Synthesis of PVP-capped trimetallic nanoparticles and their efficient catalytic degradation of organic dyes.

The study proposes a simple and efficient way to synthesize a heterogeneous catalyst that can be used for the degradation of organic dyes. A simple and fast chemical process was employed to synthesize Au: Ni: Co tri-metal nanohybrid structures, which were used as a catalyst to eliminate toxic organic dye contamination from wastewater in textile industries. The catalyst's performance was tested by degrading individual dyes as well as mixtures of dyes such as methylene blue (MB), methyl orange (MO), methyl red (MR), and Rose Bengal (RB) at various time intervals. The experimental results show the catalytic high degradation efficiency of different dyes achieving 72-90% rates in 29 s. Moreover, the material displayed excellent recycling stability, maintaining its degradation efficiency over four consecutive runs without any degradation in performance. Overall, the findings of the study suggest that these materials possess efficient catalytic properties, opening avenues toward their use in clean energy alternatives, environmental remediation, and other biological applications.

In Vitro Biofilm-Mediated Biodegradation of Pesticides and Dye-Contaminated Effluents Using Bacterial Biofilms.

Overuse of pesticides in agricultural soil and dye-polluted effluents severely contaminates the environment and is toxic to animals and humans making their removal from the environment essential. The present study aimed to assess the biodegradation of pesticides (cypermethrin (CYP) and imidacloprid (IMI)), and dyes (malachite green (MG) and Congo red (CR)) using biofilms of bacteria isolated from pesticide-contaminated soil and dye effluents. Biofilms of indigenous bacteria, i.e., Bacillus thuringiensis 2A (OP554568), Enterobacter hormaechei 4A (OP723332), Bacillus sp. 5A (OP586601), and Bacillus cereus 6B (OP586602) individually and in mixed culture were tested against CYP and IMI. Biofilms of indigenous bacteria i.e., Lysinibacillus sphaericus AF1 (OP589134), Bacillus sp. CF3 (OP589135) and Bacillus sp. DF4 (OP589136) individually and in mixed culture were tested for their ability to degrade dyes. The biofilm of a mixed culture of B. thuringiensis + Bacillus sp. (P7) showed 46.2% degradation of CYP compared to the biofilm of a mixed culture of B. thuringiensis + E. hormaechei + Bacillus sp. + B. cereus (P11), which showed significantly high degradation (70.0%) of IMI. Regarding dye biodegradation, a mixed culture biofilm of Bacillus sp. + Bacillus sp. (D6) showed 86.76% degradation of MG, which was significantly high compared to a mixed culture biofilm of L. sphaericus + Bacillus sp. (D4) that degraded only 30.78% of CR. UV-VIS spectroscopy revealed major peaks at 224 nm, 263 nm, 581 nm and 436 nm for CYP, IMI, MG and CR, respectively, which completely disappeared after treatment with bacterial biofilms. Fourier transform infrared (FTIR) analysis showed the appearance of new peaks in degraded metabolites and disappearance of a peak in the control spectrum after biofilm treatment. Thin layer chromatography (TLC) analysis also confirmed the degradation of CYP, IMI, MG and CR into several metabolites compared to the control. The present study demonstrates the biodegradation potential of biofilm-forming bacteria isolated from pesticide-polluted soil and dye effluents against pesticides and dyes. This is the first report demonstrating biofilm-mediated bio-degradation of CYP, IMI, MG and CR utilizing soil and effluent bacterial flora from Multan and Sheikhupura, Punjab, Pakistan.

Antibacterial and Cytotoxic Effects of Biosynthesized Zinc Oxide and Titanium Dioxide Nanoparticles.

