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Quince (Cydonia oblonga) is a medicinal plant and a member of family Rosaceae. It is native plant of Asia Minor and Europe. It is used in production of jam and jellies and also as a remedy of several ailments. The present study was conducted to evaluate the genetic polymorphism based on morphological and molecular traits. Different varieties of Quince were collected from different ecological zones of Khyber Pakhtunkhwa Pakistan and a total of 26 different morphological traits were recorded among studied genotypes. Based on qualitative morphological trait study, the variety collected from Tindodag was unique one with highest fruit weight (328.82 g). The lowest fruit weight (68.38 g) was recorded for Talash genotype. The Charbagh and Tindodag genotypes showed highest seed length (10.6 mm) while genotypes of Chitral was recorded as lowest (8.4 mm). Statistically, significant level of variation was noted with coefficient of variance ranged from 2.23% to 30.38%. Based on correlation analysis, fruit length had strongly correlation with fruit weight (r = 0.89**), Average Fruit width was found significant with fruit weight (r = 0.90**). Similarly, the Core Width was found strongly significant with Core Length (r = 0.95**). ANOVA analysis indicated 10 quantitative characters to be highly significant, 2 significant and 1 insignificant. Principal component analysis was also computed for the 13 quantitative traits with Eigen value of 0.48 and a total variance of 97.78%. The first principal component shows total variation of 52.52%. In PC2 the total variation was 80.15%, PC3 94.06% while in PC4 it was 97.78%. The NCBI BLAST results shows that all the genotypes have similar origin except Tindodag genotype, which shows differences in its origin. Accession number for all other genotypes is MN216014.1, while accession number of Tindodag genotype is KF861967.1. Based on this study, it can be concluded that Tindodag genotype is unique out of the studied localities. NCBI BLAST have provided further support for the drawn conclusion., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Shah et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Single nucleotide polymorphisms are the most common form of DNA alterations at the level of a single nucleotide in the genomic sequence. Genome-wide association studies (GWAS) were carried to identify potential risk genes or genomic regions by screening for SNPs associated with disease. Recent studies have shown that SCN9A comprises the NaV1.7 subunit, Na+ channels have a gene encoding of 1988 amino acids arranged into 4 domains, all with 6 transmembrane regions, and are mainly found in dorsal root ganglion (DRG) neurons and sympathetic ganglion neurons. Multiple forms of acute hypersensitivity conditions, such as primary erythermalgia, congenital analgesia, and paroxysmal pain syndrome have been linked to polymorphisms in the SCN9A gene. Under this study, we utilized a variety of computational tools to explore out nsSNPs that are potentially damaging to heath by modifying the structure or activity of the SCN9A protein. Over 14 potentially damaging and disease-causing nsSNPs (E1889D, L1802P, F1782V, D1778N, C1370Y, V1311M, Y1248H, F1237L, M936V, I929T, V877E, D743Y, C710W, D623H) were identified by a variety of algorithms, including SNPnexus, SNAP-2, PANTHER, PhD-SNP, SNP & GO, I-Mutant, and ConSurf. Homology modeling, structure validation, and protein-ligand interactions also were performed to confirm 5 notable substitutions (L1802P, F1782V, D1778N, V1311M, and M936V). Such nsSNPs may become the center of further studies into a variety of disorders brought by SCN9A dysfunction. Using in-silico strategies for assessing SCN9A genetic variations will aid in organizing large-scale investigations and developing targeted therapeutics for disorders linked to these variations., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Waheed et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Severe Acute Respiratory Syndrome Corona Virus (SARS-CoV-2) is the causative agent of COVID-19 pandemic, which has resulted in global fatalities since late December 2019. Alkaloids play a significant role in drug design for various antiviral diseases, which makes them viable candidates for treating COVID-19. To identify potential antiviral agents, 102 known alkaloids were subjected to docking studies against the two key targets of SARS-CoV-2, namely the spike glycoprotein and main protease. The spike glycoprotein is vital for mediating viral entry into host cells, and main protease plays a crucial role in viral replication; therefore, they serve as compelling targets for therapeutic intervention in combating the disease. From the selection of alkaloids, the top 6 dual inhibitory compounds, namely liensinine, neferine, isoliensinine, fangchinoline, emetine, and acrimarine F, emerged as lead compounds with favorable docked scores. Interestingly, most of them shared the bisbenzylisoquinoline alkaloid framework and belong to Nelumbo nucifera, commonly