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Isodon rugosus Wall. ex Benth (IR) is one of the ethnomedicinal important plants of Gilgit-Baltistan, Northern Pakistan. The present study aims to evaluate the micro-morphological features, phytochemical screening, and pharmacological potential of IR. SEM and LM were used as an identification tool. Five different solvents were used to prepare IR extracts. Phytochemical and antioxidant activities were determined calorimetrically. To investigate antidiabetic, α-amylase inhibition assay was adopted. Cytotoxicity was tested using a brine shrimp assay. Anti-leishmanial via MTT assay. Disc-diffusion assay was used for protein kinase inhibitory, antibacterial, and antifungal activities. Pollen was of monad, hexacolporate, and circular shape. Lophate sculpturing with outer exine pattern elevated on the rides. Seeds were oblong-ovate, small projected with smooth-rough, variously ridges/wrinkled sculpturing. Epidermis cells were irregular in shape with a slightly straight anticlinal wall. Stomata with diacytic, and unicellular glandular trichome were observed. The extracts were rich in phytochemicals, the maximum amount of phenolic and flavonoid contents was found in methanol extract (IRM) 89.76mg GAE/gm and 85.69mg QE/gm. All the extracts show substantial antioxidant activity but highest in IRM (DPPH IC50 44.51 μg/ml, total antioxidant capacity 93.60 mg AAE/g, and total reduction power of 93.44 mg AAE/g). Potential antibacterial and antifungal activities were reported for IRM. Significant protein kinase, alpha-amylase inhibition, and cytotoxic activity were revealed. Dose "dependent cytotoxic activity was exposed against Leishmania tropica (LC50 11.16 μg/mL). In conclusion, I. rugosus extracts have shown potential biological applications and should be subjected to further research work to develop further biomedical applications. [ABSTRACT FROM AUTHOR]

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Microbial proteolytic enzymes are used to hydrolyze proteins in various bio-industrial applications but most of the enzymes are not active in extreme temperature conditions, pH and salt concentrations. This increases the demand to explore enzymes to show activity at such extreme environments. Exciting findings have been described about the extremophilic bacteria isolated first time from coal mines of Orakzai Agency, Pakistan, having protease producing properties. Among various extracted isolates, ~6 isolates were purified based on showing proteolytic activity using 2% skim milk agar media at 37 °C for 24 hrs, to identify various morphological and biochemical characteristics. The isolate, IBC-2 showed maximum zonal activity (21 mm) and was identified as Pseudomonas aeruginosa (JCM 5962) using 16S rRNA gene sequencing. The strain IBC-2 exhibited significantly (P ≤ 0.05) maximum growth and protease production (335 U/ml) at 37 °C and at pH 7 after 24 hrs of incubation. The strain was also significantly (P ≤ 0.05) more active for protease production at neutral pH (435 U/ml) than at high or low pH, reflected neutral nature of the bacteria. In conclusion, the strain IBC-2 has highest protease producing ability with distinctiveness, which can be used for wide range bio-industrial applications. [ABSTRACT FROM AUTHOR]

This study investigates the pharmacological importance of selected wild mushrooms viz., Morchella esculenta (common morel), Calvatia gigantea (Giant puffball) and Astraeus hygrometricus (False earthstar) collected from South Waziristan Agency, Federally Administered Tribal Areas (FATA), Pakistan. The selected mushrooms were collected from Tehsil Makeen, Wana and Birmal of South Waziristan Agency during a sampling survey conducted in April–May 2010. The dry fruiting bodies of mushrooms were methanol extracted and evaluated for total phenolic, protein, antioxidant activity, cytotoxicity and their effects on radish seed growth. The results revealed that methanol crude extracts prepared from fruiting bodies of A. hygrometricus and C. gigantea have higher phenolic content and total antioxidant activity as well as greater brine shrimp cytotoxicity. On the other hand, M. esculenta has a high level of protein content and promoted seedling growth in radish. Antioxidant activity for A. hygrometricus, C. gigantea and M. esculenta at IC 50 values were: 9.3 ± 0.3 μg/ml, 22.2 ± 0.3 μg/ml and 18.0 ± 0.1 μg/ml, respectively. Methanolic extract of C. gigantea (10 mg/ml) reduced the seed germination and shoot length of radish by 16%. In contrast, the methanolic extract (10 mg/ml) of M. esculenta and C. gigantea enhanced the shoot length, root length and root/shoot ratio of radish. Higher LC 50 value was recorded for methanolic extract of A. hygrometricus (19.0 ± 0.3 μg/ml) followed by M. esculenta (17 ± 0.19 μg/ml), whereas methanolic extract of C. gigantea showed lower LC 50 value (16 ± 0.23 μg/ml). It is inferred from the present investigation that mushrooms collected from South Waziristan could be potential source of compounds with beneficial biological activities. [ABSTRACT FROM AUTHOR]

Bovine leukosis is an economically important disease of dairy cattle caused by the bovine leukaemia virus (BLV). The study aimed to determine the seroprevalence, haemato-biochemical effects, and risk factors pertinent to the prevalence of bovine leukosis in Holstein–Friesian purebred dairy cattle in the D.I. Khan region of Pakistan. A total of 192 sera were assayed using an enzyme-linked immunosorbent assay. Overall, 31.3% (60/192) cattle were detected as seropositive. There was a marked increase in total leukocyte count (11.29 ± 0.48×10³ /µl), lymphocytes (5.73 ± 0.42%), monocytes (0.81 ± 0.06%), haemoglobin (11.09 ± 0.46 g/dl), red blood cells (7.23 ± 0.37 ×106 /µl), and packed cell volume (31.75 ± 1.48%) in seropositive cattle. Serum biochemical parameters in seropositive cattle showed a marked increase in the liver enzymes, alanine transaminase (24.25 ± 1.03 U/l) and aspartate aminotransferase (49.33 ± 3.31 U/l), with a marked decrease in glutathione peroxidase (1365.63 ± 12.03 (U/l) and superoxide dismutase (2.14 ± 0.13 U/ml) activity. A significant association of age, pregnancy, breeding method, milk yield, and health status of seropositive animals with bovine leukosis was also recorded. The prevalence was higher in animals which were older, pregnant, artificially inseminated, low milk producers, and had a history of ailments. The current study found that bovine leukosis virus could cause changes in internal homeostasis, oxidative stress, and liver dysfunction, all of which should be considered during a control regimen. It was concluded that bovine leukosis was moderately prevalent in the D.I. Khan region in Pakistan. [ABSTRACT FROM AUTHOR]

This research purpose was focused to document the traditional ethnobotanical knowledge (TEK) associated with important wild plants and indigenous communities residing in mountainous zone of District Sudhnoti of Azad Jammu and Kashmir, Pakistan and to determine their conservation status with suggestions and recommendations to protect and propagate the rare and endangered wild flora of the area for sustainable use. The data regarding traditional ethnomedicines (TEMs) and phytogeography with conservation analysis of wild plants were collected via structured and semi structured interviews of 150 participants belonging to various professions and of both genders. Prior consent and permission were obtained from family heads and from all the participants, and the data were shared with all the informants through the local female translator guide. To validate the collected data, various microstatistical tools, such as the informant consensus factor (ICF), fidelity level (FL) and data matrix ranking (DMR) were used. The relative frequency of citations (RFC) and rank order of popularity (ROP) were also calculated to determine and authenticate the relative importance of TEMs which may be further studied in future research and used for drug discovery. In the present study, 150 locally important plants belonging to 69 families were documented, and the Rosaceae was the most dominant and prevalent family. The results indicated that many locally important plants have multiple uses such as food, medicine, fodder, shelter, ornamental and fuel. The TEMs obtained from these plants have been known to be useful for curing various infirmities such as flu, renal disorders, fever, malaria, cough, migraine, whooping cough, influenza, skin rashes, allergies, stomach aches, wounds and bruises, diabetes, tumours and joint pains. The ICF analysis revealed that renal calculus, malaria, fever, whooping cough, rheumatoid arthritis and arthritis were prevalent infirmities of the mountainous area. The FL analysis indicated the popularity of these plants used in various TEMs and among these Artemisia maritima and Berberis lycium were the most common. The DMR indicated that Pinus roxburghii was the most common species being used for multipurpose by the indigenous communities. The highest use value index was found for Indigofera linifolia. The relative frequency of citations was calculated to determine the importance of plants in traditional ethnomedicine, highlighting Morus nigra, Pinus wallichiana, and Rosa indica as significant species. The Jaccard index indicated a high level of novelty in the research which can be exploited for neo-drug discovery and drug development. In conclusion, this study has successfully documented the ethnobotanically and ethnomedicinally important plants in District Sudhnoti, Azad Jammu and Kashmir, Pakistan. The findings of this research contribute to the preservation and documentation of cultural heritage, as well as provide a foundation for further studies in ethnobotany, ethnopharmacology and biodiversity conservation efforts for sustainable provision of wild flora to the indigenous communities which will also assist in combating drasting climatic changes., (© 2024. The Author(s).)

