Chemical Profiling and Biological Activities on Nepalese Medicinal Plant Extracts and Isolation of Active Fraction of Nyctanthes arbor-tristis.

The importance of medicinal plants for the treatment of different diseases is high from the aspects of the pharmaceutical industry and traditional healers. The present study involves nine different medicinal plants, namely, Neolamarckia cadamba, Nyctanthes arbor-tristis, Pogostemon benghalensis, Equisetum debile, Litsea monopetala, Spilanthes uliginosa, Desmostachya bipinnata, Mallotus philippensis, and Phoenix humilis, collected from Chitwan district of Nepal for biochemical analysis followed by the isolation of active plant fractions from the bioactive plant extract. The methanolic extracts of roots, barks, seeds, seed cover, and the other aerial parts of plants were used for the phytochemical analysis and biological activities. The DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging assay was adopted to evaluate the antioxidant activity. Antibacterial activity was evaluated using the agar well diffusion method. The antidiabetic activity was studied by the α-amylase enzyme inhibition assay. The highest antioxidant activity was observed in extracts of Nyctanthes arbor-tristis followed by Mallotus philippensis (seed cover), Pogostemon benghalensis, Litsea monopetala, Phoenix humilis, and Neolamarckia cadamba with IC50 values of 27.38 ± 1.35, 32.08 ± 2.81, 32.75 ± 2.13, 33.82 ± 1.07, 40.14 ± 0.93, and 50.44 ± 3.75 µg/mL, respectively. The highest antidiabetic activity was observed in extracts of Phoenix humilis followed by Desmostachya bipinnata and Pogostemon benghalensis with IC50 values of 95.69 ± 6.97, 99.24 ± 12.6, and 106.3 ± 12.89 µg/mL, respectively. The mild α-amylase enzyme inhibition was found in extracts of Nyctanthes arbor-tristis, Spilanthes uliginosa Swartz, Litsea monopetala, and Equisetum debile showing IC50 values of 110.4 ± 7.78, 115.98 ± 10.24, 149.83 ± 8.3, and 196.45 ± 6.04 µg/mL, whereas Mallotus Philippensis (seed cover), Mallotus philippensis (seed), and Desmostachya bipinnata showed weak α-amylase inhibition with IC50 values of 208.87 ± 1.76, 215.41 ± 2.09, and 238.89 ± 9.27 µg/mL, respectively. The extract of Nyctanthes arbor-tristis showed high zones of inhibition against S. aureus (ATCC 25923) and E. coli (ATCC 25922) of ZOI 26 and 22 mm, respectively. The chemical constituents isolated from the active plant Nyctanthes arbor-tristis were subjected to GCMS analysis where the major chemical compounds were 11,14,17-eicosatrienoic acid and methyl ester. These results support the partial scientific validation for the traditional uses of these medicinal plants in the treatment of diabetes and infectious diseases by the people living in different communities of Chitwan, Nepal.

Phytochemistry, Biological, and Toxicity Study on Aqueous and Methanol Extracts of Chromolaena odorata.

The medicinal plant Chromolaena odorata is traditionally used by people living in different communities of Nepal and the globe against diabetes, soft tissue wounds, skin infections, diarrhea, malaria, and several other infectious diseases. The present study focuses on the qualitative and quantitative phytochemical analyses and antioxidant, antidiabetic, antibacterial, and toxicity of the plant for assessing its pharmacological potential. The extracts of flowers, leaves, and stems were prepared using methanol and distilled water as the extracting solvents. Total phenolic content (TPC) and total flavonoid content (TFC) were estimated by using the Folin-Ciocalteu phenol reagent method and the aluminum chloride colorimetric method. Antioxidant and antidiabetic activities were assessed using the DPPH assay and α-glucosidase inhibition assay. A brine shrimp assay was performed to study the toxicity, and the antibacterial activity test was performed by the agar well diffusion method. Phytochemical analysis revealed the presence of phenols, flavonoids, quinones, terpenoids, and coumarins as secondary metabolites. The methanol extract of leaves and flowers displayed the highest phenolic and flavonoid content with 182.26 ± 1.99 mg GAE/g, 128.57 ± 7.62 mg QE/g and 172.65 ± 0.48 mg GAE/g, 121.74 ± 7.06 mg QE/g, respectively. The crude extracts showed the highest DPPH free radical scavenging activity with half maximal inhibitory concentration (IC50) of 32.81 ± 5.26 µg/mL and 41.00 ± 1.10 µg/mL, respectively. The methanol extract of the leaves was found to be effective against bacterial strains such as K. pneumoniae (ZOI = 9.67 ± 0.32 mm), B. subtilis (ZOI = 15.00 ± 0 mm), and E. coli (7.3 ± 0.32 mm). The methanol extract of the flowers showed the most α-glucosidase inhibitory activity (IC50 227.63 ± 11.38 µg/mL), followed by the methanol extract of leaves (IC50 249.50 ± 0.97 µg/mL). The aqueous extract of the flowers showed the toxic effect with LC50 107.31 ± 49.04 µg/mL against the brine shrimp nauplii. In conclusion, C. odorata was found to be a rich source of plant secondary metabolites such as phenolics and flavonoids with potential effects against bacterial infection, diabetes, and oxidative stress in humans. The toxicity study showed that the aqueous extract of flowers possesses pharmacological activities. This study supports the traditional use of the plant against infectious diseases and diabetes and provides some scientific validation.

