Evaluation of the genetic structure of the urban dwelling species of Bank Myna (Acridotheres ginginisnus) using random amplified polymorphic DNA (RAPD) analysis

We used the random amplified polymorphic DNA (RAPD) technique to decipher the genetic structure of the Bank Myna (Acridotheres ginginisnus) in Pakistan. The samples were collected from four cities namely: Dera Ghazi Khan, Jahanian, Khanewal and Gujranwala. The analysis showed a high genetic diversity at species level (H = 0.318, S = 0.467) but low levels at population levels (H = 0.047, S = 0.069 to H = 0.195, S = 0.283). The four populations analyzed were genetically distant from each other and hinted the effect of urbanization role in isolating the urban dwelling species. The genetic distances between populations ranged from D = 0.2403 to D = 0.3419 and the similarity coefficient showed low range from 0.66 to 0.70. This study will help in future conservation plans and also help the understanding of the role of urbanization on fragmentation of species’ natural distribution and dispersal mechanism.Key words: Acridotheres ginginisnus, Bank Myna, random amplified polymorphic DNA (RAPD), genetic structure

Population genetic structure of Rufous-Vented Prinia (Prinia burnesii) in Pakistan

The objective of the study is to ascertain genetic variation within Rufous-vented Prinia, Prinia burnesii an endemic species, by DNA fingerprinting applying random amplified polymorphic DNA (RAPD) technique. Genetic material was obtained from three distant sites along western bank of River Indus. These sites include Chashma barrage (32°50’N, 71°20’E), in the north; Taunsa barrage (30°45’N, 70°45’E) and the southern population from Guddu barrage (28°26’N, 69°44’E). In total, 14 RAPD primers were selected to determine the genetic variability between three populations of the species. The Nei’s (1973) genetic diversity in three populations ranged from 0.190 to 0.320. The genetic distance between Populations ranged from 0.149 to 0.265. The polymorphism levels ranged from 48 to 83% in three populations and at species level, polymorphism were 93.07%. The results further indicate that they are genetically isolated populations.Keywords: Random Amplified Polymorphic DNA, Prinia, genetic variation, river Indu

Cloning and expression of Toxoplasma gondii tachyzoite P22 protein

Delay in diagnosis of Toxoplasma gondii infection in pregnant women who have been infected during the first trimester of gestation can lead to death of her fetus. Serological tests based on recombinant proteins are the main diagnosis methods for the detection of anti Toxoplasma antibody in serum samples. The aim of this study was to clone and express a gene encoding a P22 protein of T. gondii tachyzoite as using antigen for ELISA serology method. DNA was extracted from T. gondii (RH-strain) tachyzoites and PCR reaction was done using corresponding primers. The PCR product was purified, ligated to PTZ57R plasmid via T/A cloning method and subcloned into SacI and BamHI digested pET32a expression vector. Recombinant plasmid was transformed in E. coli (Bl21 DE3) and induced by 1 mM IPTG and analyzed by 12% SDS-PAGE. Expressd protein was purified by affinity chromatography and confirmed by western blot analysis. We successfully cloned and expressed T. gondii P22 protein.Key words: Toxoplasma gondii, cloning, recombinant P22

Role of Potash Alum in Hepatitis C virus Transmission at Barber's Shop

Hepatitis C virus (HCV) is the main cause of severe liver disease, including hepatocellular carcinoma, cirrhosis and end stage liver disease. In Pakistan most of HCV positive patients have history of facial/armpit shaving from barbers. 79% of barbers are rubbing Potash Alum stone on facial shaving cuts. Dark blood spots are analyzed on Potash Alum stones being used at different barber shops. The aim of the study was to check the viability of hepatitis C virus on potash alum stone being used at barber shops. Blood samples from HCV positive patients were taken and treated with 0.1, 0.2, 0.3, 0.4 and 0.5 molar concentrations of Potash Alum for different periods of time. Blood was centrifuged to isolate the serum; HCV RNA was extracted from serum and subjected to first strand synthesis and PCR. PCR fragments were confirmed by sequencing. PCR amplification was observed in all the samples, treated with different concentrations of Potash Alum, indicated that the virus remains alive on Potash Alum stone for a long period of time. Potash Alum being used by barbers on facial shaving cuts has definite role in HCV transmission in Pakistani population. Therefore use of Potash Alum stone should be banned on facial shaving cuts at barber shops

Physiological and cell ultrastructure disturbances in wheat seedlings generated by Chenopodium murale hairy root exudate.