Anaplasma spp. and Rickettsia spp. are intracellular vector-borne pathogens and harbored by a wide range of ticks and vertebrate hosts. Aim of this study was to molecularly characterize Anaplasma spp. and Rickettsia spp. in different ticks collected from livestock hosts in nine districts of Khyber Pakhtunkhwa (KP), Pakistan. In total, 862 ticks were collected from cattle, goats and sheep. Highest tick's infestation was observed on cattle 56.14% (32/57), followed by goats 45.45% (40/88), and sheep 42.05% (45/107). Rhipicephalus microplus (305/862, 35.38%) was predominant species, followed by Haemaphysalis sulcata (243/862, 28.19%), Hyalomma anatolicum (133/862, 15.42%), Haemaphysalis bispinosa (120/862, 13.92%), and Hyalomma kumari (61/862, 7.07%). A subset of 135 ticks were screened for Anaplasma spp. and Rickettsia spp. based on the amplification of partial 16 S rDNA and outer-membrane protein A (ompA) fragments, respectively. In total, 16 ticks (11.85%) were positive for Anaplasma spp. and Rickettsia spp. Obtained 16 S rDNA sequences for Anaplasma spp. detected in Ha. bispinosa and Ha. sulcata showed 99.98% identity with Anaplasma bovis, while other detected in Rh. microplus showed 99.84% identity with Candidatus Anaplasma boleense. Similarly, detected ompA sequence in Ha. sulcata showed 100% identity with Rickettsia sp. and 97.93% with Rickettsia slovaca, and another sequence detected in Rh. microplus showed 100% identity with Candidatus Rickettsia shennongii. In phylogenetic trees, these sequences clustered with corresponding species from Pakistan, China, Turkey, South Korea, South Africa, and Herzegovina. This is the first study reporting detection of A. bovis in Ha. bispinosa and Ha. sulcata, Ca. A. boleense in Rh. microplus collected from goats, and R. slovaca-like in Ha. sulcata. Our results enforce the need for regular surveillance of Rickettsiales in hard ticks infesting livestock in the region., (© 2024. The Author(s).)

Background: Sarcopenia's impact on post-liver transplant outcomes remains a subject of debate, with limited data from South Asia on its association with post-liver transplant hospital stays. This study aims to investigate sarcopenia's influence on post-transplant hospitalization duration in South Asians., Methods: In this retrospective study, patients with liver cirrhosis who underwent living-donor liver transplantation (LDLT) at Shifa International Hospital in Islamabad, Pakistan, between January 2022 and January 2023 were included. Computed tomography (CT) images were used to assess the skeletal muscle index (SMI). The areas of the psoas, erector spinae, multifidus, quadratus lumborum, rectus abdominis, transverse abdominis, and internal/external oblique muscles were quantified at the level of L3. The data were analyzed using SPSS version 29.0 (IBM)., Results: There was a total of 84 patients. Mean age was 47.4 ± 12.0 years. There were 62 (73.8%) male patients and 22 (26.2%) female patients. Hepatitis C was noted in 36 (42.9%) patients. Twenty-two (26.2%) patients had hepatocellular carcinoma. Sarcopenia was identified in 58 (69.0%) patients. No significant association was observed between sarcopenia and intensive care unit (ICU) or general floor stays. Regression analysis identified pre-transplant model for end-stage liver disease-sodium (MELD-Na) score as the sole significant factor associated with both ICU and total length of stay (P value .002; P value .009)., Conclusion: In our population, sarcopenia did not correlate with post-transplant ICU or overall hospital stay. The pre-transplant MELD-Na score emerged as the most influential predictor of length of stay. Therefore, delaying liver transplant procedures based on muscle mass estimations may not be a practical clinical approach for South Asian patients., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Baluchistan's water profile was developed by dividing it into seven zones (Northern Highlands-NH, Southern Highlands-SH, Quetta Valley-QV, Desert-D, Sibbi Plains-SP, Coastal Lasbella-CL, Coastal Gwadar-CG) based on geography, water availability, and climate of the area. A total of 106 water samples were collected from karaiz, spring water, and tube wells. Spatial distribution of EC, TDS, TH, SO 4 2- , Cl - , Na + , and K + showed an increasing trend in concentration from the highlands towards the desert and coastal zones. For anion, HCO 3 - is predominant in NH, SH, and QV, Cl - in D, CL, and CG and only SO 4 2- in SP, whereas the cationic trend in overall zones is Na + >Ca 2+ >Mg 2+ >K + . In the NH, SH, QV, and SP zones, the physicochemical parameters met the drinking water quality guidelines; however, D, CL, and CG exceeded in almost all quality parameters. Furthermore, the drinking water quality index (WQI) shows excellent to good water quality in NH, SH, QV, and D zones, while CL and CG fall in poor to unsuitable water classes. In terms of hydrogeochemical facies, maximum water samples from NH fall in Ca-Mg-HCO 3 , and SH, QV, and SP in Ca-Mg-Cl type, where major ion chemistry is controlled by rock-weathering, while D, CL, and CG fall in the NaCl type, where evaporation is dominant. Similarly, irrigation water quality parameters (EC, SAR, RSC, Na%, MH%, PI, SSP, and KR) reveal that NH, SH, QV, and SP have suitable water for irrigation, and D, CL, and CG require proper treatment. Additionally, USSL and Wilcox's diagrams indicated that NH, SH, QV, and SP have "excellent to permissible"; however, D, CL, and CG have "permissible to unsuitable" class water, requiring special management practices. Consequently, appropriate control measures and targeted water purification programmes should be implemented to protect the public health and sustainability of water resources in Baluchistan., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

End-stage renal disease (ESRD) patients are mostly managed with maintenance hemodialysis (MHD). ESRD patients on MHD also present with many complications, such as anemia, hyperparathyroidism, and hepatitis prevalence. This study depicts the real-world scenario of anemia among MHD and end-stage renal disease patients in the Pakistani population. A retrospective, multicentric, and real-world data analytical study was conducted at 4 dialysis centers in Pakistan. The study had a sample size of n  = 342 patients on maintenance hemodialysis. The data were gathered from the medical records of patients. Data analysis was performed using STATA Version 16. Statistical significance was gauged at a 0.05 level of significance. According to our results, the mean age of the patients was 45 (±15) years. Most of the patients were male ( n  = 234, 68.4%), whereas 58.1% of the patients were maintained on twice-weekly hemodialysis. The most commonly reported comorbidities were hypertension and diabetes mellitus. The frequency of dialysis ( P  < 0.01) and comorbidities ( P  = 0.009) had a significant association with anemia in MHD patients. The majority of the patients had hyperparathyroidism (52%) with anemia. Upon performing binary logistic regression, multivariate analysis displayed a similar odds value for having anemia in patients with every additional month in the duration of hemodialysis (OR 1.01, P  = 0.001), the odds of anemic patients having a positive antihepatitis-C antibody (OR 2.22, P  = 0.013), and the odds of having anemia in patients in the age category below 45 years (OR 1.93, P  = 0.013). In conclusion, the study results depict that every additional month in the duration of hemodialysis, age (<45 years), and positive anti-HCV antibody status, these variables were more likely to have anemia in our study MHD patients. While in our final multivariate model, no statistically significant association was observed between hyperparathyroidism and anemia., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2023 Humera Bukhari et al.)

