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Noncentrifugal sugar (NCS) is an unrefined, dark brown sugar containing minerals and plant secondary metabolites, unlike refined white sugar (WS). This study explored using NCS in confectionary jellies as an alternative sugar. We used different concentrations of NCS and WS to prepare low methoxyl pectin (LMP) confectionery gels characterized by their physical and rheological properties along with time-domain nuclear magnetic resonance (TD-NMR) relaxometry. The strongest LMP gel, with a hardness of 0.94 N, was achieved by substituting 25% of WS with NCS at a low CaCl 2 concentration (0.075 M). Gels with up to 50% WS replaced by NCS showed comparable hardness to standard LMP gels made solely with WS at a 0.15 M CaCl 2 concentration, attributed to NCS's unique constituents. The NCS-WS gel exhibited the shortest T 2 values (139.8 ms) and self-diffusion coefficient values (4.99 × 10-10 m 2 /s), indicating a denser, more cross-linked structure that restricted water mobility. These findings suggest NCS's complex role in affecting LMP gels' chemical and physical properties, highlighting its potential as a partial WS replacement in LMP gelation-based products, with an additional source of minerals and antioxidants., Competing Interests: The authors declare no competing financial interest., (© 2024 The Authors. Published by American Chemical Society.)

Background: Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by severe pain, inflammation, and joint deformity. Currently, it affects 1% of the population, with a projection to exceed 23 million cases by 2030. Despite significant advancements, non-steroidal anti-inflammatory drugs (NSAIDs), the first line of treatment, are associated with a range of adverse effects. Consequently, plant-based derivatives are being utilized as an effective alternative. This study evaluates the anti-inflammatory and safety profile of Colchicum luteum hydroethanolic extract (CLHE) in comparison to NSAIDs, with a focus on COX-2 and TNFα inhibition., Methods: CLHE potential was evaluated by phytochemical screening and in vitro bioactivity assays. Toxicity profile was conducted in Human Colon Epithelial Cells (HCEC) and Balb/c mice. Anti-inflammatory potential was explored in a collagen-induced arthritic (CIA) mice model. Bioactive compounds were identified computationally from GCMS data and subjected to docking and simulation studies against COX2 and TNFα., Results: CLHE demonstrated significant antioxidant (IC-50 = 6.78 µg/mL) and anti-inflammatory (IC-50 = 97.39 µg/mL) activity. It maintained 50% cell viability at 78.5 μg/µL in HCEC cells and exhibited no toxicity at a dose of 5000 mg/kg in mice. In the CIA model, CLHE significantly reduced paw swelling, arthritic scoring, C-reactive protein levels, and spleen indices, outperforming ibuprofen. Expression analysis confirmed the downregulation of COX-2, TNFα, and MMP-9. Histopathological analysis indicated the superior efficacy of CLHE compared to ibuprofen in reducing inflammation, synovial hyperplasia, and bone erosion. Computational studies identified compound-15 (CL15), (4-(4,7-dimethoxy-1,3-benzodioxol-5-yl)-2-oxo pyrrolidine-3-carboxylic acid), a non-toxic compound with strong binding affinities to COX-2 (-12.9 KJ/mol), and TNF-α (-5.8 KJ/mol)., Conclusions: The findings suggest the potential of Colchicum luteum as a safer, anti-inflammatory, and multi-targeted alternative to NSAIDs for RA treatment.

1 H fast field-cycling and time-domain nuclear magnetic resonance relaxometry studies have been performed for 15 samples of sugar of different kinds and origins (brown, white, cane, beet sugar). The extensive data set, including results for crystal sugar and sugar/water mixtures, has been thoroughly analyzed, with a focus on identifying relaxation contributions associated with the solid and liquid fractions of the systems and non-exponentiality of the relaxation processes. It has been observed that 1 H spin-lattice relaxation rates for crystal sugar (solid) vary between 0.45 s -1 and 0.59 s -1 , and the relaxation process shows only small deviations from exponentiality (a quantitative measure of the exponentiality has been provided). The 1 H spin-lattice relaxation process for sugar/water mixtures has turned out to be bi-exponential, with the relaxation rates varying between about 13 s -1 -17 s -1 (for the faster component) and about 2.1 s -1 -3.5 s -1 (for the slower component), with the ratio between the amplitudes of the relaxation contributions ranging between 2.8 and 4.2. The narrow ranges in which the parameters vary make them a promising marker of the quality and authenticity of sugar., Competing Interests: Author Leonid Grunin was employed by the company Resonance Systems GmbH. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