Chenopodium murale L. is an invasive weed species significantly interfering with wheat crop. However, the complete nature of its allelopathic influence on crops is not yet fully understood. In the present study, the focus is made on establishing the relation between plant morphophysiological changes and oxidative stress, induced by allelopathic extract. Phytotoxic medium of C. murale hairy root clone R5 reduced the germination rate (24% less than control value) of wheat cv. Nataša seeds, as well as seedling growth, diminishing shoot and root length significantly, decreased total chlorophyll content, and induced abnormal root gravitropism. The R5 treatment caused cellular structural abnormalities, reflecting on the root and leaf cell shape and organization. These abnormalities mostly included the increased number of mitochondria and reorganization of the vacuolar compartment, changes in nucleus shape, and chloroplast organization and distribution. The most significant structural changes were observed in cell wall in the form of amoeboid protrusions and folds leading to its irregular shape. These structural alterations were accompanied by an oxidative stress in tissues of treated wheat seedlings, reflected as increased level of H2O2 and other ROS molecules, an increase of radical scavenging capacity and total phenolic content. Accordingly, the retardation of wheat seedling growth by C. murale allelochemicals may represent a consequence of complex activity involving both cell structure alteration and physiological processes.This is a post-peer-review, pre-copyedit version of an article published in Protoplasma. The final authenticated version is available online at: [http://dx.doi.org/10.1007/s00709-018-1250-0

Quality-sensitive foraging by a robot swarm through virtual pheromone trails

Large swarms of simple autonomous robots can be employed to find objects clustered at random locations, and transport them to a central depot. This solution offers system parallelisation through concurrent environment exploration and object collection by several robots, but it also introduces the challenge of robot coordination. Inspired by ants’ foraging behaviour, we successfully tackle robot swarm coordination through indirect stigmergic communication in the form of virtual pheromone trails. We design and implement a robot swarm composed of up to 100 Kilobots using the recent technology Augmented Reality for Kilobots (ARK). Using pheromone trails, our memoryless robots rediscover object sources that have been located previously. The emerging collective dynamics show a throughput inversely proportional to the source distance. We assume environments with multiple sources, each providing objects of different qualities, and we investigate how the robot swarm balances the quality-distance trade-off by using quality-sensitive pheromone trails. To our knowledge this work represents the largest robotic experiment in stigmergic foraging, and is the first complete demonstration of ARK, showcasing the set of unique functionalities it provides

Defining Plasmodium falciparum Treatment in South West Asia: A Randomized Trial Comparing Artesunate or Primaquine Combined with Chloroquine or SP