known as the lotus plant. Docking analysis was conducted by considering the key active site residues of the selected proteins. The stability of the top three ligands with the receptor proteins was further validated through dynamic simulation analysis. The leads underwent ADMET profiling, bioactivity score analysis, and evaluation of drug-likeness and physicochemical properties. Neferine demonstrated a particularly strong affinity for binding, with a docking score of -7.5025 kcal/mol for main protease and -10.0245 kcal/mol for spike glycoprotein, and therefore a strong interaction with both target proteins. Of the lead alkaloids, emetine and fangchinoline demonstrated the lowest toxicity and high LD50 values. These top alkaloids, may support the body's defense and reduce the symptoms by their numerous biological potentials, even though some properties naturally point to their direct antiviral nature. These findings demonstrate the promising anti-COVID-19 properties of the six selected alkaloids, making them potential candidates for drug design. This study will be beneficial in effective drug discovery and design against COVID-19 with negligible side effects., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Shah et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

This manuscript presents high performance dual polarized eight-element multiple input multiple output (MIMO) fifth generation (5G) smartphone antenna. The design consists of four dual-polarized microstrip diamond-ring slot antennas, positioned at corners of printed circuit board (PCB). Cheap Fr-4 dielectric with permittivity 4.3 and thickness of 1.6mm is used as substrate with overall dimension of 150 × 75 × 1.6 mm3. In mobile system due to limited space mutual coupling between nearby antenna elements is an issue that distort MIMO antenna performance. Defected ground structure is used to control coupling. The defected ground structure has advantages like ease of fabrication, compact size and high efficiency as compare to other techniques. Less than 30dB coupling is achieved for adjacent elements. The -10 dB impedance bandwidth of 700 MHz is achieved for all radiating elements ranging from 3.3 GHz to 4.1 GHz. The value is about 900MHz for -6dB. The proposed antenna offers good results in terms of fundamental antenna parameters like reflection coefficient, transmission coefficient, maximum gain, total efficiency. The antenna achieved average gain more than 3.8dBi and average radiation efficiency more than 80% for single dual polarized element. The antenna provides sufficient radiation coverage in all sides. The MIMO antenna characteristics like diversity gain (DG), envelope correlation coefficient (ECC), total active reflection coefficient (TARC) and channel capacity are calculated and found according to standards. Furthermore, effect of user on antenna performance in data-mode and talk-mode are studied. Proposed design is fabricated and tested in real time. The measured results shows that proposed design can be used in future smartphones applications. The design is compared with some of the existing work and found to be the best one in many parameters and can be used for commercial use., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Fawad et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

What are the effects of parenting styles on academic performance and how unequal are these effects on secondary school students from different gender and socioeconomic status families constitute the theme of this paper. A cross-sectional and purposive sampling technique was adopted to gather information from a sample of 448 students on a Likert scale. Chi-square, Kendall's Tau-c tests and hierarchical multiple regression analyses were used to determine the extent of the relationship among the variables. Chi-square and Kendall's Tau-c (Tc) test results established that the socioeconomic status of the respondent's family explained variation in children's academic performance due to parenting style; however, no significant difference was observed in the academic performance of students based on gender. Furthermore, hierarchal multiple regression analysis established that the family's socioeconomic status, authoritative parenting, permissive parenting, the interaction of socioeconomic status and authoritative parenting, and the interaction of socioeconomic status and permissive parenting were significant predictors (P<0.05) of students' academic performance. These predictor variables explained 59.3 percent variation in the academic performance of children (R2 = 0.593). Results of hierarchal multiple regression analysis in this study ranked ordered the most significant predictors of the academic performance of children in the following order. Family socioeconomic status alone was the strongest predictor (β = 18.25), interaction of socioeconomic status and authoritative parenting was the second important predictor (β = 14.18), authoritative parenting alone was third in importance (β = 13.38), the interaction of socioeconomic status