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The present research study was aimed to investigate the efficiency of selected medicinal plants against Plasmodium vivax. Crude methanolic extracts from the seeds of leaves of Datura stramonium, Parthenium hysterophorus, Calotropis procera, and Dodonaea viscosa were prepared. In addition, Atropine was also isolated from alkaloid components of Eucalyptus obliqua to evaluate their in vitro anti-plasmodial effects. It was observed that proguanil (positive control) and Atropine displayed strong anti-plasmodial activity (94.04 and 68.02%, respectively) against P. vivax at 0.1 mg/mL concentration while the leaf extracts of other medicinal plants did not exhibit any notable anti-plasmodial activity. It was concluded that alkaloids of E. obliqua plant’s extracts were rich in anti-plasmodial compound Atropine, which exhibit a remarkable anti-plasmodial activity against P. vivax. Anti-plasmodial action of medicinal plants are attributed to these phytochemicals. In vitro studies using medicinal plant’s extracts and standardized methods will help to make more powerful and cost-effective anti-plasmodial compounds.

Malaria is an intraerythrocytic parasitic disease caused by the genus Plasmodium of which Plasmodium vivax and Plasmodium falciparum are the major species. The high cost and associated side effects of antimalarial drugs triggered research about medicinal plants to develop alternative and low-cost drugs with lesser side effects. Therefore, this study was designed to investigate the antiplasmodial activity of the Eucalyptus obliqua L’Hér. leaf extract against P. vivax and its phytochemicals in in vitro. The methanolic extract of E. obliqua was prepared and different concentrations of the crude extract and phytochemicals were used against P. vivax. The methanolic extract of E. obliqua showed profound antiplasmodial activity (LD50 0.084 mg/mL; 80.04%) at 0.1 mg/mL concentration after 24 h. Alkaloids, flavonoids, saponins, and tannins were found in the E. obliqua methanolic extract. Only alkaloids at the concentration (0.1 mg/mL) exhibited 60.93% inhibition of P. vivax. The methanolic extract of E. obliqua exhibits antiplasmodial activity in vitro. However, in vivo efficacy is an important aspect in the testing of medicinal plants against parasitic infections and should be evaluated in future.

Intellectual disability (ID) is a large group of neurodevelopmental disorders characterized by a congenital limitation in intellectual functioning (reasoning, learning, and problem solving), adaptive behavior (conceptual, social, and practical skills), originated at birth and manifested before the age of 18. By whole exome sequencing of five consanguineous Pakistani families presenting hallmark features of ID, global developmental delay, aggressive and self‐injurious behaviors, microcephaly, febrile seizures and facial dysmorphic features, we identified three novel homozygous missense variants (NM_024298.5: c.588G > T; p.Trp196Cys, c.736 T > C; p.Tyr246His and c.524A > C; p. Asp175Ala) and one rare homozygous in‐frame deletion variant (c.758_778del;p.Glu253_Ala259del) in membrane‐bound O‐acyltransferase family member 7 (MBOAT7) gene previously associated with autosomal recessive neurodevelopmental disorder. The segregation of the variants was validated by Sanger sequencing in all family members. In silico homology modeling of wild‐type and mutated proteins revealed substantial changes in the structure of both proteins, indicating a possible effect on function. The identification and validation of new pathogenic MBOAT7 variants in five cases of autosomal recessive ID further highlight the importance of this genes in proper brain function and development. [ABSTRACT FROM AUTHOR]

Introduction. This study determines the incidence of common viral and helminth coinfections with malaria in the tertiary care hospital set up in southern Khyber Pakhtunkhwa, Pakistan. Materials and Methods. The multidimensional research included malaria patients admitted to different hospitals of district Kohat during January and December 2021. Stool samples and blood were assembled from the patients. Giemsa-stained microscopy-positive samples were processed by the immunochromatography technique (ICT) to identify Plasmodium species. Common viral infections such as viral hepatitis (A, B, and C), HIV, and dengue (DENV) were analyzed by ICT kits while SARS-CoV-2 was confirmed through real-time PCR. Furthermore, the intestinal helminths were identified using the Kato-Katz thick smear method. Results. Among 1278 patients, 548 were diagnosed with malaria, 412 (75.2%) were positive for P. vivax infection, 115 (21%) for P. falciparum, and 21 (3.8%) for mixed malaria infection (P. vivax/P. falciparum), with a higher incidence among males (65.2%) than females (34.8%). Coinfection with helminths was positive in 215 (39.3%) malaria patients. The most common infections were caused by the Ascaris lumbricoides species (42.6%) followed by Enterobius vermicularis (31.7%) and hookworm. A total of 24.6% of malaria-positive cases were also coinfected with different viruses with higher frequencies of confection for HAV (8.2%) and DENV (6.2%), respectively. The patients revealed higher incidence of coinfections with P. falciparum (57%) as compared with P. vivax (39.2%) and mixed infections (3.7%). Conclusion. This study demonstrated that the study population exhibited a significant incidence of coinfections with intestinal helminth and viral malaria. [ABSTRACT FROM AUTHOR]

