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The purpose of this study was to design and engineer a simple approach to enhance the catalytic activity and thermal stability of Lipase. Lipase is an enzyme with exciting multifarious applications in the industry. In this respect, we observed that the catalytic activity of Lipase was enhanced when it was subjected to in situ synthesis of silver nanoclusters (AgNCs) within the enzyme structure. At alkaline pH, the in situ synthesis of AgNCs inside Lipase altered its activity, and the resulting nanohybrid showed enhanced enzymatic activity. Compared with the catalytic efficiency of free Lipase (72.33 min mM ), the Lipase clustered with silver nanoclusters (L-AgNCs) showed enhanced catalytic efficiency (109.22 min mM ). Thermodynamic parameters of free Lipase and L-AgNCs were determined by calculating activation energy (Ea) values to be 19.92 KJ mol-1 and 17.29 KJ mol-1, respectively. The lower activation energy of L-AgNCs designates it to be more stabilized and highly active to bind with the substrate having a higher rate of reaction instead of free Lipase. Molecular docking analysis showed that silver binds to amino acid residues of the Lipase enzyme, which was also supported by the XPS analysis. Therefore, L-AgNCs conjugate represented a novel Lipase preparation technique with attributes of high activity and stability that could be an attractive choice in diverse applications ranging from catalysis to diagnostics. The overall study proposed an innovative strategy to improve enzyme stability and catalytic activity.

Supplementary Data

Targeting the ataxia telangiectasia and RAD3-related (ATR) enzyme represents a promising anticancer strategy for tumors with DNA damage response (DDR) defects and replication stress, including inactivation of ataxia telangiectasia mutated (ATM) signaling. We report the dose-escalation portion of the phase I first-in-human trial of oral ATR inhibitor BAY 1895344 intermittently dosed 5 to 80 mg twice daily in 21 patients with advanced solid tumors. The MTD was 40 mg twice daily 3 days on/4 days off. Most common adverse events were manageable and reversible hematologic toxicities. Partial responses were achieved in 4 patients and stable disease in 8 patients. Median duration of response was 315.5 days. Responders had ATM protein loss and/or deleterious ATM mutations and received doses ≥40 mg twice daily. Overall, BAY 1895344 is well tolerated, with antitumor activity against cancers with certain DDR defects, including ATM loss. An expansion phase continues in patients with DDR deficiency.Significance:Oral BAY 1895344 was tolerable, with antitumor activity in heavily pretreated patients with various advanced solid tumors, particularly those with ATM deleterious mutations and/or loss of ATM protein; pharmacodynamic results supported a mechanism of action of increased DNA damage. Further study is warranted in this patient population.See related commentary by Italiano, p. 14.This article is highlighted in the In This Issue feature, p. 1

Intercropping provides ample scope to include two or more crops simultaneously in same piece of land thus targeting higher productivity from unit area on sustainable basis. Maize, a cereal crop of immense importance, planted in wide rows offers the possibility for adoption of intercropping. The intercropping system with maize and legume is beneficial in multiple aspects. The success of maize-legume intercropping system largely depends on choice of crops and their maturity, density, and time of planting. Advantage of maize-legume intercropping system is promoted in the form of higher yield and greater utilization of available resources, benefits in weeds control [1,2], pests and disease management [3], fixation of biological nitrogen by legumes and transfer of N to associated maize [4,5], insurance against crop failure to small holders, and control of erosion by covering a large extent of ground area [6]. Though maize-legume intercropping system exhibits limitations like less scope of farm mechanization, dependence on more human workforce, and chance of achieving less productivity from maize, the system implies more advantages for small holders in developing countries where human workforce is not a constraint.

To determine the frequency and antifungal susceptibility pattern of Candida species causing candidemia or blood stream infections (BSI) in immunocompromised patients of a bone marrow transplant unit. The study was conducted at the Department of Microbiology, Armed Forces Institute of Pathology (AFIP), Rawalpindi in collaboration with Armed Forces Bone Marrow Transplant Center (AFBMTC), Rawalpindi, from 1st June 2019 to 31st December 2019. A total 256 paired blood culture samples from patients of all ages, irrespective of gender were collected during a period of seven months. The samples were processed as recommended Clinical & Laboratory Standard Institute (CLSI) guidelines. Antifungal susceptibility by break point MICs was performed through VITEK® 2 system (version: 08.01) for Fluconazole, Itraconazole and Amphotericin B. Of the total 256 blood cultures, 46 (17.97%) were positive for Candida species. The susceptibility of C. albicans was 100% to Amphotericin B, 90% to Fluconazole and 10% to Itraconazole, while C. tropicalis showed sensitivity of 92%, 88% and 0% against Amphotericin B, Fluconazole and Itraconazole, respectively. C. parapsilosis was found sensitive to Amphotericin B while resistance to Fluconazole and Itraconazole.Frequency of candidemia among immunocompromised patients is 17.9%. C. tropicalis and C. albicans are the two most common Candida species involved in blood stream infections in our setup. Fluconazole and Amphotericin B, both were found susceptible and can be used as empirical therapy.

Field experiments were conducted in 2017-18 during main cropping seasons of the year 2017 and 2018 at Dryland (Karewa) Agriculture Research Station, SKUAST-K to determine the effect of different post and pre emergence herbicides application on weed dynamics in maize (Zea mays L.). The experiment was laid out in randomized block design with four replications and variety used was Bio-605. Five treatments Five treatments viz, Atrazine 50% WP @ 1.0 kg ha-1 as pre-emergence and Paraquat dichloride 24% SL @ 0.5 kg ha-1, 2,4-D sodium salt 58% WSC @ 1.0 kg ha-1 as post-emergence herbicides (application at 2-3 leaf stage of weeds), hand weeding as standard check and weedy check as control were used. Effect of different herbicides on weed density was found significant. in plots managed with hand weeding, no weed was recorded. The Atrazine treated plots @ 1.0 kg ai ha-1 as pre-emergence had the weed population of 5.51, 5.4, 3.73, 5.12, 4.28 and 41.4m-2 , respectively in Echinochloa spp., Eleusine indica, Digitaria sanguinalis, Amaranthus viridis, Sorghum halepense and Cyperus spp. However, the maximum population of weeds was recorded in weedy check with 21.54 m-2, 20.87 m-2, 19.16 m-2, 21.5 m-2, 16.7 m-2 and 60.7 m-2. No significant difference was observed between Paraquat dichloride 24% SL @ 0.5 kg ai ha-1 and 2,4- D Sodium salt 58% WSC @ 1.0 kg ai ha-1 except for Cyperus spp. with density of 51.37 m-2 and 43.04 m-2 respectively. The potential of the atrazine in controlling weeds thereby enhancing yield of maize was found in this study.