The research work was commenced with an aim to document plant based drug recipes used by autochthonous practitioners against liver diseases in Multan region. The data was collected by interviews and semi-structured questionnaire from 43 traditional herbal practitioners. Recorded data was evaluated using the use value, relative frequency of citation (RFC), family importance value (FIV) and family use value (FUV). Total of 69 plant species belonging to 38 families were reported to be utilized to treat various liver disorders. The most important species regarding their use value were 3.0 for Polygonum bistorta , Citrus limon . It means that highly cited plants are more important in traditional medicines but this does not mean the plants low-citation plants by respondents are less medicinal important. It may be due to unavailability of plants in the area or lack of knowledge. Most of plant parts used were leaves due to ease of collection as compared to underground parts. One the basis family use value Asteraceae as the dominating or prominent family. These plants can be further investigated for allied pharmacological studies that will affect the socio-economical condition of the local community., Competing Interests: The authors declare no conflict of interest., (© 2023 Published by Elsevier Ltd.)

Jasminum L. is the largest genus containing ~200 species found wild mostly in the tropical regions of the world. The comparative palynological study of nine Pakistani Jasminum species with SEM showed zonocolpus, trilobate, and tricolpus pollen types with simple endocolpus apertures which are plesiomorphic and conserved in the Jasminum species. The equatorial pollen view was prolate, subprolate, and perprolate with elliptic, lobate, subcircular whereas polar view was subtriangular in all species. Few characters were specific to some species like heteropolarity in Jasminum grandiflorum and foveolate exine ornamentation with rounded heterobrochate in Jasminum angulare whereas reticulate and angular homobrochate character was common in other species. The UPGMA dendrogram based on qualitative characters did not support the phylogenetic classification of the genus Jasminum as these are highly conserved. The quantitative data showed more variation in some characters whereas few characters showed little or no variation. A greater variation in pollen size was observed among the variants of same species, for example, Jasminum humile showed highly variable polar length and equatorial diameter as compared to other species. Minimum variation was observed in colpus length which divided all species in to two groups. The large lumina were specific to Jasminum nitidum and broader muri was the prominent characteristic of Jasminum angulare. Some species like Jasminum sambac and Jasminum azoricum were unable to develop true pollen due to structural or functional disabilities. So, the quantitative characters of pollen are only suitable for palynological based grouping of Jasminum species but less suitable to infer their evolutionary relationship., (© 2021 Wiley Periodicals LLC.)

The genus Jasminum L., of the family Oleaceae, includes many species occurring in the wild, or cultivated worldwide. A preliminary investigation based on inter-simple sequence repeats (ISSR) was performed to assess the genetic diversity among 28 accessions, representing nine species of Jasminum from various regions, representing a range of altitudes in Pakistan. A total of 21 ISSR primers were used, which produced 570 amplified bands of different sizes, with a mean polymorphic band percentage of 98.26%. The maximum resolving power, polymorphism information content, and index values of the ISSR markers recorded for primers 6, 16, and 19 were 0.40, 12.32, and 24.21, respectively. Based on the data of the ISSR markers, the resulting UPGMA dendrogram with the Jaccard coefficient divided the 28 accessions into two main clades. At the species level, the highest values for Shannon's information index, polymorphism percentage, effective allele number, Nei's genetic variations, and genetic unbiased diversity were found in Jasminum sambac L. and J. humile L., while the lowest were observed in J. mesnyi Hance and J. nitidum Skan. Based on Nei's unbiased genetic identity pairwise population matrix, the maximum identity (0.804) was observed between J. elongatum Willd and J. multiflorum (Burm. f.) Andrews, and the lowest (0.566) between J. nitidum Skan. and J. azoricum L. Molecular variance analysis displayed a genetic variation of 79% among the nine populations. The study was aimed to established genetic diversity in Jasminum species using ISSR markers. With the help of this technique, we were able to establish immense intra- and interspecific diversity across the Jasminum species.