This document is a correction notice for an article titled "A cross-sectional survey of hard ticks and molecular characterization of Rhipicephalus microplus parasitizing domestic animals of Khyber Pakhtunkhwa, Pakistan." The correction states that there were errors in Table 3, specifically regarding the months for the Autumn and Summer seasons. The correct months for Autumn are "Mar, Apr, and May," and for Summer are "Jun, July, and Aug." The table provides information on the prevalence of hard ticks in District Bannu among different domestic animals. The correction is authored by Muhammad Rooman, Yasir Assad, Sadia Tabassum, Samia Sultan, Sultan Ayaz, Muhammad Fiaz Khan, Shahid Niaz Khan, and Rehman Ali. [Extracted from the article]

Background. Nanotechnology finds broad applications in the field of nanomedicine, an emerging new field used for diagnosis, treatment, prevention of diseases, and improvement of health. Objectives. To synthesize silver nanoparticles (AgNPs) from Withania somnifera and Fagonia indica and to carry out their antimicrobial, insecticidal, and phytotoxic activities, a step toward the new range of nanomedicines. Methods. Silver nanoparticles were synthesized from Withania somnifera and Fagonia indica by chemical reduction method, and further biological activities of these nanoparticles were compared with crude methanolic extract, prepared through cold maceration process, at the concentration of 50 mg/ml. Results. Among all tested bacterial pathogens, crude extract of W. somnifera showed a statistically high significant inhibition zone in millimeter against Pseudomonas aeruginosa (21; p < 0.01). AgNPs showed highly significant result against Streptococcus pneumonia (14; p < 0.01). In comparison with crude extracts, AgNPs showed statistically significant (p < 0.01) results against S. pneumonia (AgNPs, 14; crude, 8.33 mm). Crude extract showed significant inhibition zone against two bacterial strains, P. aeruginosa (crude, 21; AgNPs, 11.67 mm) and Klebsiella pneumoniae (crude, 11.33; AgNPs, 8 mm). Crude extracts of F. indica showed the significant activity against Vibrio cholera (p < 0.01 ; 11.33 mm). Silver nanoparticles of F. indica exhibited the highest significant activity against Aspergillus flavus and Fusarium oxysporum while AgNPs of W. somnifera were active only against A. flavus. Extracts of W. somnifera and F. indica showed increasing phytotoxic activity with increasing concentrations. The highest significant inhibition was obtained for crude extract (46.7) and AgNPs (45.7) of F. indica at 1000 μg/ml. Insecticidal activity of crude and AgNPs of both plants showed significant inhibition against all tested insects with increasing time intervals, and the highest significant result was obtained at 72 hours with a value of p < 0.01 except T. castaneum. Conclusions. Both crude and AgNPs showed potent activity; however, in comparison, silver nanoparticles showed slightly enhanced activity. Crude and AgNPs of both plants showed good phytotoxic and insecticidal inhibition. Antimicrobial studies of AgNPs on diseases causing pathogens open a door for new antimicrobial agents and could be the answer to antibiotic resistance after further analysis. [ABSTRACT FROM AUTHOR]

Whole exome sequencing has provided significant opportunities to discover novel candidate genes for intellectual disability and autism spectrum disorders. Variants in the spectrin genes SPTAN1, SPTBN1, SPTBN2 , and SPTBN4 have been associated with neurological disorders; however, SPTBN5 gene-variants have not been associated with any human disorder. This is the first report that associates SPTBN5 gene variants (ENSG00000137877: c.266A>C; p.His89Pro, c.9784G>A; p.Glu3262Lys, c.933C>G; p.Tyr311Ter, and c.8809A>T; p.Asn2937Tyr) causing neurodevelopmental phenotypes in four different families. The SPTBN5 -associated clinical traits in our patients include intellectual disability (mild to severe), aggressive tendencies, accompanied by variable features such as craniofacial and physical dysmorphisms, autistic behavior, and gastroesophageal reflux. We also provide a review of the existing literature related to other spectrin genes, which highlights clinical features partially overlapping with SPTBN5. [ABSTRACT FROM AUTHOR]

Background: Cutaneous leishmaniasis is a neglected tropical disease caused by Leishmania spp. and transmitted by female sandflies. Terrorism and counter-insurgency military operations in Federally Administered Tribal Areas (FATA) lead to a large-scale migration of internally displaced persons (IDPs) in Khyber Pakhtunkhwa and thus, new outbreaks of several infectious diseases such as cutaneous leishmaniasis occurred. This study intended to find the prevalence of cutaneous leishmaniasis in people with cutaneous lesions suspected of having cutaneous leishmaniasis in four remote districts of Khyber Pakhtunkhwa and to assess the participant's knowledge, attitude, and practices about the infection and its control. Methods: A cross-sectional study was carried out in four remote districts of Khyber Pakhtunkhwa including Karak, Lakki Marwat, Tank, and Dera Ismail Khan (D. I. Khan) and a total of 1,674 participants were recruited using a convenience sampling technique. Results: The prevalence of cutaneous leishmaniasis among the participants with cutaneous lesions was 50.4% and the infection was comparatively more prevalent in district Karak. Among participants, 56.8% were male and mostly, 53.8% were under the age of 16 years with 52.8% living in kutcha houses and were from rural areas. Multiple skin lesions were more common, and the face was frequently affected body part. The ratio of participants with lesions older than a month was higher and the majority confronted infections with blood protozoan parasites for the first time. Most participants were unaware of the signs/symptoms of the disease, basic knowledge of the vectors, anthroponotic spread, preventive measures, secondary infections, and reservoir hosts. The use of wood/animal dung as fuel, closeness with reservoir animals, and no use of insect repellents were some of the notable risk factors. Conclusion: Cutaneous leishmaniasis is highly prevalent in the study area and a very low level of awareness was reported among the participants. This study necessitates the planning and execution of regulations and preventive programs, public health education, awareness campaigns, and disease management practices to overcome future incidence of cutaneous leishmaniasis. [ABSTRACT FROM AUTHOR]