Corynebacterium glutamicum is an authenticated microorganism that supports amino acid production consistent with dietary importance. Fermentation parameters like temperature, agitation speed, and carbon source concentration were optimized using response surface methodology. Surface response model suggested that optimal fermentation parameters including 30°C, 50 g/L glucose concentration, and shaking speed of 120 rpm furnished 14.2 g/L of glutamic acid and 5.1 g/L lysine, comparable with the predicted values. After optimizing fermentation parameters in shake flasks, the fermentation kinetics was studied in a stirred fermenter. The kinetic study revealed that the substrate consumption rate achieved a maximum level of 3.36 g/L/hour between 12 and 18 hours; afterwards, it decreased and fell to 1.9 g/L/hour. Average biomass yield over a period of 48 hours was 0.337 g/L; however, maximum biomass yield of 0.51 g/L was noted between 10 and 20 hours. Overall molar yield coefficient of CO2 ( Y C CO 2 / S ) was found to be 0.234. The molar yield coefficient of biomass ( Y C X / S ) was 0.6 in exponential phase which decreased afterwards. This study indicates that the average biomass yield over a time of 48 hours was 0.337 g/L, with a maximum yield within 10 to 20 hours obtained under optimized conditions. The molar yield coefficient of CO2 ( Y C CO 2 / S ) was found to be 0.234. The molar yield coefficient of biomass ( Y C X / S ) was 0.6 in exponential phase which decreased and fell to near zero at 48 hrs.

Effective treatment for gastrointestinal (GI) cancers is a significant unmet need. Gremlin-1, secreted by cancer-associated fibroblasts, downregulates bone morphogenetic proteins (BMP), members of the transforming growth factor β superfamily, resulting in tumor stemness, hyperproliferation, and invasiveness. Ginisortamab, a fully human IgG4P mAb, neutralizes gremlin-1, thus restoring BMP signaling. Preclinical studies showed that ginisortamab binds to gremlin-1 and has antitumor activity in several mouse models, including GI cancers. Here we report the initial findings from Part A (monotherapy dose escalation) of the first-in-human study. ONC001 (NCT04393298) is an ongoing multi-part, multicenter, nonrandomized, open-label, Phase I/II study assessing the safety, PK/PD, and antitumor activity of intravenous (IV) ginisortamab as monotherapy or in combination with selected standard of care regimens in patients (pts) with advanced solid tumors. ONC001 includes 3 dose escalation modules (Parts A-C) and a dose adaptation module (Part A1). To be eligible for Part A, pts had to be aged ≥18 years (y) with advanced disease, resistant or refractory to standard therapy, with ECOG PS ≤1. We present the primary (safety, tolerability), secondary (PK/PD), and exploratory (antitumor activity) endpoints, as well as the monotherapy RP2D from Part A. At data cutoff (June 20, 2022), 25 pts received ≥1 dose of ginisortamab (median [range] age: 64 [37-74] y; 68% male; 88% White; 36% had >3 lines of prior therapy). The most common tumor type enrolled was colorectal adenocarcinoma (44%). Ginisortamab was escalated using a modified rolling 6 design through 5 levels (100, 250, 500, 1000, and 2000 mg IV Q2W) in 28-day cycles. No DLTs, Grade ≥3 AEs related to ginisortamab, or serious related AEs were observed; the 2000 mg dose was deemed safe for further development. Temporary treatment interruption and study discontinuation due to AEs unrelated to ginisortamab occurred in 7 (28%) and 2 pts (8%), respectively. Ginisortamab PK aligned with expectations from preclinical models for an IgG4 mAb, allowing favorable dosing intervals. Confirmation of target occupancy was demonstrated by increases in total circulating gremlin-1. Antidrug antibody formation was not observed. The best overall response of stable disease was observed in 7/24 pts (29%); 4 pts had a reduction of the sum of the dimensions of their target lesions relative to baseline. Favorable safety and PK/PD data at the maximum dose of 2000 mg and preliminary antitumor activity were observed with the first-in-class anti-gremlin antibody ginisortamab. These data support further evaluation of the efficacy and safety of ginisortamab monotherapy at 2000 mg Q2W, or in combination with standard of care regimens (FOLFOX, trifluridine/tipiracil) in advanced solid tumors. Citation Format: Debashis Sarker, Sarah Blagden, Natalie Cook, Thomas R. Evans, Ruth Plummer, Angelos Angelakas, Saira Bashir, Salma El Badri, Leonidas Mavroeidis, Cienne Morton, Abdul (Moe) Muhith, Mark White, Rupert Dixon, Elodie Garric, Joanne Mann, Ivan Matthews, Udai Banerji. Phase I dose-escalation study evaluating the safety and tolerability of ginisortamab (UCB6114), a first-in-class anti-gremlin-1 monoclonal antibody (mAb), as monotherapy in advanced solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr CT094.

Some ingredients used in poultry feed formulation contain carbohydrate polymers which are difficult to digest and thus hinder nutritional feed value. Toward overcoming this limitation, exogenous enzymes have been added to poultry feed to improve its nutritive value. The present study was designed to provide first enzymatic characterization of endoglucanase (BsEgl) from the genome of B. sonorensis BD92 expressed in Pichia pastoris. Further, we tested its impact alone and in combination with a β-glucosidase (Bteqβgluc) on growth in commercial broilers as feed additive. The expressed enzyme displayed features of GH5 family and had optimum activity against carboxymethyl cellulose at pH 5 and 50 °C. The BsEgl was stable at a range of pH from 4 to 8 for 60 min and at 50 °C for 180 min. Supplementing broilers diet with BsEgl alone or in combination with Bteqβgluc resulted in better feed conversion ratio among treatments during a five weeks testing period. Moreover, meat percentage was also highest for this treatment, and all treatments with recombinant enzymes increased intestinal length in birds compared to treatment control group. Blood parameters and serum biochemistry profile showed non-significant difference among groups. These results support that recombinant cellulolytic enzymes supplement high fiber diets improve their nutritional performance.

Adequate mental and physical well-being is a defensive factor in state of mind issues, while for some women pregnancy can extend the lack of defenses to mental disorders such as melancholia. Prenatal discouragement influences about 11% of females in world nations, and sum of pregnant females on antidepressants has risen over past decade. The assessment was directed at the Sir Ganga Ram Hospital, Lahore Pakistan from 25 March 2020 to 5 May 2020. Isolation, social separation and outrageous changes in everyday life can rise danger of destitution amongst powerless populations, just like pregnant females. This is therefore fundamental to measure mental effect of COVID19 episode. Key words: coronavirus, influenza, maternal death, prematurity, infection

Targeting the ataxia telangiectasia and RAD3-related (ATR) enzyme represents a promising anticancer strategy for tumors with DNA damage response (DDR) defects and replication stress, including inactivation of ataxia telangiectasia mutated (ATM) signaling. We report the dose-escalation portion of the phase I first-in-human trial of oral ATR inhibitor BAY 1895344 intermittently dosed 5 to 80 mg twice daily in 21 patients with advanced solid tumors. The MTD was 40 mg twice daily 3 days on/4 days off. Most common adverse events were manageable and reversible hematologic toxicities. Partial responses were achieved in 4 patients and stable disease in 8 patients. Median duration of response was 315.5 days. Responders had ATM protein loss and/or deleterious ATM mutations and received doses ≥40 mg twice daily. Overall, BAY 1895344 is well tolerated, with antitumor activity against cancers with certain DDR defects, including ATM loss. An expansion phase continues in patients with DDR deficiency. Significance: Oral BAY 1895344 was tolerable, with antitumor activity in heavily pretreated patients with various advanced solid tumors, particularly those with ATM deleterious mutations and/or loss of ATM protein; pharmacodynamic results supported a mechanism of action of increased DNA damage. Further study is warranted in this patient population. See related commentary by Italiano, p. 14. This article is highlighted in the In This Issue feature, p. 1