The CRISPR-Cas9 system has recently evolved as a powerful mutagenic tool for targeted genome editing. The impeccable functioning of the system depends on the optimal design of single guide RNAs (sgRNAs) that mainly involves sgRNA specificity and on-target cleavage efficacy. Several research groups have designed algorithms and models, trained on mammalian genomes, for predicting sgRNAs cleavage efficacy. These models are also implemented in most plant sgRNA design tools due to the lack of on-target cleavage efficacy studies in plants. However, one of the major drawbacks is that almost all of these models are biased for considering only coding regions of the DNA while excluding ineffective regions, which are of immense importance in functional genomics studies especially for plants, thus making prediction less reliable. In the present study, we evaluate the on-target cleavage efficacy of experimentally validated sgRNAs designed against diverse ineffective regions of Arabidopsis thaliana genome using various statistical tests. We show that nucleotide preference in protospacer adjacent motif (PAM) proximal region, GC content in the PAM proximal seed region, intact RAR and 3 rd stem loop structures, and free accessibility of nucleotides in seed and tracrRNA regions of sgRNAs are important determinants associated with their high on-target cleavage efficacy. Thus, our study describes the features important for plant sgRNAs high on-target cleavage efficacy against ineffective genomic regions previously shown to give rise to ineffective sgRNAs. Moreover, it suggests the need of developing an elaborative plant-specific sgRNA design model considering the entire genomic landscape including ineffective regions for enabling highly efficient genome editing without wasting time and experimental resources., Competing Interests: The authors declare there are no competing interests., (©2021 Malik et al.)

In plants, F-box proteins (FBPs) constitute one of the largest superfamilies of regulatory proteins. Most F-box proteins are shown to be an integral part of SCF complexes, which carry out the degradation of proteins and regulate diverse important biological processes. Anthers and pollen development have a huge importance in crop breeding. Despite the vast diversity of FBPs in Arabidopsis male reproductive organs, their role in anther and pollen development is not much explored. Moreover, a standard nomenclature for naming FBPs is also lacking. Here, we propose a standard nomenclature for naming the FBPs of Arabidopsis thaliana uniformly and carry out a systematic analysis of sperm cell-specific FBP gene, i.e., 3p.AtFBP113 due to its reported high and preferential expression, for detailed functional annotation. The results revealed that 3p.AtFBP113 is located on the small arm of chromosome and encodes 397 amino acid long soluble, stable, and hydrophilic protein with the possibility of localization in various cellular compartments. The presence of the C-terminal F-box associated domain (FBA) with immunoglobulin-like fold anticipated its role in protein binding. Gene ontology based functional annotation and tissue-specific gene co-expression analysis further strengthened its role in protein binding and ubiquitination. Moreover, various potential post/co-translational modifications were anticipated and the predicted tertiary structure also showed the presence of characteristic domains and fold. Thus, the outcomes of the study will be useful in developing a better understating of the function of 3p.AtFBP113 during the process of pollen development, which will be helpful for targeting the gene for manipulation of male fertility that has immense importance in hybrid breeding., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2020 Malik, Gul, Amir, Munir, Babar, Bakhtiar, Hayat, Paracha, Khalid and Alipour.)

Rheumatoid arthritis (RA) is a multifactorial disease which is complicated by apoptosis resistance. Autophagy is one of the key mechanisms which are involved in the development of resistance to apoptosis as well as to the standard therapies against RA. Aberration in autophagy and apoptosis homeostasis results in the development of oxidative stress thus complicates the pathogenesis of RA. In the given study, tomorou, an indigenous herb of Hunza-Nagar Valley, has been evaluated for its pro-apoptotic, anti-inflammatory, and anti-rheumatic activity. Several major classes of bioactive phytochemicals including steroids, terpenoids, phenols, flavonoids, and essential oils have been detected in the aqueous and ethyl acetate extracts of tomorou through phytochemical analysis. Plant extracts depicted enhanced free radical scavenging activity through di-phenyl-2-picryl hydrazyl hydrate (DPPH) assay and ameliorated the symptoms of arthritis in collagen induced arthritic (CIA) mice model. Moreover, the 6 week extract treatment resulted in the reduction of IL-6 serum levels thus making it an effective anti-inflammatory agent. Upregulation of microtubule-associated proteins light chain 3b (LC3b) and downregulation of UNC51-like kinase 1 (ULK-1) in arthritic mice proposed a ULK-1 independent non-canonical autophagy pathway. Treatment with extracts upregulated the expression of caspase 3 which in turn inhibited the activity of LC3b thus altering the autophagy pathway. However, ULK-1 expression was restored to normal in aqueous extract treated group whereas it was upregulated in ethyl acetate extract treated group. On the other hand, a novel LC3b-independent autophagy pathway was observed in mice treated with ethyl acetate extract due to ULK-1 upregulation. Despite of significantly high IL-6 levels, the arthritic symptoms waned off which suggested the participation of IL-6 in LC3b-independent autophagy pathway in the extract prepared in ethyl acetate. Conclusively, the study established pro-apoptotic, antioxidant, anti-inflammatory and anti-rheumatic activity of tomorou and suggested an intricate autophagy pathway shift., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© The Author(s) 2019.)