Background: Understanding the genetic diversity of Plasmodium species through polymorphic studies can assist in designing more effective control strategies of malaria like new drug formulation and development of a vaccine. Pakistan is moderate endemic for Plasmodium falciparum , but little is known about the genetic diversity of this parasite. This study aimed to investigate the molecular diversity of P. falciparum based on msp-1 and msp-2 genes in the malaria-endemic regions of Khyber Pakhtunkhwa, Pakistan. Methods: A total of 199/723 blood samples, tested positive by microscopy for falciparum malaria, were collected from four districts (Dera Ismail Khan, Karak, Mardan, and Peshawar) of Khyber Pakhtunkhwa. Nested PCR amplification technique was employed to target block 2 of msp-1 and the central domain of msp-2 genes, including their respective allelic families K1, MAD20, RO33, FC27, and 3D7/IC, and to detect the extent of genetic diversity of P. falciparum clinical isolates. Results: Among the 199 microscopy-positive P. falciparum samples, a total of 192 were confirmed using PCR. Ninety-seven amplicons were observed for msp - 1 and 95 for msp-2. A total of 33 genotypes, 17 for msp-1 (eight K1, six MAD20, and three RO33) and 16 for msp-2 (nine FC27 and seven 3D7/IC), were identified. The specific allelic frequency of the K1 family was higher (44.3%) than that of MAD20 (33.0%) and RO33 (23.0%) for msp-1 , while the FC27 allelic family was dominant (60.0%) compared with 3D7/IC (40.0%) for msp-2. No polyclonal infection was observed in msp-1 and msp-2. The expected heterozygosity was 0.98 and 0.97 for msp-1 and msp-2 , respectively. Conclusion: It was concluded that the P. falciparum populations are highly polymorphic, and diverse allelic variants of msp-1 and msp-2 are present in Khyber Pakhtunkhwa, Pakistan. [ABSTRACT FROM AUTHOR]

Background: In tropical and subtropical countries, tick infestation causes major public health problems and considerable financial losses to the livestock industry. This study was aimed to assess the species composition of richness and analyze the phylogeny of Rhipicephalus microplus in the District Bannu of Khyber Pakhtunkhwa, Pakistan. Methods: Collected ticks were identified morphologically and DNA extracted from R. microplus was amplified and subjected to sequencing. Results: A total of 3,600 animals were examined among them 1,494 animals were found to be infested with ticks, including 669 cows, 476 buffaloes, 163 goats, and 186 sheep (p = 0.001). Tick infestation was significantly high (43.58%) in animals of age group (<1 year) (p-value = 0.027). Female animals were more (44.05%) infested with ticks than males (34.43%) (p = 0.001). The intensity of infestation was significantly higher in summer (77.49%) (p = 0.001). A total of 5,557 ticks were collected comprising three genera and six species. R. microplus was predominantly prevalent (n = 1,474; 26.52%), followed by Rhipicephalus annulatus (n = 1,215; 21.86%), Hyalomma anatolicum (n = 1,139; 20.49%), Hyalomma marginatum (n = 1,086; 19.54%), and Rhipicephalus turanicus (n = 761; 13.69%), while the least common was Haemaphysalis aciculifer (n = 80; 1.43%) (p = 0.001). Morphologically identified R. microplus species were also analyzed genetically by using two genetic markers 16S ribosomal RNA (16S rRNA) and internal transcribed spacer 2 (ITS2) genes. The phylogenetic study revealed that R. microplus is genetically diversified and clustered in clade B with R. microplus species from China, India, and Pakistan. Conclusion: Ticks infestation was significantly correlated with various factors including age, sex, season, and animal type. R. microplus genetically resembled species reported from India and China. However, major knowledge gaps concerning various species of ticks exist and many areas are still unexplored in Pakistan. Therefore, it is necessary to explore the epidemiological and molecular aspects of various tick species in other regions of southern Khyber Pakhtunkhwa. [ABSTRACT FROM AUTHOR]

Cystic echinococcosis (CE) is a zoonotic helminthiasis caused by different species of the genus Echinococcus, and is a major economic and public health concern worldwide. Synthetic anthelmintics are most commonly used to control CE, however, prolonged use of these drugs may result in many adverse effects. This study aims to discuss the in vitro/in vivo scolicidal efficacy of different medicinal plants and their components used against Echinococcus granulosus. Google Scholar, ScienceDirect, PubMed and Scopus were used to retrieve the published literature from 2000–2020. A total of 62 published articles met the eligibility criteria and were reviewed. A total of 52 plant species belonging to 22 families have been reported to be evaluated as scolicidal agents against E. granulosus worldwide. Most extensively used medicinal plants against E. granulosus belong to the family Lamiaceae (25.0%) followed by Apiaceae (11.3%). Among various plant parts, leaves (36.0%) were most commonly used. Essential oils of Zataria multiflora and Ferula asafetida at a concentration of 0.02, and 0.06 mg/ml showed 100% in vitro scolicidal activity after 10 min post application, respectively. Z. multiflora also depicted high in vivo efficacy by decreasing weight and size while also causing extensive damage to the germinal layer of the cysts. Plant-based compounds like berberine, thymol, and thymoquinone have shown high efficacy against E. granulosus. These plant species and compounds could be potentially used for the development of an effective drug against E. granulosus, if further investigated for in vivo efficacy, toxicity, and mechanism of drug action in future research. [ABSTRACT FROM AUTHOR]

