Naveeda Akhtar Qureshi

Development of mannose-anchored thiolated amphotericin B nanocarriers for treatment of visceral leishmaniasis.

Aim: Our goal was to improve treatment outcomes for visceral leishmaniasis by designing nanocarriers that improve drug biodistribution and half-life. Thus, long-acting mannose-anchored thiolated chitosan amphotericin B nanocarrier complexes (MTC AmB) were developed and characterized. Materials & methods: A mannose-anchored thiolated chitosan nanocarrier was manufactured and characterized. MTC AmB was examined for cytotoxicity, biocompatibility, uptake and antimicrobial activities. Results: MTC AmB was rod shaped with a size of 362 nm. MTC AmB elicited 90% macrophage viability and 71-fold enhancement in drug uptake compared with native drug. The antileishmanial IC50 for MTC AmB was 0.02 μg/ml compared with 0.26 μg/ml for native drug. Conclusion: These studies show that MTC can serve as a platform for clearance of Leishmania in macrophages.

Synthesis, in vitro potential and computational studies on 2-amino-1,4-dihydropyrimidines as multitarget antibacterial ligands

In this study, we have investigated small multitargeted molecules containing 2-aminopyrimidine scaffold that may further act as precursor for developing more potent antibacterials. An efficient route to 2-amino-1,4-dihydropyrimidines by using ultrasound irradiation as the energy source was developed. In silico density functional theory calculations illustrated that tin chloride-mediated Biginelli reaction to produce 2-amino-1,4-dihydropyrimidines has energetics quite accessible under the reaction conditions. Calculated minimum inhibitory concentrations against the various bacterial strains showed that compounds 3 and 11 displayed comparable in vitro activity to ciprofloxacin in Staphylococcus aureus strains and reduced potency in Escherichia coli strains. Further, we investigated in silico ADMET profiling of synthesized compounds in order to understand the mechanism of action that help in explaining in vitro results. Lead compounds 3, 6, and 11 are predicted to have acceptable pharmacokinetic/drug-like properties. Data mining and computational analysis were employed to derive compound promiscuity phenomenon. All the compounds were found nonsubstrate towards various aminergic G-protein coupled receptors, ion-channels, kinase inhibitor, nuclear receptor ligand, protease inhibitor, and enzyme inhibitor. Compound 3 was further investigated by in silico binding to different antibacterial targets. Binding energy data revealed that that these compounds have the ability to bind with other bacterial targets. Hence, combined in silico and in vitro studies shed insights into the mechanism of synthesis and antibacterial activity of 2-amino-1,4-dihydropyrimidines. Results of this study are promising and can be used for further investigation by medicinal chemists to explore their chemical functionalization and in vivo studies.

Design, synthesis, antibacterial activity and docking study of some new trimethoprim derivatives

In present study, nineteen novel trimethoprim (TMP) derivatives were designed, synthesized and evaluated for their antibacterial potential. Hydroxy trimethoprim 2 (HTMP) was synthesized by following the demethylation of 4-methoxy group at trimethoxy benzyl ring of TMP. Structure-activity relationship (SAR) studies were explored on HTMP by incorporating various substituents leading to the identification of some new compounds with improved antibacterial activities. The results revealed that the introduction of benzyloxy (4a-e) and phenyl ethanone (5a-e) group at 4-position of dimethoxy benzyl ring leads to overall increase in the antibacterial activity. The most potent antibacterial compound discovered is benzyloxy derivative 4b with MIC value of 5.0μM against Staphylococcus aureus and 4.0μM against Escherichia coli strains higher than the standard TMP (22.7μM against S. aureus and 55.1μM against E. coli). Substitution at 4-NH2 group was not tolerated and the resulting Schiff base derivatives 3a-h demonstrated very little or no antibacterial activity in the tested concentration domain. We further performed exploratory docking studies on dihydrofolate reductase (DHFR) to rationalize the in vitro biological data and to demonstrate the mechanism of antibacterial activity. For the ability to cross lipophilic outer membrane, logP was computed. It was found that the compounds possessing high hydrophobicity have high activity against E. coli.

New endemic focus of cutaneous leishmaniasis in Pakistan and future epidemics threats

Abstract Objective To get a clear epidemiological picture of cutaneous leishmaniasis in a new endemic focus, in order to address the most probable epidemic threats and its spread to the neighboring regions. Methods A total of 870 clinically diagnosed patients were included in the study after taking consent and filling questionnaires from each individual. The field Giemsa-stained slides and cultures were examined microscopically. The data was organized in tables and graphs and analyzed using statistical package SPSS V 21. Results The ratio of male was higher than females (499:371), cutaneous and mucocutaneous cases were 96.00% and 4.00%, and only 25.20% patients used bed nets. A total of 57.10% had dirty places near their homes. Similar cases in the neighborhood and in families were 48.50% and 67.10% respectively. Only 87.35 % of Patients in the age range of 1–20 years received treatment. Up to five lesions per individual were noticed and 72.20% of them had symptomatic secondary infections. The lesions were observed on different sites of the body. Conclusions Leishmaniasis is neglected in Landi Kotal Khyber Agency, the reasons being prevailing security situations and political sensitivity. North Atlantic Treaty Organization forces in Afghanistan are in contact with Afghanis and with local people of Landi Kotal for logistic purposes. They may act as carriers in co-spreading of the species. There are potential risks of cross border transmission, with the probability of future epidemics in the surrounding regions.