and permissive parenting stood at fourth place in importance (β = 11.46), and permissive parenting was fifth (β = 9.2) in influencing academic performance of children in the study area. Children who experienced authoritative parenting and were from higher socioeconomic status families perform better as compared to children who experienced authoritarian and permissive parenting and were from low socioeconomic status families., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Ali et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Fasciola gigantica, responsible for the zoonotic disease fasciolosis, pose a great threat to the livestock and human health worldwide. The triclabendazole (TCBZ) has been used for decades as a broad spectrum anthelmintic to control this perilous disease but the emergence of resistance in flukes against TCBZ has prompted researchers across the world to explore for new drugs and antigenic targets. World Health Organization has strongly recommended the utilization of neurobiologically significant biomolecules as new drug/antigenic targets because of their significant role in the physiology of parasites. Monoamine Oxidase (MAO) is an important neurobiological enzyme which catabolizes aminergic neurotransmitters thus preventing prolonged excitation of neurons and in non-neuronal cells it prevents cellular toxicity due to accumulation of toxic monoamines. Owing to the important role of MAO in the survival and perpetuation of parasites, multipronged approaches were undertaken for the characterization of MAO-A in F. gigantica. The activity of MAO was found to be 1.5 times higher in the mitochondrial samples than the whole homogenate samples. The adult worms of the F. gigantica appeared to possess both the isoforms of MAO i.e., MAO-A and MAO-B. The zymographic studies revealed strong enzyme activity in its native state as assessed through prominent dark bands at 250KDa in the zymogram. The enzyme was also found to be highly immunogenic as revealed by high antibody titer at 1:6400 dilution. The immunogenicity of MAO-A enzyme was further established in the Western Blots in which a strong band of 50KDa was distinctly evident. Despite ubiquitous presence of MAO in F. gigantica some regions like tegumental surface and intestinal caecae displayed strong immunofluorescence as compared to other regions. The detection of MAO-A in the F. gigantica samples in Dot-Blot assay indicate a great potential of this molecule for the immunodiagnostics of fasciolosis, particularly in the field conditions. The enzyme activity was sensitive to the specific inhibitor clorgyline in a concentration dependant manner, particularly in the late incubation period. The zymographic results also exhibited similar trend. The strong intensity of spots in Dot-blots indicate high immunogenicity of the MAO protein. The intensity of bands/spots in the samples of worms treated with clorgyline also declined, clearly indicating that the tropical liver fluke possesses prominent MAO-A activity., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Beg et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Wheat (Triticum aestivum L.) is an important staple food crop for one third of global population and important crop for securing future food security. Rapid changes in the climate on global scale could be a threat for future food security. This situation urges plant breeders to explore the genetic potential of existing wheat germplasm. This study screened forty diverse wheat genotypes for their yield under two different agroclimatic conditions, i.e., Layyah and Dera Ghazi Khan, Pakistan. Data relating to plant height, peduncle length, flag leaf area, spike length, number of spikelets, number of grains per spike, thousand grain weight, chlorophyll content and grain yield were recorded. The tested wheat genotypes significantly differed for grain yield and related traits. Grain yield was positively correlated with plant height, spike length, spike number, flag leaf length, number of grains per spike, and 1000-grain weight. Biplot obtained from the cluster analysis by Euclidean method grouped the studied genotypes in 3 different groups. The genotypes exhibited 10.77% variability within quadrants, whereas 72.36% variability was recorded between quadrants according to clustering. Dendrogram grouped the tested genotypes into two main clusters. The main cluster "I" comprised of 2 genotypes, i.e., 'Seher-2006' and 'AS-2002'. The cluster "II" contained 38 genotypes based on Euclidian values. Genotypes within same cluster had smaller D2 values compared to those belonging to other clusters. The genetic relationships of genotypes provide useful information for breeding programs. Overall, the results revealed that genotypes 'Line 9733', 'Bhakar-2002', 'Line A9' and 'SYN-46' had better yield and yield stability under climatic conditions of southern Punjab. Therefore, these genotypes could be recommended for general cultivation in the study region., Competing Interests: The authors have declared that no competing interests exist.

