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Objectives were to determine the effects of 3 different levels of dietary cation-anion difference (DCAD) fed during the last 22 d of gestation to pregnant nulliparous cows on pre- and postpartum acid-base balance, mineral metabolism, and health responses. In all, 132 pregnant nulliparous Holstein cows were enrolled at 250 (248-253) d of gestation, blocked by genomic merit of energy-corrected milk yield, and assigned randomly to diets varying in DCAD: +200 (P200, n = 43), -50 (N50, n = 45), or -150 (N150, n = 44) mEq/kg of dry matter. Dietary treatments were fed until calving, after which cows received the same lactation diet for the first 100 d postpartum. Urine and blood were sampled throughout the prepartum period and in the first weeks postpartum, and urine was assessed for pH, whereas blood was analyzed for gases, measures of acid-base balance, minerals, and metabolites. Calcium (Ca) and magnesium (Mg) retention and phosphorus (P) digestibility were evaluated in the last week of gestation and first week of lactation. Incidence of diseases was evaluated for the first 100 d postpartum. Data are presented in sequence as P200, N50, N150 (LSM ± SEM). Reducing the DCAD reduced urine (8.17 vs. 6.50 vs. 5.51 ± 0.11) and blood pH (7.442 vs. 7.431 vs. 7.410 ± 0.004) and induced a state of compensated metabolic acidosis with a reduction in blood HCO3- (28.4 vs. 26.7 vs. 24.9 ± 0.3 mM) and partial pressure of CO2 (41.8 vs. 40.1 vs. 39.1 ± 0.4 mmHg) prepartum. Reducing the DCAD linearly increased blood ionized Ca (iCa; 1.224 vs. 1.243 vs. 1.259 ± 0.008 mM) and serum total Ca (tCa; 2.50 vs. 2.53 vs. 2.56 ± 0.02 mM) prepartum, blood iCa on the day of calving, and serum Mg in the first days postpartum. Reducing the DCAD linearly increased the apparent absorption of Ca (12.9 vs. 19.0 vs. 20.9 ± 1.4 g/d) and Mg (7.0 vs. 9.9 vs. 10.4 ± 1.4 g/d) prepartum, but apparent retention of both Ca (13.9 g/d) and Mg (3.4 g/d) did not differ with treatment. Treatment did not affect digestibility of P pre- or postpartum or retention of Ca or Mg postpartum. Treatment did not affect the incidence or prevalence of subclinical hypocalcemia, hepatic composition, or the prevalence of fatty liver. Reducing the DCAD had a quadratic effect on incidence of fever (46.5 vs. 17.6 vs. 33.9 ± 7.0%), uterine diseases (36.3 vs. 25.6 vs. 46.0 ± 7.3%), and morbidity (41.4 vs. 28.1 vs. 55.6 ± 7.3%). Feeding a diet with -50 mEq/kg of dry matter promoted moderate changes in acid-base balance, altered mineral metabolism, and benefited health of nulliparous cows; however, further reducing the DCAD to -150 mEq/kg negated the benefits to health.