Analysis of Phenolic and Flavonoid Content, α-Amylase Inhibitory and Free Radical Scavenging Activities of Some Medicinal Plants.

In Nepal, about 700 plant species have been reported to be used for the primary care of different diseases. However, many of them have not been studied yet for their scientific evidence. The main aim of this study is the quantitative analysis of flavonoids and phenolic content, antioxidant, and antidiabetic activities of the extracts of four different medicinal plants, namely, Pogostemon benghalensis, Aleuritopteris bicolor, Crateva unilocularis, and Rungia pectinata growing in Nepal. The methanol extracts of plant samples were prepared by the hot percolation method using the Soxhlet apparatus. The phytochemicals of the plant extracts were analysed by colour differentiation methods using different analytical reagents. The phenolic content was estimated by using Folin-Ciocalteu's phenol reagent and the flavonoid was estimated by the aluminium chloride colorimetric method. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay was used to evaluate the antioxidant potential. The α-amylase enzyme inhibition activity was performed to evaluate the antidiabetic activity of plant extracts. The amount of total phenolics and flavonoids content was found to be the highest in Pogostemon benghalensis (169.43 ± 3.58 mg GAE/g and 65.2 ± 2.0 mg QE/g), respectively, which also showed the most potent free radical scavenging activity (IC50 35.92 ± 0.65 µg/mL). The extract of Aleuritopteris bicolor showed the highest α-amylase inhibitory activity (IC50 651.58 ± 10.32 µg/mL) whereas Crateva unilocularis and Pogostemon benghalensis exhibited moderate activity. The extract of Rungia pectinata showed the least activity towards α-amylase inhibition. Some of the medicinal plants selected in this study showed high TPC and TFC values with potent biological activities. To the best of our knowledge, these medicinal plants have the least exposure to their biological activities, and the results provide scientific evidence for the traditional uses of these plants against diabetes and infectious diseases. However, a detailed study can be performed in these plants to isolate the active chemical compounds and to evaluate in vivo pharmacological activities to know the active drug candidates for the future drug development process.

Phytochemical Analysis and In Vitro Antioxidant and Antibacterial Activity of Different Solvent Extracts of Beilschmiedia roxburghiana Nees Stem Barks.

Plants have long been considered as a basis of medicines for different indigenous cultures around the globe. They continue as a prominent source of important phytoconstituents which exhibit significant biological activities. In this study, we performed the phytochemical screening, estimation of total phenolic and flavonoids, antioxidants, and antimicrobial activities of the stem bark of Beilschmiedia roxburghiana Nees using different solvents. The total phenolic and total flavonoid contents ranged from 106.73 ± 1.62 mg GAE/g and 99.32 ± 0.66 mg QE/g (methanol extract) to 65.59 ± 1.79 mg GAE/g and 29.98 ± 0.90 mg QE/g (n-hexane extract), respectively. The maximum 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity with a half-maximal inhibitory concentration (IC50) of 39.86 ± 3.69 µg/mL was observed for methanol extract followed by aqueous (IC50 = 43.55 ± 6.16 µg/mL), ethyl acetate (IC50 = 44.30 ± 5.88 µg/mL), dichloromethane (IC50 = 71.50 ± 4.70 µg/mL), and the lowest activity was observed for n-hexane extract. The disc diffusion method revealed that the ethyl acetate extract exhibited relatively higher activity against Salmonella typhi (ZOI = 13 mm), and moderate activities against Shigella sonnei, Acinetobacter baumannii, Klebsiella pneumoniae, and Staphylococcus aureus (ZOI = 12 mm). The methanol and aqueous extracts showed nearly parallel and the n-hexane and dichloromethane extracts exhibited mild antibacterial activities. The results indicated that the polarity index of the extracting solvents amplified the biological activities of the extract. The study is helpful to support the validity of the traditional application of the plant as natural medicine.