Artemisia L. is among the most diverse and medicinally important genera of the plant family Asteraceae. Discrepancies arise in the taxonomic classification of Artemisia due to the occurrence of multiple polyploidy events in separate lineages and its complex morphology. The discrepancies could be resolved by increasing the genomic resources. A. scoparia is one of the most medicinally important species in Artemisia . In this paper, we report the complete chloroplast genome sequence of Artemisia scoparia . The genome was 151,060 bp (base pairs), comprising a large single copy (82,834 bp) and small single copy (18,282 bp), separated by a pair of long inverted repeats (IRa and IRb: 24,972 bp each). We identified 114 unique genes, including four ribosomal RNAs, 30 transfer RNAs, and 80 protein-coding genes. We analysed the chloroplast genome features, including oligonucleotide repeats, microsatellites, amino acid frequencies, RNA editing sites, and codon usage. Transversion substitutions were twice as frequent as transition substitutions. Mutational hotspot loci included ccsA-ndhD , trnH-psbA , ndhG-ndhI , rps18-rpl20 , and rps15-ycf1 . These loci can be used to develop cost-effective and robust molecular markers for resolving the taxonomic discrepancies. The reconstructed phylogenetic tree supported previous findings of Artemisia as a monophyletic genus, sister to the genus Chrysanthemum , whereby A. scoparia appeared as sister to A. capillaris .

An improved active packaging system was developed for fresh fruits using silver nanoparticles (AgNPs) coupled with calcium alginate (Ca-ALG). For the synthesis of AgNPs aqueous, ethanol and methanol extracts of Artemisia scoparia (AS) were used. These AgNP's were characterized using UV-Vis, SEM, EDS, AFM, FTIR and gel electrophoresis. Ethanol extract of AS (ASE) produced AgNPs with smallest size in comparison to aqueous AS (ASA) and methanol extract of AS (ASM). AgNPs synthesized from ASE had a size range of 12.0-23.3 nm and were tested on Human Corneal Epithelial Cells to evaluate their cytotoxicity. At 0.05 ng/mL of AgNP's concentration, no toxic effects were observed on the evaluated cell line. Therefore, 0.05 ng/mL of AgNPs mixed with edible coating of Ca-ALG were applied on strawberries and loquats as active coating to increase their shelf life. Significant improvement was observed in the quality parameters of strawberries and loquats such as microbial analysis, acidity loss, soluble solid content loss, weight loss and quality decay. Ca-ALG coating incorporated with AgNPs enhanced the shelf life of strawberries and loquats in comparison to no treatment and simple Ca-ALG coatings. This study provides an insight to food industry to extend the shelf life of fresh fruits using AgNP's formulated coatings., (Copyright © 2019. Published by Elsevier B.V.)

Background: Diversification patterns in the Himalayas have been important to our understanding of global biodiversity. Despite recent broad-scale studies, the most diverse angiosperm genus of the temperate zone- Carex L. (Cyperaceae), with ca. 2100 species worldwide-has not yet been studied in the Himalayas, which contains 189 Carex species. Here the timing and phylogenetic pattern of lineage and ecological diversification were inferred in this ecologically significant genus. We particularly investigated whether priority, adaptation to ecological conditions, or both explain the highly successful radiation of the Kobresia clade (ca. 60 species, of which around 40 are present in the Himalayas) of Himalayan Carex., Methods: Phylogenetic relationships were inferred using maximum likelihood analysis of two nuclear ribosomal DNA (nrDNA) regions (ITS and ETS) and one plastid gene ( mat K); the resulting tree was time-calibrated using penalized likelihood and a fossil calibration at the root of the tree. Biogeographical reconstruction for estimation of historical events and ancestral ranges was performed using the dispersal-extinction-cladogenesis (DEC) model, and reciprocal effects between biogeography and diversification were inferred using the geographic state speciation and extinction (GeoSSE) model. Climatic envelopes for all species for which mapped specimen data available were estimated using climatic data from WORLDCLIM, and climatic niche evolution was inferred using a combination of Ornstein-Uhlenbeck models of shifting adaptive optima and maximum likelihood inference of ancestral character states under a Brownian motion model., Results: The Himalayan Carex flora represents three of the five major Carex clades, each represented by multiple origins within the Himalayas. The oldest Carex radiation in the region, dating to ca. 20 Ma, near the time of Himalayan orogeny, gave rise to the now abundant Kobresia clade via long-distance dispersal from the Nearctic. The Himalayan Carex flora comprises a heterogeneous sample of diversifications drawn from throughout the cosmopolitan, but mostly temperate, Carex radiation. Most radiations are relatively recent, but the widespread and diverse Himalayan Kobresia radiation arose at the early Miocene. The timing and predominance of Kobresia in high-elevation Himalayan meadows suggests that Kobresia may have excluded other Carex lineages: the success of Kobresia in the Himalayas, in other words, appears to be a consequence largely of priority, competitive exclusion and historical contingency., Competing Interests: The authors declare there are no competing interests. Pedro Jiménez-Mejías is an Academic Editor for PeerJ.