Objectives were to determine the effects of 3 levels of dietary cation-anion difference (DCAD) fed prepartum to nulliparous cows on productive and reproductive performance. We enrolled 132 pregnant nulliparous Holstein cows at 250 (248-253) d of gestation in a randomized block design. Cows were blocked by genomic merit of energy-corrected milk yield and assigned randomly to diets varying in DCAD, +200 (P200; n = 43), -50 (N50; n = 45), or -150 (N150; n = 44) mEq/kg of dry matter (DM). Dietary treatments were fed during the last 22 d of gestation and, after calving, postpartum cows received the same lactation diet. Productive performance was evaluated for the first 14 wk of lactation, and reproduction was assessed until 305 d postpartum. Intake of DM prepartum decreased linearly (results presented in sequence as least squares means ± standard error of the mean, P200 vs. N50 vs. N150) with a reduction in DCAD (9.0 vs. 8.9 vs. 8.4 ± 0.1 kg/d), which resulted in linear decreases in net energy balance (0.34 vs. 0.20 vs. -0.36 ± 0.20 Mcal/d), body weight change (1.1 vs. 0.8 vs. 0.3 ± 0.1 kg/d), and mean body weight (652 vs. 649 vs. 643 ± 2 kg) prepartum. Treatment did not affect yield of colostrum (6.3 vs. 5.8 vs. 5.1 ± 0.6 kg) or the contents or yields of fat, protein, lactose, IgG, Ca, or Mg in colostrum. Intake of DM (19.4 vs. 19.2 vs. 19.0 ± 0.2 kg/d), yields of milk (36.6 vs. 36.7 vs. 35.8 ± 0.6 kg/d) or energy-corrected milk (36.7 vs. 36.3 vs. 35.9 ± 0.5 kg/d), feed efficiency (1.93 vs. 1.92 vs. 1.93 ± 0.03 kg of energy-corrected milk per kilogram of DM intake), and content and yield of milk components did not differ among treatments during the first 14 wk of lactation. Prepartum DCAD did not affect the cumulative milk yield by 305 d of lactation (9,653 vs. 10,005 vs. 9,918 ± 196 kg). Of the 132 cows, 40 P200, 45 N50, and 43 N150 received at least 1 artificial insemination (AI), and treatment did not affect pregnancy per AI at first (32.5 vs. 35.6 vs. 37.2%) or all AI (30.6 vs. 33.9 vs. 40.2%), although reducing the DCAD increased the proportion of cows pregnant by 305 d postpartum (76.7 vs. 88.9 vs. 93.2%) without altering the rate of pregnancy. Collectively, manipulating the DCAD of prepartum diets, from +200 to -150 mEq/kg of DM, fed to late gestation nulliparous cows did not affect subsequent lactation productive performance, but may have provided some benefit to reproduction, which warrants further confirmation.

Application of green manures substitutes use of chemical fertilizers, improves soil health, prevents soil degradation also ensures food security. Two years’ field experiment was laid out to determi...

Successful development of a chemoprophylaxis against SARS-CoV-2 could provide a tool for infection prevention implementable alongside vaccination programmes. Camostat and nafamostat are serine protease inhibitors that inhibit SARS-CoV-2 viral entry in vitro but have not been characterised for chemoprophylaxis in animal models. Clinically, nafamostat is limited to intravenous delivery and while camostat is orally available, both drugs have extremely short plasma half-lives. This study sought to determine whether intranasal dosing at 5 mg/kg twice daily was able to prevent airborne transmission of SARS-CoV-2 from infected to uninfected Syrian golden hamsters. SARS-CoV-2 viral RNA was above the limits of quantification in both saline- and camostat-treated hamsters 5 days after cohabitation with a SARS-CoV-2 inoculated hamster. However, intranasal nafamostat-treated hamsters remained RNA negative for the full 7 days of cohabitation. Changes in body weight over the course of the experiment were supportive of a lack of clinical symptomology in nafamostat-treated but not saline- or camostat-treated animals. These data are strongly supportive of the utility of intranasally delivered nafamostat for prevention of SARS-CoV-2 infection and further studies are underway to confirm absence of pulmonary infection and pathological changes.

Currently nitazoxanide is being assessed as a candidate therapeutic for SARS-CoV-2. Unlike many other candidates being investigated, tizoxanide (the active metabolite of nitazoxanide) plasma concentrations achieve antiviral levels after administration of the approved dose, although higher doses are expected to be needed to maintain these concentrations across the dosing interval in the majority of patients. Here an LC-MS/MS assay is described that has been validated in accordance with Food and Drug Administration (FDA) guidelines. Fundamental parameters have been evaluated, and these included accuracy, precision and sensitivity. The assay was validated for human plasma, mouse plasma and Dulbeccos Modified Eagles Medium (DMEM) containing varying concentrations of Foetal Bovine Serum (FBS). Matrix effects are a well-documented source of concern for chromatographic analysis, with the potential to impact various stages of the analytical process, including suppression or enhancement of ionisation. Therefore, a robustly validated LC-MS/MS analytical method is presented capable of quantifying tizoxanide in multiple matrices with minimal impact of matrix effects. The validated assay presented here was linear from 15.6ng/mL to 1000ng/mL. Accuracy and precision ranged between 102.2% and 113.5%, 100.1% and 105.4%, respectively. The presented assay here has applications in both pre-clinical and clinical research and may be used to facilitate further investigations into the application of nitazoxanide against SARS-CoV-2.