Nanotechnology is a rapidly developing field of research that studies materials having dimensions of less than 100 nanometers. It is applicable in many areas of life sciences and medicine including skin care and personal hygiene, as these materials are the essential components of various cosmetics and sunscreens. The aim of the present study was to synthesize Zinc oxide (ZnO) and Titanium dioxide (TiO2) nanoparticles (NPs) by using Calotropis procera (C. procera) leaf extract. Green synthesized NPs were characterized by UV spectroscopy, Fourier transform infrared (FTIR), X-ray diffraction (XRD), and Scanning Electron Microscopy (SEM) to investigate their structure, size, and physical properties. The antibacterial and synergistic effects of ZnO and TiO2 NPs along with antibiotics were also observed against bacterial isolates. The antioxidant activity of synthesized NPs was analyzed by their α-diphenyl-ß-picrylhydrazyl (DPPH) radical scavenging activity. In vivo toxic effects of the synthesized NPs were evaluated in albino mice at different doses (100, 200, and 300 mg/kg body weight) of ZnO and TiO2 NPs administered orally for 7, 14, and 21 days. The antibacterial results showed that the zone of inhibition (ZOI) was increased in a concentration-dependent manner. Among the bacterial strains, Staphylococcus aureus showed the highest ZOI, i.e., 17 and 14 mm against ZnO and TiO2 NPs, respectively, while Escherichia coli showed the lowest ZOI, i.e., 12 and 10 mm, respectively. Therefore, ZnO NPs are potent antibacterial agents compared to TiO2 NPs. Both NPs showed synergistic effects with antibiotics (ciprofloxacin and imipenem). Moreover, the DPPH activity showed that ZnO and TiO2 NPs have significantly (p > 0.05) higher antioxidant activity, i.e., 53% and 58.7%, respectively, which indicated that TiO2 has good antioxidant potential compared to ZnO NPs. However, the histological changes after exposure to different doses of ZnO and TiO2 NPs showed toxicity-related changes in the structure of the kidney compared to the control group. The current study provided valuable information about the antibacterial, antioxidant, and toxicity impacts of green synthesized ZnO and TiO2 NPs, which can be influential in the further study of their eco-toxicological effects.

Efficacy of rice husk and halophilic bacterial biofilms in the treatment of saline water.

Water salinity causes less production of agricultural productivity, low economic returns, soil destructions, less sustainability, and reduction in the germination rate. The current study was aimed to understand the combined potential of halophilic bacteria and rice husk in treating water salinity. In total, 10 halophilic bacterial isolates were isolated from Khewra Mines, Pakistan. Bacterial isolates were characterized by biochemical tests. 16S rRNA gene sequencing identified the isolate SO 1 as Bacillus safensis (accession number ON203008) being the promising halophilic bacteria tolerating upto 3 M NaCl concentration. Next, rice husk was used as carbon source for bacterial biofilm formation, growth and propagation. For saline water treatment, the experimental setting comprising glass wool, rice husk and artificial sea water (3 M) was set. B. safensis biofilm was developed in test samples to desaline the saline water containing 3 M NaCl concentration. Following NaCl decline, flame photometric analysis was used to check the desalination extent of treated saline water. Results showed decreased sodium level in sea water in the presence of rice husk and glass wool. The eluted water used for the germination of Zea mays seeds showed improved growth performance. Also, decreased photosynthetic pigments (chlorophyll "a" = 18.99, and chlorophyll "b" = 10.65), sugar contents (0.7593), and increased carotenoid (1526.91), protein contents (0.4521) were noted compared to control. This eco-friendly approach for bioremediation of salt-affected soils to optimize crop yields under stress through halophilic bacteria and rice husk may overcome the problem of the reduced yield of cash crops/agriculture and water shortage by salinity.

Antimicrobial Efficacy of Biogenic Cobalt and Copper Nanoparticles against Pathogenic Isolates.