Infertility is a multifactorial disorder that affects approximately 12% of couples of childbearing ages worldwide. Few studies have been conducted to understand the genetic causes of infertility in depth. The synaptonemal complex (SC), which is essential for the progression of meiosis, is a conserved tripartite structure that binds homologous chromosomes together and is thus required for fertility. This study investigated genetic causes of infertility in a Pakistani consanguineous family containing two patients suffering from non-obstructive azoospermia (NOA). We performed whole-exome sequencing, followed by Sanger sequencing, and identified a novel pathogenic variant (c.7G > A [p.D3N]) in the SC coding gene C14orf39, which was recessively co-segregated with NOA. In silico analysis revealed that charges on wild-type residues were lost, which may result in loss of interactions with other molecules and residues, and a reduction in protein stability occurred, which was caused by the p.D3N mutation. The novel variant generated the mutant protein C14ORF39 D3N , and homozygous mutations in C14orf39 resulted in NOA. The transcriptome profile of C14ORF39 shows that it is specifically expressed in early brain development, which suggests that research in this area is required to study other functions of C14ORF39 in addition to its role in the germline. This research highlights the conserved role of C14orf39/SIX6OS1 in assembly of the SC and its indispensable role in facilitating genetic diagnosis in patients with infertility, which may enable the development of future treatments., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Pollen micromorphology is not only used to check the functional and structural evolution in plants but also to solve the taxonomic problem related to the classification of plants. Therefore, keeping in view the significance of pollen traits, selected taxa of the subfamily Caesalpiniaceae was collected from different geographical regions of Pakistan. The species were then analyzed under both light microscopy and scanning electron microscopy techniques to investigate the importance of micromorphological characters of pollen in the identification and classification of species. Great variation was recorded in equatorial shape, surface ornamentation, tectum, polar diameter, equatorial diameter, and exine thickness. However, little variation was observed in pollen type, polar shape, and fertility of pollen. The equatorial shape of five types was observed: prolate, prolate-spheroidal, spheroidal-subprolate, subspheroidal-prolate, and subspheroidal. Four types of surface ornamentation, psilate, granulate, clavate, and perforate, were recorded. Tectum of five types, intactate, reticulate regulate, medium reticulate, tactate, and striate, was observed. Sexine was thicker than nexine in all studied species. The largest polar diameter was observed in Caesalpinia pulcherrima 64.1 μm while the smallest in Parkinsonia aculeata 26.1 μm. The largest equatorial diameter was found in C. pulcherrima of 70.25 μm whereas the smallest in P. aculeata 27.57 μm. All the pollens analyzed were tricolporate. All studied species have a fertility ratio of more than 90%. A taxonomic key was developed to show the variation in pollen features and delimit species for the correct identification. In conclusion, the pollen traits were found useful to define species boundaries at various taxonomic ranks and will strengthen the taxonomy of this subfamily. Besides, this study also explored the palynological traits and their implication in the taxonomy of the subfamily Caesalpiniaceae., (© 2021 Wiley Periodicals LLC.)

Shigella infection (shigellosis) is an intestinal disease caused by a shigella isolates belongs to a family Enterobacteriacea. Watery diarrhea, abdominal pain and tenesmus are the prominent symptoms of shigella infection. The present study was designed to determine period prevalence and antimicrobial susceptibility of Shigella species recovered from stool specimens obtained from diarrheal paediatric patients under 5 years of age. This cross-sectional study was carried out for a period of six months (Jan to June, 2016). All Shigella isolates were identified based on colony morphology, microscopic characteristics, and biochemical characteristics. After applying Kirby Baur disc diffusion method only 22 (18.96%) stool specimens were found positive for Shigella isolates among the 116 stool specimens. The isolates were also found susceptible to Levofloxacin (72.72%), Azithromycin (59.09%), and Cefotaxime (40.90%). However, the said isolates were resistant to Lincomycin (100%) and Penicillin-G (100%), followed by Amoxicillin (95.45%) and Oxacillin (95.45%). The chi-square test was used to check the close association among antimicrobial agents used and as highly significant (p-value < 2.2e-16). Based on antimicrobial susceptibility findings, Levofloxacin, Azithromycin and Cefotoxime were found effective for the control of shigellosis.

This study investigated the concentration of radon ( 222 Rn) in hot springs water. For this purpose, 222 Rn concentration was measured using the RAD7 (Durridge Company, USA) in the water of hot springs located in Tata Pani, Gilgit (n = 4), and Garam Chashma, Chitral (n = 6), northern Pakistan. Water samples from the springs (background, n = 3) were also collected and analyzed for 222 Rn concentration 40-50 km away from the hot springs in Gilgit and Chitral, northern Pakistan, to be used as background/reference concentration. The determined 222 Rn in hot springs water surpassed the threshold of maximum contamination level (MCL, 11.1 Bq/L) set by the United States Environmental Protection Agency (US-EPA) in 100% samples collected from Tata Pani, Gilgit, and Garam Chashma, Chitral sites. Soil 222 Rn along with the hot springs exhibited a decreasing trend with increasing distance. 222 Rn concentration in hot springs water was used to calculate the exposure doses of human health through ingestion and inhalation pathways. The total effective dose for human (E WT ) of 222 Rn contaminated water consumption was 626 μSv/a in the Tata Pani, Gilgit and 34.7 μSv/a in the Garam Chashma, Chitral. Results revealed that hot springs water in the Tata Pani, Gilgit had surpassed the threshold limit (100 μSv/a) set by the World Health Organization (WHO). This study concluded that hot springs water should be avoided for drinking and other domestic uses., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

According to IPCC Annual Report (AR-5), environmental impact assessment of any product prototype is recommended before its large-scale commercialization; however, no environmental profile analysis of any biodiesel prototype has been conducted in Pakistan. Therefore, objective of this study was to conduct a comprehensive life cycle assessment (LCA), water footprint and cumulative energy demand (CED) of biodiesel production from Jatropha curcas L. (JC) seeds oil in Pakistan. A cradle-to-gate LCA approach was applied for 400 liter (L) JC biodiesel produced in Pakistan. JC biodiesel production chain was divided into three stages i.e., 1). cultivation of JC crop 2). crude oil extraction from JC seeds and 3). crude oil conversion to biodiesel. Primary data for all the stages were acquired through questionnaire surveys, field visits and measurements in the field. Potential environmental impacts were calculated in SimaPro v.9.2 software using Eco-indicator 99 methodology. Results showed that crude oil extraction stage accounted for highest emissions (77%) to the overall environmental impact categories evaluated, followed by oil conversion stage (21%) and JC cultivation stage (02%), respectively. The three stages of JC biodiesel production chain are major contributor to ecotoxicity with a contribution of 57% to this impact category. Higher contribution to ecotoxicity was due to agrochemicals used in the JC cultivation. Similarly, fossil fuels impact category was responsible for 38% of overall environmental impacts. In addition, water footprint of JC biodiesel production chain was 2632.54 m3/reference unit. Cumulative energy required for 400L JC biodiesel production chain was 46745.70 MJ in Pakistan. Fossil diesel consumption, synthetic fertilizers use and purchased electricity were major hotspot sources to environmental burdens caused by JC biodiesel production in Pakistan. By performing sensitivity analysis at 20% reduction of the baseline values of fossil diesel used, synthetic fertilizers and purchased electricity, a marked decrease in environmental footprint was observed. It is highly recommended that use of renewable energy instead of fossil energy would provide environmental benefits such as lower greenhouse gases and other toxic emissions as compared to conventional petroleum fuels. It is also recommended that JC as a biofuel plant, has been reported to have many desired characteristics such as quick growth, easy cultivation, drought resistance, pest and insect resistance, and mainly great oil content in JC seeds (27-40%). Therefore, JC plant is highly recommended to Billion Tree Afforestation Project (BTAP) for plantation on wasteland because it has multipurpose benefits., Competing Interests: The authors have declared that no competing interests exist.