The control of COVID-19 across the world requires the formation of a range of interventions including vaccines to elicit an immune response and immunomodulatory or antiviral therapeutics. Here, we demonstrate the nanoparticle formulation of a highly insoluble drug compound, niclosamide, with known anti SARS-CoV-2 activity as a cheap and scalable long-acting injectable antiviral candidate.

Creatinine clearance is an important tool to describe the renal elimination of drugs in pharmacokinetic evaluations and clinical practice. In critically ill patients, unstable kidney function invalidates the steady state assumption underlying equations such as Cockcroft-Gault. While measured creatinine clearance (mCrCL) is often used in non-steady state situations, it assumes that observed data is error-free, neglecting frequently occurring errors in urine collection. In contrast, compartmental non-linear mixed effects models of creatinine allow to describe dynamic changes in kidney function while explicitly accounting for a residual error associated with observations. Based on 530 serum and 373 urine creatinine observations from 138 critically ill patients, a one compartment creatinine model with zero-order creatinine generation rate (CGR) and first-order creatinine clearance (CrCL) was evaluated. An autoregressive approach for inter-occasion variability provided a distinct model improvement compared to a classical approach (ΔAIC -49.0). Fat-free mass (FFM), plasma urea concentration, age and liver transplantation were significantly related to CrCL, while weight and sex were linked to CGR. The model-based CrCL estimates were superior to standard approaches to estimate CrCL (or glomerular filtration rate) including Cockcroft-Gault, mCrCL, four-variable modification of diet in renal disease (MDRD), six-variable MDRD and chronic kidney disease epidemiology collaboration (CKD-EPI) as a covariate to describe cefepime and meropenem pharmacokinetics in terms of objective function value. In conclusion, a dynamic model of creatinine kinetics provides the means to estimate actual CrCL despite dynamic changes in kidney function, and it can easily be incorporated into population pharmacokinetic evaluations.

Background The aim of this study was to identify sources of variability including patient gender and body surface area (BSA) in pharmacokinetic (PK) exposure for high-dose methotrexate (MTX) continuous infusion in a large cohort of patients with hematological and solid malignancies. Methods We conducted a retrospective PK analysis of MTX plasma concentration data from hematological/oncological patients treated at the University Hospital of Cologne between 2005 and 2018. Nonlinear mixed effects modeling was performed. Covariate data on patient demographics and clinical chemistry parameters was incorporated to assess relationships with PK parameters. Simulations were conducted to compare exposure and probability of target attainment (PTA) under BSA adjusted, flat and stratified dosing regimens. Results Plasma concentration over time data (2182 measurements) from therapeutic drug monitoring from 229 patients was available. PK of MTX were best described by a three-compartment model. Values for clearance (CL) of 4.33 [2.95–5.92] L h− 1 and central volume of distribution of 4.29 [1.81–7.33] L were estimated. An inter-occasion variability of 23.1% (coefficient of variation) and an inter-individual variability of 29.7% were associated to CL, which was 16 [7–25] % lower in women. Serum creatinine, patient age, sex and BSA were significantly related to CL of MTX. Simulations suggested that differences in PTA between flat and BSA-based dosing were marginal, with stratified dosing performing best overall. Conclusion A dosing scheme with doses stratified across BSA quartiles is suggested to optimize target exposure attainment. Influence of patient sex on CL of MTX is present but small in magnitude.

BackgroundSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been declared a global pandemic by the World Health Organisation and urgent treatment and prevention strategies are needed. Many clinical trials have been initiated with existing medications, but assessments of the expected plasma and lung exposures at the selected doses have not featured in the prioritisation process. Although no antiviral data is currently available for the major phenolic circulating metabolite of nitazoxanide (known as tizoxanide), the parent ester drug has been shown to exhibitin vitroactivity against SARS-CoV-2. Nitazoxanide is an anthelmintic drug and its metabolite tizoxanide has been described to have broad antiviral activity against influenza and other coronaviruses. The present study used physiologically-based pharmacokinetic (PBPK) modelling to inform optimal doses of nitazoxanide capable of maintaining plasma and lung tizoxanide exposures above the reported nitazoxanide 90% effective concentration (EC90) against SARS-CoV-2.MethodsA whole-body PBPK model was constructed for oral administration of nitazoxanide and validated against available tizoxanide pharmacokinetic data for healthy individuals receiving single doses between 500 mg – 4000 mg with and without food. Additional validation against multiple-dose pharmacokinetic data when given with food was conducted. The validated model was then used to predict alternative doses expected to maintain tizoxanide plasma and lung concentrations over the reported nitazoxanide EC90in >90% of the simulated population. Optimal design software PopDes was used to estimate an optimal sparse sampling strategy for future clinical trials.ResultsThe PBPK model was validated with AAFE values between 1.01 – 1.58 and a difference less than 2-fold between observed and simulated values for all the reported clinical doses. The model predicted optimal doses of 1200 mg QID, 1600 mg TID, 2900 mg BID in the fasted state and 700 mg QID, 900 mg TID and 1400 mg BID when given with food, to provide tizoxanide plasma and lung concentrations over the reportedin vitroEC90of nitazoxanide against SARS-CoV-2. For BID regimens an optimal sparse sampling strategy of 0.25, 1, 3 and 12h post dose was estimated.ConclusionThe PBPK model predicted that it was possible to achieve plasma and lung tizoxanide concentrations, using proven safe doses of nitazoxanide, that exceed the EC90for SARS-CoV-2. The PBPK model describing tizoxanide plasma pharmacokinetics after oral administration of nitazoxanide was successfully validated against clinical data. This dose prediction assumes that the tizoxanide metabolite has activity against SARS-CoV-2 similar to that reported for nitazoxanide, as has been reported for other viruses. The model and the reported dosing strategies provide a rational basis for the design (optimising plasma and lung exposures) of future clinical trials of nitazoxanide in the treatment or prevention of SARS-CoV-2 infection.

The objectives of the present study were to evaluate the effects of photoperiodicity, gauge (G) of ovum pick-up (OPU) needle, and two methods of follicular wave emergence on follicular turn-over, oocyte recovery (OR), quality of the oocytes (OQ), and early in-vitro developmental competence of embryos in Nili-Ravi buffaloes (n = 20). In 1st experiment, buffaloes (n = 12; 4 buffaloes/season) were randomly assigned to optimize the OPU’s (n) either with 17 G or 18 G needle in one of the following seasons: 1) peak breeding season (PBS; Sep–Nov; n = 31), 2) transition breeding season (TBS; Dec–Feb; n = 32), and 3) low breeding season (LBS; Apr–June; n = 32). During 2nd experiment, buffaloes (n = 8) were enrolled randomly in a 2 × 2 cross-over design to compare the two methods of wave emergence either using follicular ablation (FA; n = 4), or synchronization protocol (CIDR-EB; n = 4) during PBS. In FA method, the ovarian follicles were aspirated (week -1), and subsequently repeated OPU’s (n = 55) were performed for 7 weeks. However, in CIDR-EB synchronized buffaloes, a progesterone device (CIDR) was inserted in the anterior vagina and a single dose of estradiol benzoate (2 mg) and prostaglandin (150 μg) were administered i.m. on d 1. The CIDR was removed on d 7 and repeated OPU’s (n = 56) were performed. In both experiments, a 7-d resting period was provided between each OPU session. Data on the follicular turn-over, OR, OQ, and early stages of embryonic development were analyzed with mixed models using the PROC MIXED and GLIMMIX procedures of SAS. Results revealed that the number of medium sized follicles (LSM ± SEM) and rate of OR (%) were greater (P 0.05) of season and G of needle on OQ in buffaloes. In experiment 2, the number of small (3.50 ± 0.63 vs.2.69 ± 0.60) sized follicles was higher (P 0.05) in FA and CIDR-EB treated buffaloes, respectively. Similarly, rates of in-vitro maturation (66% vs. 59%), cleavage (42% vs. 53%), 4-cell (27% vs. 32%), 8-cell staged embryos (23% vs. 25%), and morula (13% vs. 8%) did not differ (P > 0.05) between FA and CIDR-EB methods of follicular wave emergence, respectively. Taken together, it is concluded that peak or transition breeding seasons are suitable to perform OPU using 17 G needle for maximum follicular turn-over, and OR in buffaloes. However, the two fundamental methods of synchronization of wave emergence resulted in similar efficiency for OPU derived in-vitro embryo production in Nili-Ravi buffaloes.

