Cucumis melo var. momordica as a Potent Antidiabetic, Antioxidant and Possible Anticovid Alternative: Investigation through Experimental and Computational Methods.

Diabetes mellitus is a typical life threatening of disease, which generate due to the dysfunction of ß cells of pancreas. In 2014, WHO stated that 422 million people were infected with DM. The current pattern of management of diabetes included synthetic or plant based oral hypoglycemic drugs and insulin but drug resentence is become a very big issues in antidiabetic therapy. Thus, it's very earnest to discover now medication for this disease. Now the days, it is well acknowledged that diabetic patients are more prone towards covid and related complications. Thus, medical practitioners reformed the methodology of prescribing medication for covid infected antidiabetic therapy and encouraging the medication contains dual pharmacological properties. It is also well know that polyphenols specifically hold a significant role in oxidative stress and reduced the severity of many inflammatory diseases. Cucumis melo has rich history as ethano-pharmacological use in Indian subcontinent. The fruit and seed are well-known for the treatment of various diseases due to the presence of phenolics. Therefore, in this study, the combined mixture of flower and seeds were used for the extraction of polyphenolic rich extract and tested for antidiabetic activity through the antioxidant and in vivo experiments. The antioxidant potential measurement exhibited that the selected plant extract has the significant competence to down-regulate oxidative stress (DPPH scavenging IC50 at 60.7±1.05 µg/mL, ABTS IC50 at 62.15±0.50 µg/mL). Furthermore, the major polyphenolic phyto-compounds derived from the Cucumis melo were used for in silico anticovid activity, docking, and complementarity studies. The anticovid activity prognosis reflected that selected phyto-compounds amentoflavone and vanillic acid have optimal possibility to interact with 3C-like protease and through this moderate anticovid activity can be exhibit. The docking experiments established that the selected compounds have propensity to interact with protein tyrosine phosphatase 1B, 11ß-Hydroxysteroid dehydrogenase, superoxide dismutase, glutathione peroxidase, and catalase ß-glucuronidase receptor. In vivo experiments showed that 500 mg/kg, Cucumis melo extract ominously amplified body weight, plasma insulin, high-density lipoprotein levels, and biochemical markers. Furthermore, extract significantly downregulate the blood glucose, total cholesterol, triglycerides, low-density lipoprotein, and very low-density lipoprotein.

Genetic diversity and population structure of critically endangered Dactylorhiza hatagirea (D. Don) Soo from North-Western Himalayas and implications for conservation.

Dactylorhiza hatagirea (D. Don) Soo is medicinally important herb, which is widely used in ayurveda, unani, and folk/traditional medicine system to cure diseases. Due to its immense ethno-botanical properties, the trade of D. hatagirea is estimated to be USD 1 billion/year in India. Unfortunately, due to overexploitation of the herb from the wild, has resulted in dwindling of its populations in their natural habitats, which has led to its critically endangered status. Molecular genetic studies are still scarce in D. hatagirea, therefore, in current study, genetic diversity and population structure analysis was carried out of 10 populations (48 individuals) collected from three cold desert regions (2527 m-3533 m amsl) of Himachal Pradesh. Mean observed heterozygosity (Ho) and expected heterozygosity (He) was recorded 0.185 and 0.158. The maximum values for Fst (fixation index) and Nm (gene flow) were recorded 0.945 at locus KSSR14 and 1.547 at locus KSSR 4 respectively. Mean genetic differentiation (Fst) coefficient was estimated to 0.542. Overall, low levels of genetic diversity was recorded in the populations of D. hatagirea, might be due to habitat specificity (alpine meadows ecosystem; humid laden undulating habitat), restricted distribution and high anthropogenic activities. However, two populations viz., Bathad and Rangrik were recorded with high diversity and largest number of private alleles, stipulates that these populations might have high evolutionary significance and response to selection. Dendrogram analysis revealed that the populations of D. hatagirea were clustered into four major clusters, which was supported by Bayesian based STRUCTURE predictions. Clustering pattern of majority individuals of different populations revealed consistency with their geographic origin. Outcomes of current study reveals the status of genetic diversity and population structure of endangered D. hatagirea, which can be futuristically utilised for appropriate planning of conservation strategies.