Human Parainfluenza virus (HPIV) causes lower respiratory tract infections (LRTI) mostly in young children. Respiratory viral infections may decline T cells in circulation and display enhanced pathogenicity. This study is aimed to analyze T cells alterations due to HPIV in children with LRTIs. Children (N = 152) with bronchitis or pneumonia, admitted in tertiary care hospitals were included in the study. Respiratory samples (throat or nasopharyngeal swabs) were taken and HPIV genotypes (1–4) were analyzed through RT-PCR. Peripheral blood T cells, CD3+, CD4+, CD8+, and CD19+, were analyzed in confirmed HPIV positive and healthy control group children through flow cytometry. The positivity rate of HPIV was 24.34% and the most prevalent genotype was HPIV-3 (20.40%). HPIV-1 and HPIV-2 were detected in 0.66% and 02% children respectively. The T lymphocyte counts were observed significantly reduced in children infected with HPIV-3. CD4+ cell (1580 ± 97.87) counts did not change significantly but the lowest CD8+ T cell counts (518.5 ± 74.00) were recorded. Similarly, CD3+ and CD19 cell ratios were also reduced. The CD4/CD8 ratio was significantly higher (3.12 ± 0.59) in the study population as compared to the control group (2.18 ± 0.654). Changes in the count of CD8+ T cells were more pronounced in patients with bronchiolitis and pneumonia. It is concluded that CD8+ T cells show a reduced response to HPIV-3 in children with severe LRTIs suggesting a strong association of these cells with disease severity. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii (T. gondii) is a protozoan parasite that infects all warm-blooded animals including domesticated birds and humans. Birds normally get infected by ground feeding and human beings contract the disease by consumption of undercooked chicken meat. This study aimed to analyze seroprevalence and DNA of T. gondii in chickens (domesticated and broiler) and to assess possible transfer to humans by review of available literature from Pakistan. Blood from and tissues from domesticated and broilers chickens were analyzed for Toxo-IgM/IgG and Toxoplasma DNA through ELISA and PCR respectively. Furthermore, research articles published during 1990–2019 on the prevalence of T. gondii in humans from Pakistan, were analyzed to assess the possible infection burden in the area in connection to transmission from chickens. The overall prevalence of IgM and IgG for T. gondii was 17.83% and 8.8% respectively in the study areas. Significant seroprevalence was found in domesticated chickens than broilers. In domesticated chickens, the prevalence was high in age ≥ 2 years. Toxoplasma DNA was detected in tissues with an overall prevalence of 10.84%. Higher prevalence was observed in liver (10.50%) than heart (9.5%) and muscles (7.11%). Only 4.78% broiler and 2.38% domesticated chickens were positive for both IgM and DNA, 1.2% domesticated and 1.30% broilers were positive for IgG and DNA, while 2.98% domesticated and 2.17% broilers were positive for IgM, IgG, and DNA. Available literature showed that 25.8% of human beings were infected with T. gondii in Pakistan. The prevalence was 20.64% in male and 26.82%in the female. The rate of infections increases with age and high (37.36%) was found in humans of age range 40 to 60 years. A high prevalence of T. gondii is found in both domesticated and broiler chickens in the study area. Moreover, the literature survey indicates that a high seroprevalence of T. gondii is present in human beings of Pakistan. It is concluded that the high prevalence of T. gondii in humans may be associated with the parasite transmission through infected chicken's meat in Pakistan. [ABSTRACT FROM AUTHOR]

Objective: Studies on diabetic foot ulcers (DFU) involving a representative sample of patients in Pakistan are scarce. This study aimed to determine baseline characteristics of infected DFUs in patients hospitalised at the Pakistan Institute of Medical Sciences Islamabad. Method: In this cross-sectional study, carried out during May 2015 and June 2016, foot ulcer characteristics of patients with DFUs were investigated and documented. From infected DFUs, aerobic bacterial pathogens were isolated, identified and evaluated for antimicrobial susceptibility. Results: A total of 214 patients were recruited to the study, 62.6% of which were male, 90.2% were aged ≥40 years, 76.2% had type 1 diabetes and 78.5% had poor glycaemic control at time of presentation to hospital. Most patients had grade 3/moderate ulceration (based on the Wagner and International Working Group on the Diabetic Foot/Infectious Diseases Society of America criteria). Over half of the DFUs (57.9%) were of ≤3 months' duration and 70.1% were ≥3 cm2. Of the patients with deep infection grade ulcers, 26.6% underwent amputation, accounting for their prolonged hospital stay (≥20 days). Significant differences were observed between patients with type 1 and type 2 diabetes with DFUs in relation to gender (p≤0.0001), ulcer size (p=0.0421) and duration of hospital stay (p=0.0253). The most significant predictors for lower extremity amputation were osteomyelitis (p=0.0114), retinopathy (p=0.0001) and neuropathy (p=0.0001. Piperacillin/tazobactam was found to be an effective antibiotic against the most commonly isolated Staphylococcus non-aureus (35.48%), Pseudomonas aeruginosa (22.26%), and Staphylococcus aureus (20.96%) species indentified in the DFU infections. Conclusion: The findings of this study may be helpful in the optimal management and appropriate treatment of patients with infected DFUs. [ABSTRACT FROM AUTHOR]