Use of crumb rubber to replace fine aggregates has opened up new avenues to explore the trends to develop more robust and eco-friendly construction material (Rubcrete) when compared to conventional concrete. Research trends indicate an increase in toughness in concrete with the use of crumb rubber, and the addition of polypropylene fibres counters the development of microcracks. The brittle nature of concrete can be explained with respect to the fracture energy required to open unit area of the crack surface. The paper highlights the results of experiments carried out to ascertain the significance of polypropylene fibres on the fracture energy of M40 grade rubcrete. Using a notched three-point bending beam (3PBB) specimen based on RILEM TC 50 FMC, cracks in rubcrete are subjected to fracture under Mode I loading. Variation in the fracture energy was studied for the polypropylene fibre-reinforced rubcrete with the addition of fibres up to 0.3% of total volume of the mix starting from 0.1% with an increment of 0.1%. Analysis of results for toughness and energy absorption characteristics showed that the specimens reinforced with polypropylene fibres have lesser brittle behaviour than that of conventional concrete specimens and optimum results were obtained for the addition of 0.3% polypropylene fibres.

The crude ethanolic extract of Chrozophora prostrata (Cp.Cr) was tested using in vivo and ex vivo assays for its possible bronchodilatory effects in order to validate its medicinal use in respiratory disorders, like asthma and cough. Cp.Cr exhibited dose-dependent inhibition of carbachol (CCh)-induced bronchospasm in anesthetized rats, similar to aminophylline. When tested on guinea-pig tracheal preparations, Cp.Cr caused relaxation of both CCh (1 μM) and high K+ (80 mM)-induced contractions with comparable potencies, similar to papaverine, a dual inhibitor of phosphodiesterse (PDE) and Ca+2 influx. Pre-treatment of the tracheal tissues with Cp.Cr resulted in potentiation of the inhibitory effect of isoprenaline on CCh-induced contractions, like that caused by papaverine indicative of PDE inhibitory activity, which was confirmed when Cp.Cr concentration dependently (1 and 3 mg/mL) increased intracellular cAMP levels of the tracheal preparations, like papaverine. Cp.Cr shifted concentrationresponse curves of Ca+2 constructed in guinea-pig tracheal preparation towards right with suppression of the maximum response, similar to both verapamil and papaverine. These data indicate bronchodilator activity of Chrozophora prostrata mediated possibly through dual inhibition of PDE and Ca+2 influx, thus, showing therapeutic potential in asthma with effect enhancing and side-effect neutralizing potential Copyright © 2016 John Wiley & Sons, Ltd.