Gametophyte Development Needs Mitochondrial Coproporphyrinogen III Oxidase Function.

Tetrapyrrole biosynthesis is one of the most essential metabolic pathways in almost all organisms. Coproporphyrinogen III oxidase (CPO) catalyzes the conversion of coproporphyrinogen III into protoporphyrinogen IX in this pathway. Here, we report that mutation in the Arabidopsis (Arabidopsis thaliana) CPO-coding gene At5g63290 (AtHEMN1) adversely affects silique length, ovule number, and seed set. Athemn1 mutant alleles were transmitted via both male and female gametes, but homozygous mutants were never recovered. Plants carrying Athemn1 mutant alleles showed defects in gametophyte development, including nonviable pollen and embryo sacs with unfused polar nuclei. Improper differentiation of the central cell led to defects in endosperm development. Consequently, embryo development was arrested at the globular stage. The mutant phenotype was completely rescued by transgenic expression of AtHEMN1 Promoter and transcript analyses indicated that AtHEMN1 is expressed mainly in floral tissues and developing seeds. AtHEMN1-green fluorescent protein fusion protein was found targeted to mitochondria. Loss of AtHEMN1 function increased coproporphyrinogen III level and reduced protoporphyrinogen IX level, suggesting the impairment of tetrapyrrole biosynthesis. Blockage of tetrapyrrole biosynthesis in the AtHEMN1 mutant led to increased reactive oxygen species (ROS) accumulation in anthers and embryo sacs, as evidenced by nitroblue tetrazolium staining. Our results suggest that the accumulated ROS disrupts mitochondrial function by altering their membrane polarity in floral tissues. This study highlights the role of mitochondrial ROS homeostasis in gametophyte and seed development and sheds new light on tetrapyrrole/heme biosynthesis in plant mitochondria.

Promoter Trapping and Deletion Analysis Show Arabidopsis thaliana APETALA2 Gene Promoter Is Bidirectional and Functions as a Pollen- and Ovule-Specific Promoter in the Reverse Orientation.

The Arabidopsis thaliana promoter trap mutant Bitrap-112 expressing green fluorescent protein (GFP) gene in the ovules was found to carry transferred DNA (T-DNA) insertion at -309 position of the APETALA2 (AP2) gene. Bitrap-112 line did not show phenotype associated with the AP2 mutation, suggesting that T-DNA insertion did not interrupt the AP2 promoter. Further, head-to-head orientation of GFP and AP2 genes indicated that the AP2 promoter could be bidirectional. A detailed deletion analysis of the upstream sequences of the AP2 gene was done to identify the promoter. GUS assay of transgenic A. thaliana plants carrying various AP2 upstream fragments fused to the uidA gene showed that ~200-bp 5' UTR sequences are capable of driving gene expression at low levels in vegetative tissues whereas inclusion of further upstream sequences (~300 bp) enhanced uidA expression comparable to native AP2 expression levels in various tissues including ovules. In the reverse orientation, the 519-bp AP2 upstream fragment was found to drive gene expression in immature ovules and pollen. Absence of antisense transcripts corresponding to the sequences upstream of AP2 gene in wild-type A. thaliana plants suggests that promoter trapping has uncovered a cryptic promoter, which in reverse orientation is capable of driving gene expression in ovules and anthers.

Genetic and biochemical diversity among Valeriana jatamansi populations from Himachal Pradesh.