Cellulases and xylanases have high industrial demand due to their paramount importance in biological processes. The present study was aimed to evaluate the fiber degrading potential of Bacillus spp. isolated from buffalo digestive system. A total of fourteen isolates from rumen and eight isolates from dung were screened on carboxymethyl cellulose (CMC; 1%) agar plates, showing clear zone of CMC hydrolysis. All screened isolates were confirmed by targeting the 16S rRNA gene, sequencing and phylogenetic analysis. The enzyme activity index (EAI) of all screened isolates was calculated and it was observed that BR28 (Bacillus subtilis), BR96 (Bacillus amyloliquefaciens), BD69 (Bacillus tequilensis) and BD92 (Bacillus sonorensis) exhibit 2.11, 2.05, 2.56 and 2.45 EAI, respectively. The isolates with high EAI (>2) were selected for further enzyme production studies. The results of enzyme activities showed that BD92 had higher endoglucanse (carboxymethyl-cellulase, CMCase, 240.76±4.12 U/L) and avicelase (153.56±7.28 U/L) activities after 72 h of incubation. However, BR96 showed highest xylanase (3379.27±10.58 U/L) activity than others isolates and previously reported studies. This study provides a helpful insight into the identification of Bacillus spp. from the buffalo digestive system with higher production of cellulases and xylanases for their application in the animal feed industry.

Objective: To recognize occurrence also danger issues of hemorrhoids and fissures throughout pregnancy and afterwards childbirth. Population: The overall 290 pregnant females trailed up till 1 month after delivery. Methods: Females remained inspected 4 times via pregnancy and after delivery; those that developed perianal diseases were compared with those that did not. Main outcome measures Incidence, time and risk factors of hemorrhoids and fissures. Our current research was led at Sir Ganga Ram Hospital, Lahore from April 2018 to March 2019. Results: In altogether, 127 (45.7%) females established perianal illness:1.7% in initial trimester, 64% throughout 3rd trimester, 35.2% afterwards delivery and 4.4% 1 month after delivery; 118 (41.8%) females remained analyzed by hemorrhoids, seven (2.6%) with hemorrhoids and anal fissure and two (0.73%) with anal fissure. Ninety-nine (80.5%) women had vaginal delivery and 26 (19.6%) women had undergone caesarean section. Multivariate analysis identified personal history of perianal diseases (odds ratio [OR] 11.93; 95% confidence interval [96% CI] 2.18–65.30), constipation (OR 18.98; 95% CI 7.13–50.54), straining throughout delivery for extra than 21 mins (OR 28.77; 96% CI 5.01–226.24) and birthweight of newborn >3800 g (OR 18.98; 96% CI 4.28–97.50) as substantial analysts of hemorrhoids and anal fissures throughout pregnancy and perinatal phase. Conclusions: Hemorrhoids and fissures remain mutual throughout last trimester of pregnancy and one month afterwards delivery, through constipation, individual past of hemorrhoids or fissures, birth weight of new >3806 g, straining throughout delivery for extra than 21 mins being self-sufficiently related risk factors. Keywords: Anal fissure, hemorrhoids, pregnancy, prospective study, risk.

3103 Background: CDC7, a protein with key roles in regulating cell-cycle progression is often over-expressed in malignant cells, particularly those with TP53 mutations. LY3143921, an orally administered ATP-competitive CDC7 inhibitor, demonstrated favorable pre-clinical anti-cancer activity in colorectal cancer (CRC) and squamous non-small cell lung cancer (sqNSCLC), particularly in TP53 mutant models. Methods: Phase Ia (dose escalation) recruited patients (pts) with advanced solid tumors enriched for malignancies associated with TP53 mutation. Pts received LY3143921 OD or BD continuously on a 21-day schedule, using an accelerated 3+3 escalation design, starting at 30 mg OD. Phase Ib recruited pts with CRC or sqNSCLC treated continuously at RP2D, or pts with other advanced tumors treated at RP2D on days 1-3 every 7 days. Radiological assessment was performed every 2 cycles initially. Pts in phase Ib could consent to pre- and on-treatment skin +/- tumor biopsies. Primary objectives: assess safety/tolerability and determine MTD and RP2D of LY3143921. Secondary objectives: evaluate preliminary efficacy and pharmacokinetic (PK) profile of LY3143921. Exploratory objective: correlate efficacy to baseline molecular/genetic alterations, including TP53 mutation and measure markers including pMCM2 in pre- and on- treatment tumor and skin samples. Results: 68 pts were recruited and 67 treated (38 phase Ia, 29 phase Ib). Most frequent drug-related CTCAEs (all grades): nausea (75%), orthostatic hypotension (50%), vomiting (47%), fatigue (45%) & diarrhea (44%). Grade 3-4 LY3143921 related AEs occurred in 17 pts. In phase Ia 8 DLTs occurred in 5 pts (G3 nausea, vomiting, fatigue & hyponatraemia and G2 diarrhea, anorexia & lethargy). RP2D was 360 mg BD (continuous non-fasted dosing schedule). 37 pts were evaluable for radiological response with no complete or partial responses seen, and stable disease (SD) was observed in 24 pts (65%). In phase Ia 3 pts achieved long term SD of 1, 2.5 and 3+ years duration. For evaluable pts treated in phase Ib, SD was seen in 8/12 CRC pts, 1/2 sqNSCLC pts and 2/2 pts treated with the intermittent schedule (median duration 15 weeks, range 6-18+). 2 pts remain on-study. Recruitment ceased due to lack of radiological response according to RECIST. Dose-dependent increases in LY3143921 exposure (Cmax & AUC0-24) were seen. IHC analyses of skin biopsies demonstrated reductions in pMCM2, indicating on-target activity of LY3143921. Pre-clinical testing of combination with standard of care agents is ongoing. Additional PD and PK data will be presented. Conclusions: LY3143921 is well tolerated, exhibits dose-dependent increases in plasma exposure and demonstrates evidence of target inhibition. Significant monotherapy clinical activity was not observed; further analyses should investigate potential predictive response biomarkers and rational combination approaches. Clinical trial information: NCT03096054.