Introduction Amoxicillin (AMO) is the most consumed of the aminopenicillins. It is not regularly detected in water in-line for the lack of a standardized, sensitive and selective monitoring system that meets required limits. However, wastewater treatment plants cannot remove all antibiotics and thus release them into the environment, either without modification, or as metabolites [1]. This among others increases the risk of introducing antibiotic resistance, thus posing a serious threat to public health. Analytical method have been used as the detection techniques include mass spectrometry, chromatography or their combination. However, they suffer from some limitations mainly relating to sample preparation and stability [2]. Hence, it is necessary to design sensors for straightforward screening measurements. For this purpose, molecular imprinting is a technique to design robust receptor materials for any given analyte, thus mimicking natural recognition. The reusability of imprinted materials has a crucial role in developing applications that are reliable, economical and sustainable [3]. Experimental To synthesize MIP thin films against AMO, we utilized methacrylic acid (MAA) as the functional monomer, and ethylene glycol dimethacrylate (EGDMA) as the cross-linker and thermally pre-polymerized them at 60 °C for 30 minutes. Then, we deposited them on quartz crystal microbalance (QCM) electrodes by spin coating and hardened overnight. The non-imprinted (NIP) sample was prepared in the same manner without template. QCM measurements served to characterize AMO MIP films according to their sensitivity and selectivity, respectively. Results and Conclusion Figure 1 shows the results of sensitivity assessment at three different AMO concentrations. It clearly reveals concentration-dependent sensor signals of the system. Evidently, the MIP yield substantially higher frequency shifts at all concentrations than the corresponding NIP. For instance, at 1000ppm AMO concentration the MIP exhibits a frequency shift of -540 Hz while NIP signal reaches only around -90 Hz. Thus, frequency shifts on MIP-coated electrodes against NIP-coated ones reveals more than five times higher sensitivity, which is strongly indicative of successful imprinting and thus the incorporation of AMO molecules into recognition sites. Additionally, sensor signals are fully reversible and reproducible. Furthermore, selectivity is a key issue for any sensor: Figure 2 shows the outcome of selectivity studies toward PenicillinV (PenV) and PenicillinG (PenG), which demonstrate selectivity factors of around 1.3 each. This is even more remarkable given how similar the molecular structures of those three compounds are: they differ by an amino-group and hydroxyl-group; howover the core-structures are totally identical. Finally, sensor characteristics reveal limit of detection at 15ppm. Acknowledgement Authors gratefully acknowledge funding of this work by the Austrian Research Promotion Agency (FFG) through project AquaNOSE (Grant agreement no. 864893). References [1] A.G.Ayankojo, J.Reut, A.Öpik, A.Furchner, V.Syritski, Hybrid molecularly imprinted polymer for amoxicillin detection, Biosensors and Bioelectronics 118 (2018) 102–107. doi:10.1016/j.bios.2018.07.042. [2] P.T.Phuong-Hoa, S.Managaki, N.Nakada, H.Takada, A.Shimizu, D.Hong-Anh, P. Hung-Viet, S.Suzuki, Antibiotic contamination and occurrence of antibiotic-resistant bacteria in aquatic environments of northern Vietnam, Science of The Total Environment 409 (2011) 2894-2901. doi:10.1016/j.scitotenv.2011.04.030. [3] W.Cuypers, P.A.Lieberzeit, Combining Two Selection Principles: Sensor Arrays Based on Both Biomimetic Recognition and Chemometrics, Frontiers in Chemistry 6 (2018) 268-278. doi:10.3389/fchem.2018.00268. Figure 1

A recent flow cytometry-based in vivo mutagenicity assay involves the hemizygous phosphatidylinositol class A (Pig-a) gene. Pig-a forms the catalytic subunit of N-acetylglucosaminyltransferase required for glycophosphatidylinositol (GPI) anchor biosynthesis. Mutations in Pig-a prevent GPI-anchor synthesis resulting in loss of cell-surface GPI-linked proteins. The aim of the current study was to develop and validate an in vitro Pig-a assay in L5178Y mouse lymphoma cells. Ethyl methanesulfonate (EMS)-treated cells (186.24-558.72 µg/ml; 24 h) were used for method development and antibodies against GPI-linked CD90.2 and stably expressed CD45 were used to determine GPI-status by flow cytometry. Antibody concentration and incubation times were optimised (0.18 µg/ml, 30 min, 4 °C) and Zombie Violet™ (viability marker; 0.5%, 30 min, RT) was included. The optimum phenotypic expression period was 8 days. The low background mutation frequency of GPI-deficiency [GPI(-)] in L5178Y cells (0.1%) constitutes a rare event, thus flow cytometry acquisition parameters were optimised; 104 cells were measured at medium flow rate to ensure a CV ≤ 30%. Spiking known numbers of GPI(-) cells into a wild-type population gave high correlation between measured and spiked numbers (R2 0.999). We applied the in vitro Pig-a assay to a selection of well-validated genotoxic and non-genotoxic compounds. EMS, N-ethyl-N-nitrosourea and 4-nitroquinoline-N-oxide dose dependently increased numbers of GPI(-) cells, while etoposide, mitomycin C, and a bacterial-specific mutagen did not. Cycloheximide and sodium chloride were negative. Sanger sequencing revealed Pig-a mutations in the GPI(-) clones. In conclusion, this in vitro Pig-a assay could complement the in vivo version, and follow up weak Ames positives and late-stage human metabolites or impurities.