The present study aimed to investigate the efficacy of traditionally used anti-mastitis plants ( Allium sativum, Bunium persicum, Oryza sativa and Triticum aestivum ) in northwest Pakistan against bacterial pathogens. Selected plants were phytochemically screened for Alkaloids, Flavonoids, and Saponins and checked for in vitro antibacterial activity at concentration of 50 mg/ml against S. aureus, E. coli and K. pneumoniae by agar well diffusion method. Minimum inhibitory concentration and minimum bactericidal concentration was determined against multidrug resistant bacteria using tube dilution method. All extracts were found to significantly inhibit (p < 0.01, p < 0.05) the activity against bacterial strains examined. Among phytochemicals, alkaloids of all tested antimastitis plants produced significantly higher inhibition zones against bacteria. The minimum inhibitory concentration and minimum bactericidal concentration of phytochemicals and crude methanolic extracts against tested bacterial strains ranged between 12.5–50 mg/ml and 25–50 mg/ml, respectively. Medicinal plants traditionally used against mastitis are therapeutically active against bacterial pathogens. A. sativum and B. persicum were found to be potential candidate species for the development of novel veterinary drugs with low cost and fewer side effects. [ABSTRACT FROM AUTHOR]

Toxoplasmosis is the major cause of abortion and stillbirths among women of childbearing age. The aim of this study is to determine chronic Toxoplasma gondii infection at various trimesters of pregnancy and to identify various possible routes of transmission and contamination in North-West Pakistan. A total of 733 blood samples were randomly obtained from antenatal/maternity centres. A detail questionnaire was designed to analyse the clinical history of the subjects tested for specific anti-Toxoplasma IgG antibodies by using immunofluorescence assay. Overall toxoplasmosis was detected in 18.41% of the studied population. Anti-Toxoplasma IgG was found 7.17% in first trimester with highest prevalence 31.28% in third trimester. Increase in the prevalence of parasite was seen with rise in age highest found in higher age group (P ˂ 0.05). As compared to district Lower Dir and Swat, Upper Dir had high rate 33.03% of chronic infection. Consumption of undercooked meat was also found 45.22% related with the prevalence of infection. A significant relation was observed with level of education (P ˂ 0.05) and prevalence of toxoplasmosis. An association was observed between prevalence of T. gondii infection and various risk factors, i.e. consumption of raw eggs, vegetables, contact with cats, cattle, soil, water sources and use of unpasteurized milk. This study shows high prevalence of the parasite in the studied area. Pregnant women in contact with cats were found more at risk. Education of childbearing age women, personal hygiene and contact with soil was also found important possible risk factors in the spread of infection. [ABSTRACT FROM AUTHOR]

In this study, five different localities viz., Dergai, Warsak, Dera Ismail Division, Agriculture Research Institute Tarnab and Peshawar University of Khyber-Pakhtun-Khwa province, were surveyed for plant parasitic nematodes. Sixteen species of plant parasitic nematodes out of which 11 are being reported for the first time in this region, and 4 previously reported in Pakistan. The highest numbers of nematode species were found in Dera Ismail Khan, followed by Dergai, Warsak and Agriculture Research Institute Tarnab. [ABSTRACT FROM AUTHOR]

Background: Plasmodium vivax is one of the widespread human malarial parasites accounting for 75% of malaria epidemics. However, there is no baseline information about the status and nature of genetic variation of Plasmodium species circulating in various parts of Pakistan. The present study was aimed at observing the molecular epidemiology and genetic variation of Plasmodium vivax by analysing its merozoite surface protein-3α (msp-3α) and merozoite surface protein-3β (msp-3β) genes, by using suballele, species-specific, combined nested PCR/RFLP detection techniques. Methods: A total of 230 blood samples from suspected subjects tested slide positive for vivax malaria were collected from Punjab, Sindh, Khyber Pakhtunkhwa, and Balochistan during the period May 2012 to December 2013. Combined nested PCR/RFLP technique was conducted using Pvmsp-3α and Pvmsp-3β genetic markers to detect extent of genetic variation in clinical isolates of P. vivax in the studied areas of Pakistan. Results: By PCR, P. vivax, 202/230 (87.82%), was found to be widely distributed in the studied areas. PCR/RFLP analysis showed a high range of allelic variations for both msp-3α and msp-3β genetic markers of P. vivax, i.e., 21 alleles for msp-3α and 19 for msp-3β. Statistically a significant difference (p ≤ 0.05) was observed in the genetic diversity of the suballelic variants of msp-3α and msp-3β genes of P. vivax. Conclusion: It is concluded that P. vivax populations are highly polymorphic and diverse allelic variants of Pvmsp-3α and Pvmsp-3β are present in Pakistan. [ABSTRACT FROM AUTHOR]