In Vitro and In Silico Studies for the Identification of Potent Metabolites of Some High-Altitude Medicinal Plants from Nepal Inhibiting SARS-CoV-2 Spike Protein.

Despite ongoing vaccination programs against COVID-19 around the world, cases of infection are still rising with new variants. This infers that an effective antiviral drug against COVID-19 is crucial along with vaccinations to decrease cases. A potential target of such antivirals could be the membrane components of the causative pathogen, SARS-CoV-2, for instance spike (S) protein. In our research, we have deployed in vitro screening of crude extracts of seven ethnomedicinal plants against the spike receptor-binding domain (S1-RBD) of SARS-CoV-2 using an enzyme-linked immunosorbent assay (ELISA). Following encouraging in vitro results for Tinospora cordifolia, in silico studies were conducted for the 14 reported antiviral secondary metabolites isolated from T. cordifolia-a species widely cultivated and used as an antiviral drug in the Himalayan country of Nepal-using Genetic Optimization for Ligand Docking (GOLD), Molecular Operating Environment (MOE), and BIOVIA Discovery Studio. The molecular docking and binding energy study revealed that cordifolioside-A had a higher binding affinity and was the most effective in binding to the competitive site of the spike protein. Molecular dynamics (MD) simulation studies using GROMACS 5.4.1 further assayed the interaction between the potent compound and binding sites of the spike protein. It revealed that cordifolioside-A demonstrated better binding affinity and stability, and resulted in a conformational change in S1-RBD, hence hindering the activities of the protein. In addition, ADMET analysis of the secondary metabolites from T. cordifolia revealed promising pharmacokinetic properties. Our study thus recommends that certain secondary metabolites of T. cordifolia are possible medicinal candidates against SARS-CoV-2.

Sensitive quantification of coixol, a potent insulin secretagogue, in Scoparia dulcis extract using high-performance liquid chromatography combined with tandem mass spectrometry and UV detection.

Diabetes is a major global health problem which requires new studies for its prevention and control. Scoparia dulcis, a herbal product, is widely used for treatment of diabetes. Recent studies demonstrate coixol as a potent and nontoxic insulin secretagog from S. dulcis. This study focuses on developing two quantitative methods of coixol in S. dulcis methanol-based extracts. Quantification of coixol was performed using high-performance liquid chromatography-tandem mass spectrometry (method 1) and high-performance liquid chromatography-ultraviolet detection (method 2) with limits of detection of 0.26 and 11.6 pg/µL, respectively, and limits of quantification of 0.78 and 35.5 pg/µL, respectively. S. dulcis is rich in coixol content with values of 255.5 ± 2.1 mg/kg (method 1) and 220.4 ± 2.9 mg/kg (method 2). Excellent linearity with determination coefficients >0.999 was achieved for calibration curves from 10 to 7500 ng/mL (method 1) and from 175 to 7500 ng/mL (method 2). Good accuracy (bias < -8.6%) and precision (RSD < 8.5%) were obtained for both methods. Thus, they can be employed to analyze coixol in plant extracts and herbal formulations.

Potent Insulin Secretagogue from Scoparia dulcis Linn of Nepalese Origin.

Ethno-botanical inspired isolation from plant Scoparia dulcis Linn. (Sweet Broomweed) yielded six compounds, coixol (1), glutinol (2), glutinone (3), friedelin (4), betulinic acid (5), and tetratriacontan-1-ol (6). There structures were identified using mass and 1D- and 2D-NMR spectroscopy techniques. Compounds 1-6 were evaluated for their insulin secretory activity on isolated mice islets and MIN-6 pancreatic ß-cell line, and compounds 1 and 2 were found to be potent and mildly active, respectively. Compound 1 was further evaluated for insulin secretory activity on MIN-6 cells. Compound 1 was subjected to in vitro cytotoxicity assay against MIN-6, 3T3 cell lines, and islet cells, and in vivo acute toxicity test in mice that was found to be non-toxic. The insulin secretory activity of compounds 1 and 2 supported the ethno-botanic uses of S. dulcis as an anti-diabetic agent.