Endo-glucanase (cellulase) and xylanase have high industrial demand due to their vast application in industrial processes. This study reports statistical based experimental optimization for co-production of endo-glucanase and xylanase from Bacillus sonorensis BD92. Response surface methodology (RSM) involving central composite design (CCD) with full factorial experiments (23) was applied to elucidate the components that significantly affect co-production of endo-glucanase and xylanase. The optimum co-production conditions for endo-glucanase and xylanase were as follows: carboxymethyl cellulose (CMC) 20 g/L, yeast extract 15 g/L, and time 72 h. The maximum endo-glucanase and xylanase production obtained was 1.46 and 5.69 U/mL, respectively, while the minimum endo-glucanase and xylanase production obtained was 0.66 and 0.25 U/mL, respectively. This statistical model was efficient because only 20 experimental runs were necessary to assess the highest production conditions, and the model accuracy was very satisfactory as coefficient of determination (R2) was 0.95 and 0.89 for endo-glucanase and xylanase, respectively. Further, potential application of these enzymes for saccharification of lignocellulosic biomass (wheat bran, wheat straw, rice straw, and cotton stalk) was also investigated. The results revealed that the biomass was susceptible to enzymatic saccharification and the amount of reducing sugars (glucose and xylose) increased with increase in incubation time. In conclusion, Bacillus sonorensis BD92 reveals a promise as a source of potential endo-glucanase and xylanase producer that could be useful for degrading plant biomass into value-added products of economic importance using precise statistically optimized conditions.

Herein we report the development of a core-shell-like Co Fe 2 O 4 − BaTi O 3 multiferroic nanocomposite (1:1 wt ratio) for their enhanced magnetoelectric coupling and energy storage density by the wet chemical route. Rietveld refinement analysis of the XRD pattern verified the formation of cubic spinel ( Co Fe 2 O 4 ) and tetragonal perovskite ( BaTi O 3 ) structures. Whereas the SEM and EDX analysis confirmed the formation of core-shell-like morphology in the composite. The temperature dependence of dielectric permittivity showed two distinct peaks corresponding to the constituent phases´ structural phase transitions. However, the frequency dependence of low-temperature peak revealed an abnormal relaxor-type behavior, which is elucidated due to intrinsic structural defects that might cause alteration in the octahedron arrangement in BaTi O 3 . Impedance spectroscopy analysis was utilized to calculate the activation energy (Ea) of respective phases, which was found to be>1 eV that confirms the dominance of doubly ionized oxygen vacancies in governing the thermally stimulated conduction process. It was found that the prepared composite exhibited high dielectric permittivity (~2700) and moderate values of saturation magnetization (20 emu/g) and polarization (6.2 μC/cm2) along with low remnant polarization (3 μC/cm2) at room temperature. Moreover, we also observed enhanced energy storage efficiency (67%) and magnetoelectric coefficient (0.18 V/cm Oe), which are explained due to strong interfacial coupling and reduced leakage current, making this composition promising for multifunctional device applications.

To compare vitamin D levels with bone mineral density, serum calcium, phosphorous and alkaline phosphatase.The cross-sectional multicentre study was conducted at Pakistan Health Research Council centres in Islamabad, Lahore, and Karachi,and comprised subjects coming for either vitamin D testing or bone mineral density examination. Information related to demography, height/weight, skin colour, smoking, use of sun screen, daily milk intake, sun exposure and exercise was taken along with biochemical tests like serum calcium and phosphorous, alkaline phosphatase, and 25 hydroxy vitamin D.Bone mineral density was done using dual-energy X-ray absorptiometry scan. SPSS 15.0 was used for data analysis.Of the 291 subjects, 100(34.3%) each were enrolled from Islamabad and Lahore, while 91 (31.2%) were from Karachi. Overall, 245(84%) had insufficient vitamin D which was significantly associated with age and dark skin colour (p0.05 each). Besides, 137(48%) cases had a reduced bone density, and there was a significant difference between age groups, gender and skin colour (p0.05 each).Vitamin D deficiency did not have any effect on bone mineral density (p0.05) Serum calcium and phosphorus levels were also similar regardless of vitamin D level (p0.05).Vitamin D deficiency did not have any direct impact on bone mineral density and serum calcium, phosphorous and alkaline phosphatase.

This descriptive observational study was conducted at Pathology lab Jinnah hospital, Lahore, January 2013 to October 2015. We include all the food workers in different restaurants, slaughters, inn, recreation centre and bakeries of Lahore for the purpose to identify the salmonella infection, we used Typhoid IgGI/IgM Rapid test device methods (Serum/Plasma), is a lateral flow chromatographic immunoassay technique were calculated. We analysed 1025 individuals, 62 (6%) were found positive for the said diseases out of which 14 (1.3%) IgG positive, 18 (1.8%) IgG IgM positive and 30 (2.9%) were IgM positive. All the individuals who were found positive were without signs and symptoms. Salmonellosis continues to be an important global cause of infectious intestinal disease and spread readily by means of food personnel, zoonotic and contaminated equipments and surface. The screening of the individuals who are silent carriers, help in controlling the spread of salmonella infection in the public.

3007 Background: The ATR kinase is a key regulator of the DNA damage response (DDR) machinery, activated by DNA damage and replication stress. BAY 1895344 is a novel, potent, and selective ATR inhibitor with anti-tumor activity in preclinical models with DDR defects. Methods: Pts with advanced metastatic solid tumors resistant or refractory to standard treatment, with and without DDR defects, received BAY 1895344 BID, 3 days (d) on/4 d off continuously in 3-weekly cycles. Results: As of December 20, 2018, 18 pts with colorectal (4), breast (3), prostate (2), and ovarian (2) cancers were enrolled across 6 cohorts (5 mg, 10 mg, 20 mg, 40 mg, 60 mg, and 80 mg BID). Median prior lines of treatment was 5. No dose-limiting toxicities (DLTs) were reported in the 5-40 mg cohorts. 2/3 pts had DLTs in the 80 mg cohort (grade [G] 4 neutropenia, G4 neutropenia and G4 thrombocytopenia) and 2/7 had DLTs in the 60 mg cohort (G4 neutropenia, G2 fatigue). 40 mg BID 3 on/4 off was defined as the maximum tolerated dose. Most common treatment-emergent adverse events included anemia, neutropenia, nausea, and fatigue. Pharmacokinetics appeared dose proportional. Pharmacodynamic analyses showed modulation of pH2AX and/or pKAP1 in paired tumor biopsies at exposures associated with preclinical anti-tumor activity. In 13 pts with and without DDR defects treated at dose levels ≥40 mg BID, the objective response rate was 30.7%, including 2/2 pts at 40 mg (appendix and urothelial cancer), 1/8 pts at 60 mg (breast), and 1/3 pts at 80 mg (endometrial). All responders had ATM protein loss of expression and/or ATM mutation; median treatment duration was 347 d (range 293-364 d). A BRCA1-mutant, olaparib-resistant ovarian cancer pt (60 mg) had a CA125 response and stable disease >10 months. 41 additional pts have been enrolled in ongoing expansion cohorts in cancers with DDR defects (prostate, breast, gynecologic, colorectal) or ATM protein loss (all comers) with responses observed. Conclusions: The ATR inhibitor BAY 1895344 is tolerated at biologically active doses with anti-tumor activity against cancers with certain DDR defects, including ATM protein loss. Clinical trial information: NCT03188965.

