Application of microscopy and spectroscopy in investigating anti-cancer potential of Achyranthes aspera L. leaves.

The genus Achyranthes belong to the family Amaranthaceae which constitutes an important group of herbs and shrubs with immense medicinal value. The present research work was conducted to investigate the anticancer potential of Achyranthes aspera L. leaves by focusing on the antioxidant, aniproliferative and antimitotic activities of leaf extracts. Plant extraction was carried out by soxhelt method with different solvents. Phytochemical characterization of the plants extracts using chemical methods identified the presence of cardiac glycosides, saponins, coumarins, proteins, tannins, flavonoids and triterpenes. Alkaloid was present in methanolic and ethanolic extract. High performance liquid chromatography showed presence of different concentration of myricetin, quercetin and kaempferol in different extracts with the highest concentration of myricetin (84.53 µg/mL) in n-butanolic extract. The extracts were then tested for antioxidant activity using 2,2-diphenylpicrylhydrazyl (DPPH) radical scavenging assay by spectrophotometric method. In DPPH radical scavenging assay, antioxidant activity of A. aspera ranged between 79.78 ± 0.034% and 58.63 ± 0.069%. Highest antioxidant activity was observed for methanolic extract and lowest for acetone. Antimitotic activity was determined by using Allium cepa assay in which microscopic investigation was carried out to observe normal and abnormal phases of mitosis. In this assay, n-butanolic extract had highest antimitotic activity with minimum mitotic index at 2 mg/mL (57 ± 0.0351%). The plant extracts also caused chromosomal and mitotic aberrations which were clearly observed under 40× and 100× magnification of compound microscope. Antiproliferative activity was determined by using yeast cell model in which light microscope with hemocytometer was used for cell counting. In case of Antiproliferative activity, the ethyl acetate extract of A. aspera had highest antiproliferative activity with lowest cell viability (22.14 ± 0.076%) at highest extract concentration (2 mg/mL) while methanol extract of A. aspera had highest antiproliferative activity with lower cell viability (24.24 ± 0.057%) at lowest extract concentration (0.25 mg/mL). The results of the study indicated that the leaves extract of A. aspera have strong potential to be used as a source of anti-cancer agent. RESEARCH HIGHLIGHTS: Achyranthes aspera L. leaves have various phytochemicals which contribute to its medicinal properties Various extracts of the leaves of A. aspera L. possess antioxidant, antimitotic and antiproliferative potential The results of the study indicated that the leaves extract of A. aspera have strong potential to be used as a source of anti-cancer agent.

Termitomyces pakistanensis, a new mushroom species from Pakistan based on scanning electron microscopy and ITS-rDNA barcoding.

Termitomyces pakistanensis sp. nov. is a member of an edible genus generally distributed in Asia and Europe. This species has been described as new species based on its different morphology, and scanning electron microscopy (SEM) of basidiospores. The novelty and degree of endemism is confirmed by analyzing the genetic variation of the internal transcribed spacer regions (ITS1-5.8 S-ITS2) of the ribosomal DNA gene, a universal fungal marker. The evolutionary affinities of new species is also evaluated with Asian and European species by phylogenetic analysis based on ITS sequences. In our phylogenetic analysis, this genus is found monophyletic comprising of two monophyletic sub clades: Clade I, Microcarpus, with small sized fruiting bodies generally less than 5 cm without pseudorrhiza and Clade II, Macrocarpus, with large sized fruiting bodies generally more than 5 cm having pseudorrhiza. All collections of Pakistani species clustered independently in Microcarpus clade showing their endemic genetic makeup as it is clustering independently. A comprehensive description, photographs of the basidiocarps and Scanning electron microscopy (SEM) micrographs of spores are provided. RESEARCH HIGHLIGHTS: It has a new species from Pakistan to world based on the scanning electron microscopy and further confirmed by DNA barcoding. The exact shape and size of basidiospores of this novel species is first time introduced by using SEM analysis. This genus is rarely described from Pakistan. This paper has introduced a two clade, Microcarpus and macrocarpus, in the world for this genus.