Synthesis of silver nanoparticles assisted by aqueous root and leaf extracts of Rhus chinensis Mill and its antibacterial activity.

In the present study, silver nanoparticles were synthesized using aqueous root and leaf extracts of Rhus chinensis Mill. This study aimed to undertake the green synthesis of silver nanoparticles utilizing plant extracts in an eco-friendly, cost-effective, and more efficient manner with its antibacterial application. The prepared silver nanoparticles (AgNPs) were characterized by using different techniques. Such as ultraviolet-visible spectroscopy (UV-Vis), Fourier transform infrared spectroscopy (FT-IR), x-ray diffraction spectroscopy (XRD), field emission-scanning electron microscopy (FE-SEM), and energy dispersive X-ray (EDX). The color changes from yellowish to reddish brown can be visualized and it indicates the formation of silver nanoparticles. The UV-Vis absorption peak shown by the synthesized AgNPs assisted by root and leaf extract was at 443 nm and 440 nm respectively. The functional group present in plants' secondary metabolites may act as capping and stabilizing agents, indicated by the shifting and disappearing of the peak in the plant extracts and the extracts-assisted synthetic nanoparticles. The crystallite size of synthesized AgNPs assisted by the root and leaf extracts of Rhus cinensis was found to be 11.01 nm and 13.39 nm respectively, while with the help of FE-SEM image the shape and particle size of synthesized AgNPs root and leaf extract was found spherical with particle diameter of 54.40 nm and 30.89 nm respectively. The presence of an intense silver component was confirmed by EDX analysis which showed an intense peak at around 3 Kev and other elements like Cl, O, C, and N were also reported in synthesized AgNPs. Both the plant extracts assisted synthesized AgNPs showed higher zones of inhibition (ZOI) against both the Gram-positive and Gram-negative bacteria. The results of the study indicate the potential benefit of synthesized silver nanoparticles using Rhus chinensis root and leaf extracts for biomedical purposes.

Chemical analysis and biological activities on solvent extracts from different parts of Rhus chinensis mill.

The present research is focused on the preparation of a variety of solvent extracts using different parts of Rhus chinensis Mill. for the estimation of phytochemicals and to perform biological activities. The highest phenolic and flavonoid contents were recorded as 141.48 ± 0.67 mg GAE/g in methanol root extract and 54.34 ± 0.28 mg QE/g in ethyl acetate root extract. Ethyl acetate root extract displayed an IC50 of 7.83 ± 0.18 µg/mL in the DPPH assay. TPC and TFC were found moderately correlated with antioxidant activity. The root and leaf extracts showed antibacterial activities comparable to those of standard drugs against the gram-positive and gram-negative bacteria. The MIC and MBC for root extract against Staphylococcus aureus were 7.8125 mg/mL and 15.625 mg/mL respectively. Similarly, the MIC and MBC for leaf extract against Escherichia coli were 15.625 mg/mL and 31.25 mg/mL respectively.

Smartphone microscopic method for imaging and quantification of microplastics in drinking water.

Analysis of microplastics in drinking water is often challenging due to smaller particle size and low particle count. In this study, we used a low cost and an easy to assemble smartphone microscopic system for imaging and quantitating microplastic particles as small as 20 µm. The system consisted of a spherical sapphire ball lens of 4 mm diameter attached to a smartphone camera as a major imaging component. It also involved pre-concentration of the sample using ZnCl2 solution. The spike recovery and limit of detection of the method in filtered distilled and deionized water samples (n = 9) were 55.6% ± 9.7% and 34 particles/L, respectively. Imaging performance of the microscopic system was similar to a commercial bright field microscopic system. The method was further implemented to examine microplastic particles in commercial bottled and jar water samples (n = 20). The particles count in bottled and jar water samples ranged from 0-91 particles/L to 0-130 particles/L, respectively. In both sample types, particles of diverse shape and size were observed. The particles collected from water samples were further confirmed by FTIR spectra (n = 36), which found 97% of the particles tested were made of plastic material. These findings suggested that the smartphone microscopic system can be implemented as a low-cost alternative for preliminary screening of microplastic in drinking water samples. RESEARCH HIGHLIGHTS: Ball lens based smartphone microscopic method was used for microplastic analysis. Particles of diverse shape and size were found in bottle and jar water samples.