2025 Background: BAL101553 (prodrug of BAL27862) is a novel TCC that promotes tumor cell death by modulating the spindle assembly checkpoint. BAL27862 is a lipophilic, small molecule shown in rodents to penetrate the brain (brain/plasma ratio around unity), with promising antitumor activity in orthotopic preclinical GBM models as monotherapy or in combination with radiotherapy (RT) with or without temozolomide. In this ongoing study (NCT02490800, CDI-CS-002), daily oral BAL101553 was initially examined in solid-tumor patients, with an MTD of 16 mg/d and DLTs of G4 hyponatremia and G2 hallucinations (Lopez 2018, JCO 36, 2018, suppl. A2530). Subsequently the study was expanded by including a separate cohort of patients with progressive or recurrent GBM or high-grade glioma (Ingles Garces 2017, JCO 35, 2018, suppl. TPS2601). Methods: Patients with histologically-confirmed GBM or high-grade glioma, with progressive or recurrent disease after prior RT with/without chemotherapy, received once-daily oral BAL101553 (28-day cycles) in a 3+3 dose-escalation design to determine the maximum tolerated dose (MTD). Adverse events were assessed by CTCAE v4.03 grade (G), and tumor response by RANO every two cycles. Pharmacokinetics (PK) were evaluated on Day 1 of Cycles 1 and 2. Results: In the ongoing study, 23 pts (13M/10F; median age 50 y), median (min–max) number of prior regimens = 2 (1–5), received doses of 8, 15, 20, 25 or 30 mg oral BAL101553 once daily. One DLT of reversible G2 depression and fatigue occurred at 20 mg. Both mean Cmax and AUC increased with dose between 8 and 30 mg. The PK exposure in GBM patients was lower than for solid tumor patients, in particular at 20 and 25 mg. At 25 mg/d (n = 3), one patient with IDH-mutated GBM had a partial response (63% area reduction per RANO) and continues on study > 8 months, and another patient had stable disease for 5 months. At 15–20 mg/d, stable disease was observed in 3/10 patients. Conclusions: The current data in patients with GBM or high-grade glioma suggest that BAL101553 is well tolerated at dose levels above the MTD established in patients with advanced solid tumors, and shows indications of clinical activity. Clinical trial information: 02490800.

TPS3167 Background: CDC7 is a protein with key roles in DNA replication initiation, the intra-S-phase checkpoint and M-phase completion. CDC7 is over-expressed in malignant compared to non-malignant cells, particularly those with TP53 mutations, making it an attractive therapeutic target. LY3143921 hydrate is an orally administered ATP-competitive CDC7 inhibitor. Pre-clinical studies in colorectal cancer (CRC) and squamous non-small cell lung cancer (sqNSCLC) demonstrate favourable anti-cancer activity, particularly in squamous NSCLC and in CRC with TP53 null and missense mutations. We hypothesise that solid tumours mutated in TP53 will be sensitive to LY3143921 therapy. Methods: This is a first-in-human, phase I trial of LY3143921 hydrate (LY3143921) monotherapy given twice daily, continuously on a 21 day schedule until disease progression, patient (pt) withdrawal or unacceptable toxicity (NCT03096054). Eligible pts have histologically proven advanced/metastatic solid tumours for which no further standard therapy exists and WHO PS 0-1. Pts have regular clinical assessment and tumour imaging every 2 cycles. Phase Ia (dose escalation) is recruiting in a 3+3 design following 3 initial single patient cohorts (starting dose 30 mg OD), enriching for patients with malignancies associated with p53 mutations (CRC, sqNSCLC, high grade serous ovarian, squamous cell oesophageal, squamous cell head & neck, urothelial, pancreatic and triple negative breast cancer). Recruitment to cohort 6 (180 mg BD) is ongoing. On determination of the maximum tolerated dose (MTD) and recommended phase II dose and schedule (RP2D), 2 expansion cohorts (≤ 25 pts each) of patients with CRC and sqNSCLC will be evaluated. Primary objectives: assess safety and tolerability of LY3143921, determine MTD and RP2D. Secondary objectives: evaluate preliminary efficacy and PK profile of LY3143921. All pts will have archival tumour tissue retrospectively analysed, while patients in phase Ib will also have pre- and on-treatment tumour biopsies. Evaluation of potential predictive and pharmacodynamic biomarkers including p53 mutations, phosphorylated MCM2, cyclin B1 and molecular subgroups of target tumours will be included. Clinical trial information: NCT03096054.

Objective: The objective of this study was to see treatment outcome of morbidly adherent Placenta.Methodology: This descriptive case series was carried out in Obstetrics and Gynae Unit-2 Lady Willingdon Hospital Lahore from January 2013 to December 2014. In this period 127 females between 20 to 45 years with any parity were enrolled in this study. All the patients were diagnosed for Placenta Accreta, Increta and Percreta by color flow Doppler. Operative delivery was carried out in all. Cesarean Hysterectomy with or without Placental separation, conservative measures (uterine artery ligation, tamponade and hemostatic sutures) and Internal iliac ligation were used. The outcome measures recorded were total blood loss, neonatal birth weight, mean gestational age at delivery, neonatal Apgar score, intra and postoperative complications and maternal mortality. Non-probability, sampling technique was used. The results were analyzed using SPSS version 20. Results: The mean age of subjects in this study was 26.77 ± 3.17 years. Gestational age at diagnosis and at surgery was 34.83 ± 1.94 weeks and 36.19 ± 1.557 weeks respectively. According to previous history of cesarean section, 60 (47.3%) had 1 previous C-section, 38 (29.9%) had 2 previous C-section and rest of 29 (22.9%) had ≥ 3 C-section. The diagnosis was made on Doppler Ultrasonography (USG) and the patients had surgery under emergency as well as elective conditions. We found that 96 (75.5%) females had Accreta, 27 (21.26%) had Increta and only 4 (3.15%) females had Percreta. Placental separation was done in 63 (49.6%) followed by Cesarean Hysterectomy were done in 36 (57%), and conservative measures like tamponade, haemostatic sutures in Placental bed and Uterine artery ligation in the rest i.e. 27 (43%). Non-separation of Placenta followed by direct Cesarean Hysterectomy was done in 64 (50.3%), and Internal iliac ligation was done in 30 (23%). During surgery the average blood loss was 2.97 ± 1.36 L. According to neonatal outcome Apgar score at 5 minutes was 5.69 ± 1.446 and weight of new baby was 2.85±0.30 kg. According to final outcome 83 (65.35%) females were successfully discharged, 29 (22.83%) had bladder rupture occurred in 29 (22.83%) while maternal mortality occurred in 15 (11.81%) only.Conclusion: An increasing number of patients with morbidly adherent Placenta is coming up along with the rising incidence of C-Section. The key to the management of this condition is prompt diagnosis and referral to well-equipped hospitals having multidisciplinary input. Conservative measures with uterine sparing are worth trying in selected cases.