This study reports on a simple approach for the fabrication of an electrode modified with biocompatible C-dot wrapped ZnO nanoparticles for selective photoelectrochemical monitoring of H 2 O 2 released from living cells. The biocompatibility of the ZnO nanoparticles was confirmed through in-vitro cellular testing using the MTT assay on Huh7 cell lines. The ZnO nanoparticles wrapped with dopamine-derived C-dots possess numerous catalytically active sites, excessive surface defects, good electrical conductivity, and efficient separation ability of photo-induced electrons and holes. These properties offer highly sensitive and selective non-enzymatic photo-electrochemical monitoring of H 2 O 2 released from HeLa cells after stimulation with N-formylmethionyl-leucyl-phenylalanine. The sensor has a wide linear range (20-800 nM), low detection limit (2.4 nM), and reliable reproducibility, this implying its suitability for biological and biomedical applications. Graphical abstract Schematic of the fabrication of ZnO nanoparticles by using a plant extract as a reducing agent. Wrapping of ZnO with C-dots enhances the photoelectrocatalytic efficacy. Sensitive and selective photoelectrochemical monitoring of H 2 O 2 released from cancer cells is demonstrated.

Skin wound healing is a multistage phenomenon that is regulated by cell-cell interplay and various factors. Endogenous serotonin is an important neurotransmitter and cytokine. Its interaction with the serotonin 1A receptor (5-HTR1A) delivers downstream cellular effects. The role of serotonin (5-hydroxytryptamine, 5-HT) and the 5-HT1A receptor has been established in the regeneration of tissues such as the liver and spinal motor neurons, prompting the investigation of the role of 5-HT1A receptor in skin healing. This study assessed the role of 5-HT1A receptor in excisional wound healing by employing an excisional punch biopsy model on 5-Ht1a receptor knockout mice. Post-harvest analysis revealed 5-Ht1a receptor knockout mice showed impaired skin healing, accompanied by a greater number of F4/80 macrophages, which prolongs the inflammatory phase of wound healing. To further unravel this phenomenon, we employed the 5-HT1A receptor agonist [(R)-(+)-8-Hydroxy-DPAT hydrobromide] as a topical cream treatment in an excisional punch biopsy model. The 5-HT1A receptor agonist treated group showed a smaller wound area, scar size, and improved neovascularization, which contributed to improve healing outcomes as compared to the control. Collectively, these findings revealed that serotonin and 5-HT1A receptor play an important role during the healing process. These findings may open new lines of investigation for the potential treatment alternatives to improve skin healing with minimal scarring.

Artemisinin and synthetic derivatives of dihydroartemisinin are known to possess various biological activities. Post-functionalization of dihydroartemisinin with triazole heterocycles has been proven to lead to enhanced therapeutic potential. By using our newly developed triazolization strategy, a library of unexplored fused and 1,5-disubstituted 1,2,3-triazole derivatives of dihydroartemisinin were synthesized in a single step. All these newly synthesized compounds were characterized and evaluated for their anti-HIV (Human Immunodeficiency Virus) potential in MT-4 cells. Interestingly; three of the synthesized triazole derivatives of dihydroartemisinin showed activities with half maximal inhibitory concentration (IC 50 ) values ranging from 1.34 to 2.65 µM.

Cannabis sativa (C.sativa) is well-known for its medicinal, industrial and recreational use. However, allergies in relation to Cannabis sativa (C.sativa) are rarely reported. C. sativa is one of the common weeds found in Pakistan and its pollen grains are common in spring and fall season. Although categorized as an aeroallergen, there are limited number of reports regarding allergenic potential in C. sativa. Therefore, the current study is aimed at exploring the IgE- binding potential among the C. sativa pollen in local pollen allergic patients. Initial screening of C. sativa sensitized individuals was carried out by dot blot from the sera of pollen allergic patients. Proteins from the pollen grains were extracted and resolved on 10% gel. Eight bands were visible on gel however only one protein fragment i.e. of 14KDa size was found to bind to IgE as analyzed through protein gel blot analysis. Strong IgE affinity of a 14 kDa protein fragment from C. sativa pollen extract suggests its allergenic potential. Further study is required to find the exact nature of this protein fragment.