Dietary phenolic compounds as promising therapeutic agents for diabetes and its complications: A comprehensive review.

In the middle of an ever-changing landscape of diabetes care, precision medicine, and lifestyle therapies are becoming increasingly important. Dietary polyphenols are like hidden allies found in our everyday meals. These biomolecules, found commonly in fruits, vegetables, and various plant-based sources, hold revolutionary potential within their molecular structure in the way we approach diabetes and its intimidating consequences. There are currently numerous types of diabetes medications, but they are not appropriate for all patients due to limitations in dosages, side effects, drug resistance, a lack of efficacy, and ethnicity. Currently, there has been increased interest in practicing herbal remedies to manage diabetes and its related complications. This article aims to summarize the potential of dietary polyphenols as a foundation in the treatment of diabetes and its associated consequences. We found that most polyphenols inhibit enzymes linked to diabetes. This review outlines the potential benefits of selected molecules, including kaempferol, catechins, rosmarinic acid, apigenin, chlorogenic acid, and caffeic acid, in managing diabetes mellitus as these compounds have exhibited promising results in in vitro, in vivo, in silico, and some preclinical trials study. This encompassing exploration reveals the multifaceted impact of polyphenols not only in mitigating diabetes but also in addressing associated conditions like inflammation, obesity, and even cancer. Their mechanisms involve antioxidant functions, immune modulation, and proinflammatory enzyme regulation. Furthermore, these molecules exhibit anti-tumor activities, influence cellular pathways, and activate AMPK pathways, offering a less toxic, cost-effective, and sustainable approach to addressing diabetes and its complications.

Metabolome Mining of Curcuma longa L. Using HPLC-MS/MS and Molecular Networking.

Turmeric, Curcuma longa L., is a type of medicinal plant characterized by its perennial nature and rhizomatous growth. It is a member of the Zingiberaceae family and is distributed across the world's tropical and subtropical climates, especially in South Asia. Its rhizomes have been highly valued for food supplements, spices, flavoring agents, and yellow dye in South Asia since ancient times. It exhibits a diverse array of therapeutic qualities that encompass its ability to combat diabetes, reduce inflammation, act as an antioxidant, exhibit anticancer properties, and promote anti-aging effects. In this study, organic extracts of C. longa rhizomes were subjected to HPLC separation followed by ESI-MS and low-energy tandem mass spectrometry analyses. The Global Natural Product Social Molecular Networking (GNPS) approach was utilized for the first time in this ethnobotanically important species to conduct an in-depth analysis of its metabolomes based on their fragments. To sum it up, a total of 30 metabolites including 16 diarylheptanoids, 1 diarylpentanoid, 3 bisabolocurcumin ethers, 4 sesquiterpenoids, 4 cinnamic acid derivatives, and 2 fatty acid derivatives were identified. Among the 16 diarylheptanoids identified in this study, 5 of them are reported for the first time in this species.

Quantification of Phenolic and Flavonoid Content, Antioxidant Activity, and Proximate Composition of Some Legume Seeds Grown in Nepal.

This study was carried out to evaluate some legume seeds growing in Nepal for their proximate composition, quantification of total phenolic (TPC) and flavonoid (TFC) contents, and in vitro, antioxidant and antidiabetic activities. These included legume grains such as chickpeas (Cicer arietinum), pea (Pisum sativum), mung bean (Vigna mungo), lima bean (Phaseolus lunatus), broad bean (Vicia faba), lentil (Lens culinaris), soybean (Glycine max), and common bean (Phaseolus vulgaris). The legume seeds were ground to make the flour which was extracted with methanol. The phenolic and flavonoid content was estimated by Folin-Ciocalteu's phenol and aluminum chloride colorimetric methods. The in vitro antioxidant and antidiabetic activity was evaluated by using DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical scavenging and α-amylase enzyme inhibition assay. The different legumes showed considerable variations in their phenolic contents (30.64 ± 1.50 mg·GAE/g to 46.65 ± 1.25 mg·GAE/g legume seeds). Similarly, the total flavonoid contents showed 135.5 ± 10.88 mg·QE/g to 191.7 ± 8.73 mg·QE/g legume seeds. The in vitro antioxidant activity was evaluated in IC50 which ranged from 31.60 ± 0.06 µg/mL to 69.74 ± 0.89 µg/mL. The α-amylase inhibition was evaluated in IC50 which ranged from 217.38 µg/mL to 425.75 µg/mL as compared to the standard acarbose of 52.76 µg/mL. This study suggested that legumes are good sources of proteins, carbohydrates, and fats mainly for vegetarian people. The selection of the right legume species could be a good source of natural antioxidants and antidiabetics for nutraceutical uses and the beneficial effects of legumes from human health perspectives. Legume seeds growing in Nepal could be used as a sustainable and cheap meat alternative and are considered the most important food source.