Background: Diabetes mellitus is a common disease effecting the lifestyles of majority world population. In this research work, we have embarked the potential role of crude extracts and isolated compounds of Notholirion thomsonianum for the management diabetes mellitus., Methods: The crude extracts of N. thomsonianum were initially evaluated for α-glucosidase, α-amylase and antioxidant activities. The compounds were isolated from the activity based potent solvent fraction. The structures of isolated compounds were confirmed with NMR and MS analyses. The isolated compounds were tested for α-glucosidase, α-amylase, protein tyrosine phosphatase 1B (PTP1B) and DPPH activities. The molecular docking studies were carried out to find the binding interactions of isolated compounds for α-glucosidase, α-amylase and PTP1B., Results: Initially, we screened out crude extracts and subfractions of N. thomsonianum against different in-vitro targets. Among all, Nt.EtAc was observed a potent fraction among all giving IC 50 values of 67, 70, < 0.1, 89 and 16 μg/mL against α-glucosidase, α-amylase, DPPH, ABTS and H 2 O 2 respectively. Three compounds (Nt01, Nt02 and Nt03) were isolated from Nt.EtAc of N. thomsonianum. The isolated compounds Nt01, Nt02 and Nt03 exhibited IC 50 values of 58.93, 114.93 and 19.54 μM against α-glucosidase, while 56.25, 96.54 and 24.39 μM against α-amylase respectively. Comparatively, the standard acarbose observed IC 50 values were 10.60 and 12.71 μM against α-glucosidase, α-amylase respectively. In PTP1B assay, the compounds Nt01, Nt02 and Nt03 demonstrated IC 50 values of 12.96, 36.22 and 3.57 μM in comparison to the standard ursolic acid (IC 50 of 3.63 μM). The isolated compounds also gave overwhelming results in DPPH assay. Molecular docking based binding interactions for α-glucosidase, α-amylase and PTP1B were also encouraging., Conclusions: In the light of current results, it is obvious that N. thomsonianum is potential medicinal plant for the treatment of hyperglycemia. Overall, Nt.EtAc was dominant fraction in all in-vitro activities. Three compounds Nt01, Nt02 and Nt03 were isolated from ethyl acetate fraction. The Nt03 specifically was most potent in all in-vitro assays. The molecular docking studies supported our in-vitro results. It is concluded that N. thomsonianum is a rich source of bioactive antidiabetic compounds which can be further extended to in-vivo based experiments., (© 2021. The Author(s).)

Background: Shiga toxin-producing E. coli (STEC) are important foodborne pathogens that causing serious public health consequences worldwide. The present study aimed to estimate the prevalence ratio and to identify the zoonotic potential of E. coli O157 isolates in slaughtered adult sheep, goats, cows and buffaloes., Materials and Methods: A total of 400 Recto-anal samples were collected from two targeted sites Rawalpindi and Islamabad. Among them, 200 samples were collected from the slaughterhouse of Rawalpindi included sheep (n = 75) and goats (n = 125). While, 200 samples were collected from the slaughterhouse of Islamabad included cows (n = 120) and buffalos (n = 80). All samples were initially processed in buffered peptone water and then amplified by conventional PCR. Samples positive for E. coli O157 were then streaked onto SMAC media plates. From each positive sample, six different Sorbitol fermented pink-colored colonies were isolated and analyzed again via conventional PCR to confirm the presence of rfbE O157 gene. Isolates positive for rfbE O157 gene were then further analyzed by multiplex PCR for the presence of STEC other virulent genes (sxt1, stx2, eae and ehlyA) simultaneously., Results: Of 400 RAJ samples only 2 (0.5%) showed positive results for E. coli O157 gene, included sheep 1/75 (1.33%) and buffalo 1/80 (1.25%). However, goats (n = 125) and cows (n = 120) found negative for E. coli O157. Only 2 isolates from each positive sample of sheep (1/6) and buffalo (1/6) harbored rfbE O157 genes, while five isolates could not. The rfbE O157 isolate (01) of sheep sample did not carry any of STEC genes, while the rfbE O157 isolate (01) of buffalo sample carried sxt1, stx2, eae and ehlyA genes simultaneously., Conclusion: It was concluded that healthy adult sheep and buffalo are possibly essential carriers of STEC O157. However, rfbE O157 isolate of buffalo RAJ sample carried 4 STEC virulent genes, hence considered an important source of STEC infection to humans and environment which should need to devise proper control systems., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Lamioideae comprised the second-largest subfamily in Lamiaceae. Although considerable progress has recently been made in the taxonomic study of Lamioideae, the subfamily remains one of the most poorly investigated subfamily in Lamiaceae. Therefore, the present study was designed with the aim to document the pollen micromorphology of some selected Lamioideae taxa and its taxonomic significance from Pakistan. Pollen micromorphological features were observed using scanning electron microscopy. The pollen grains are monad, tricolpate, radially/bilateral symmetrical. The pollen grains were small to medium-sized having oblate, oblate/subspheroidal, and subspheroidal shape. Exine sculpturing was observed as reticulate, microreticulate, and bireticulate. The colpus surface ornamentation was found as verrucate, gemmate, scabrate, and psilate. There was a considerable variation between the species in the micromorphology, that is, the coarseness of the reticulum, thickness of the muri comprising the reticulum and the number of secondary lumina per primary lumen. Hence, this study documented the pollen morphology of some selected taxa of the subfamily Lamioideae from Pakistan and strengthens the taxonomic identification of subfamily based on pollen characters, which helps in the correct identification, discrimination of the species of Lamioideae at generic and species level., (© 2020 Wiley Periodicals LLC.)

The family Scrophulariaceae consists of taxonomically complex genera and species. The delimitation of the taxa within this family is always challenging. In this paper, we studied leaf epidermis anatomical characteristics and its taxonomic significance of four species belonging to four genera of the family Scrophulariaceae collected from northern Pakistan. The species were examined under light and scanning electron microscopes (LM and SEM). Qualitative and quantitative foliar epidermal anatomical features were examined for both adaxial and abaxial surfaces. Qualitative characters like epidermal cell shape, epidermal cell cover, anticlinal wall, trichomes type, stomata type and stomata position were examined. Quantitative characters like the length and width of leaf epidermis, stomata, stomatal pore, subsidiary cell and trichomes for both adaxial and abaxial surfaces were studied and measured. Stomatal index within the species and between the species was found to be different on adaxial and abaxial surfaces. Diacytic stomata and glandular trichomes on epidermis were only found in Anticharis glandulosa while rest of the taxa has anomocytic type stomata and dendroid trichomes on both surfaces. Based on the micromorphological characters, we did principal component analysis (PCA), and cluster analysis for the species delimitation and identification. A taxonomic key has been provided to delimit and identify the studied taxa based on foliar epidermal characters. The aim of the present research was to elucidate the micromorphological characters to distinguish the studied taxa for taxonomic purposes., (© 2020 Wiley Periodicals LLC.)

This study investigated variation in the keratin-associated proteins gene, KRTAP6-3, in 5 Pakistani sheep breeds/crosses using polymerase chain reaction-single strand confirmation polymorphism (PCR-SSCP) analysis. Different banding patterns were revealed, including previously described patterns and a novel pattern (named variant H). The amplified PCR product of the novel banding pattern was directly sequenced, and a synonymous nucleotide variation c.51T>C was revealed. Among the wool traits assessed, a strong correlation (r = 0.929; P < 0.001) was observed between fibre diameter standard deviation (FDSD) and coefficient of variation of fibre diameter (CVFD), between FDSD and medullation (r = 0.720; P < 0.001), between FDSD and medullation standard deviation (MeSD) (r = 0.734; P < 0.001), between MeSD and coefficient of variance of medullation (CVMed), (r = 0.903, P < 0.001), and between CVFD and medullation (r = 0.660), CVFD and MeSD (r = 0.786; P < 0.001), CVFD and CVMed (r = 0.701; P < 0.001) and medullation and MeSD (r = 0.771; P < 0.001). Variant B was found to be associated (P = 0.018) with CVFD; the presence of B being associated with a higher CVFD, than in its absence (41.08 ± 3.98 versus 36.34 ± 3.08). Variant C was associated with CVMed (P = 0.040), where sheep with C had a lower CVMed than sheep where it was absent. Variation in KRTAP6-3 was found to affect fibre diameter related traits of wool.