Valeriana jatamansi Jones is an important medicinal plant that grows wild in Himachal Pradesh, India. Molecular and biochemical diversity among 13 natural populations from Himachal Pradesh was assessed using RAPD and GC-MS to know the extent of existing variation. A total of seven genetically diverse groups have been identified based on RAPD analysis which corroborated well with the analysis based on chemical constituents. The essential oil yield ranged from 0.6% to 1.66% (v/w). A negative correlation between patchouli alcohol and viridiflorol, the two major valued constituents, limits the scope of their simultaneous improvement. However, other few populations like Chamba-II and Kandi-I were found promising for viridiflorol and patchouli alcohol, respectively. The analysis of chemical constitution of oil of the populations from a specific region revealed predominance of specific constituents indicating possibility of their collection/selection for specific end uses like phytomedicines. The prevalence of genetically diverse groups along with sufficient chemical diversity in a defined region clearly indicates the role of ecology in the maintenance of evolution of this species. Sufficient molecular and biochemical diversity detected among natural populations of this species will form basis for the future improvement.

Fast ultrasound-assisted extraction followed by capillary gas chromatography combined with nitrogen-phosphorous selective detector for the trace determination of tebuconazole in garlic, soil and water samples.

A fast and an efficient ultrasound-assisted extraction technique using a lower density extraction solvent than water was developed for the trace-level determination of tebuconazole in garlic, soil and water samples followed by capillary gas chromatography combined with nitrogen-phosphorous selective detector (GC-NPD). In this approach, ultrasound radiation was applied to accelerate the emulsification of the ethyl acetate in aqueous samples to enhance the extraction efficiency of tebuconazole without requiring extra partitioning or cleaning, and the use of capillary GC-NPD was a more sensitive detection technique for organonitrogen pesticides. The experimental results indicate an excellent linear relationship between peak area and concentration obtained in the range 1-50 µg/kg or µg/L. The limit of detection (S/N, 3 ± 0.5) and limit of quantification (S/N, 7.5 ± 2.5) were obtained in the range 0.2-3 and 1-10 µg/kg or µg/L. Good spiked recoveries were achieved from ranges 95.55-101.26%, 96.28-99.33% and 95.04-105.15% in garlic, Nanivaliyal soil and Par River water, respectively, at levels 5 and 20 µg/kg or µg/L, and the method precision (% RSD) was ≤5%. Our results demonstrate that the proposed technique is a viable alternative for the determination of tebuconazole in complex samples.

Purification and characterization of Mn-peroxidase from Musa paradisiaca (banana) stem juice.

Mn-peroxidase (MnP), a biotechnologically important enzyme was purified for the first time from a plant source Musa paradisiaca (banana) stem, which is an agro-waste easily available after harvest of banana fruits. MnP was earlier purified only from the fungal sources. The enzyme was purified from stem juice by ultrafiltration and anion-exchange column chromatography on diethylamino ethylcellulose with 8-fold purification and purification yield of 65%. The enzyme gave a single protein band in SDS-PAGE corresponding to molecular mass 43 kDa. The Native-PAGE of the enzyme also gave a single protein band, confirming the purity of the enzyme. The UV/VIS spectrum of the purified enzyme differed from the other heme peroxidases, as the Soret band was shifted towards lower wavelength and the enzyme had an intense absorption band around 250 nm. The K(m) values using MnSO4 and H2O2 as the substrates of the purified enzyme were 21.0 and 9.5 microM, respectively. The calculated k(cat) value of the purified enzyme using Mn(II) as the substrate in 50 mM lactate buffer (pH 4.5) at 25 degrees C was 6.7s(-1), giving a k(cat)/K(m) value of 0.32 microM(-1)s(-1). The k(cat) value for the MnP-catalyzed reaction was found to be dependent of the Mn(III) chelator molecules malonate, lactate and oxalate, indicating that the enzyme oxidized chelated Mn(II) to Mn(III). The pH and temperature optima of the enzyme were 4.5 and 25 degrees C, respectively. The enzyme in combination with H2O2 liberated bromine and iodine in presence of KBr and KI respectively. All these enzymatic characteristics were similar to those of fungal MnP. The enzyme has the potential as a green brominating and iodinating agent in combination with KBr/KI and H2O2.

Genetic structure of Indian valerian (Valeriana jatamansi) populations in western Himalaya revealed by AFLP.