Salinity in soil and water is one of the environmental factors that severely hinder the crop growth and production particularly in arid and semi-arid regions. A pot experiment was conducted to investigate the impact of salinity levels (1.5 dS m-1, 3.5 dS m-1, 7.5 dS m-1 and 11.5 dS m-1) on emergence, growth and biochemical traits of moringa landraces under completely randomized design having three replications. Four landraces of Moringa oleifera (Faisalabad black seeded moringa [MFB], Patoki black seeded moringa [MPB], Faisalabad white seeded moringa [MFW] and Rahim Yar Khan black seeded moringa [MRB]) were selected for experimentation. All the salinity levels significantly affected the emergence parameters (time to emergence start, time to 50% emergence, mean emergence time, emergence index and final emergence percentage) of moringa landraces. However, 1.5 dS m-1 and 3.5 dS m-1 were found more favorable. Higher salinity levels (7.5 dS m-1 and 11.5 dS m-1) significantly minimized the root surface area, root projected area, root volume and root density as compared to 1.5 dS m-1, 3.5 dS m-1. Number of branches, leaves, leaflets and leaf length were also adversely affected by 7.5 dS m-1 and 11.5 dS m-1. Maximum seedling fresh and dry weights, and seedling length were recorded at 1.5 dS m-1 followed by 3.5 dS m-1. Chlorophyll a and b contents, carotenoids and membrane stability index were also observed highest at salinity level of 1.5 dS m-1. In case of moringa landraces, MRB performed better regarding emergence attributes, growth parameters, and biochemical analysis followed by MFW as compared to MFB and MPB. Moringa landraces i.e. MRB and MFW were found more tolerant to salinity stress as compared to MFB and MPB., Competing Interests: The authors have declared that no competing interests exist.

Wheat is an important crop, used as staple food in numerous countries around the world. However, wheat productivity is low in the developing world due to several biotic and abiotic stresses, particularly drought stress. Non-availability of drought-tolerant wheat genotypes at different growth stages is the major constraint in improving wheat productivity in the developing world. Therefore, screening/developing drought-tolerant genotypes at different growth stages could improve the productivity of wheat. This study assessed seed germination and seedling growth of eight wheat genotypes under polyethylene glycol (PEG)-induced stress. Two PEG-induced osmotic potentials (i.e., -0.6 and -1.2 MPa) were included in the study along with control (0 MPa). Wheat genotypes included in the study were 'KLR-16', 'B6', 'J10', '716', 'A12', 'Seher', 'KTDH-16', and 'J4'. Data relating to seed germination percentage, root and shoot length, fresh and dry weight of roots and shoot, root/shoot length ratio and chlorophyll content were recorded. The studied parameters were significantly altered by individual and interactive effects of genotypes and PEG-induced osmotic potentials. Seed germination and growth parameters were reduced by osmotic potentials; however, huge differences were noted among genotypes. A reduction of 32.83 to 53.50% was recorded in seed germination, 24.611 to 47.75% in root length, 37.83 to 53.72% in shoot length, and 53.35 to 65.16% in root fresh weight. The genotypes, 'J4', 'KLR-16' and 'KTDH-16', particularly 'J4' better tolerated increasing osmotic potentials compared to the rest of the genotypes included in the study. Principal component analysis segregated these genotypes from the rest of the genotypes included in the study indicated that these can be used in the future studies to improve the drought tolerance of wheat crop. The genotype 'J4' can be used as a breeding material to develop drought resistant wheat genotypes., Competing Interests: The authors have declared that no competing interests exist.

Conyza sumatrensis (Retz.) E. H. Walker is an obnoxious weed, emerging as an invasive species globally. Seed germination biology of four populations of the species stemming from arid, semi-arid, temperate, and humid regions was determined in this study. Seed germination was recorded under six different environmental cues (i.e., light/dark periods, constant and alternating day and night temperatures, pH, salinity, and osmotic potential levels) in separate experiment for each cue. Populations were main factor, whereas levels of each environmental cue were considered as sub-factor. The impact of seed burial depths on seedling emergence was inferred in a greenhouse pot experiment. Seed germination was recorded daily and four germination indices, i.e., seed germination percentage, mean germination time, time to reach 50% germination, and mean daily germination were computed. Tested populations and levels of different environmental cues had significant impact on various seed germination indices. Overall, seeds stemming from arid and semi-arid regions had higher seed germination potential under stressful and benign environmental conditions compared to temperate and humid populations. Seed of all populations required a definite light period for germination and 12 hours alternating light and dark period resulted in the highest seed germination. Seed germination of all populations occurred under 5-30°C constant and all tested alternate day and night temperatures. However, the highest seed germination was recorded under 20°C. Seeds of arid and semi-arid populations exhibited higher germination under increased temperature, salinity and osmotic potential levels indicating that maternal environment strongly affected germination traits of the tested populations. The highest seed germination of the tested populations was noted under neutral pH, while higher and lower pH than neutral had negative impact on seed germination. Arid and semi-arid populations exhibited higher seed germination under increased pH compared to temperate and humid populations. Seed burial depth had a significant effect on the seedling emergence of all tested populations. An initial increase was noted in seedling emergence percentage with increasing soil depth. However, a steep decline was recorded after 2 cm seed burial depth. These results indicate that maternal environment strongly mediates germination traits of different populations. Lower emergence from >4 cm seed burial depth warrants that deep burial of seeds and subsequent zero or minimum soil disturbance could aid the management of the species in agricultural habitats. However, management strategies should be developed for other habitats to halt the spread of the species., Competing Interests: The authors have declared that no competing interests exist.