Background: Gestational diabetes mellitus is characterized by high blood glucose levels during pregnancy. The reported prevalence of GDM is 7% worldwide while 8% in Pakistan. About 19.5% of the women with GDM develop post-partum metabolic syndrome. Methods: A prospective cohort study carried out at tertiary-care hospital in Lahore from Jan 2013 - Dec 2014 to determine the incidence of Mets in women with GDM. After informed consent, women with and without GDM were enrolled during 28th week of pregnancy. Demographic and personal data were recorded. Presence of Mets was evaluated 6 months after delivery measuring their blood pressure, anthropometry, fasting blood sugar (FBS) and triglycerides. Cumulative incidence of Mets, means of blood pressure, anthropometric measures were calculated. Comparisons of general characteristic and component of metabolic syndrome were done by student t-test. Results: 102 women with GDM and 104 without GDM were followed. The Cumulative Incidence of Mets was 33% in GDM while 15.4% in non-GDM (P-value = 0.002). GDM group were significantly older than non-GDM group (P-Value Conclusion: The study highlights that GDM group has more risk of developing Mets. They should be monitored as high risk for other cardiovascular diseases along with diabetes after delivery. Keywords: metabolic syndrome, gestational diabetes mellitus, postpartum period

Bacillary dysentery, common in developing countries, is usually caused by Shigella species. A major problem in shigellosis is the rapid emergence of multidrug-resistant strains. This is the first detailed molecular study on drug resistance of Shigella isolates from the Faisalabad region of Pakistan. Ninety-five Shigella isolates obtained after screening of 2500 stool samples were evaluated for in vitro resistance to commonly used antimicrobial agents; the presence or absence of 20 of the most relevant drug resistance genes; and the prevalence of integrons 1, 2, and 3. Shigella flexneri was found to be the most prevalent and most resistant species. Collectively, high resistance was found towards ampicillin (96.84%), tetracycline (93.68%), streptomycin (77.89%), and chloramphenicol (72.63%). Significant emerging resistance was detected towards the modern frontline drugs ciprofloxacin (12.63%), cefradine (17.89%), ceftriaxone (20.00%), cefoperazone (22.10%), and cefixime (28.42%). Prevalence rates for blaTEM, blaCTX-M, gyrA, gyrB, qnrS, aadA1, strAB, tetA, tetB, catA, and catP were 78.94%, 12.63%, 20.00%, 21.05%, 21.05%, 67.36%, 42.10%, 12.63%, 53.68%, 33.68%, and 25.26%, respectively. Class 2 integrons (42.10%) were more common in the local isolates. Simultaneous detection of class 1 and 2 integrons in some isolates and a rapidly emerging resistance to modern frontline drugs are the major findings of this study.

P63 is a gene product required in cell cycle regulation which plays vital roles in tumor differentiation. Aims of the present study were to assess the frequency, pattern, sensitivity and specificity of two p63 protein clones P63 4A4 and P63 4A4+Y4A3 in squamous cell carcinomas (SCCs). Thirty cases of head and neck region SCC diagnosed on the basis of HandE staining were examined along with 60 cases of head and neck region biopsies other than squamous cell carcinoma, negative on HandE staining, were taken as control. Immunostaining was performed on slides according to the Thermo Scientific UltraVision LP detection System. P63 4A4+Y4A3 clone is more sensitive 96.6% in comparison to 86% in P63 4A4 with having greater NPV of 98.3%. The results signify the importance of P63 4A4+Y4A3 marker over the old markers and may be used as a confirmatory marker of squamous cell carcinoma.

Background: There is a paucity of lung specific biomarkers to diagnose exacerbations of chronic obstructive pulmonary disease (COPD) and to track their progression. Surfactant protein D (SP-D) is a pulmonary collectin regulating the innate immunity of the lung and its serum expression is perturbed in COPD. However, it is not known whether serum levels change during exacerbations. We sought to determine whether serum SP-D levels are raised in COPD exacerbations.Objectives: To determine whether or not patients with exacerbations have elevated serum SP-D levels compared with asymptomatic controls, stable disease.Study design: case control study.Methods: We measured serum SP-D levels from patients with stable COPD (n= 14), patients experiencing acute exacerbations (n= 13) and in control subjects (n= 54) using a specific immunoassay and compared the levels using analysis of variance.Results: Serum SP-D levels were significantly increased in patients who experienced an acute exacerbation (227 ± 120 ng/mL) compared to patients with stable disease (151 ± 83 ng/mL) or control subjects (128 ± 65 ng/mL;p= 0.003). Serum SP-D levels were also found to be inversely related to various lung function parameters including FEV1/FVC% predicted.Conclusions: Our study suggests that serum SP-D levels are increased in patients during exacerbations and may be a potential diagnostic biomarker for COPD exacerbations.

To assess the counter-irritant and anti-inflammatory activity of Rumexvesicarius in dermatological use.The animal-based experimental study was conducted at the Royal Institute of Medical Sciences, Multan, Pakistan, in November 2014. Sodium lauryl sulfate, phenol, histamine and sandpaper irritation models were used. Irritation was induced by the clockwise frictional movement of fine sandpaper to the ear of rabbits and then applying sodium lauryl sulfate, histamine and phenol single topical application onto the ear of the rabbit. The counter-irritant effect was determined by calculating the mean decrease in redness and erythema with those of control and standard dexamethasone.There were 20 rabbits in the study with a mean weight of 1.50±0.033 kg. R. vesicarius (100 and 150mg/mL) showed excellent counter-irritant effect when compared with control and standard groups. Both the doses depicted counter-irritant effect with the highest inhibition (94.42%) in sandpaper group, followed by sodium lauryl sulphate (90%), phenol (94.23%) and histamine (88.46%) irritation models respectively.Methanol leaf extract of R. vesicarius countered the effect of irritation in experimental animals. It showed significant effect in terms of dose and counter-irritancy time.

To assess knowledge and practices related to dengue management among physicians.The cross-sectional study was conducted at hospitals in Islamabad, Lahore, Faisalabad, Peshawar, Quetta and Karachi between June and December 2012Physicians from public and private sectors filled a self-administered questionnaire about dengue knowledge and its management practices. A maximum score of 100 was assigned to the knowledge portion. Data was analysed using SPSS 15.A total of 400 subjects participated in the study; 200(50%) each from public and private hospitals. Of them, 223(56%) were males; 268(67%) were in the 21-30 years age bracket. The highest score was recorded in Quetta 67 followed by 65 in Karachi, 62 in Lahore, Faisalabad, Peshawar and 59 in Islamabad. Of the total, 200 (50%) were not aware that leucopenia is a criterion for diagnosing probable dengue. Similarly 140 (35%) did not know the criteria for diagnosing dengue haemorrhagic fever and warning signs of severe dengue. Total of 204 (51%) were not aware of the criteria for discharging of the admitted cases. There was no significant difference between dengue knowledge of the physicians belonging to public and private sectors (p0.05).Quite a large number of physicians lacked knowledge of probable diagnosis of dengue and appropriate time to discharge the patients.

