in anaesthetized rats caused hypotension and tachycardia. DPJ 955 or DPJ 890 alone produced a fall in mean arterial pressure and bradycardia in a dose-dependent manner. Administration of isoprenaline to anaesthetized rats pre-treated with test compounds significantly blocked both the tachycardial and hypotensive responses induced by isoprenaline. The test compounds shifted the concentration response curves of isoprenaline towards the right for isolated rat right atrial preparations, rat uterus and rat colon, indicating beta(1), beta(2) and beta(3) adrenoreceptor blockade, respectively. The selectivity ratio for beta(1)/beta-adrenoreceptors to DPJ 955 and DPJ 890 was 64.6 and 83.2, respectively. DPJ 890 was more potent in blocking beta(1)-adrenoreceptors and was more selective towards beta(1) receptors than to other beta-adrenoreceptor subtypes. In conclusion, DPJ 955 and DPJ890 have beta-adrenoreceptor blocking activity with high selectivity for the beta(1)-adrenoreceptor subtype.

Study of beta-adrenoreceptor antagonistic activity of DPJ 904 in rats.

beta-Adrenoreceptor antagonistic activity of a newly synthesized compound was evaluated in vivo by measuring the mean arterial blood pressure and heart rate of urethane-anesthetized rats treated with isoprenaline. In vitro beta(1)-, beta(2)- and beta(3)-antagonism was studied using isolated rat right atria, isolated rat uterus and isolated rat colon preparations, respectively, in comparison to isoprenaline response. DPJ 904 (1, 3 and 10 mg/kg, i.v.) produced dose-dependent hypotensive and bradycardia response in anesthetized rat. DPJ 904 (1, 3 and 10 mg/kg, i.v.) significantly inhibited both the tachycardial effects and hypotensive response induced by isoprenaline. DPJ 904-antagonized isoprenaline induced positive chronotropic effects of isolated rat right atria and a uterine relaxant effect indicating beta(1)- and beta(2)-blockade. The parallel shift to the right of the concentration-response curve of isoprenaline in the presence of DPJ 904 in KCl (30 mmol/l) induced contraction of the rat colon suggesting that DPJ 904 also possessed beta(3)-adrenoreceptor antagonistic activity. The selectivity to beta(1)-adrenoreceptor was nearly 20.5 times greater than to beta(2)-adrenoreceptor. The present study indicates that DPJ 904 possesses beta-adrenoreceptor antagonistic activity with slightly more affinity to the beta(1)-adrenoreceptor subtype.

Efficacy of different organophosphate and synthetic pyrethroid insecticides to the larvae of malaria vector Anopheles stephensi, Liston.

Evaluation of the potency of four organophosphate (OP) (Malathion, Fenthion, Fenitrothion and Temephos) and two synthetic pyrethroid (SP) compounds (cypermethrin and deltamethrin) was carried out against larvae of Anopheles stephensi, the major malaria vector in district Bikaner (Rajasthan). Different concentrations supplied by WHO were used for determining the percent mortalities. LC50 values as calculated by probit analysis were 1.2740, 0.0600, 0.0405, 0.0046, 0.0019 and 0.0016 mg/l respectively for the above six insecticides. Both SP compounds were found more toxic followed by temephos among the OP compounds. Malathion was found the least toxic while fenthion and fenitrothion were in between these two extremes.

Insecticide susceptibility status of Aedes aegypti to DDT and dieldrin in desert and non-desert parts of Rajasthan.

Seasonal prevalence and insecticide susceptibility tests were carried out on the adults of Aedes aegypti, the vector of dengue and dengue hemorrhagic fever(DF/DHF), in three desert (Bikaner, Jaisalmer and Jodhpur) and three non-desert (Alwar, Ajmer and Jaipur) districts of Rajasthan. Among the three species viz. Ae. aegypti, Ae. vittatus and Ae. w-albus encountered during the survey the former was the most prevalent species. Studies were carried out both in rural and urban areas against different concentrations of two organochlorines (DDT and Dieldrin). In rural areas resistance has been observed with DDT and dieldrin both in desert and non-desert parts while an intermediate resistance has been observed in the urban areas with both the insecticides which required further verification. Resistance was more pronounced in rural areas as compared to urban areas. LC50 along with regression equation and Chi-square values have been determined for both the insecticides.

Multiple isoforms of protein kinase C in lymphocytes and airway smooth muscle of guinea pig.

The isoenzyme pattern of protein kinase C (PKC) in lymphocytes and airway smooth muscles (ASM) was examined by Western blot using commercially available monoclonal antibodies. The results showed the presence of PKC alpha, beta, gamma, epsilon, eta, mu and zeta in lymphocytes and PKC alpha, gamma, epsilon, eta and zeta in ASM. The unexpected feature was the presence of PKCgamma in both lymphocytes and ASM of guinea pigs. Expression of this PKC isoform is usually restricted to tissues in the central nervous system or spinal cord. Expression of PKC delta, theta, lambda and tau was not detected in either lymphocytes or ASM.