Theileria annulata is a tick-borne hemoprotozoan parasite responsible for tropical theileriosis in the bovine population, which causes substantial economic losses to the livestock sector. The present study has investigated, characterized, and shaped epidemiologic and phylogenetic profiles of T. annulata infection in the cattle population of central Khyber Pakhtunkhwa, Pakistan. A total of 600 blood samples were collected from cattle. Microscopy and PCR (18S rRNA taxonomic marker) assays were performed to detect T. annulata infection in cattle from the study area. The overall relative prevalence rates of T. annulata in the examined cattle population were 12.8% (microscopy) and 23.7% (PCR). District-wise analysis (microscopy/PCR) showed that cattle from district Mardan were found more infected (16.0%/28.0%), as compared to cattle from district Charsadda (13.5%/25.5%) and district Peshawar (9.0%/17.5%). Based on host demographic and ecological parameters analysis, theileriosis was found to be higher in young, female, crossbred, freely grazing, tick-infested, and irregular/no acaricides treated cattle. The univariate logistic analysis showed that host age, tick infestation, acaricides use, and feeding method were significant risk factors (P<0.05) whereas multivariate analysis indicated that host age, gender, tick infestation, acaricidal application, and feeding method were potential risk factors (P<0.05) for tropical theileriosis in the cattle population. Phylogenetic and sequence analysis showed that T. annulata 18S rRNA isolates shared homology and phylogeny with other isolates from Asia and Europe. This study has addressed the epidemiology and phylogeny of T. annulata circulating in bovid in the study area where gaps were still present. These findings will serve as a baseline and will facilitate future large-scale epidemiological investigations on tropical theileriosis in the cattle population at a national level., Competing Interests: All the authors have declared that they have no conflict of interest.

Vehicular emissions cause heavy metal pollution and exert negative impacts on environment and roadside vegetation. Wild plants growing along roadsides are capable of absorbing considerable amounts of heavy metals; thus, could be helpful in reducing heavy metal pollution. Therefore, current study inferred heavy metal absorbance capacity of some wild plant species growing along roadside. Four different wild plant species, i.e., Acacia nilotica L., Calotropis procera L., Ricinus communis L., and Ziziphus mauritiana L. were selected for the study. Leaf samples of these species were collected from four different sites, i.e., Control, New Lahore, Nawababad and Fatehabad. Leaf samples were analyzed to determine Pb2+, Zn2+, Ni2+, Mn2+ and Fe3+ accumulation. The A. nilotica, Z. mauritiana and C. procera accumulated significant amount of Pb at New Lahore site. Similarly, R. communis and A. nilotica accumulated higher amounts of Mn, Zn and Fe at Nawababad and New Lahore sites compared to the rest of the species. Nonetheless, Z. mauritiana accumulated higher amounts of Ni at all sites compared with the other species included in the study. Soil surface contributed towards the uptake of heavy metals in leaves; therefore, wild plant species should be grown near the roadsides to control heavy metals pollution. Results revealed that wild plants growing along roadsides accumulate significant amounts of heavy metals. Therefore, these species could be used to halt the vehicular pollution along roadsides and other polluted areas., Competing Interests: No competing interest exist.