Extended spectrum beta lactamase (ESBL) producing members of the family Enterobacteriaceae are a major health problem in the hospitals and community. These enzymes confer bacterial resistance to penicillins, first, second and third generation cephalosporins and aztreonam. The prevalence of ESBLs varies from organism to organism and is increasing day by day. The study was designed to test typhoidal salmonellae for production of ESBL. One hundred and fifty eight (158) isolates of typhoidal salmonellae; Salmonella Typhi (n=126), Salmonella Paratyphi A (n=26) and Salmonella Paratyphi B (n=6) were collected from different hospitals of Lahore and Gujranwala. The isolates were identified morphologically, biochemically (API-20E) and serologically (BD Difco, USA). ESBL production has been tested by three methods; CLSI screening method, double disk diffusion synergistic method and CLSI phenotypic confirmatory method. CLSI screening method detected twenty two (22) strains as ESBL producers. However, when tested by the CLSI confirmatory method and by disk diffusion synergistic method, none proved to be ESBL producer. Based on our study, we concluded that the extended spectrum beta lactamase enzyme does not exist in tested clinical isolates of typhoidal salmonellae, however the isolates suspected to be ESBL by phenotypic methods must be subjected further for molecular analysis. Key words: ESBL, typhoidal salmonellae, isolate.

Lead is a non-essential element reduced plant growth and development that may cause multifarious disturbance in physiological, biochemical and structural integrity of plants. SA is an efficient signal molecule induces systemic resistance responses that control local defense reactions in plants. To evaluate the effect of SA on photosynthetic activity of Pb affected plants therefore a pot experiment was conducted on barley genotype Juo- 93 and Juo- 87 in the Old Botanical Garden, University Of Agriculture, Faisalabad, Pakistan. Three treatment levels (0, 100 and 200 µM) of lead sulphate (PbSO4) were applied in thrice replication with or without SA (0.5 mM) along half strength of Hoagland’s solution till the termination of experiment. The experiment was arranged in a completely randomized design. During course of study growth and pigments modulations were recorded. The result indicated that growth parameters such as root and shoot length, leaf area, fresh weight of shoot and root, dry weight of shoot and root were reduced under lead toxicity. Pb stress damaged the photosynthetic pigments such as Chlorophyll a, chlorophyll b, total chlorophyll and carotenoids but chlorophyll a/b was increased under Pb stress whereas exogenous application of SA alleviated the negative effect of lead toxicity. Juo- 93 showed more tolerance to Pb toxicity as compared to Juo-87.

This study was undertaken to validate the medicinal use of Chrozophora prostrata in constipation. The crude extract of C prostrata produced laxative effect in mice at 100 and 300 mg/kg, similar to carbachol and castor oil. In spontaneously contracting rabbit jejunum tissues, C. prostrata (0.01-3 mg/mL) exhibited atropine-sensitive spasmogenic effect, which was reproducible in guinea-pig ileum. Interestingly, at high concentrations, C. prostrata showed concentration-dependent inhibitory effect on rabbit jejunum, suggesting the presence of accompanied spasmolytic activity. When tested on induced contractions, C. prostrata completely relaxed low K+-induced contraction, while partially inhibited high K+, similar to cromakalim. Tetraethylammonium pretreatment abolished the relaxant effect of C. prostrata on low K+-induced contractions while glibenclamide caused partial inhibition, suggesting the involvement of some non-specific and ATP-dependent K+ channels. These data show that C. prostrata possesses laxative effect at low doses mediated through cholinergic action followed by the spasmolytic activity at high doses mediated possibly through K+ channel activation.