Effect of feeding protein deficient diet on phospholipid turnover and protein kinase C mediated protein phosphorylation in rat brain.

Feeding of protein deficient diet is known to alter the transmembrane signalling in brain of rat by reducing total protein kinase C (PKC) activity. Phospholipid metabolism regulates the activation of PKC through generation of second messengers and the extent of PKC activation accordingly influences the magnitude of phosphorylation of its endogenous substrate proteins. Thus it was speculated that ingestion of protein deficient diet may modify the turnover rate of membrane phospholipids and magnitude of phosphorylation of endogenous substrate proteins of PKC. The experiments were conducted on rats fed on three different types of laboratory prepared diets viz. casein (20% casein), deficient (4% protein, rice flour as source of protein) and supplemented (deficient diet supplemented with L-lysine and DL-threonine) for 28 days. The metabolism of phosphoinositides (PIs) and phosphatidyl choline (PC) was studied by equilibrium labeling with [3H] myo inositol and [14C methyl] choline chloride respectively. The phosphorylation of endogenous substrate proteins of PKC was studied by using 32P-gamma-ATP followed by SDS-PAGE and autoradiography. The results suggest that in deficient group, there is an increased incorporation of [3H] myo inositol in PIs and inositol phosphate pool in comparison to the casein group. The phosphatidyl inositol (PI) turnover reduced, although there was a marginal increase in the phosphatidyl inositol monophosphate (PIP) and phosphatidyl inositol bis phosphate (PIP2). Supplementation of diet showed a reversal of the pattern towards control to a considerable extent. In the deficient group, PC metabolism showed an increased incorporation of [14C methyl] choline in choline phospholipids but decreased incorporation in phosphoryl choline in comparison with the casein group. The increase in total PC contents was significant but marginal in residue contents. The turnover rate of PC increased only marginally and that of residue declined. Supplementation of diet reduced the total contents of PC and residue, but the turnover rate of PC and residue remained still higher. Phosphorylation of endogenous proteins showed four different proteins of 78, 46, 33 and 16 kDa to be the substrates of PKC in casein group. In deficient group, phosphorylation of these proteins increased markedly while supplementation of diet had a reversing effect rendering the values to be intermediate between casein and the supplemented group. The changes in phospholipid metabolism and in phosphorylation of endogenous substrate proteins of PKC suggest that dietary protein deficiency causes alterations in transmembrane signalling mechanism in rat brain. These effects are partially reversed by improving the quality of proteins in the diet.

Predictors of asthma severity in the elderly: results of a community survey in Northeast England.

A number of risk factors for the development and severity of asthma in childhood are known. Particularly, there is information on allergens, excessive use of beta2- agonists, and indoor environmental pollutants. Similar information on elderly patients is lacking. We examined the risk factors for current asthma and for the severity of asthma in 95 elderly subjects (>65 years old) compared to 274 elderly subjects with obstructive spirometry who did not have asthma as defined by the following criteria: symptoms of episodic wheeze, cough, or chest tightness and forced expiratory volume in 1 sec/vital capacity (FEV1/VC) <70% with >15% or 200 mL reversibility in FEV1 to 200 microg salbutamol given from a metered-dose inhaler. The severity of airflow limitation was graded on the basis of the FEV1/VC ratio as mild (60%-70%), moderate (40%-60%), and severe (<40%). Asthma history was collected using the Medical Research Council respiratory questionnaire and a follow-up postal questionnaire. Data were analyzed using multiple logistic regression and the overall goodness-of-fit of the model was checked using the Hosmer-Lemeshow (HL) statistic. History of allergy (to one or more of the following allergens: cat, house dust, or grass or tree pollen) (odds ratio [OR] 25; 95% confidence interval [CI] 13-51; p = 0.0001) and history of childhood wheeze (OR 8; 95% CI 4-9; p = 0.004) were strong predictors of current asthma. Duration of wheezing, smoking history, indoor heating, history of working in coal mines, and sex were not predictors (HL 6.75, degrees of freedom [df] = 8, p = 0.56). Use of >4 puffs of salbutamol/ day (OR 5.3; 95% CI 2-14; p = 0.005), more than 10 years of asthma symptoms (OR 4.2; 95% CI 4.1-36.2; p = 0.0001), and >500 mL reversibility in FEV1 (OR 4.2; 95% CI 1.2-14.3; p = 0.05) were independent predictors of moderate to severe asthma. History of atopy was the strongest predictor of asthma in the elderly population studied. Indoor heating, presence of pets at home, sex, smoking history, and history of working in coal mines were not predictors of asthma. The severity of asthma as assessed by measurement of airflow limitation was related to the frequency of use of beta2-agonists, duration of symptoms of asthma, and increased reversibility of FEV1 to beta2-agonist.