Rice (Oryza sativa L.) feeds to two-third of the global population by serving as staple food. It is the main export commodity of several countries; thus, contributes towards foreign exchange earnings. Unfortunately, average global rice yield is far below than its genetic potential. Low nitrogen (N) use efficiency (NUE) is among the major reasons for low average yield. Current study evaluated the impact of nitrogen fertilizer application methods (conventional and deep placement) on growth, yield-related traits, chlorophyll contents, photosynthesis rate, agronomic N-use efficiency (ANUE), partial factors productivity of applied N (PFP) and economic returns of two different transplanted rice varieties (Basmati-515 and Super-Basmati). Fertilizer application methods significantly affected allometry, yield-related traits, chlorophyll contents, photosynthesis rate, ANUE, PFP and economic returns. Deep placement of N-fertilizer (DPNF) observed better allometric traits, high chlorophyll contents, photosynthesis rate, ANUE, PFP, yield attributes and economic returns compared to conventional application of N-fertilizer (CANF). Similarly, Basmati-515 had better allometric and yield-related traits, chlorophyll contents, photosynthesis rate, ANUE, PFP and economic returns than Super-Basmati. Regarding interactions among N-fertilizer application methods and rice varieties, Basmati-515 with DPNF resulted in higher chlorophyll contents, photosynthesis rate, ANUE, PFP, allometric and yield related traits and economic returns than CANF. The lowest values of these traits were observed for Super-Basmati with no application of N-fertilizer. Both varieties had better yield and economic returns with DPNF compared to CANF. It is concluded that DPNF improved yield, ANUE and economic returns; therefore, should be opted to improve productivity of transplanted fine rice. Nonetheless, lower nitrogen doses need to be tested for DPNF to infer whether it could lower N use in rice crop., Competing Interests: The authors have read the journal’s policy and have the following competing interests: AK and MB are paid employees of Agricultural Research Ltd. There are no patents, products in development or marketed products associated with this research to declare. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Numerous cropping systems of the world are experiencing the emergence of new weed species in response to conservation agriculture. Conyza stricta Willd. is being a newly emerging weed of barley-based cropping systems in response to conservational tillage practices. Seed germination ecology of four populations (irrigated, rainfed, abandoned and ruderal habitats) was studied in laboratory and greenhouse experiments. The presence/absence of seed dormancy was inferred first, which indicated seeds were non-dormant. Seed germination was then recorded under various photoperiods, constant and alternating day/night temperatures, and pH, salinity and osmotic potential levels. Seedling emergence was observed from various seed burial depths. Seeds of all populations proved photoblastic and required 12-hour light/dark period for germination. Seeds of all populations germinated under 5-30°C constant temperature; however, peak germination was recorded under 17.22-18.11°C. Nonetheless, the highest germination was noted under 20/15°C alternating day/night temperature. Ruderal and irrigated populations better tolerated salinity and germinated under 0-500 mM salinity. Similarly, rainfed population proved more tolerant to osmotic potential than other populations. Seeds of all populations required neutral pH for the highest germination, whereas decline was noted in germination under basic and alkaline pH. Seedling emergence was retarded for seeds buried >2 cm depth and no emergence was recorded from >4 cm depth. These results add valuable information towards our understanding of seed germination ecology of C. stricta. Seed germination ability of different populations under diverse environmental conditions suspects that the species can present severe challenges in future if not managed. Deep seed burial along with effective management of the emerging seedlings seems a pragmatic option to manage the species in cultivated fields. However, immediate management strategies are needed for rest of the habitats., Competing Interests: The authors have declared that no competing interests exist.