Biogenic synthesis of cobalt (Co) and copper (Cu) nanoparticles (NPs) was performed using the bacterial strains Escherichia coli and Bacillus subtilis. Prepared NPs were confirmed by a color change to maroon for CoNPs and green for CuNPs. The NPs characterization using FTIR showed the presence of functional groups, i.e., phenols, acids, protein, and aromatics present in the Co and CuNPs. UV-vis spectroscopy of E. coli and B. subtilis CuNPs showed peaks at 550 and 625 nm, respectively. For E. coli and B. subtilis CoNPs, peaks were observed at 300 nm and 350 nm, respectively. Antibacterial and antifungal activity of B. subtilis and E. coli Co and CuNPs was determined at 100 mg/mL concentration against two bacterial strains at 5, 2.5, and 1.5 mg/mL against fungal two strains F. oxysporum and T. viridi, respectively. B. subtilis CuNPs showed significantly higher inhibition zones (ZOI=25.7-29.7 mm) against E. coli and B. subtilis compared to other biogenic NPs. Likewise, B. Subtilis CuNPs showed lower MIC (4.3 ± 6.3) and MBC (5.3 mg/mL) values against both tested isolates. Antifungal activity of B. subtilis and E. coli CuNPs and CoNPs showed a concentration-dependent decrease in ZOI. Among all biogenic NPs, B. subtilis CoNPs showed the highest ZOI (25-30 mm) against F. oxysporum followed by E. coli CuNPs with maximum ZOI (20-27 mm) against T. viridi. Again, B. subtilis CoNPs and E. coli CuNPs showed lowest MIC and MFC values against both fungal isolates. In conclusion, the current study showed that biogenically synthesized B. subtilis Cu or CoNPs can be used as effective antimicrobial agents due to their potential antibacterial and antifungal potential.

Analysis of the growth performance, stress, profile of fatty acids and amino acids and cortisol in Tilapia (Oreochromis niloticus), cultured at high stocking density using in-pond raceway system.

In-pond raceway system technology (IPRS) was introduced in Pakistan in 2019 as solution for sustainable aquaculture approach by effectively increasing production, reducing pollution and facilitating feed and pond management. Fingerlings of GIFT Tilapia (Oreochromis niloticus) (n = 16,500 in each raceway, initial weight = 32.00 ± 1.26 g) were stocked in June 2019 in two IPRS raceways (area of each raceway = 220 m3) for 171 days until harvested on November 30, 2019. Fingerlings stocked in traditional earthen ponds (area of each pond = 6167 m3) were studied as control (n = 3000 in each pond, initial weight = 32.00 ± 1.26 g). Average harvested biomass from raceways was 57.33 kg/m3 with an average daily weight gain per fish of 4.47 g per day. On the other hand, average harvested biomass from control ponds was 0.38 kg/m3 with an average daily weight gain per fish of 4.60 g per day. Average feed conversion ratio (FCR) in both raceways and control ponds was recorded as 1.25 and 1.24, respectively. Overall survival rate in both raceways and control ponds was above 99%. No sign of any disease was noted at any stage in both study groups. Crude protein and fats contents did not reduce in any raceway despite of high stocking density and sharp seasonal changes. Profile of essential and non-essential amino acids were found to be upto nutritional requirements of adult human. High levels of n-3 and n-6 fatty acids in fish collected from raceways as compared to those in traditional earthen pond show that muscle quality was not compromised due to high stocking density in small area. Return on investment excluding capital cost was 47.05 which implies that IPRS technology can be economically feasible with further modifications.

In vitro activity of Nigella sativa against antibiotic resistant Salmonella enterica.