Protozoidal activities of Eucalyptus cammeldulensis, Dalbergia sissoo and Acacia arabica woods and their different parts on the entozoic flagellates of Heterotermes indicola and Coptotermes heimi

Different parts of three woods of Eucalyptus cammeldulensis , Dalbergia sissoo and Acacia arabica were analyzed for their toxicity potentials against two species of termites ( Heterotermis indicola and Coptotermis heimi ). Termite workers were allowed to feed on 2 g complete wood powder of plant species and their parts, including; bark, sapwood and heartwood. Samples of flagellates were collected after each 24 h from the termites’ gut and they showed a significant variation in their mortality rate as per the wood species and their parts used in the experiments. After six days, mortality rates in flagellates were 100% with all wood parts of E. cammeldulensis , whereas it was 87.2, 47.61 and 100% with bark, sapwood and heartwood of D. sissoo respectively. However, in the case of A. Arabica , only bark inflicted 44.5% mortality on the flagellates in termites on the 6th day. It is revealed from the results that different woods or their specific parts have some specific toxic compounds that inflicted varying degree of toxicity on enteric flagellates of termites. Considering the toxigenic nature of different woods and their respective parts, the three woods; E. cammeldulensis , D. sissoo and A. arabica and their parts barks, sapwoods and heartwoods were analyzed for the presence of water soluble constituents such as lignin, benzene-ethanol soluble components and alpha cellulose contents. However, it is highly recommended that such protozoicidal compounds should be isolated, purified and biochemically characterized in order to apply them as commercial products for the control of pest like termites, which cause a huge damage to woody plants, and their products. Keywords: Bark, sapwood, heartwood, Eucalyptus cammeldulensis , Dalbergia sissoo , Acacia arabica , termite flagellates

A preliminary survey on prevalence and knowledge about different aspects of somnambulism in Buner District of Khyber Pakhtunkhwa, Pakistan

PurposePresent study was aimed to investigate the prevalence and knowledge about different aspects of somnambulism in general population of Buner District, KP, Pakistan, during December 2015–November 2016.MethodsData was collected through convenient sampling technique which was comprised of structured and detailed questionnaire. Collected data regarding different actions performed by sleepwalkers were adjusted through a weighted variable to generalize results. Univariate and multivariate logistic regression was used to evaluate association between variables related to somnambulism. Their odds ratios (ORs) were reported with their corresponding 95% CIs.ResultsIn total, 11,881 individuals were surveyed, of which 448 (4%) suffered with sleep disorder. Females 256/448 (57%) were more prevalent as compared to males 192/448 (43%). Prevalence of somnambulism among children was 38% whereas 94% sufferers reported no effect on their growth. Majority of respondents reported that main cause of sleepwalking is stress and tension. Somnambulism was taken normal and mostly amusing disorder by sufferers and their families.ConclusionSleepwalking is not age specific and serious disorder. Sleepwalkers need special care during their episode due to unconsciousness.

Comparative analysis of egg adapted vaccines and salinomycin against coccidiosis in chicks

Avian coccidiosis is an intestinal protozoan parasitic disease of genus Eimeria. Wide use of anticoccidial drugs has resulted in development of drug resistant strains. Current study is based upon the development of egg adapted vaccines; gametocytes, formalin inactivated and formalin inactivated sonicated gametocytes against coccidiosis. Day old chicks (n = 90) were divided into six groups (A-F) with triplicate. On 5th day of age, three groups were orally given developed vaccines (0.2 ml) while on 21st day groups (A-E) were challenged orally with mixed Eimeria spp (60-70,000 oocysts/chick). The comparative effect of vaccines and drug salinomycin was evaluated on basis of different parameters. Blood, liver and caecal tissues were collected on 5th, 7th and 15th day post infection for indirect hemagglutination test, biochemical analysis and histopathology. Significantly higher body weight gain, feed consumption, mild bloody diarrhea with lowest oocyst count and survival rate of 100% was recorded for gametocytes vaccinated group (P < 0.05). Indirect hemagglutination test showed maximum antibodies titer (IgG) in gametocytes vaccinated group. Present investigation revealed that gametocyte vaccines was significantly effective in control of coccidiosis by inducing strong protection in immune chicks contained high level of antibodies that resisted heavy dose of challenge as compared to anticoccidial drug salinomycin.