In all over the world, herbal drugs are usually adulterated with similar species or varieties due to incorrect identification. Most of herbal products devoid purity and quality, therefore an attempt was carried out to identify plant species and authenticate its herbal drug products from Mentha suaveolens. Microscopy tools provide an excellent platform to identify plants at species level. In this study, microscopic and pharmacokinetic parameters of M. suaveolens were observed. Plant species were collected from high diverse areas of Northern Pakistan. Macro and micro-morphology including palynology and anatomical features were analyzed to study M. suaveolens. Species characteristics were studied, while implementing microscopic techniques for the delimitation and identification of the species. Traditionally Mentha species are used to cure several diseases that is, digestive disorders, respiratory disorders. Micromorphology (stem, leaves, flowers structure, length etc.), palynology (shape, size of pollen etc.), and anatomical characters (types of stomata, epidermal cell shape, and trichomes) were studied. Micromorphology and anatomical characters were of great interest and significance to discuss the taxonomy of the species. Taxonomic characters were studied to characterize and authenticate the species. The aim of the present study is to observe in detail the taxonomic identification of the species in term of morphology, palynology, and foliar epidermal anatomy for the correct identification along with their medicinal uses in the area., (© 2019 Wiley Periodicals, Inc.)

Aim: Neuronal ceroid lipofuscinosis (NCLs) are the most common neurodegenerative disorders, with global incidence of 1 in 100,000 live births. NCLs affect central nervous system, primarily cerebellar and cerebral cortices. Juvenile neuronal ceroid lipofuscinosis (JNCL), also known as Batten disease, is the most common form of NCLs. JNCL is primarily caused by pathogenic mutations in CLN3 gene, which encodes a transporter transmembrane protein of uncertain function. The 1.02 kb deletion is the most common mutation in CLN3 that results in frame shift and a premature termination leading to nonfunctional protein. Here, we invetigated a large consanguineous family consisting of four affected individuals with clincal symptoms suggestive of Juvenile neuronal ceroid lipofuscinosis. Materials and methods: We conducted clinial and radilogical investigation of the family and performed NGS based Gene Panel sequencing comprising of five hundred and forty five candidate genes to characterize it at genetic level. Results: We identified a novel homozygous c.181&#95;183delGAC mutation in the CLN3 gene seggregating witht the disorder in the family. The mutation induces in-frame deletion, deleting one amino acid (p.Asp61del) in CLN3 protein. The deleted amino acid aspartic acid plays an important role as general acid in enzymes active centers as well as in maintaining the ionic character of proteins. Conclusion: Our finding adds to genetic variability of Juvenile neuronal ceroid lipofuscinosis associated with CLN3 gene and a predicted CLN3 protein interacting domain site.

Spore morphology of Thelypteridaceae species growing in Malakand Division, Northern Pakistan, was studied using both light microscopy and scanning electron microscopy. The taxa are Christella dentata and Glaphyropteridopsis erubescens in the subfamily Thelypteridoideae, and Phegopteris connectilis, Pseudophegopteris pyrrhorhachis, and Pseudophegopteris levingei in the subfamily Phegopteridoideae. The studied species exhibit differences in spore size, exospore thickness, color, and ornamentation. Spores of the studied species are monolete and medium-sized, and shape is ellipsoidal in both polar and equatorial views. The average measurement of the polar diameter ranges from 27 μm to 31 μm, whereas in the equatorial direction it varied from 20 μm to 40 μm. The exospore thickness ranges from 1.2 μm to 2.4 μm. Reticulate, laevigate with microgranules, cristate, and coarsely echinate surface ornamentation are observed among the species. Multivariate analysis including unweighted pair group method with arithmetic mean and principal component analysis was used for the grouping and discrimination of species and genera., (© 2019 Wiley Periodicals, Inc.)

Seed micromorphological and macromorphological characteristics of 12 taxa of Caesalpiniaceae from Pakistan have been studied, using light and scanning electron microscopy, to investigate the importance of seed coat features as a taxonomic tool. Great variations have been observed in color, shape, dimension, and seed surface pattern among the different genera of the family. A taxonomic key was prepared for the studied taxa. Six types of seed shapes were observed; circular, elliptical, irregular, oblong, oval, and ovoid. All examined seeds were hard except fragile seeds of Haematoxylom campechianum. Eight types of surface ornamentation have been noticed that include levigate, lugose, papillate, reticulate, reticulate irregular, reticulate regular, rhombus, and rogues. Majority of the taxa has been observed with thick ornamentation wall but thin ornamentation wall has also been recorded in few species. Fracture line of the various patterns was present in all taxa except genus Bauhinia. Three types of texture crudeness; coarse, medium, and fine have been recorded. Both micromorphological and macromorphological characters of seed are very fruitful in identification and classification of Caesalpiniaceae., (© 2019 Wiley Periodicals, Inc.)

The present study is insight into pollen morphology for characterizing species and their utility in the taxonomic separation of certain taxa of subfamily Nepetoideae (Lamiaceae) from Pakistan. The pollen micromorphology of 11 species of the Nepetoideae was analyzed and documented using light microscopy and scanning electron microscopy (SEM) for both qualitative and quantitative characteristics. Most species have hexazonocolpate pollen grains but trizonocolpate and tetrazonocolpate pollen with circular and oval amb were also rarely observed in Mentha spicata. The basic pollen shape in most of the studied species was subspheroidal but prolate grains were also observed in M. spicata, S. coccinea, and S. plebeia. The exine sculpturing of Nepetoideae pollen was taxonomically very informative particularly at subfamily level. Observations of exine sculpturing with SEM revealed various types of pollen grains: reticulate, bireticulate, microreticulate, perforate, aerolate, and gammate. The bireticulate type further subdivided into three subtypes based on the number of secondary lumina in each primary lumen and is characterized by varying characteristics of the secondary reticulum and primary muri. A significant variation was observed in colpus surface ornamentation. The maximum polar diameter was found in O. americanum (58 ± 5.8 μm) and the maximum equatorial diameter observed in O. basilicum (50.25 ± 1.37 μm). Pollen features of the studied species were discussed and compared based on the current taxonomical concepts. The results showed that pollen traits of the subfamily Nepetoideae was found significant to classify the taxa. Furthermore, pollen features provide additional evidence to distinguish macromorphologically similar taxa from each other., (© 2019 Wiley Periodicals, Inc.)

Oculocutaneous albinism (OCA) is an autosomal-recessive disorder of a defective melanin pathway. The condition is characterized by hypopigmentation of hair, dermis, and ocular tissue. Genetic studies have reported seven nonsyndromic OCA genes, among which Pakistani OCA families mostly segregate TYR and OCA2 gene mutations. Here in the present study, we investigate the genetic factors of eight consanguineous OCA families from Pakistan. Genetic analysis was performed through single-nucleotide polymorphism (SNP) genotyping (for homozygosity mapping), whole exome sequencing (for mutation identification), Sanger sequencing (for validation and segregation analysis), and quantitative PCR (qPCR) (for copy number variant [CNV] validation). Genetic mapping in one family identified a novel homozygous deletion mutation of the entire TYRP1 gene, and a novel deletion of exon 19 in the OCA2 gene in two apparently unrelated families. In three further families, we identified homozygous mutations in TYR (NM&#95;000372.4:c.1424G > A; p.Trp475*), NM&#95;000372.4:c.895C > T; p.Arg299Cys), and SLC45A2 (NM&#95;016180:c.1532C > T; p.Ala511Val). For the remaining two families, G and H, compound heterozygous TYR variants NM&#95;000372.4:c.1037-7T > A, NM&#95;000372.4:c.1255G > A (p.Gly419Arg), and NM&#95;000372.4:c.1255G > A (p.Gly419Arg) and novel variant NM&#95;000372.4:c.248T > G; (p.Val83Gly), respectively, were found. Our study further extends the evidence of TYR and OCA2 as genetic mutation hot spots in Pakistani families. Genetic screening of additional OCA cases may also contribute toward the development of Pakistani specific molecular diagnostic tests, genetic counseling, and personalized healthcare., (© 2019 John Wiley & Sons Ltd/University College London.)