Salmonellosis is a major food-borne disease worldwide and antimicrobial resistance in Salmonella is a public health problem. Phytochemicals are alternative therapeutics to treat antibiotic resistant Salmonella. Biochemically identified Salmonella enterica of human and poultry origin (n = 10) were confirmed by polymerase chain reaction. Susceptibility to antibiotics was determined by Kirby Bauer disc diffusion method. In-vitro anti-salmonella activity of N. sativa essential oil and extracts (aqueous and methanol) was determined against antibiotic resistant isolates by well diffusion and micro broth dilution method. Cytotoxic potential of N. sativa was observed by MTT assay. In S. eneterica the highest resistance (100%) was detected against nalidixic acid and ampicillin followed by oflaxacin (80%), tetracycline, co-trimoxazole and amoxicillin (60%), ciprofloxacin (40%) and gentamicin (20%). Methanol extract of N. sativa produced zone of inhibition from 35 ±â€¯1.00 to 17 ±â€¯1.00 with mean MIC value ≥562.5 ±â€¯384.1 µg/mL. Essential oil showed antibacterial activity with zone of inhibition from 20 ±â€¯1.00 to 14 ±â€¯1.00 mm and mean MIC value ≥1000.0 ±â€¯322.7 µg/mL. Aqueous extract had no anti-salmonella activity. MTT results showed more than 50 percent cell survival at concentrations >625 and >1250 µg/mL for methanol extract and essential oil of N. sativa respectively; concentrations less than cytotoxic values required for anti-salmonella activity. It was concluded that N. sativa had in-vitro activity against S. enetrica and can be used as therapeutic.

Detection of Peste des Petits Ruminants Viral RNA in Fecal Samples of Goats after an Outbreak in Punjab Province of Pakistan: A Longitudinal Study.

Peste des petits ruminants (PPR) is a highly contagious viral disease of domestic and wild small ruminants and thus has serious socioeconomic implications. In Pakistan, during the year 2012-2013, estimated losses due to PPR were worth Rs. 31.51 billions. Close contact between infected and susceptible animals is an important route of transmission of PPR. Therefore, carrier animals play an important role in unnoticed transmission of PPR. The objective of the study was to investigate the detection of PPR virus in goats recovered from PPR. A suspected PPR outbreak was investigated and confirmed as PPR after analysing appropriate samples collected from infected animals using rRT-PCR. A longitudinal study was conducted over the period of 16 weeks to ascertain the detection of PPR virus (PPRV) in faecal samples of recovered goats. Ninety-six (96) faecal samples from each sampling were collected at 4, 8, 12, and 16 weeks after the outbreak. Faecal samples were analysed using rRT-PCR. Of 96 from each sampling a total of 46, 37, 29, and 25 samples were positive for PPR viral genome at 4, 8, 12, and 16 weeks, respectively, after recovery. Attempts were made for the isolation of PPR virus on Vero cells, but results were negative. These results indicated the detection of PPR viral RNA up to 16 weeks after infection. Therefore, these results may help in the future epidemiology of PPR virus shedding and possible role as source of silent infection for healthy animals especially when there is no history of any outbreak in nearby flock or area.

Adult cancer clinical trials that fail to complete: an epidemic?

The number and diversity of cancer therapeutics in the pipeline has increased over the past decade due to an enhanced understanding of cancer biology and the identification of novel therapeutic targets. At the same time, the cost of bringing new drugs to market and the regulatory burdens associated with clinical drug development have progressively increased. The finite number of eligible patients and limited financial resources available to evaluate promising new therapeutics represent rate-limiting factors in the effort to translate preclinical discoveries into the next generation of standard therapeutic approaches. Optimal use of resources requires understanding and ultimately addressing inefficiencies in the cancer clinical trials system. Prior analyses have demonstrated that a large proportion of trials initiated by the National Cancer Institute (NCI) Cooperative Group system are never completed. While NCI Cooperative Group trials are important, they represent only a small proportion of all cancer clinical trials performed. Herein, we explore the problem of cancer clinical trials that fail to complete within the broader cancer clinical trials enterprise. Among 7776 phase II-III adult cancer clinical trials initiated between 2005-2011, we found a seven-year cumulative incidence of failure to complete of approximately 20% (95% confidence interval = 18% to 22%). Nearly 48000 patients were enrolled in trials that failed to complete. These trials likely contribute little to the scientific knowledge base, divert resources and patients from answering other critical questions, and represent a barrier to progress.

Susceptibility and irritability of adult forms of main malaria vectors against insecticides used in the indoor residual sprays in Muzaffargarh District, Pakistan: a field survey.