To assess the economic burden of dengue infection by calculating cost per patient and disability adjusted life years lost.The cross-sectional study was conducted in Islamabad, Lahore, Faisalabad and Karachi from July 2012 to March 2013. Residential addresses and telephonic numbers of dengue patients were taken from the records of Pakistan Institute of Medical Sciences, Islamabad, Mayo and Ganga Ram Hospital, Lahore, Civil Hospital, Karachi, and Allied Hospital, Faisalabad. A total of 250 dengue confirmed cases - 50 from each hospital - were randomly selected. Information regarding duration of illness and out-of-pocket expenses were collected to estimate the direct cost, while indirect cost (number of work days missed by the patient) was calculated from disability adjusted life years using Murray's formula.Overall, there were 162(65%) men and 88(35%) with a mean age of 30.4±13.5years. More than half 138(55%) were below 30 years of age. Socio-economically, 145(58%) belonged to low, 70(28%) middle and 35(14%) to high socioeconomic groups. Of the total, 210(84%) cases had dengue fever followed by 32(12.8%) dengue haemorrhagic fever and 8(3.2%) dengue shock syndrome cases. Average duration of illness was 32±7.1 days. Overall direct cost per patient was Rs.35,823 (US$358) and average pre-hospitalisation, hospitalisation and post-hospitalisation was Rs.6154, Rs.21,242 and Rs.8,427 respectively. The overall disability adjusted life years per million population was 133.76.Although the government had provided free treatment for dengue in public-sector hospitals, still patients had to pay Rs.21,242 during hospital stay, resulting in substantial burden which needs to be addressed.

Background Dengue epidemic in Lahore (2011) resulted in hundreds of deaths and affected thousands. As most of the studies were focused on its diagnosis and treatment, scanty data is available on associated diseases/co-morbidities in these patients that could have contributed to a higher mortality. There were no local guidelines available on recording, reporting and management of these co-morbidities. The objective of this study was to analyze the initial presentations of dengue cases and to estimate the frequency of co-morbidities in dengue patients. Methods Data of 556 dengue cases was retrieved from 2 major public sector tertiary-care hospitals for patients who were admitted during 2011 epidemic and a case record analysis was done. Data was retrieved from patient’s information reports which included demography, signs and symptoms and the laboratory investigations. In addition verbal autopsy of deceased cases was also done from their relatives using standardized WHO verbal autopsy form after making modifications as per needed. Results Of 556 cases studied, 390 (70%) were males. The mean age was 36 years and 30% of the cases were between 20-29 years. Average duration of the hospital stay was 6 days. Out of the total, 435 (78%) were dengue fever (DF) cases followed by dengue hemorrhagic fever (DHF) in 95 (17%) and dengue shock syndrome (DSS) in 26 (4%) cases. A total of 40 cases died and among them 17 were diagnosed with DSS, 13 DF and 10 DHF. Further the verbal autopsy from relatives of deceased cases showed 29 (60%) deceased had co-morbid diseases which included hypertension, diabetes etc. DSS was common in patients who had hypertension (27) either alone or associated with other illnesses. Conclusions Co-morbidities with dengue infection were seen in 60% deceased cases indicating the reasons for high dengue related complications and death.

The fixed-dose combination containing the antihypertensive agent amlodipine and the statin, atorvastatin, is the first combination of its kind designed to treat two risk factors for cardiovascular disease (CVD), i.e., hypertension and dyslipidemia. In this study, blood pressure and lipid lowering effects of combination of amlodipine and atorvastatin were evaluated in uncontrolled hypertensive patients.Thirty patients both male and female in the age group 35-60 years attending the Hypertensive Clinic of PMRC FJMC suffering from uncontrolled hypertension were selected. Baseline blood pressure was checked after half hour rest in sitting and standing position using mercury sphygmomanometer. Blood sample was collected from all patients after overnight fasting for assessment of serum cholesterol, triglycerides, LDL and HDL cholesterol levels. They were prescribed with fixed dose combination of 5 mg amlodipine and 10 mg atorvastatin. Patients were followed for their blood pressure measurement after every 4 weeks up to 12 weeks. At the end of 12 weeks their fasting blood sample was taken again for determination of serum cholesterol, triglyceride, LDL and HDL cholesterol levels.Systolic blood pressure after 4, 8 and 12 weeks was significantly lower at all intervals from baseline. When systolic blood pressure after 8 and 12 weeks was compared with 4 weeks, the effect was again significant (p=0.024, p=0.002 respectively).There was no significant reduction seen in 8 versus 12 weeks (p=0.493). Diastolic blood pressure at 4, 8 and 12 weeks was significantly lower from baseline. Diastolic blood pressure after 4 and 8 weeks when compared with 8 and 12 weeks was not significantly low (p=0.99 and 0.91 respectively). Lipid profile of the patients was significantly reduced from baseline after twelve weeks of fixed dose combination of treatment (p0.000).Combination therapy proved to be effective in controlling hypertension and dyslipidemia than single pill. It also improved patient's compliance. It is suggested that polypill should be prescribed instead of multiple drugs.

The objective of this work was the phylogenetic characterization of local clinical isolates of uropathogenic E. coli with respect to drug resistance. A total of 59 uropathogenic E. coli responsible for community acquired urinary tract infections were included in this study. A triplex PCR was employed to segregate each isolate into four different phylogenetic groups (A, B1, B2 and D). Drug resistance was evaluated by disc diffusion method. The drugs used were ampicillin, aztreonam, cefixime, cefoperazone, ceftriaxone, cephradine among β-lactam group; amikacin, gentamicin, and streptomycin among aminoglycosides; nalidixic acid and ciprofloxacin from quinolones; trimethoprim-sulfomethoxazole, and tetracycline. Among 59 uropathogenic E. coli isolates majority belonged to phylogenetic group B2 (50%) where as 19% each belonged to groups A and B1, and 12% to group D. All the isolates were multiple drug resistant (MDR). Most effective drugs against Group A, B1, and B2 were gentamicin, amikacin and cefixime; ceftriaxone and quinolones; and ceftriaxone and amikacin, respectively. Group D isolates were found to be highly resistant to all drugs. Our results have shown emergence of MDR isolates among uropathogenic E. coli with dominance of phylogenetic group B2. However, it was found that group D isolates were though less frequent, more drug resistant as compared with group B2. Groups A and B1 were relatively uncommon. Amikacin, ceftriaxone and gentamicin were the most effective drugs in general.