Morphological characterization and biological control of Alitropus typus (Isopoda: Aegidae)

The Alitropus typus infestation has a serious influence on fish farming in Pakistan. The present study focuses on the external morphology and control of highly infested ectoparasitic isopod Alitropus typus. Sixteen morphological parameters of the whole body were measured and statistically analyzed for mean, standard error and coefficient of variation by using the student t test. Non‐significant variations were observed in the size of pereopods which depict the significance of swimming and attached to its host. For biological control, different concentrations (1000, 5000 and 10 000 ppm) of a leaf extract of four plants (i.e. Euphorbia helioscopia, Ajuga bracteosa, Cannabis sativa and Callistemon citrinus) were tested against A. typus, and the mortality rates were recorded after 12, 24 and 36 h. One way Anova and Tukey tests were used to analyze the results. The recorded percentage mortality was in the order of E. helioscopia > A. bracteosa > C. sativa, whereas C. citrinus did not showed any toxicity. Concentrations of biochemical components like carbohydrates (mg/g), proteins (mg/g), and lipids (mg/dL) of treated isopods were estimated by the phenol‐sulfuric acid process, Lowrys method and biochemistry analyzer, respectively. The protein contents of the insects tested had decreased markedly as compared to control rates, and this might be due to insecticidal stress caused by the extracts. In another biocontrol experiment, crabs were introduced along with isopods into an aquarium. It was then noted that one crab consumes an average of seven isopods/week and can act as a scavenger for dead organisms.

Prevalence of Leishmania tropica in school boys of khyber agency, FATA near Pak-Afghan border.

In Pakistan leishmaniasis occurs periodically throughout the year and various out breaks are reported frequently. In continuation of our research on this neglected disease, the aim of present study is to explore: (1) the prevalence of cutaneous leishmaniasis in school boys; (2) Leishmania species identification in order to epidemiology and dynamics of the disease; (3) Identification of risk factors for Leishmaniasis especially for CL. The data was collected in August 2014 The experimental strategy involved a questionnaire for data collection and along with clinical diagnosis of 134 out of 9368 students for incidence of Leishmania spp. in 7 square kilometres area in the schoolboys at Tehsil Landi Kotal, District Khyber Agency, FATA Pakistan. The parasitological and molecular diagnosis of clinically suspected cutaneous leishmaniasis cases were performed using microscopical examination of Giemsa-stained smears of lesion exudates and minicircle kDNA semi nested PCR, respectively. Microscopy (x=1000) positive cases were 84/134 (62.6%) and 50/134 (37.4%) slides did not showed any presence of amastigotes of Leishmania spp. The samples were amplified using kDNA semi nested PCR and confirmed the presence of L. Tropica (Ac.no KT 985473). PCR positive cases were 97/134 (72.4%) and 37(27.6%) were negative. The prevalence of L. tropica in school boys was 1.4% (134/9368) in the total population studied (n=9368). The parasite prevalence might be greater as only male students were considered in the study due to ethical and social issues and limitations.

Prevalence and distribution of human Plasmodium infection in Federally Administrative Tribal Areas of Pakistan.

About 3.6 million Pashtun and over 1.5 million immigrants from Afghanistan live in the federally administered tribal areas (FATA) on the border between Pakistan’s Khyber Pakhtunkhwa Province and southern Afghanistan. Although malaria cases are common in FATA, no detailed studies have yet been performed to reveal the actual status of malaria in the local population and epidemiological data are insufficient to elucidate the actual incidence. A malariometric survey of 691 patients of all ages and genders in seven agencies (districts) in FATA was carried out in 2013 using whole blood samples. Microscopically confirmed positive species were subjected to nested-PCR for the reconfirmation and detection of four species of Plasmodium causing human malaria. Of the 626 PCR positive cases, 81.1% were P. vivax, 13.8% P. falciparum and 4.9% mixed species containing both P. vivax and P. falciparum. P. malariae and P. ovale and were not found in any analysis. Sixty-five microscopic positive samples were identified as negative by PCR. The incidence of P. vivax ranged from 10.4% in Orakzai Agency to 22.8% in North Waziristan Agency. The prevalence of P. falciparum ranged from 1.3% in Orakzai Agency to 4.7% in North Waziristan, and Khyber Agency had the highest prevalence of 1.7% of mixed species. In FATA, P. vivax and P. falciparum are the main causative agents of malaria, while mixed species infections are also prevalent with varying transmission intensities. In addition, Estimates of malaria incidence shows that variation in the incidence, frequency and species composition of malarial parasites is high.