Potential medicinal plants involved in inhibiting 3CLpro activity: A practical alternate approach to combating COVID-19.

At present, a variety of vaccines have been approved, and existing antiviral drugs are being tested to find an effective treatment for coronavirus disease 2019 (COVID-19). However, no standardized treatment has yet been approved by the World Health Organization. The virally encoded chymotrypsin-like protease (3CLpro) from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which facilitates the replication of SARS-CoV in the host cells, is one potential pharmacological target for the development of anti-SARS drugs. Online search engines, such as Web of Science, Google Scholar, Scopus and PubMed, were used to retrieve data on the traditional uses of medicinal plants and their inhibitory effects against the SARS-CoV 3CLpro. Various pure compounds, including polyphenols, terpenoids, chalcones, alkaloids, biflavonoids, flavanones, anthraquinones and glycosides, have shown potent inhibition of SARS-CoV-2 3CLpro activity with 50% inhibitory concentration (IC50) values ranging from 2-44 µg/mL. Interestingly, most of these active compounds, including xanthoangelol E (isolated from Angelica keiskei), dieckol 1 (isolated from Ecklonia cava), amentoflavone (isolated from Torreya nucifera), celastrol, pristimerin, tingenone and iguesterin (isolated from Tripterygium regelii), tannic acid (isolated from Camellia sinensis), and theaflavin-3,3'-digallate, 3-isotheaflav1in-3 gallate and dihydrotanshinone I (isolated from Salvia miltiorrhiza), had IC50 values of less than 15 µg/mL. Kinetic mechanistic studies of several active compounds revealed that their mode of inhibition was dose-dependent and competitive, with Ki values ranging from 2.4-43.8 µmol/L. Given the significance of plant-based compounds and the many promising results obtained, there is still need to explore the phytochemical and mechanistic potentials of plants and their products. These medicinal plants could serve as an effective inexpensive nutraceutical for the general public to help manage COVID-19.

Structural and Evolutionary Adaptations of Nei-Like DNA Glycosylases Proteins Involved in Base Excision Repair of Oxidative DNA Damage in Vertebrates.

Oxidative stress is a type of stress that damages DNA and can occur from both endogenous and exogenous sources. Damage to DNA caused by oxidative stress can result in base modifications that promote replication errors and the formation of sites of base loss, which pose unique challenges to the preservation of genomic integrity. However, the adaptive evolution of the DNA repair mechanism is poorly understood in vertebrates. This research aimed to explore the evolutionary relationships, physicochemical characteristics, and comparative genomic analysis of the Nei-like glycosylase gene family involved in DNA base repair in the vertebrates. The genomic sequences of NEIL1, NEIL2, and NEIL3 genes were aligned to observe selection constraints in the genes, which were relatively low conserved across vertebrate species. The positive selection signals were identified in these genes across the vertebrate lineages. We identified that only about 2.7% of codons in these genes were subjected to positive selection. We also revealed that positive selection pressure was increased in the Fapy-DNA-glyco and H2TH domain, which are involved in the base excision repair of DNA that has been damaged by oxidative stress. Gene structure, motif, and conserved domain analysis indicated that the Nei-like glycosylase genes in mammals and avians are evolutionarily low conserved compared to other glycosylase genes in other "vertebrates" species. This study revealed that adaptive selection played a critical role in the evolution of Nei-like glycosylase in vertebrate species. Systematic comparative genome analyses will give key insights to elucidate the links between DNA repair and the development of lifespan in various organisms as more diverse vertebrate genome sequences become accessible.

Ethnobotanical study of Mandi Ahmad Abad, District Okara, Pakistan.