In southern Punjab, Pakistan, Muzaffargarh District is known to have insecticide-resistant Anopheles and drug-resistant Plasmodium spp. In this part of the country, five anopheline mosquitoes, Anopheles stephensi Liston, Anopheles culicifacies Giles, Anopheles fluviatilis James, Anopheles superpictus Grassi, and Anopheles subpictus Grassi (Diptera: Culicidae) are known as malaria vectors. Among these, An. culicifacies is the primary and An. stephensi is the secondary malaria vector. Outbreaks of malaria usually occur after rainy episodes. We conducted field surveys to collect field strains of An. culicifacies and An. stephensi mosquitoes from different areas of Muzaffargarh District. We determined susceptibility and irritability levels of their adult stages to the discriminative dose of different insecticides. For this purpose, we used World Health Organization's established criteria for assessment. Mortality was calculated after 1 h exposure and for 24 h recovery period for various insecticides. An. stephensi was found to be significantly resistant to dichlorodiphenyltrichloroethane (DDT, an organochlorine), dieldrin (a chlorinated hydrocarbon), and malathion (organophosphorus), with lethal times (LT50) of 83.17, 52.48, and 37.53, respectively. However, the species was significantly sensitive to permethrin, deltamethrin (pyrethroids), and fenitrothion (organophosphate) with LT50 of 2.85, 2.34, and 13.18, respectively. Among these, permethrin showed more promising results against adult An. stephensi. When analyzed for irritancy, we found that among pyrethroids, permethrin was the most irritant insecticide for both An. stephensi and An. culicifacies. DDT and dieldrin showed least irritancy with 0.42 +/- 0.08 and 0.77 +/- 0.12 takeoffs per minute per adult, respectively, against An. stephensi. The mean number of takeoffs per minute per adult with permethrin showed significant irritancy for permethrin when compared with DDT. Based on this study, we conclude that the use of organochlorine (DDT) and chlorinated hydrocarbon (dieldrin) should not be reintroduced in Malaria Control Programme in Pakistan until there is enough evidence to do so at any stage in future, and the use of pyrethroids should continue, with preference to permethrin for better control of malariaby indoor residual spraying.

Safety and efficacy of addition of VEGFR and EGFR-family oral small-molecule tyrosine kinase inhibitors to cytotoxic chemotherapy in solid cancers: a systematic review and meta-analysis of randomized controlled trials.

BACKGROUND: The approach of combining cytotoxic chemotherapy with oral small molecule tyrosine kinase inhibitors (TKIs) has been explored in a large number of randomized trials, in a variety of tumor. We performed a systematic review and meta-analysis to evaluate the safety and efficacy of this therapeutic approach. PATIENTS AND METHODS: PubMed and the ASCO databases were searched up to March 2013. We included randomized trials in which the FDA approved vascular endothelial growth factor receptor (VEGFR) or epidermal growth factor receptor-family (EGFR)-targeted TKI in combination with chemotherapy was compared with chemotherapy alone in patients with any type of solid cancer. The endpoints included safety [fatal adverse events (FAEs), treatment discontinuation, any severe (grade 3 or 4) adverse events (AEs), and individual severe AEs] and efficacy [progression-free survival (PFS), and overall survival (OS)]. The pooled relative risk (RR) or hazard ratio (HR) were calculated. RESULTS: A total of 16,011 patients from 43 trials were included. Compared with chemotherapy alone, the addition of a TKI significantly increased the risk of FAEs (RR, 1.63; 95% CI, 1.32-2.01), treatment discontinuation (RR, 1.80; 95% CI, 1.58-2.06), and any severe AE (RR, 1.25; 95% CI, 1.16-1.36). The addition of a TKI was associated with a significant improvement in PFS (HR, 0.82; 95% CI, 0.76-0.89), but not OS (HR, 0.99; 95% CI, 0.95-1.03). CONCLUSIONS: It is important for physicians to weigh the risk of toxicity versus the modest PFS benefit associated with chemotherapy plus TKI in patients with solid cancers.

Adverse event reporting in cancer clinical trial publications.