In our environment, various naturally occurring radionuclides are present (both underground and overground) in several places, which results in lifelong human exposure. The radiation dose received by human beings from the radiation emitted by these naturally occurring radionuclides is approximately 87%. Exposure to radiation poses radiological health hazards. To assess the human health hazards from radiation, the concentration of these naturally occurring radionuclides are measured in soil (used for cultivation), building materials (soil, bricks, sand, marble, etc.), water and dietary items, worldwide. The available literature revealed that numerous studies related to the subject have been carried out in Pakistan. Most of these studies measured the radioactivity concentrations of primordial [uranium (238U), thorium (232Th), radium (226Ra) and potassium (40K)] and anthropogenic [cesium (137Cs)] radionuclide in soil samples (used for cultivation), fertilizers, building materials (i.e. bricks, rocks, sand, soil, marble, etc.), as well as water and dietary items, using a sodium iodide detector or high purity germanium. An effort was made in 2008 to compile these studies as a review article. However, since then, considerable studies have been undertaken and reported in the literature. Therefore, the main objective of the present article is to provide a countrywide baseline data on radionuclide levels, by overviewing and compiling the relevant studies carried out in Pakistan.

Palyno-anatomical study of monocots taxa using Light and Scanning Electron Microscopy (SEM) was first time conducted with a view to evaluating their taxonomic significance. Studied plants were collected from different eco-climatic zones of Pakistan ranges from tropical, sub-tropical, and moist habitats. The aim of this study is to use palyno-anatomical features for the correct identification, systematic comparison, and investigation to elucidate the taxonomic significance of these features, which are useful to taxonomists for identifying monocot taxa. A signification variation was observed in quantitative and qualitative characters by using the standard protocol of light microscopy (LM) and SEM. Epidermal cell length varied from maximum in Allium griffthianum (480 ± 35.9) μm at the adaxial surface to minimum in Canna indica (33.6 ± 8.53) μm on abaxial surface. Maximum exine thickness was observed in Canna indica (4.46) μm and minimum in Allium grifthianum (0.8) μm. Variation was observed in shape and exine ornamentation of the pollen, shape of the epidermal cell, number, size, and type of stomata, guard cell shape, and anticlinal wall pattern. Based on these palyno-anatomical features a taxonomic key was developed, which help in the discrimination of studied taxa. In conclusion, LM and SEM pollen and epidermal morphology is explanatory, significant, and can be of special interest for the plant taxonomist in the correct identification of monocots taxa., (© 2019 Wiley Periodicals, Inc.)

In this study, comparative morphology, foliar anatomy and palynology of Spergula fallax and Spergula arvensis (Caryophyllaceae) were studied using multiple microscopic techniques. Genus Spergula includes worldwide five species, while in Flora of Pakistan the genus has two species. In this research, the comparative morphological, anatomical, and palynological characters of the two Pakistani Spergula species were studied. We examined some distinguishing morphological features, in both species, such as plant size, habitat, leaf morphological characters, inflorescences, flowers outer whorls, sepals and petals, and flowers number. These characters species were studied analyzing their comparative systematic significant. The foliar anatomical features also provided distinctive characters as the epidermal cell shape, the wall of the epidermal cell, lobes per cell. The differences in quantitative characters were also examined. The palynological characters showed difference in echini arrangement, echini density, and numbers of pore. Quantitative characters were variations in size of polar, equatorial, exine thickness, pore length, and width and P/E ratio. The multiple microscopic techniques provided sufficient evidence about the systematics of the genus Spergula. Based on morphological, anatomical, and palynological characters, analytical keys were developed for the identification and distinction of the species S. fallax and S. arvensis., (© 2019 Wiley Periodicals, Inc.)

In present study, multiple microscope techniques were used for the systematics identification of the species Asplenium dalhousiae. The plant was collected from different phytogeographical and its natural habitat of Pakistan, where it shows higher diversity. Morphology, foliar epidermal anatomy, and spore morphological characters of the species were studied in detailed using multiple microscopic techniques through light microscopy (LM) and scanning electron microscopy (SEM). LM and SEM were used for the systematics identification of the species. Traditionally, the species is used in the ailment of many diseases, so the spore morphology, anatomical features, and morphological characters are relevant to describe the species taxonomy. The importance of multiple methods of taxonomic study (e.g., documentation and morphological characteristics) for characterizing herbs are important step in systematic certification to maintain the efficacy of herbal medicines. The aim of the present study is to examine the morphological, anatomical, and spore morphology of the species A. dalhousiae in more detailed for the correct taxonomic identification and their medicinal validation from Pakistan., (© 2019 Wiley Periodicals, Inc.)

The present study is insights into foliar epidermal anatomy for characterizing clades, and their utility in taxonomic segregation of certain species of Pteridaceae from Northern Pakistan. The leaf epidermal anatomy of 10 species of Pteridaceae representing four genera were examined using light and scanning electron microscope. A micromorphological matrix was constructed for eight qualitative and 12 quantitative characters. unweighted pair group method with arithmetic means and principal components analysis statistical analysis were performed to test the validity of foliar epidermal anatomical features as method of separating species and genera, and phylogenetic clusters among species are constructed using qualitative and quantitative traits. The qualitative characters described here are shape of epidermal cells, stomata, guard cell and subsidiary cells, anticlinal wall pattern, and trichomes types which is helpful in defining groups within Pteridaceae. In addition, the size of stomata, guard cells, subsidiary cells, stomatal pore epidermal cells, and trichomes are quantitatively analyzed. All species have hypostomatic leaves. Two types of stomata were observed in studied species, anomocytic and polocytic. Anomocytic stomata were observed in three genera namely: Adiantum, Onychium, and Chielanthes whereas Pteris can be discriminated from other genera by its polocytic stomata. On the basis of multivariate analysis present study does provides sufficient information on the taxonomic importance of foliar anatomy which validate its efficacy in species and genera discrimination. From result obtained here it is further possible to use leaf micromorphologic data in ferns phylogeny and providing basis for future taxonomic delimitation in other taxa., (© 2018 Wiley Periodicals, Inc.)

Information on prevalence of soil-transmitted helminth (STH) infections among school children is scarce in Pakistan. This study was aimed to investigate the prevalence of soil-transmitted helminth in school children of three districts in, Khyber Pakhtunkhwa, Pakistan. A total of 300 stool samples were examined from August 2015 to August 2016 using direct smear (Normal saline and Lugol's Iodine solution) and the concentration methods. One hundred and eighty seven (62.3%) pupils were found infected with soil-transmitted helminths. One hundred and forty five (77.5%) were infected with single parasite and forty two (22.4%) with multiple infections. Ascaris lumbricoides 125 (66.4%), Trichuris trichura 50 (26.5%) and Ancylostoma duedenale 13 (6.91%) were detected. The children above 8 years in age were more parasitized than below 8 years (p=0.7832; P>0.05). Males were found more parasitized than females (p=0.9315; P>0.05). Children in lower Dir district were found more infected followed by Swat and upper Dir (P< 0.0001; p<0.05). No significant relationship was found among the examined and that of infected children for ages and sex in all the districts. Malakand division is an area with poor hygiene located in temperate zone near the border of Afghanistan and China. The prevalence of reported nematode parasites here compared with the same studies is unexpectedly high. These types of studies should continue time to time to know the hazardous nature of such parasitic infections for the betterment of the human health.

The pollen morphology of 11 salt tolerant plant species of family Amaranthaceae from the salt range of Northern Punjab, Pakistan has been studied. The palyno-morphological characters were examined using light and scanning electron microscope. The examined all salt tolerant species have a slight difference in size but have similarity in shape, pore ornamentation, and polarity. The observed morphological characters of pollen grains were pollen symmetry, size, shape, pore ornamentation, pore size, number of pores, exine thickness, polar and equatorial diameter and, P/E ratio. Apolar type of pollens has been observed in all species. Shape of pollens was spheroidal. Exine sculpturing of pollen grains was scabrate (six spp), microechinate (four spp), and microechinate-scabrate (one spp). Different pori numbers were observed in different species. The pantoporate aperturate and sunken pore ornamentation have been reported in all species. A pollen taxonomic key was developed using examined morphological characters for the accurate identification of halophytic taxa. The high fertility and low sterility of pollens confirmed that the selected halophytes are well-established in the salt region. The findings highlight the taxonomic significance of pollen morphology in correct identification and differentiation of salt tolerant plant species., (© 2019 Wiley Periodicals, Inc.)