Multidrug resistance (MDR) to chemotherapeutic drugs remains one of the major impediments to the treatment of cancer. Discovery and development of drugs that can prevent and reverse the acquisition of multidrug resistance constitute a foremost challenge in cancer therapeutics. In this work, we screened a library of 1,127 compounds with known targets for their ability to overcome Pgp-mediated multidrug resistance in cancer cell lines. We identified four compounds (CHIR-124, Elesclomol, Tyrphostin-9 and Brefeldin A) that inhibited the growth of two pairs of parental and Pgp-overexpressing multidrug-resistant cell lines with similar potency irrespective of their Pgp status. Mechanistically, CHIR-124 (a potent inhibitor of Chk1 kinase) inhibited Pgp activity in both multidrug-resistant cell lines (KB-V1 and A2780-Pac-Res) as determined through cell-based Pgp-efflux assays. Other three inhibitors on the contrary, were effective in Pgp-overexpressing resistant cells without increasing the cellular accumulation of a Pgp substrate, indicating that they overcome resistance by avoiding efflux through Pgp. None of these compounds modulated the expression of Pgp in resistant cell lines. PIK-75, a PI3 Kinase inhibitor, was also determined to inhibit Pgp activity, despite being equally potent in only one of the two pairs of resistant and parental cell lines. Strong binding of both CHIR-124 and PIK-75 to Pgp was predicted through docking studies and both compounds inhibited Pgp in a biochemical assay. The inhibition of Pgp causes accumulation of these compounds in the cells where they can modulate the function of their target proteins and thereby inhibit cell proliferation. In conclusion, we have identified compounds with various cellular targets that overcome multidrug resistance in Pgp-overexpressing cell lines through mechanisms that include Pgp inhibition and efflux evasion. These compounds, therefore, can avoid challenges associated with the co-administration of Pgp inhibitors with chemotherapeutic or targeted drugs such as additive toxicities and differing pharmacokinetic properties., Competing Interests: The authors have declared that no competing interests exist.

The digenetic trematode Fasciola gigantica is a parasite of great agricultural and economic importance. Along with Fasciola hepatica, F. gigantica incurs huge economic losses to the agricultural sector. Because of unavailability of an effective and commercial vaccine, the earliest diagnosis of the disease is the only way to control the disease. The conventional coprological techniques are able to detect the disease only after the parasites get matured and starts releasing their eggs with the faeces of host, therefore prepatent infection remain undiagnosed. The alternative method is by serological tests that uses circulatory antigens. Despite high sensitivity, their reliability is quite low because of the common antigens shared between different helminth parasites. To overcome this, investigation was shifted to identify the copro-antigens which could be more sensitive and reliable. In the present study, we tried to identify some of the immunodominant proteins from the Excretory Secretory (ES) product of F. gigantica which can be further characterized and used for early detection of infection and also as drug and vaccine candidates. The ES products of F. gigantica were collected and used for raising the polyclonal antibody in rabbit. The polypeptide profile was generated as well as immunogenic polypeptides were identified. The Source of ES antigen was immunolocalized using confocal microscopy and dot blot assay was performed to diagnose field infection. The polypeptide profile of ES products revealed a total of 24 polypeptides out of which 12 immunogenic polypeptides were identified by western blotting. Confocal micrographs showed the immunolocalization of antigens in the intestinal caecae, vitalline glands, gonads as well as in the tegument of the worm. The dot blot assay confirmed the utility of ES products for the detection of field infection. Subsequently, cross reactivity was found negative with Gigantocotyle explanatum; an amphitome parasite of same habitat. However, the cross reactivity with other helminths needs to be worked out.

The current study presents the application of fluorescence spectroscopy for the identification of cow and buffalo milk based on β-carotene and vitamin-A which is of prime importance from the nutritional point of view. All samples were collected from healthy animals of different breeds at the time of lactation in the vicinity of Islamabad, Pakistan. Cow and buffalo milk shows differences at fluorescence emission appeared at band position 382 nm, 440 nm, 505 nm and 525 nm both in classical geometry (right angle) setup as well as front face fluorescence setup. In front face fluorescence geometry, synchronous fluorescence emission shows clear differences at 410 nm and 440 nm between the milk samples of both these species. These fluorescence emissions correspond to fats, vitamin-A and β-carotene. Principal Component Analysis (PCA) further highlighted these differences by showing clear separation between the two data sets on the basis of features obtained from their fluorescence emission spectra. These results indicate that classical geometry (fixed excitation wavelength) as well as front face (synchronous fluorescence emission) of cow and buffalo milk nutrients could be used as fingerprint from identification point of view. This same approach can effectively be used for the determination of adulterants in the milk and other dairy products.