Molecular Docking and Dynamics Simulation of Several Flavonoids Predict Cyanidin as an Effective Drug Candidate against SARS-CoV-2 Spike Protein.

The in silico method has provided a versatile process of developing lead compounds from a large database in a short duration. Therefore, it is imperative to look for vaccinations and medications that can stop the havoc caused by SARS-CoV-2. The spike protein of SARS-CoV-2 is required for the viral entry into the host cells, hence inhibiting the virus from fusing and infecting the host. This study determined the binding interactions of 36 flavonoids along with two FDA-approved drugs against the spike protein receptor-binding domain of SARS-CoV-2 through molecular docking and molecular dynamics (MD) simulations. In addition, the molecular mechanics generalized Born surface area (MM/GBSA) approach was used to calculate the binding-free energy (BFE). Flavonoids were selected based on their in vitro assays on SARS-CoV and SARS-CoV-2. Our pharmacokinetics study revealed that cyanidin showed good drug-likeness, fulfilled Lipinski's rule of five, and conferred favorable toxicity parameters. Furthermore, MD simulations showed that cyanidin interacts with spike protein and alters the conformation and binding-free energy suited. Finally, an in vitro assay indicated that about 50% reduction in the binding of hACE2 with S1-RBD in the presence of cyanidin-containing red grapes crude extract was achieved at approximately 1.25 mg/mL. Hence, cyanidin may be a promising adjuvant medication for the SARS-CoV-2 spike protein based on in silico and in vitro research.

Catunaregam spinosa (Thunb.) Tirveng: A Review of Traditional Uses, Phytochemistry, Pharmacological Activities, and Toxicological Aspects.

Catunaregam spinosa (Thunb.) Tirveng. (Syn. Randia dumetorum (Retz.) Lam.), belonging to the Rubiaceae family, is distributed in south Asian countries. It is used as a traditional medicine to treat gastrointestinal and hepatic problems and as an anti-inflammatory and antimicrobial agent. The main aim of this review is to collect and analyze the available scientific information on traditional uses, phytochemistry, and pharmacological activities of C. spinosa. The scientific information related to C. spinosa was collected from various resources and databases such as SciFinder, Scopus, PubMed, and other databases. C. spinosa was found to be an important crude drug of the traditional medicinal systems such as Ayurveda. It was found to be used by the people of India as an alternative medicine, while the fruit of this plant was found to be used in dietary regimens as well. Active phytochemicals such as catunarosides, randianin, and several other saponins and triterpenoids possess various pharmacological activities such as anti-inflammatory, hepatoprotective, antibacterial, and immunomodulatory activities. Many studies have been performed to isolate the active compounds; however, there is a need for more activity-guided isolation studies. Various in vitro studies showed promising results but there are not many studies related to mechanism of actions using animal models. Hence, future studies on C. spinosa should focus on correlating the traditional uses with active phytoconstituents and modern pharmacological activities.

α-Amylase Inhibitory Activity of Catunaregam spinosa (Thunb.) Tirveng.: In Vitro and In Silico Studies.