The Shiga toxin-producing Escherichia coli (STEC) is an emerging foodborne pathogen. The proportion of cases attributed to STEC in an episode of diarrhea in the Faisalabad region of Pakistan was investigated. In addition, as increase in Shiga toxin (Stx) release after exposure to various antimicrobial agents is widely reported, we also elucidated the in vitro effects of three commonly used antibiotics (ampicillin, gentamicin, and cefotaxime) on Stx release. Isolation and detection of STEC was done using enzyme-linked immunosorbent assay and polymerase chain reaction, followed by phenotypic characterization. In vitro Stx release from isolated STEC was determined using enzyme-linked immunosorbent assay, and Stx-induced verocytotoxicity was quantified using cytotoxicity detection assay. STEC was detected in 5 (21.7%) of 23 patients. Exposure to minimum inhibitory concentration (MIC) of ampicillin, gentamicin, and cefotaxime resulted in a considerable decrease in toxin release and level of cytotoxicity in most of the STEC isolates when compared with control (without antibiotic exposure). Exposure to sub-MIC of ampicillin resulted in a relative increase in Stx release and cytotoxicity (p

Escherichia coli are one of the leading causes of infection in wounds. Emerging multiple drug resistance among E. coli poses a serious challenge to antimicrobial therapy for wounds. This study was conducted to ascertain a baseline profile of antimicrobial resistance in E. coli isolates infecting surgical wounds. A total of 64 pus samples from hospitalized patients were screened and 29 (45.3%) were found to have E. coli, which were identified biochemically and confirmed by molecular methods. Using the disc diffusion method, antimicrobial resistance was observed toward tetracycline (100%), cefradine (100%), nalidixic acid (93.1%), ampicillin (86.2%), gentamicin (86.2%), cefixime (82.8%), ceftriaxone (82.8%), aztreonam (82.8%), ciprofloxacin (75.9%), streptomycin (72.4%), cefoperazone (65.5%), chloramphenicol (58.6%) and amikacin (58.6%). In an effort to find relevant genes, 11 different genes were targeted by PCR. Among these, the mutated gyrA gene was found to be the most prevalent (82.8%), followed by the TEM (72.4%), catP (68.9%), catA1 (68.9%), tetB (62.1%), blt (58.6%), bla(CTX-M-15) (27.6%), bla(TEM) (20.7%), bla(OXA) (17.2%), tetA (17.2%) and aadA1 (13.8%) genes. The presence of integrons was also studied among these isolates. The prevalence of class 1 integrons was the highest (44.8%), followed by class 2 (27.6%). Three (10.3%) isolates carried both class 1 and class 2 integrons (first report from E. coli infecting wounds). The high incidence of integrons points toward their facilitation for carriage of antimicrobial resistance genes; however, in nearly 37% isolates, no integrons were detected, indicating the significance of alternative mechanisms of gene transfer. Another salient finding was that all isolates were multidrug-resistant E. coli.

Background: Drug resistance is a major problem in Escherichia coli isolated from surgical wound infections. In this study, we evaluated relationship between phylogenicity and drug resistance. Methodology: A total of 29 multi-drug resistant (MDR) E. coli isolates of known drug resistance genes and integron profile were selected for the present study. Triplex PCR was conducted for phylogenetic classification of these isolates into four established phylogenetic groups: A, B1, B2 and D. Statistical analysis was done to determine the association of different drug resistance genes and integrons with the phylogenetic groups. Results: Most of the isolates (44.8%) belonged to phylogenetic group A followed by group B2 and D (24.1% each) and group B1 (6.9%). Conclusions: There is a definitive relationship between drug resistance and various phylogenetic groups of E. coli infecting wounds. A shift towards phylogenetic group A might be observed with an increasing drug resistance profile.

The present paper describes the field evaluation of local gametocyte vaccine and its comparative efficacy with commercial anticoccidial vaccine, LivaCox, used in breeder and broiler flocks in Pakistan. Humoral immune response in vaccinated and control chickens was monitored by enzyme-linked immunosorbent assay. Results demonstrated significantly elevated antibody titres in vaccinated groups as compared to control groups conducted both in Laboratory and field experiments. Significantly (P0.01) higher antibody titres in local gametocyte-vaccinated group as compared to LivaCox-vaccinated chickens were recorded. Splenic cell migration inhibition assay was used to detect the cell-mediated immune (CMI) response, and results were expressed in terms of per cent migration index. Lower per cent migration index in LivaCox-vaccinated chickens indicated the higher CMI response, as compared to local gametocyte-vaccinated chickens, although the difference was statistically non-significant (P0.05). Results of the challenge studies in laboratory experiments revealed significantly higher (P0.05) oocyst count in LivaCox-vaccinated group as compared to local gametocyte-vaccinated chickens.Maximum protection (75%) against mixed species of genus Eimeria was recorded in chickens vaccinated with gametocyte vaccines as compared to LivaCox-vaccinated group. The mean body weight gains in chickens vaccinated with local gametocyte vaccine were significantly better (P0.05) than in chickens vaccinated with LivaCox vaccine, both in laboratory and field experiments. Majority of the chickens (70-72%) in control group demonstrated severe lesions (3.0-4.0), while 20-26% chickens showed moderate lesions (2.0). On the other hand, local gametocyte- and LivaCox-immunized chickens developed 78% and 85% mild to moderated lesions (1.0-2.0), respectively. Results of the present study provide a probable explanation for cross-protection induced by Eimeria tenella gametocyte vaccines against other species of genus Eimeria.

Background: Diabetic nephropathy is the leading cause of end-stage renal disease globally and contributes to the total disease burden. The study was carried to estimate the occurrence of microalbuminuria in type II diabetics and to find its association with blood pressure, duration of disease and anthropometric measurements. Findings: The cross-sectional study was conducted for six months in Pakistan Medical Research Council Research Centre, Fatima Jinnah Medical College, Lahore. A total of 91 subjects who met the inclusion criteria were selected. A comprehensive questionnaire including age, education, socio-economic status, medical history, blood pressure and anthropometric measurements was used as a study tool. Random blood sample was used for the estimation of plasma glucose, blood urea and serum creatinine, while first morning urine sample was collected for the qualitative screening of microalbuminuria. Data analysis was done using Statistical Package for Social Sciences version 15 (SPSS-15). The mean age of 91 selected subjects was 50 ± 9 years and male to female ratio was 2:1. 80.2% subjects were diabetic hypertensive and remaining 19.8% had only diabetes. The overall prevalence of microalbuminuria was 58.2%. Prevalence of microalbuminuria among males was 30.2% and among females 69.8%. Correlation analysis showed that plasma glucose level, duration of diabetes and systolic blood pressure correlated with microalbuminuria at P

Background Uropathogenic E.coli (UPEC) are among major pathogens causing urinary tract infections. Virulence factors are mainly responsible for the severity of these emerging infections. This study was planned to investigate the distribution of virulence genes and cytotoxic effects of UPEC isolates with reference to phylogenetic groups (B2, B1, D and A) to understand the presence and impact of virulence factors in the severity of infection in Faisalabad region of Pakistan. Methods In this study phylogenetic analysis, virulence gene identification and cytotoxicity of 59 uropathogenic E.coli isolates obtained from non-hospitalized patients was studied. Results Among 59 isolates, phylogenetic group B2 (50%) was most dominant followed by groups A, B1 (19% each) and D (12%). Isolates present in group D showed highest presence of virulence genes. The prevalence hlyA (37%) was highest followed by sfaDE (27%), papC (24%), cnf1 (20%), eaeA (19%) and afaBC3 (14%). Highly hemolytic and highly verotoxic isolates mainly belonged to group D and B2. We also found two isolates with simultaneous presence of three fimbrial adhesin genes present on pap, afa, and sfa operons. This has not been reported before and underlines the dynamic nature of these UPEC isolates. Conclusions It was concluded that in local UPEC isolates from non-hospitalized patients, group B2 was more prevalent. However, group D isolates were most versatile as all were equipped with virulence genes and showed highest level of cytotoxicity.