Background: The structural and functional differences between hepatitis B virus (HBV) genotypes are the mainstay to severity, complications, treatment and possibly vaccination against the virus. This study was conducted to determine the HBV genotypes in HBsAg positive patients of Afghanistan as no such large scale data available previously. Methods: Two hundred and fourteen HBsAg-positive patients were included in this study. All patients were anti-HCV and anti-HIV negative. All the samples were confirmed for HBV DNA with nested PCR while HBV DNA positive samples were subjected to type specific PCR for HBV genotyping (A-F). Results: Of the total samples, 168 (78.5%) were males and 46 (21.49%) females, aged ranged between 18 to 71 years. This study demonstrated that genotype D (35.67%) is the predominant genotype circulating in Afghani's population. Genotype C was observed in 32.16% followed by genotype A (19.30%), and genotype B (7.02%) while 6.07% of the individuals were not typed. Conclusion: This study has shown a heterogeneous distribution of HBV genotypes. Further more, extensive studies are required to investigate genetic and geographical divergence and characteristics of the virus in the country, as no such large sample sized study has been carried out so far in this country. [ABSTRACT FROM AUTHOR]

Background: Hepatitis B Virus (HBV) may progress to serious consequences and increase dramatically beyond endemic dimensions that transmits to or from health care workers (HCWs) during routine investigation in their work places. Basic aim of this study was to canvass the safety of HCWs and determine the prevalence of HBV and its possible association with occupational and non-occupational risk factors. Hepatitis B vaccination coverage level and main barriers to vaccination were also taken in account. Results: A total of 824 health care workers were randomly selected from three major hospitals of Peshawar, Khyber Pakhtunkhwa. Blood samples were analyzed in Department of Zoology, Kohat University of Science and Technology Kohat, and relevant information was obtained by means of preset questionnaire. HCWs in the studied hospitals showed 2.18% prevalence of positive HBV. Nurses and technicians were more prone to occupational exposure and to HBV infection. There was significant difference between vaccinated and non-vaccinated HCWs as well as between the doctors and all other categories. Barriers to complete vaccination, in spite of good knowledge of subjects in this regard were work pressure (39.8%), negligence (38.8%) un-affordability (20.9%), and unavailability (0.5%). Conclusions: Special preventive measures (universal precaution and vaccination), which are fundamental way to protect HCW against HBV infection should be adopted. [ABSTRACT FROM AUTHOR]

Background: Hepatitis C virus (HCV) genotype 3a is known to show comparatively better response to combination therapy than genotype 1 and 4. Mutations within NS5A gene of HCV have earlier been implicated with response to interferon (IFN) therapies in chronic HCV patients among various populations. As response to therapy are available in different populations because of the ethnic and viral factors and there was no study available on the phenomenon of resistivity to IFN. Results: Chronic HCV 3a infected Pakistani patients were kept on IFN-α and ribavirin therapy for six months. NS5A gene of HCV was amplified and sequenced in the case of all the patients prior to therapy and the sequences were analysed for mutations. Out of the total 27 patients, 20 (74.07%) were observed with sustained virological response (SVR), 4 (14.81%) patients were non responder (NR) while 3 (11.11%) patients exhibited in end of treatment response (ETR). Three (3/20) (15%) SVR patients and two (2/3) ETR patients had mutations (ranging from I-V amino acids) within the NS5A ISDR regions. While the rest of the SVR patients (85%) and the NR had no mutations at ISDR region when compared with HCV K3a ISDR. Conclusions: Mutations within the NS5A gene of HCV 3a genotype may not influence the outcome of combination therapy in Pakistani populations. [ABSTRACT FROM AUTHOR]

Background: Studies of the molecular epidemiology and risk factors for hepatitis C virus (HCV) in health care workers (HCWs) of Peshawar, Khyber Pakhtunkhwa region are scarce. Lack of awareness about the transmission of HCV and regular blood screening is contributing a great deal towards the spread of hepatitis C. This study is an attempt to investigate the prevalence of HCV and its possible association with both occupational and nonoccupational risk factors among the HCWs of Peshawar. Results: Blood samples of 824 HCWs, aged between 20-59 years were analysed for anti-HCV antibodies, HCV RNA and HCV genotypes by Immunochromatographic tests and PCR. All relevant information was obtained from the HCWs with the help of a questionnaire. The study revealed that 4.13% of the HCWs were positive for HCV antibodies, while HCV RNA was detected in 2.79% of the individuals. The most predominant HCV genotype was 3a and 2a. Conclusion: A program for education about occupational risk factors and regular blood screening must be implemented in all healthcare setups of Khyber Pakhtunkhwa province in order to help reduce the burden of HCV infection. [ABSTRACT FROM AUTHOR]