α-Amylase is an enzyme involved in the breaking down of large insoluble starch molecules into smaller soluble glucose molecules. Catunaregam spinosa (Thunb.) Tirveng. (syn. Randia dumetorum (Retz.) Lam., Family: Rubiaceace) has been used as traditional medicine for the treatment of gastrointestinal problems, skin diseases, and diabetes. In this context, we studied the in vitro α-amylase inhibiting properties of methanol extracts of leaves and bark of C. spinosa. The methanol extract of bark was further fractionated into hexane, dichloromethane and ethyl acetate, and water-soluble fractions, and their α-amylase inhibitory activity was evaluated. In silico molecular docking and ADMET analysis of several compounds previously reported from the bark of C. spinosa were also performed. The in vitro α-amylase inhibition activity assay of the dichloromethane fraction of extract of bark (IC50: 77.17 ± 1.75 µg/mL) was more potent as compared to hexane and ethyl acetate fractions. The in silico molecular docking study showed that previously reported compounds from the stem bark such as balanophonin, catunaregin, ß-sitosterol, and medioresinol were bounded well with the active catalytic residue of porcine pancreatic α-amylase indicating better inhibition. The ADMET analysis showed the possible drug-likeness and structure-activity relationship of selected compounds. These compounds should be studied further for their potential α-amylase inhibition in animal models.

Assessing volatile organic compound level in selected workplaces of Kathmandu Valley.

Volatile organic compounds (VOCs) are one of the major contributors to poor indoor air quality. Due to advancements in sensor technologies, continuous if not regular monitoring total VOC (TVOC) and or some specific VOC in potential high risk workplaces is possible even in resource limited settings. In this study, we implemented a portable VOC sensor to measure concentration of TVOC and formaldehyde (HCHO) in six types of potential high risk workplaces (n = 56 sites) of Katmandu Valley. For comparison, concentration was also measured in immediate surroundings (n = 56) of all the sites. To get preliminary information on safety practices, a survey study was also conducted. The mean TVOC and HCHO concentration in the sites ranged from 1.5‒8 mg/m3 and <0.01-5.5 mg/m3, respectively. The indoor: outdoor TVOC and HCHO ratio (I/O) was found to be significantly higher (I/O > 1.5 and p < 0.05) in 34 (~61%) and 47 sites (∼84%), respectively. A strong positive correlation between HCHO and TVOC concentration was observed in furniture industry (R = 0.91) and metal workshops (R = 0.98). Interestingly, we found TVOC and HCHO concentration higher than WHO safe limit in ∼64% and ∼32% sites, respectively. A rough estimate of chronic daily intake (CDI) of formaldehyde showed that CDI is higher than WHO limit in four sites. These findings suggested that indoor air quality in the significant number of the workplaces is poor and possible measures should be taken to minimize the exposure.

Coixol amplifies glucose-stimulated insulin secretion via cAMP mediated signaling pathway.

Recently, we reported the role of coixol (6-methoxy-2(3H)-benzoxazolone), an alkaloid from Scoparia dulcis, in glucose-dependent insulin secretion; however, its insulin secretory mechanism(s) remained unknown. Here, we explored the insulinotropic mechanism(s) of coixol in vitro and in vivo. Mice islets were batch incubated, perifused with coixol in the presence of agonists/antagonists, and insulin secretion was measured by ELISA. Intracellular cAMP levels were measured using enzyme immunoassay. K+- and Ca2+-currents were recorded in MIN6 cells using whole-cell patch-clamp technique. The in vivo glucose tolerance and the insulinogenic index were evaluated in diabetic rats treated with coixol at 25 and 50 mg/kg, respectively. Coixol, unlike sulfonylurea, enhanced insulin secretion in batch incubated and perifused islets at high glucose, with no effect at basal glucose concentrations. Coixol showed no pronounced effect on the inward rectifying K+- and Ca2+-currents in whole-cell patch recordings. Moreover, coixol-induced insulin secretion was further amplified in the depolarized islets. Coixol showed an additive effect with forskolin (10 µM)-induced cAMP level, and in insulin secretion; however, no additive effect was observed with isobutylmethylxanthine (IBMX, 100 µM)-induced cAMP level, nor in insulin secretion. The PKA inhibitor H-89 (50 µM), and Epac2 inhibitor MAY0132 (50 µM) significantly inhibited the coixol-induced insulin secretion (P < 0.01). Furthermore, insulin secretory kinetics revealed that coixol potentiates insulin secretion in both early and late phases of insulin secretion. In diabetic animals, coixol showed significant improvement in glucose tolerance and on fasting blood glucose levels. These data suggest that coixol amplifies glucose-stimulated insulin secretion by cAMP-mediated signaling pathways.