INTRODUCTION: Antimalarial resistance has led to a global policy of artemisinin-based combination therapy. Despite growing resistance chloroquine (CQ) remained until recently the official first-line treatment for falciparum malaria in Pakistan, with sulfadoxine-pyrimethamine (SP) second-line. Co-treatment with the gametocytocidal primaquine (PQ) is recommended for transmission control in South Asia. The relative effect of artesunate (AS) or primaquine, as partner drugs, on clinical outcomes and gametocyte carriage in this setting were unknown. METHODS: A single-blinded, randomized trial among Afghan refugees in Pakistan compared six treatment arms: CQ; CQ+(single-dose)PQ; CQ+(3 d)AS; SP; SP+(single-dose)PQ, and SP+(3 d)AS. The objectives were to compare treatment failure rates and effect on gametocyte carriage, of CQ or SP monotherapy against the respective combinations (PQ or AS). Outcomes included trophozoite and gametocyte clearance (read by light microscopy), and clinical and parasitological failure. FINDINGS: A total of 308 (87%) patients completed the trial. Failure rates by day 28 were: CQ 55/68 (81%); CQ+AS 19/67 (28%), SP 4/41 (9.8%), SP+AS 1/41 (2.4%). The addition of PQ to CQ or SP did not affect failure rates (CQ+PQ 49/67 (73%) failed; SP+PQ 5/33 (16%) failed). AS was superior to PQ at clearing gametocytes; gametocytes were seen on d7 in 85% of CQ, 40% of CQ+PQ, 21% of CQ+AS, 91% of SP, 76% of SP+PQ and 23% of SP+AS treated patients. PQ was more effective at clearing older gametocyte infections whereas AS was more effective at preventing emergence of mature gametocytes, except in cases that recrudesced. CONCLUSIONS: CQ is no longer appropriate by itself or in combination. These findings influenced the replacement of CQ with SP+AS for first-line treatment of uncomplicated falciparum malaria in the WHO Eastern Mediterranean Region. The threat of SP resistance remains as SP monotherapy is still common. Three day AS was superior to single-dose PQ for reducing gametocyte carriage. TRIAL REGISTRATION: ClinicalTrials.gov NCT00959517

A comparison of four fibrosis indexes in chronic HCV: Development of new fibrosis-cirrhosis index (FCI)

<p>Abstract</p> <p>Background</p> <p>Hepatitis C can lead to liver fibrosis and cirrhosis. We compared readily available non-invasive fibrosis indexes for the fibrosis progression discrimination to find a better combination of existing non-invasive markers.</p> <p>Methods</p> <p>We studied 157 HCV infected patients who underwent liver biopsy. In order to differentiate HCV fibrosis progression, readily available AAR, APRI, FI and FIB-4 serum indexes were tested in the patients. We derived a new fibrosis-cirrhosis index (FCI) comprised of ALP, bilirubin, serum albumin and platelet count. FCI = [(ALP × Bilirubin) / (Albumin × Platelet count)].</p> <p>Results</p> <p>Already established serum indexes AAR, APRI, FI and FIB-4 were able to stage liver fibrosis with correlation coefficient indexes 0.130, 0.444, 0.578 and 0.494, respectively. Our new fibrosis cirrhosis index FCI significantly correlated with the histological fibrosis stages F0-F1, F2-F3 and F4 (r = 0.818, p < 0.05) with AUROCs 0.932 and 0.996, respectively. The sensitivity and PPV of FCI at a cutoff value < 0.130 for predicting fibrosis stage F0-F1 was 81% and 82%, respectively with AUROC 0.932. Corresponding value of FCI at a cutoff value ≥1.25 for the prediction of cirrhosis was 86% and 100%.</p> <p>Conclusions</p> <p>The fibrosis-cirrhosis index (FCI) accurately predicted fibrosis stages in HCV infected patients and seems more efficient than frequently used serum indexes.</p

Electronic medical record in cardiology: a 10-year Italian experience

SummaryObjectives:the aim of this study was to report a ten years experience in the electronic medical record (EMR) use. An estimated 80% of healthcare transactions are still paper-based.Methods:an EMR system was built at the end of 1998 in an Italian tertiary care center to achieve total integration among different human and instrumental sources, eliminating paper-based medical records. Physicians and nurses who used EMR system reported their opinions. In particular the hospital activity supported electronically, regarding 4,911 adult patients hospitalized in the 2004- 2008 period, was examined.Results:the final EMR product integrated multimedia document (text, images, signals). EMR presented for the most part advantages and was well adopted by the personnel. Appropriateness evaluation was also possible for some procedures. Some disadvantages were encountered, such as start-up costs, long time required to learn how to use the tool, little to no standardization between systems and the EMR technology.Conclusion:the EMR is a strategic goal for clinical system integration to allow a better health care quality. The advantages of the EMR overcome the disadvantages, yielding a positive return on investment to health care organization