Background: Haemonchus contortus is an important pathogenic nematode parasite and major economic constraint of small ruminants in tropics and subtropics regions. This review is an attempt to systematically address the; (a) efficacy of different plants against H. contortus by in vitro and in vivo proof; (b) toxicology, mechanism of action, and active phyto-compounds involve in anti-haemonchiasis activity; (c) and comparative analysis of plant species evaluated both in vitro and in vivo. Methods: Online databases (Google Scholar, PubMed, Scopus, and ScienceDirect) were searched and published research articles (1980–2020) were gathered and reviewed. Results: A total of 187 plant species were reported belonging to 59 families and 145 genera with Asteraceae and Fabaceae being frequently used. Out of the total plant species, 171 species were found to be evaluated in vitro and only 40 species in vivo. Twenty-four species were commonly evaluated for in vitro and in vivo anti-haemonchiasis activity. Among the reported assays, egg hatching test (EHT) and fecal egg count reduction (FECR) were the most widely used assays in vitro and in vivo , respectively. Moreover, sheep were the frequently used experimental model in vivo. After comparative analysis, Lachesiodendron viridiflorum , Corymbia citriodora , Calotropis procera , and Artemisia herba-alba were found highly effective both in vitro and in vivo. L. viridiflorum inhibited enzymatic activities and metabolic processes of the parasite and was found to be safe without toxic effects. C. citriodora was moderately toxic in vivo , however, the plant extract produced promising nematicidal effects by causing muscular disorganization and changes in the mitochondrial profile. Additionally, C. procera and A. herba -alba despite of their high anti-haemonchiasis activity were found to be highly toxic at the tested concentrations. C. procera caused perforation and tegumental disorganization along with adult worm paralysis. Nineteen compounds were reported, among which anethole and carvone completely inhibited egg hatching in vitro and significantly reduced fecal egg count, decreased male length, and reproductive capacity of female in vivo. Conclusion: This review summarized different medicinal plants owing to nematicidal activities against H. contortus eggs, larvae, and adult worms. Plants like L. viridiflorum, C. citriodora, C. procera, and A. herba-alba , while compounds anethole and carvone having promising nematicidal activities and could be an alternative source for developing novel drugs after further investigation. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) continues a persistent challenge to health and development worldwide. A total of 153 sputum samples were collected from Khyber Teaching Hospital Peshawar Khyber Pakhtunkhwa, from patients suspected for pulmonary TB and were studied under microscope. Of the total samples collected, 49 (32.02%) were positive for TB. Female (33.33%) were more infected than male (30.66%). High occurrence (68.96%) was recorded in age group 10-20 year, followed by 21-40 years (35.29%) and the lowest was recorded (15.06%) in individuals with age more than 40 years. It is concluded that pulmonary TB is still prevalent in district Peshawar, Khyber Pakhtunkhwa Pakistan. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance In remote areas, medicinal plants have an imperative role in curing various livestock's ailments. In Pakistan, people residing in remote areas including South Waziristan Agency and Bajaur Agency depend on traditional herbal remedies for treating their domestic animals. Medicinal plants are an important part of the medical system in these Agencies. The prime goal of the current study is to explore the ethnoveterinary practices in the two regions and discuss cross-cultural consensus on the use of medicinal plants. In this study, we have given detailed description on the ethnoveterinary usage of certain medicinal plants and their recipes. Moreover, we have also elaborated the ethnoveterinary potential of certain plants in relation to their ethnomedicinal, pharmacological and phytochemicals reports. Methodology Fieldwork comprised of two fields surveys conducted at South Waziristan Agency and Bajaur Agency. A total of 75 informants from South Waziristan Agency and 80 informants from Bajaur Agency were interviewed with the help of semi-structured questionnaires. Use reports (URs) were recorded for all the documented taxa. Data were quantitatively analyzed by using informant consensus factor (F ic ) index in order to find out information homogeneity provided by the informants. To analyze the cross-cultural consensus, the recorded data were tabulated as well as shown by Venn diagram. Results Overall, 94 medicinal plant taxa were recorded in the comparative analysis. Out of these, most of the plants species (72 species) were used at Bajaur Agency than South Waziristan Agency (37 species). Cross-cultural analysis showed that only 15 medicinal plants were used in common by the indigenous communities in both Agencies, which indicates a low interregional consensus with regard to the ethnoveterinary practices of medicinal plants. Apiaceae was the dominant family in both regions by representing maximum number of plant species (11 species). Gastro intestinal complexities were common in both regions having higher F ic values (above 90). Moreover, the current investigation reported new ethnoveterinary uses of medicinal plants from South Waziristan Agency, which were Sideroxylon mascatense, Raphanus sativus, Salix babylonica, Solanum nigrum, Sophora mollis, Taraxacum campylodes and Tulipa stellata . On the other hand from Bajaur Agency, Boerhavia erecta, Celtis australis , Chamaecyparis obtusa var. obtuse , Eryngium biehersteinianum , Gossypium arboreum , Narcissus tazetta , Opuntia littoralis , Streblus asper were reported with new ethnoveterinary uses. Conclusions The current study has an important contribution towards the preservation of indigenous plants’ based knowledge. Several plants are carrying important ethnoveterinary uses being practiced by the local people mostly against the gastrointestinal disorders in both regions. Importantly, the cross-cultural approach has reported some new traditional uses of plants against livestock's diseases. Hence, this is an opportunity to investigate such plants phytopharmacologically and toxicologically for the discovery of new drug sources. [ABSTRACT FROM AUTHOR]