Background: Cassia angustifolia Vahl. (commonly known as senna makkai or cassia senna), native to Saudi Arabia, Egypt, Yemen and also extensively cultivated in Pakistan, is a medicinal herb used traditionally to cure number of diseases like liver diseases, constipation, typhoid, cholera etc. This study was conducted to evaluate the in-vitro antimicrobial, antioxidant and anticancer assays and phytochemical constituents of aqueous and organic extracts of C. angustifolia leaves., Methods: The antimicrobial activities of C. angustifolia aqueous and organic (methanol, ethanol, acetone, ethyl acetate) extracts were investigated by the disk diffusion method. These extracts were further evaluated for antioxidant potential by the DPPH radical scavenging assay. Anticancer activities of the extracts were determined by the MTT colorimetric assay. The total phenolic and flavonoid contents of C. angustifolia extracts were evaluated by the Folin-Ciocalteu method and aluminum chloride colorimetric assay, respectively. The structures of the bioactive compounds were elucidated by NMR and ESI-MS spectrometry., Results: Bioactivity-guided screening of C. angustifolia extracts, led to the isolation and identification of three flavonoids quercimeritrin (1), scutellarein (2), and rutin (3) reported for the first time from this plant, showed significant anticancer activity against MCF-7 (IC 50, 4.0 μg/μL), HeLa (IC 50, 5.45 μg/μL), Hep2 (IC 50, 7.28 μg/μL) and low cytotoxicity against HCEC (IC 50, 21.09 μg/μL). Significant antioxidant activity was observed with IC 50 2.41 μg/mL against DPPH radical. Moreover, C. angustifolia extracts have the potential to inhibit microbial growth of E. cloacae, P. aeruginosa, S. mercescens and S. typhi., Conclusion: C. angustifolia extracts revealed the presence of quercimeritrin (1), scutellarein (2), and rutin (3), all known to have useful bioactivities including antimicrobial, antioxidant and anticancer activities.

Hepatitis C Virus (HCV) is the most prevalent human pathogen in Pakistan and is the major cause of liver cirrhosis and hepatocellular carcinoma in infected patients. It has shifted from being hypo-endemic to being hyper-endemic. There was no information about the origin and evolution of the local variants. Here we use newly developed phyloinformatic methods of sequence analysis to conduct the first comprehensive investigation of the evolutionary and biogeographic history in unprecedented detail and breadth. Considering evolutionary rate and molecular-clock hypothesis in context, we reconstructed the spatiotemporal spread of HCV in the whole territory of its circulation using a combination of Bayesian MCMC methods utilizing all sequences available in GenBank. Comparative analysis were performed and were addressed. Whole genome and individual gene analysis have shown that sub-types 1a, 1b and 3a are recognized as epidemic strains and are distributed globally. Here we confirm that the origin of HCV 3a genotypes is in South Asia and HCV has evolved in the region to become stably adapted to the host environment., Competing Interests: The authors have declared that no competing interests exist.

Background: The present study is intended to compare and document the therapeutic flora, their remedial use, and the traditional knowledge used frequently by the residents of the Cholistan desert and Pothwar (Potohar) Plateau of Punjab, Pakistan. The old endemic remedies of these areas are diminishing due to lack of qualitative and quantitative research., Methods: The data was generated by unstructured-interviews, informal meetings, open-ended conversations and group discussions with local people and traditional health healers of the study area. Reported literature was also utilized., Results: The study recorded a list of various medicinal plants used as traditional medicines by local people. Total 86 numbers of plant species belonging to 38 families and 67 plant species belonging to 29 families have been reported in the Pothwar and Cholistan respectively. Only 10.5% of similar plant species were present in the studied areas., Conclusion: The investigation revealed that the local people of study areas inherit a rich traditional knowledge but there is great danger of losing this wealth of knowledge in the near future. Documentation of the knowledge exclusively from desert area of Cholistan, Pakistan is unique information in its nature. The study presents the undocumented knowledge worth recognition that will not only help in conservation of medicinal plant species but will highlight the pharmacological capacity for improved human healthcare regarding many common ailments.