Fasciolosis an economically important global disease of ruminants in the temperate and tropical regions, caused by Fasciola hepatica and F. gigantica, respectively, also poses a potential zoonotic threat. In India alone it causes huge losses to stakeholders. Anthelmintics including triclabendazole have been used to control this menace but the emerging resistance against the available compounds necessitates identification of novel and alternative therapeutic measures involving plant derived natural compounds for their anthelmintic potential. Thymoquinone (T) and curcumin (C), the active ingredients of Nigella sativa and Curcuma longa respectively have been used as antiparasitic agents but the information on their flukicidal effect is very limited. Adult flukes of F. gigantica were in vitro exposed to different concentrations of thymoquinone and curcumin separately for 3h at 37+ 1°C. A significant (p<0.05) reduction in the worm motility at 60 μM concentration of both T and C was observed though all the worms remained alive after 3h exposure, whereas the effect on egg shedding was statistically insignificant. Pronounced tegumental disruptions and erosion of spines in the posterior region and around the acetabulum was evident. A significant (p<0.05) decrease in glutathione-S-transferase and superoxide dismutase activity and reduced glutathione (GSH) level was observed, while protein carbonylation increased differentially. A significant inhibition of CathepsinL (CatL) gene expression in thymoquinone treated worms was also evident. Further, in silico molecular docking of T and C with CatL revealed a stronger interaction of curcumin with the involvement of higher number of amino acids as compared to thymoquinone that could be more effective in inhibiting the antioxidant enzymes of F. gigantica. It is concluded that both the compounds understudy will decrease the detoxification ability of F. gigantica, while inhibition of CatL will significantly affect their virulence potential. Thus, both thymoquinone and curcumin appeared to be promising anthelmintic compounds for further investigations., Competing Interests: The authors have declared that no competing interests exist.

Proteases are widely used to remove affinity and solubility tags from recombinant proteins to avoid potential interference of these tags with the structure and function of the fusion partner. In recent years, great interest has been seen in use of the human rhinovirus 3C protease owing to its stringent sequence specificity and enhanced activity. Like other proteases, activity of the human rhinovirus 3C protease can be affected in part by the buffer components and additives that are generally employed for purification and stabilization of proteins, hence, necessitate their removal by tedious and time-consuming procedures before proteolysis can occur. To address this issue, we examined the effect of elution buffers used for common affinity based purifications, salt ions, stability/solubility and reducing agents, and detergents on the activity of the human rhinovirus 3C protease using three different fusion proteins at 4°C, a temperature of choice for purification of many proteins. The results show that the human rhinovirus 3C protease performs better at 4°C than the frequently used tobacco etch virus protease and its activity was insensitive to most of the experimental conditions tested. Though number of fusion proteins tested is limited, we expect that these finding will facilitate the use of the human rhinovirus 3C protease in recombinant protein production for pharmaceutical and biotechnological applications.

Helminth parasites of veterinary importance cause huge revenue losses to agrarian economy worldwide. With the emergence of drug resistance against the current formulations, there is a need to focus on the alternative approaches in order to control this menace. In the present study, biocompatible zinc oxide nanoparticles (ZnO NPs) were used to see their in vitro effect on the biliary amphistomes, Gigantocotyle explanatum, infecting Bubalus bubalis because these nanoparticles are involved in generation of free radicals that induce oxidative stress, resulting in disruption of cellular machinery. The ZnO NPs were synthesized by using egg albumin as a biotemplate and subsequently characterized by Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), X-ray Diffraction and Spectrophotometrical, which showed that ZnO NPs were highly purified wurtzite type polycrystals, with a mean size of 16.7 nm. When the parasites were treated with lower concentrations (0.004% and 0.008%) of the ZnO NPs, the worms mounted a protective response by stimulating the antioxidant system but the treatment of G. explanatum with 0.012% ZnO NPs produced significant inhibition of the antioxidant enzymes like superoxide dismutase (SOD) (p< 0.05) and glutathione S- transferase (GST) (p<0.01), while the level of malondialdehyde (MDA), a lipid peroxidation marker, was significantly (p< 0.01) elevated. SEM and histopathology revealed pronounced tegumental damage showing the disruption of surface papillae and the annulations, particularly in the posterior region near acetabulum. The under expression of a number of polypeptides, loss of worm motility in a time dependent manner, further reflect strong anthelmintic potential of ZnO NPs. It can be concluded that the anthelmintic effect might be due to the production of reactive oxygen species that target a variety of macromolecules such as nucleic acid, protein and lipids which are involved in different cellular processes.