Cystic echinococcosis (CE) is a neglected zoonotic disease prevalent in Pakistan, but the genetic diversity of the cestode is largely unexplored in the country. This study investigated the molecular epidemiology of CE infecting the livestock population of the Malakand division, Khyber Pakhtunkhwa, Pakistan. A total of 1,200 livestock, including buffaloes, cattle, goats, and sheep, were examined for echinococcosis from November 2017-2018 at different slaughterhouses in the Malakand division. Hydatid cysts were collected from different organs, and hydatid cyst fluid (HCF) was examined microscopically and used for DNA extraction. The LSU (rrnl) and NAD1 genes were amplified and sequenced. The overall prevalence of CE was 17% (204/1,200), including cows (21.7%), buffaloes (17.4%), goats (10%), and sheep (9.6%). The infection was relatively more prevalent among males (17%) than females (16.9%) and animals of older age (>5 years) ( p = 0.710). Liver (63.2%) and lungs (25%) were more affected as compared to kidneys (6.8%) and heart (4.9%). HCF analysis indicated that 52.0% of the cysts were sterile and (48.0%) were fertile. Sequencing and phylogenetic analyses confirmed 80.0% of the isolates as Echinococcus granulosus sensu stricto (G1-G3) in all animal species, while Echinococcus equinus (G4) and Echinococcus ortleppi (G5) were present in buffaloes. The present study concluded that CE is prevalent in the livestock population of Malakand. Besides E. granulosus s. s. (G1-G3), E. ortleppi genotype (G5) and E. equinus (G4) in livestock were also reported., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Khan, Basharat, Khan, Jamal, Rahman, Shah, Khan, Ali, Khan and Ali.)

Medicinal plants have been used from ancient time against different infectious diseases caused by microorganisms across the globe. The present review represents different medicinal plants of Pakistan used traditionally for the treatment of variety of ailments caused by Pseudomonas aeruginosa, their in-vitro activities against P. aeruginosa and phytochemistry. These plants were extracted with different solvents that showed good in-vitro activities against P. aeruginosa, due to the presence of active phytoconstituents including alkaloids, terpenoids etc. Among all the solvents used for extraction process, alcoholic extracts were mostly preferred in Pakistan. However, non-alcoholic solvents like ethyl acetate and chloroform also showed good anti-P. aeruginosa activities. Statistically, increase in concentration (mg/ml) of ethyl acetate and chloroform extracts significantly increase (p=0.000 and p= 0.046) inhibitory activity against P. aeruginosa. This review provides scientific validation of the traditional knowledge in using medicinal plants for the treatment of different diseases caused by this bacterium. Reported Pakistani medicinal plants contain variety of phytochemical compounds that could be very useful in the production of new drugs with fewer side effects on living system compared to some allopathic drugs. This review also provides baseline information for future research studies on the phytochemistry of unexplored plants. Further research studies should be carried out on non-alcoholic extracts that could be helpful in the extraction new compounds, which could lead to the development of some novel drugs in the pharmaceutical industries of Pakistan.

Background: Aurora kinases (Aurora-A, B and C) belong to a family of conserved serine/threonine kinases which are key regulators of cell cycle progression. Aurora-A and Aurora-B are expressed in somatic cells and involved in cell cycle regulation while aurora-C is meiotic chromosome passenger protein. As Aurora kinase C is rarely expressed in normal somatic cells and has been found over expressed in many cancer lines. It is suggested that Aurora-C-T191D is not hyperactive mutant., Result: Aurora-C-T191D variant form was investigated and compared with wild type. The overexpression of Aurora-C-T191D was observed that it behaves like Aurora-C wild type (aurC-WT). Both Aurora-C-T191D and aurC-WT induce abnormal cell division resulting in centrosome amplification and multinucleation in transiently transfected cells as well as in stable cell lines. Similarly, Aurora-C-T191D and aurC-WT formed foci of colonies when grown on soft agar, indicating that a gain of Aurora-C activity is sufficient to transform cells. Furthermore, we reported that NIH-3 T3 stable cell lines overexpressing Aurora-C-T191D and its wild type partner induced tumour formation when injected into nude mice, demonstrating the oncogenic activity of enzymatically active Aurora kinase C. Interestingly enough tumour aggressiveness was positively correlated with the rate of kinase activity, making Aurora-C a potential anti-cancer therapeutic target., Conclusion: These findings proved that Aurora C-T191D is not hyperactive but is constitutively active mutant.

Aim: High prevalence of Hepatitis C virus (HCV) has been reported among the dialysis patients throughout the world. No serious efforts were taken to investigate HCV in patients undergoing hemodialysis (HD) treatment who are at great increased risk to HCV. HCV genotypes are important in the study of epidemiology, pathogenesis and reaction to antiviral therapy. This study was performed to investigate the prevalence of active HCV infection, HCV genotypes and to assess risk factors associated with HCV genotype infection in HD patients of Khyber Pakhtunkhwa as well as comparing this prevalence data with past studies in Pakistan., Methods: Polymerase chain reaction was performed for HCV RNA detection and genotyping in 384 HD patients. The data obtained was compared with available past studies from Pakistan., Results: Anti HCV antibodies were observed in 112 (29.2%), of whom 90 (80.4%) were HCV RNA positive. In rest of the anti HCV negative patients, HCV RNA was detected in 16 (5.9%) patients. The dominant HCV genotypes in HCV infected HD patients were found to be 3a (n = 36), 3b (n = 20), 1a (n = 16), 2a (n = 10), 2b (n = 2), 1b (n = 4), 4a (n = 2), untypeable (n = 10) and mixed (n = 12) genotype., Conclusion: This study suggesting that i) the prevalence of HCV does not differentiate between past and present infection and continued to be elevated ii) HD patients may be a risk for HCV due to the involvement of multiple routes of infections especially poor blood screening of transfused blood and low standard of dialysis procedures in Pakistan and iii) need to apply infection control practice.