This study hypothesized that native people have unique traditional knowledge of plant resources in the rural areas and basic objective was the documentation of this valuable inheritance. Ethnobotanical data was collected from a remote rural area of Mandi Ahmad Abad, Union council number NA-144 Tehsil Depalpur District Okara, Pakistan. A total of 94 informants were randomly interviewed to collect data about local names of plant species, mode of administration, recipes and ailments, and ethnobotanical uses through semi-structured questionnaire, interviews and group discussion methods. The collected data was statistically analyzed by calculating use value (UV), frequency of citation (FC), relative frequency of citation (RFC), factor of informant consensus (FIC), family importance value, and relative importance (RI). This study is also compared with ethnobotanical literature by using Jaccard's index (JI) for similarity analysis. A total of 126 species belonging to 52 families were documented. The Poaceae (13spp.), Leguminosae (12spp.), Solanaceae (10spp.) and Cucurbitaceae (10spp.) were dominant families. Highest used value (UV = 0.22) was obtained for Azadirachta indica. The minimum used value (UV) was showed by Alhagi maurorum, Eclipta prostrata, Hibiscus rosa-sinensis, Solanum virginianum and Trianthema potulacastrum (UV = 0.01). Hepatitis, stomach ulcer, bowel disorders, urinary problems, psoriasis, cancer, and leucoderma were the most treated ailments with ICF value of 1, followed by leucorrhea (ICF: 0.89), and vomiting (ICF: 0.86). The highest Jaccard's similarity index value (JI = 0.329) showed that plant species reported in our study was more similar with Arid regions of Northern Punjab, Pakistan. This novel ethnobotanical report concluded that traditional knowledge about use of medicinal plants is decreasing due to allopathic medicines. Immediate steps should be taken for conservation and documentation of traditional knowledge of plants especially those having medicinal properties.

Light microscopy and scanning electron microscopy of bacterial colonies isolated from ras malai samples from different markets of Lahore, Pakistan.

The current study investigates the total bacterial contamination in various packed and unpacked ras malai samples of 14 different localities of Lahore, Pakistan. The bacterial colonies such as Bacillus sp. and Gamella sp. were isolated from ras malai samples and grown on agar-broth media under sterile environmental conditions. Serial dilution technique was used to compose the replicates to get a viable count of bacteria in the samples. Results indicated that in case of packed ras malai samples, maximum bacterial count was observed in Sample 1 (422 × 10-2 to 402 × 10-6 ) and minimum bacterial count was in Sample 4 (21 × 10-2 to 9.3 × 10-6 ). For unpacked ras malai samples, maximum bacterial count was in Sample 3 (200.3 × 10-2 to 181.3 × 10-6 ) and minimum bacterial count was observed in Sample 1 (110 × 10-2 to 90.4 × 10-6 ). It was concluded that the marketed samples contain more bacterial count as compared to the standard sterilization values. Such products could possibly become the cause of many health problems in children.

A global perspective on the biology, impact and management of Chenopodium album and Chenopodium murale: two troublesome agricultural and environmental weeds.

Chenopodium album and C. murale are cosmopolitan, annual weed species of notable economic importance. Their unique biological features, including high reproductive capacity, seed dormancy, high persistence in the soil seed bank, the ability to germinate and grow under a wide range of environmental conditions and abiotic stress tolerance, help these species to infest diverse cropping systems. C. album and C. murale grow tall and absorb nutrients very efficiently. Both these species are allelopathic in nature and, thus, suppress the germination and growth of native vegetation and/or crop plants. These weed species infest many agronomic and horticultural crops and may cause > 90% loss in crop yields. C. album is more problematic than C. murale as the former is more widespread and infests more number of crops, and it also acts as an alternate host of several crop pests. Different cultural and mechanical methods have been used to control these weed species with varying degrees of success depending upon the cropping systems and weed infestation levels. Similarly, allelopathy and biological control have also shown some potential, especially in controlling C. album. Several herbicides have been successfully used to control these species, but the evolution of wide-scale herbicide resistance in C. album has limited the efficacy of chemical control. However, the use of alternative herbicides in rotation and the integration of chemicals and biologically based control methods may provide a sustainable control of C. album and C. murale.