Valeriana jatamansi Jones is a natural tetraploid species indigenous to the Indian Himalaya. To assess its genetic diversity and population structure, we analyzed six natural populations from the western Himalayan region using amplified fragment length polymorphism. An analysis of molecular variance found that 93% of the genetic variation of V. jatamansi was within populations and 7% among populations. The correlation between genetic and geographic distances (r = 0.14) was not significant. Though the populations are well separated, the lack of distinct genetic variation between populations may be due to either recent rapid fragmentation from a wide and continuous area resulting in genetically similar populations or wide dispersal of seed by wind, since the follicles are feathery. Polyploidy may be the reason for the lack of genetic impoverishment due to fragmentation.

No-patch 23-gauge vitrectomy under topical anesthesia: a pilot study.

A pilot study was designed to evaluate the safety and efficacy of 23-gauge vitrectomy under topical anesthesia. Five eyes of five patients underwent 23-gauge sutureless vitrectomy under topical anesthesia with a pledget soaked in 0.5% proparacaine hydrochloride anesthetic, for vitreous hemorrhage (four eyes), epiretinal membrane (one eye). Subjective pain and discomfort were graded using a visual analogue chart from 0 (no pain or discomfort) to 4 (severe pain and discomfort). At the end of surgery no patch was applied and patients were given dark glasses. Patients underwent an immediate postoperative assessment, followed by next day and one week postoperative evaluation. Four patients had Grade 0 pain during the surgery. One patient had Grade 1 pain during the placement and withdrawal of the micro cannulas. The surgical outcomes were favorable. 23-gauge vitrectomy under topical anesthesia is safe and effective in selected cases. Further study is recommended to validate the outcome of this study.

Quality control of herbal medicines by using spectroscopic techniques and multivariate statistical analysis.

Herbal medicines play an important role in modern human life and have significant effects on treating diseases; however, the quality and safety of these herbal products has now become a serious issue due to increasing pollution in air, water, soil, etc. The present study proposes Fourier transform infrared spectroscopy (FTIR) along with the statistical method principal component analysis (PCA) to identify and discriminate herbal medicines for quality control. Herbal plants have been characterized using FTIR spectroscopy. Characteristic peaks (strong and weak) have been marked for each herbal sample in the fingerprint region (400-2000 cm(-1)). The ratio of the areas of any two marked characteristic peaks was found to be nearly consistent for the same plant from different regions, and thus the present idea suggests an additional discrimination method for herbal medicines. PCA clusters herbal medicines into different groups, clearly showing that this method can adequately discriminate different herbal medicines using FTIR data. Toxic metal contents (Cd, Pb, Cr, and As) have been determined and the results compared with the higher permissible daily intake limit of heavy metals proposed by the World Health Organization (WHO).

Molluscicides from some common medicinal plants of eastern Uttar Pradesh, India.

Many aquatic snails act as intermediate hosts for the larvae of trematodes, Fasciola hepatica and Fasciola gigantica, which cause the diseases fascioliasis and schistosomiasis. The WHO has tested several thousands of synthetic compounds for the control of the snail host. Although effective, these molluscicides have so far not proved themselves to be entirely satisfactory. With a growing awareness of environmental pollution, efforts are being made to discover molluscicidal products of plant origin. Being products of biosynthesis, these are potentially biodegradable in nature. Several groups of compounds present in various plants have been found to be toxic to target organisms at acceptable doses ranging from <1 to 100 ppm. Common medicinal plants, i.e. Thevetia peruviana, Alstonia scholaris (Family; Apocynaceae), Euphorbia pulcherima and Euphorbia hirta (Family; Euphorbiaceae), have potent molluscicidal activity against freshwater snails. The toxicological actions of Thevetia peruviana may be due to the presence of apigenin-5-methyl ether (flavonoid) and triterpenoid glycosides, while a number of alkaloids (pseudo-akuammigine in addition to betulin, ursolic acid and beta-sitosterol), steroids and triterpenoids are present in Alstonia scholaris and the diterpenoids, pulcherrol, beta-sitosterol, hentriacontane, ellagic acid and beta-amyrin are present in Euphorbia hirta and in Euphorbia pulcherima. Although, at present very little literature is available on the control of vector snails through plant origin pesticides, an attempt has been made in this review to assemble all the known information on molluscicidal properties of common medicinal plants of eastern Uttar Pradesh, India, which might be useful for the control of harmful snails.