PURPOSE: Reporting adverse events is a critical element of a clinical trial publication. In 2003, the Consolidated Standards of Reporting Trials (CONSORT) group generated recommendations regarding the appropriate reporting of adverse events. The degree to which these recommendations are followed in oncology publications has not been comprehensively evaluated. METHODS: A review of citations from PubMed, Medline, and Embase published between Jan 1, 2009 and December 31, 2011, identified eligible randomized, controlled phase III trials in metastatic solid malignancies. Publications were assessed for 14 adverse event-reporting elements derived from the CONSORT harms extension statement; a completeness score (range, 0 to 14) was calculated by adding the number of elements reported. Linear regression analysis identified which publication characteristics associated with reporting completeness. RESULTS: A total of 175 publications, with data for 96,125 patients, were included in the analysis. The median completeness score was eight (range, three to 12). Most publications (96%) reported only adverse events occurring above a threshold rate or severity, 37% did not specify the criteria used to select which adverse events were reported, and 88% grouped together adverse events of varying severity. Regression analysis revealed that trials without a stated funding source and with an earlier year of publication had significantly lower completeness scores. CONCLUSION: Reporting of adverse events in oncology publications of randomized trials is suboptimal and characterized by substantial selectivity and heterogeneity. The development of oncology-specific standards for adverse event reporting should be established to ensure consistency and provide critical information required for medical decision-making.

Risk of hematologic toxicities in patients with solid tumors treated with everolimus: a systematic review and meta-analysis.

We performed a systematic review and meta-analysis of hematologic toxicities associated with everolimus, an oral mammalian target of rapamycin (mTOR) inhibitor. Eligible studies included phase II and III trials of patients with solid tumors on 10mg of everolimus daily describing events of neutropenia, thrombocytopenia, anemia or lymphopenia. The incidence of everolimus-associated all-grade and high-grade (Grade 3-4) hematologic toxicities were, respectively: neutropenia: 21.7% and 3.6%; thrombocytopenia: 36.0% and 4.7%; anemia: 61.2% and 8.4% and lymphopenia: 40.9% and 14.9%. Everolimus was associated with an increased risk of all-grade neutropenia (RR=2.24, [95% CI 1.51-3.32]), all-grade (RR=9.19, [95% CI 4.51-18.70]) and high-grade (RR=7.46, [95% CI 2.58-21.61]) thrombocytopenia, all-grade (RR=1.58, [95% CI 1.25-1.99]) and high-grade (RR=3.92, [95% CI 1.46-10.52]) anemia and all-grade (RR=1.72, [95% CI 1.50-1.97]) and high-grade (RR=2.70, [95% CI 1.86-3.93]) lymphopenia.

Risk of hematologic toxicities in cancer patients treated with sunitinib: a systematic review and meta-analysis.

BACKGROUND: The incidence and risk of unique toxicities associated with a multi-targeted tyrosine kinase inhibitor sunitinib, such as hypertension and thromboembolic events, have been previously reported. However, the incidence and risk of hematologic toxicities have been less well characterized. We performed an up-to-date meta-analysis of trials to evaluate the risk of sunitinib-related hematologic toxicities. METHODS: We searched Medline and the American Society of Clinical Oncology online database of meeting abstracts up to July 2012 for relevant clinical trials. Eligible studies included phase II and III trials and expanded access programs of sunitinib that reported adequate safety data profile reporting neutropenia, thrombocytopenia or anemia. The summary incidence, relative risk (RR) and 95% confidence intervals (CIs) were calculated. RESULTS: A total of 8,526 patients from 60 trials of sunitinib as a single agent revealed that the incidence of sunitinib-associated all-grade and high-grade (Grades 3 and 4) hematologic toxicities were, respectively: neutropenia: 42.1% and 12.8%; thrombocytopenia: 44.7% and 10.7% and anemia: 50.4% and 6.2%. Sunitinib-treated patients (2667 subjects from 10 randomized trials) had a significantly increased risk of all-grade (RR=3.58; 95% CI, 1.71-7.49) and high-grade (RR=3.32; 95% CI, 1.60-6.90) neutropenia, all-grade (RR=4.59; 95% CI, 2.76-7.63) and high-grade (RR=5.84; 95% CI, 2.22-15.41) thrombocytopenia and all-grade anemia (RR=1.15; 95% CI, 1.00-1.31). CONCLUSIONS: Sunitinib is associated with a significant increase in the risk of developing all-grade and high-grade neutropenia and thrombocytopenia and all-grade anemia compared with control.