Foliar epidermal features were based on the micromorphology of trichomes types, epidermal cells and stomatal complex. Even though each feature has its own limited taxonomic value but collectively these characteristics may be systematically important especially for the discrimination and identification of complex and problematic taxa. The systematics significance of nonglandular (NGTs) and glandular trichomes (GTs), stomatal complex and epidermal cells of Lamiaceous flora were analyzed by using the light microscopy (LM) and scanning electron microscopy (SEM). Variations on the observed epidermal appendages were divided into two basic types: glandular and nonglandular. GTs can be divided into subtypes: sessile capitate, subsessile capitate, and barrel and sunken. NGTs were also divided into subtypes: dendritic, stellate, conical, falcate, simple and 1-6 cells long having granulate and smooth surface ornamentation. NGTs were the most dominant features of both adaxial and abaxial surfaces of all observed taxa. Vitex negundo, Isodon rugosus, Colebrookea oppositifolia, and Marrubium vulgare could be demarked because of their twisted like appearance of NGTs at the abaxial surface. The Lamiaceae had both hypostomatic and amphistomatic leaf. Stomata were observed as diacytic, anisocytic, and anomocytic. Epidermal cells were found to be irregular, isodiametric, and rectangular. Based on these characters a taxonomic key was developed to delimit the closely related taxa. Distribution and morphology of the foliar epidermal trichomes through SEM highlight an important taxonomic tool used by the taxonomists as an aid to the correct identification of problematic Lamiaceae taxa., (© 2019 Wiley Periodicals, Inc.)

Current research carried out in Pakistan is the first report on spermoderm ornamentation of eight species of tribe Acacieae (Mimosoidae) by using scanning electron microscopic techniques representing two genera, Fedherbia and Acacia were examined. Different spermoderm ornamentation were observed, described and discussed for their taxonomic importance. Seeds surfaces of the studied tribe possess novel variations in macro and micro morphology. Great variations were observed in both qualitative and quantitative characters of seeds. Seeds shape was oblong, ovate to elliptical and spermoderm ornamentation was levigate, rugose, polygonal and discoid, colliculate, and papillose type. These variations in the spermoderm ornamentation can be used as an aid in identification and classification of the members of tribe Acacieae., (© 2018 Wiley Periodicals, Inc.)

Pollen micro-morphological features have proven to be helpful for the plant taxonomists in the identification and classification of plants. The utilization of this plantmayhelpfulin the areas of lignocellulosic conversion to biofuels and diversify application toward biomass. The current study was planned with the aim to evaluate the pollen features of complex Ranunculaceous flora of District Chitral, Northern Pakistan using both scanning electron microscopy (SEM) and Light Microscope (LM) for their taxonomic importance. Pollens of 18 Ranunculaceous species belonging to 6 genera were collected from different localities of the research area. SEM and LM were used to examine both qualitative and quantitative micro-morphological features. Sculptring of the sexine include; Scabrate, psilate, echinate, verrucate, perforate gemmate, and reticulate and so forth. Shape of the pollens was sub-spheroidal, spheroidal, prolate, subprolate and oblate and so forth. Type of pollen was ranged from mono to tricolpate and tricolporate. Quantitative characters include length/width of the pollen, colpus, exine thickness, and P/E ratio. Based on these micro-morphological features a taxonomic key was prepared for the fast and correct identification. RESEARCH HIGHLIGHT: Study of the pollen micro-morphological features of Ranunculaceous species by SEM and LM. Analysing both qualitative and quantitative characters of the pollens. Preparation of taxonomic key based on micro-morphological features for the correct and fast identification., (© 2018 Wiley Periodicals, Inc.)

Dryopteris and Polystichum are the 2 complex taxonomic genera of Dryopteridaceae. The comparative foliar epidermal anatomy of 12 species of both genera from Pakistan were studied using standard protocols of light microscopy (LM) and scanning electron microscopy (SEM). The objective of which was systematic comparison and investigation to elucidate the taxonomic importance of foliar micromorphology, which may be useful to taxonomists for identifying complex Dryopteridaceae taxa. Principal component analysis and UPGMA clustering analysis were performed to test the validity of leaf anatomical features as method of separating species and genera. The foliar epidermal anatomy described here is a good source of taxonomic characters in both groups that can help genera and species delimitation. This is the first report on leaf micromorphology in most of these species. Observation of foliar anatomy showed that stomata are only present on the abaxial surface; i.e., leaves of all species are hypostomatic. The shapes of epidermal cells in all studied species are irregular. The anticlinal walls are strongly lobed, irregular wavy and elongated wavy. However, substantial variation in epidermal cell size and other stomatal features were observed on both upper and lower surfaces in all investigated species. Two types of stomata were observed in all studied species. The presence of polocytic stomata in Dryopteris and staurocytic stomata in Polystichum are the important characters for the segregation of these genera. Elongate elliptic stomatal shape, narrow kidney shaped guard cells and broad elliptic shaped stomatal pores are diagnostic for all five species of Dryopteris selected. On the other hand, size and number of epidermal cells, lobes per cell, stomatal size, subsidiary cell size, stomatal pore size and stomatal index are the key features for species differentiation in Polystichum. An identification key was developed in order to apply the foliar anatomical characters in the discrimination of the species studied., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The Pollen morphology of subfamily Caryophylloideae (Caryophyllaceae) from different phytogeographical region of Pakistan has been evaluated. In this research, 16 species belong to 6 genera of subfamily Caryophylloideae have been studied using light (LM) and scanning electron microscopy (SEM) for both qualitative and quantitative characters. Different palyno-morphological features were observed including; pollen ornamentation, pore ornamentation, echini arrangement, echinidensity, number of pori, size of polar and equatorial diameter, P/E ratio, exine thickness, and size of pore were studied. The palyno-morphological characters of subfamily Caryophylloideae have taxonomically significant in identification and delimitation of species. Two pollen types, i.e., subspheroidal (15 species) and prolate (one species Vaccaria hispanica) were observed. Microechinate-punctate and microechinate-perforate pollen ornamentation were examined. Two pollen types of pori, i.e., prominent (5 species) and sunken (11 species) ornamentation have been reported. Three types of echini arrangement have been reported irregular, regular and rather regular, while three type of echinidensity (i.e., medium, dense and sparse) were observed. Pori numbers were found different in different species range from 5 in Silene apetala to 19-35 in Silene vulgaris. Based on different palyno-morphological characters, taxonomic key was develop for quick and accurate identification. The quantitative data were processed using SPSS software for average, minimum, maximum, and standard error., (© 2018 Wiley Periodicals, Inc.)

Objective: To determine the correlation of serum calcium with severity of acute ischaemic stroke., Methods: This cross-sectional study was conducted at the Khyber Teaching Hospital, Peshawar, Pakistan, from January to September 2013, and comprised patients who had suffered ischaemic stroke. All patients aged above 18 years who had been diagnosed to have suffered ischaemic stroke through history, physical examination and computed tomography scan, and who were admitted within the first 72 hours of the onset of the illness were included. Serum calcium was obtained within 78 hours of the onset of stroke and the severity of stroke was assessed at the same time using the National Institute of Health Stroke score., Results: Of the 138 patients, 71(51.4%) were women and 67(48.6%) men. The overall mean age was 61.09±11.93 years (range: 34-100 years). The mean National Institute of Health Stroke score was 17.77±7.73 (range: 2-35). Serum calcium of all patients was measured and adjusted for serum albumin level. The mean serum calcium level was 8.82±0.69 mg/dl (range: 6.84-10.48). Bivariate correlation was calculated for continuous data of serum calcium and National Institute of Health Stroke score. The overall Pearson's correlation coefficient was r= -0.237 (p=0.005)., Conclusions: Lower serum calcium levels may be associated with more severe clinical findings at the onset of stroke.