Molluscicidal evaluation of three common plants from India.

Mortality caused by the aqueous extract of latex of Thevetia peruviana, Alstonia scholaris and Euphorbia pulcherrima against two harmful freshwater snails, Lymnaea acuminata and Indoplanorbis exustus, is reported. Latices of all the plants at high doses were also lethal to freshwater fish Channa punctatus, which shares the habitat with these snails, but within 24 h, LC90 of snail L. acuminata did not cause any mortality to fishes in a mixed population of snail and fish. Therefore, these plant extracts may eventually be of great value for the control of harmful aquatic snails and other molluscan pests.

Toxic effect of stem bark and leaf of Euphorbia hirta plant against freshwater vector snail Lymnaea acuminata.

The aqueous stem bark and leaf extracts of plant Euphorbia hirta (family-Euphorbiaceae) have potent molluscicidal activity. Sub-lethal doses (40% and 80% of LC50) of aqueous stem bark and leaf extracts of this plant also significantly (P<0.05) alter the levels of total protein, total free amino acid, nucleic acids (DNA and RNA) and the activity of enzyme protease and acid and alkaline phosphatase in various tissues of the vector snail Lymnaea acuminata in time and dose dependent manner. Euphorbia hirta (family-Euphorbiaceae) commonly known as Dudhi, is a common medicinal plant of India, which is used in variety of diseases i.e. cough, asthma, colic, dysentery, genito urinary diseases.

2-hydroxymethyl-4-[5-(4-methoxyphenyl)-3-trifluoromethyl-1H-1-pyrazolyl]-1-benzenesulfonamide (DRF-4367): an orally active COX-2 inhibitor identified through pharmacophoric modulation.

Analogs of 1,5-diarylpyrazoles with a novel pharmacophore at N1 were designed, synthesized and evaluated for the in-vitro cyclooxygenase (COX-1/COX-2) inhibitory activity. The variations at/around position-4 of the C-5 phenyl ring in conjunction with a CF3 and CHF2 groups at C-3 exhibited a high degree of potency and selectivity index (SI) for COX-2 inhibition. The in-vivo evaluation of these potent compounds with a few earlier ones indicated the 4-OMe-phenyl analog and the 4-NHMe-phenyl analog with a CF3, and the 4-OEt-phenyl analog with a CHF2 group at C-3 to possess superior potency than celecoxib. In addition to its impressive anti-inflammatory, antipyretic, analgesic and anti-arthritic properties, compound (DRF-4367) was found to possess an excellent pharmacokinetic profile, gastrointestinal (GI) safety in the long-term arthritis study and COX-2 potency in human whole blood assay. Thus, compound was selected as an orally active anti-inflammatory candidate for pre-clinical evaluation.

Toxic effect of two common Euphorbiales latices on the freshwater snail Lymnaea acuminata.

The aqueous and serially purified latex extracts of plants Euphorbia pulcherima and Euphorbia hirta (family Euphorbiaceae) have potent molluscicidal activity. Sub-lethal doses (40 and 80% of LC(50)) of aqueous and partially purified latex extracts of both the plants also significantly alter the levels of total protein, total free amino acid, nucleic acid (DNA and RNA) and the activity of enzyme protease and acid and alkaline phosphatase in nervous tissue of the snail Lymnaea acuminata in time and dose dependent manner. E. pulcherima and E. hirta are common medicinal plants of family Euphorbiaceae. E. pulcherima is useful for a variety of conditions, such as rheumatism, snakebite, asthma, obstipation, and skin-diseases. While, E. hirta is also used in cough, asthma, colic, dysentery, and genito urinary diseases.