Problems encountered in bicistronic IRES-GFP expression vectors employed in functional analyses of GC-induced genes.

Our laboratory has developed a series of Gateway(®) compatible lentiviral expression systems for constitutive and conditional gene knock-down and over-expression. For tetracycline-regulated transgenic expression, we constructed a lentiviral "DEST" plasmid (pHR-TetCMV-Dest-IRES-GFP5) containing a tetracycline-responsive minimal CMV promoter, followed by an attP site-flanked DEST cassette (for efficient cloning of cDNAs by "Gateway(®)" recombination cloning) and green fluorescent protein (GFP) driven by an internal ribosomal entry site (IRES).This lentiviral bicistronic plasmid allows immediate FACS identification and characterization of successfully transfected cell lines. Although this system worked well with several cDNAs, we experienced serious problems with SLA, Bam and BMF. Particularly, we cloned the cDNA for human SLA (Src-like adapter), a candidate gene in GC-induced apoptosis, into this plasmid. The resulting construct (pHR-TetCMV-SLA-IRES-GFP5) was transfected into HEK 293-T packaging cells to produce viral particles for transduction of CEM-C7H2-2C8 cells. Although the construct produced many green fluorescent colonies at the HEK 293-T and the CEM-C7H2-2C8 level, we could not detect any SLA protein with α-SLA antibody from corresponding cell lysates. In contrast, the antibody readily detected SLA in whole cell lysate of HEK 293-T cells transfected with a GST-flagged SLA construct lacking IRES-GFP. To directly address the potential role of the IRES-GFP sequence, we cloned the SLA coding region into pHR-TetCMV-Dest, a vector that differs from pHR-TetCMV-Dest-IRES-GFP5 just by the absence of the IRES-GFP cassette. The resulting pHR-TetCMV-SLA construct was used for transfection of HEK 293-T cells. Corresponding lysates were assayed with α-SLA antibody and found positive. These data, in concert with previous findings, suggest that the IRES-GFP cassette may interfere with translation of certain smaller size cDNAs (like SLA) or generate fusion proteins and entail defective virus production in an unpredictable manner.

Morphological and molecular characterisation of Echinococcus granulosus in livestock and humans in Punjab, Pakistan.

Cystic echinococcus poses an important economic and public health problem to Pakistan. Our study determined the prevalence and genotypes of Echinococcus present in domestic livestock and humans in Punjab, Pakistan. Out of 39,738 animals examined, 6.67% of animals were found infected. The prevalence and fertility of hydatid cysts was highest in camels (prevalence 17.29%; proportion fertile 95%), followed by sheep (prevalence 7.52%; proportion fertile 86.4%), buffalo (prevalence 7.19%; proportion fertile 84.3%), goats (prevalence 5.48%; proportion fertile 79.09%) and cattle (prevalence 5.18%; proportion fertile 75.25%). Phylogenetic analysis of the cytochrome oxidase-1 gene revealed that the common sheep strain (G1) and buffalo strain (G3) are cycling among livestock in Punjab and that these strains are highly adapted to goats, camels and cattle. Both human cysts were found to belong to the common sheep strain (G1) of E. granulosus, reinforcing this strain has the most potential for zoonotic transfer. Both morphological and molecular results support earlier studies suggesting that Echinococcus of sheep and buffalo origin is phenotypically and genetically similar which adds further evidence to support its recognition as one species viz, Echinococcus granulosus sensu stricto.