Crop Production under Drought and Heat Stress: Plant Responses and Management Options.

Abiotic stresses are one of the major constraints to crop production and food security worldwide. The situation has aggravated due to the drastic and rapid changes in global climate. Heat and drought are undoubtedly the two most important stresses having huge impact on growth and productivity of the crops. It is very important to understand the physiological, biochemical, and ecological interventions related to these stresses for better management. A wide range of plant responses to these stresses could be generalized into morphological, physiological, and biochemical responses. Interestingly, this review provides a detailed account of plant responses to heat and drought stresses with special focus on highlighting the commonalities and differences. Crop growth and yields are negatively affected by sub-optimal water supply and abnormal temperatures due to physical damages, physiological disruptions, and biochemical changes. Both these stresses have multi-lateral impacts and therefore, complex in mechanistic action. A better understanding of plant responses to these stresses has pragmatic implication for remedies and management. A comprehensive account of conventional as well as modern approaches to deal with heat and drought stresses have also been presented here. A side-by-side critical discussion on salient responses and management strategies for these two important abiotic stresses provides a unique insight into the phenomena. A holistic approach taking into account the different management options to deal with heat and drought stress simultaneously could be a win-win approach in future.

A systematic review on ethnomedicines of anti-cancer plants.

Cancer is a serious health problem and the second leading cause of death around the globe. Present review is an attempt to provide utmost information based on ethno-pharmacological and toxicological aspects of anti-cancer plants of the world. A total of 276 articles published in English journals and containing maximum ethnomedicinal information were reviewed using several data sources such as; Google scholar, Web of Science, Scopus, PubMed and floras of different countries. A total of 199 anti-cancer plants were recorded in present review and results indicated that traditional medicines are mostly being use in developing countries for cancer treatment. Traditionally and scientifically skin and breast cancer types gained more focus. Seventy plants were reportedly analyzed for in-vitro activities while 32 plants were having in-vivo reports. Twenty nine pure compounds (mostly phenolic) were reportedly isolated from anti-cancer plants and tested against different cancer cell lines. Inspite having better efficiency of ethnomedicines as compared to synthetic drugs, several plants have also shown toxic effects on living system. Therefore, we invite researchers attention to carry out detailed ethno-pharmacological and toxicological studies on un-explored anti-cancer plants in order to provide reliable knowledge to the patients and develop novel anti-cancer drugs. Copyright © 2017 John Wiley & Sons, Ltd.

Biology and management of two important Conyza weeds: a global review.

Weed management is one of the prime concerns for sustainable crop production. Conyza bonariensis and Conyza canadensis are two of the most problematic, noxious, invasive and widespread weeds in modern-day agriculture. The biology, ecology and interference of C. bonariensis and C. canadensis have been reviewed here to highlight pragmatic management options. Both these species share a unique set of biological features, which enables them to invade and adapt a wide range of environmental conditions. Distinct reproductive biology and an efficient seed dispersal mechanism help these species to spread rapidly. Ability to interfere strongly and to host crop pests makes these two species worst weeds of cropping systems. These weed species cause 28-68 % yield loss in important field crops such as soybean and cotton every year. These weeds are more prevalent in no-till systems and, thus, becoming a major issue in conservation agriculture. Cultural practices such as crop rotations, seed rate manipulation, mulching, inter-row tillage and narrow row spacing may provide an effective control of these species. However, such methods are not feasible and applicable under all types of conditions. Different herbicides also provide a varying degree of control depending on crop, agronomic practices, herbicide dose, application time and season. However, both these species have evolved resistance against multiple herbicides, including glyphosate and paraquat. The use of alternative herbicides and integrated management strategies may provide better control of herbicide-resistant C. bonariensis and C. canadensis. Management plans based on the eco-biological interactions of these species may prove sustainable in the future.