Background: Ethnomedicinally Rumex hastatus D. Don has been used since long for various ailments especially in neurological disorders. The reported data and the importance of Rumex genus demonstrate the vital medicinal value of R. hastatus., Methods: In the current investigational study, isolation of essential oil and its antioxidant and anticholinesterase assays were performed. The essential oil of R. hastatus was analyzed by GC-MS for the first time. The essential oil was evaluated for anticholinesterase and antioxidant assays. The anticholinesterase assay was conducted at various concentrations (62.5 to 1000 μg/ml) against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Similarly, the antioxidant potential was determined using DPPH and ABTS free radicals., Results: The GC-MS analysis of essential oil showed 123 components. The result recorded for the anticholinesterase assays demonstrated a marked potential against AChE and BChE with IC50 values of 32.54 and 97.38 μg/ml respectively which were comparable with the positive control i.e., galanthamine (AChE, IC50 = 4.73 μg/ml and BChE, IC50 = 11.09 μg/ml). The antioxidant assays against DPPH and ABTS free radicals also exhibited significant scavenging potential with IC50 values of 3.71 and 6.29 μg/ml respectively, while for ascorbic acid the IC50 value was <0.1 μg/ml against both free radicals., Conclusions: Based on the current investigational studies, it may be concluded that R. hastatus is an effective source of essential oil's components having anticholinesterase and antioxidant potentials, which after subjecting to drug development may lead to novel drug candidates against neurodegenerative disorders.

Malaria is wide spread in poor world and its burden has been assessed by the enumeration of malarial parasites in blood of patients. This study was designed to find a relationship between social structure, and spread of malaria in Khyber agency. The average parasite density was 2050 parasite/μl in Khyber Agency. Due to economic and social setup most of the people have habit of sleeping in open air thus playing role in high malaria prevalence and Plasmodium vivax remains the prevalent species. Genetic study performed on 110 Blood samples showed less genetic diversity for both Plasmodium vivax and Plasmodium falciparum. Eight alleles were distinguished both for Pvmsp 3α and Pvmsp 3β in total of 20 and 39 amplified samples of P. vivax respectively. Out of 17 samples amplified for P. falciparum 11 showed genotype K1 and 10 for MAD at Pfmsp-1 while 14 alleles were identified for 3D7/1C and two for FC27 of corresponding families of Pfmsp-2 gene. This shows that Plasmodium parasites are not genetically diverse in Khyber agency.

Toxoplasmosis is spread through contamination of water sources and results in morbidity globally. In the current study 300 water samples were processed by polymerase chain reaction (PCR) for detection of Toxoplasma gondii. The overall prevalence in different water sources was 6.6% (17/300). Among different water sources the highest prevalence was recorded in drain water at 7% (7/100), followed by tube well water at 7.5% (3/40) and open well water at 5% (5/100) ,and the lowest was recorded in tap water at 3.33% (2/60). The highest prevalence was recorded in summer. Evidence indicates that cleaning and filtration need to be adopted to avoid the health hazards of waterborne zoonotic parasites.

Background: Medicinal plants are used for the treatment of different diseases in almost all cultures. Teucrium species grow wildly at different geographical locations around the world. Teucrium stocksianum is used in folk medicine for the treatment of diarrhea, cough, jaundice and abdominal pain. Scientific study on Teucrium stocksianum shows that it possesses anthelmintic, cytotoxic and antispasmodic activity. The aim of our present study is to identify the chemical composition and antinociceptive potential of the essential oil extracted from Teucrium stocksianum bioss., Method: Essential oil (EO) from the aerial parts of Teucrium stocksianum were extracted by hydrodistillation process. The qualitative and quantitative composition of essential oil was determined with Gas chromatography/Mass spectrometer. Antinociceptive activity was determined by acetic acid induced writhing method. Percent inhibition of writhes of the test concentration was determined by comparing it with that of control. Tween-80 emulsion 2.5% (5 ml/kg b.w) was used as a control while Diclofenic sodium 50 mg/kg (b.w) was used as a standard drug., Results: The chromatogram of the essential oil of Teucrium stocksianum shows differences both qualitatively and quantatively from essential oil composition reported in other countries. Hydrodistillation of Teucrium stocksianum yielded 0.4% (v/w), pale yellowish oil on dry basis. A total of 26 chemicals were identified by GC-MS accounting for 90.28% of the oil. The major components of essential oil were δ-cadinene (12.92%), α-pinene (10.3%), myrcene (8.64%), β-caryophyllene (8.23%), germacrene D (5.18%) and limonene (2.36%). Essential oil of Teucrium stocksianum has shown outstanding antinociceptive activity. It has been observed that increase in percent writhe inhibition (PWI) occurred from 20-80 mg/kg (b.w) and maximum writhe inhibition has been noted at a concentration of 80 mg/kg (b.w), but PWI decreased at 160 mg/kg, which may be due to some toxic effect of higher dose. ED50 value for Teucrium stocksianum was calculated as 31.5 ± 1.72415 mg/kg (b.w)., Conclusion: Our results indicate that there is a lot of variation in the composition of essential oil of Teucrium stocksianum boiss, which may be due to different climatic and experimental conditions. Secondly, the essential oil possesses strong antinociceptive activity and could be used in analgesic preparations especially for topical use.

Malaria is one of the serious diseases threatening human health in Pakistan and contributes to a large proportion of the total malaria deaths in South Asia. However, little is known about the nature and extent of genetic diversity of the malarial parasites circulating in Pakistan. This study was designed to assess the infection status of Plasmodium and the genetic diversity of Plasmodium vivax and Plasmodium falciparum by analyzing msp-3α, msp-3β and msp-1, msp-2 genes respectively using allele specific nested PCR and RFLP assays. For this purpose, 130 field isolates were collected from the individuals who exhibited clinical symptoms associated with malaria in the Kohat region of Khyber Pakhtoonkhwa (KPK), Pakistan. Among 130 blood samples collected, P. vivax was detected in 105/130 (80.8%) and P. falciparum in 21/130 (16.2%). Mixed infections with both parasites were detected in 4/130 (3%) of the isolates. A large number of distinguishable alleles were found for msp genetic markers: 10 alleles for msp-3α and seven for msp-3β with one mixed infection in case of msp-3β. The genotyping of P. falciparum showed that K1+MAD20 mixed genotype was dominant in msp-1 and FC27 in msp-2. The results collectively suggest that P. vivax and P. falciparum populations in this region are highly polymorphic and mixed infections are prevalent.

Objective: To determine the frequency and presentation of mammalian bite injuries to the head and neck region seen in a plastic surgery unit., Design: Observational study. PATIENTS AND DURATION OF STUDY: The Plastic and Reconstructive Surgery Unit, PGMI, HMC, Peshawar from July 1999 to June 2003., Patients and Methods: During the study period 40 patients presented with mammalian bite injuries to the head and neck region. Information about patient's demographics, biting animal, circumstances of sustaining injury, duration of injury, location and extent of injury, treatment administered, cosmetic outcome and complications were noted., Results: The mammalian bite injuries accounted for 10% of patients managed within the study period for soft tissue injuries to the orofacial region. Among the 40 cases studied, 3 patients were females and 37 were males. Age ranged from 2.5 years to 71 years with the mean of 23 years. The duration of injury varied from 1 hour to 1 year (median 3+2 days). Mammals implicated were humans, dogs, horse, donkey, bear and wolf. All of the 40 patients presented with head and neck injury. Auricular injury was seen in 10 patients, lip injuries in 10 patients, nasal injuries in 10 patients, cheek injuries in 6 patients and scalp injuries in 4 patients. In 70% (28) of cases avulsion of tissue (type-III) was noted. Patients presenting earlier for treatment achieved better cosmesis than the patients presenting late in the course of their injury., Conclusion: Mammalian bite injuries to the head and neck region can result in facial disfigurement with distressing physical and psychological consequences. Early management of such complex injuries usually guarantees satisfactory outcome.