Thymus linearis Extracts Ameliorate Indices of Metabolic Syndrome in Sprague Dawley Rats.

Materials and Methods: The extract library (n-hexane (NH), ethyl acetate (EA), methanol (M), distilled water (DW), and combined extract (CE)) was standardized using in vitro phytochemical, antioxidant, and α-amylase inhibition assays, after which the protective effect of selected "hit," i.e., CE against metabolic syndrome, was determined in vivo, using rats fed a high-fat diet supplemented with additional cholesterol administration. CE was administered to Sprague Dawley rats in high dose as 100 mg/kg in carboxymethyl cellulose (CMC) (1 ml; 0.75% in DW) and low-dose group as 50 mg/kg in CMC (0.5 ml; 0.75% in DW). After 10 weeks, the effects of CE on insulin resistance, lipid metabolism, nonalcoholic fatty liver disease (NAFLD), oxidative stress, and genotoxicity were assessed through histological, biochemical, and hematological investigations. Results: Phytochemical analysis including RP-HPLC analysis of the extracts showed that flavonoids and phenolics (myricetin, kaempferol, and apigenin), previously known to be effective against obesity and diabetes, are present in the extracts. Antioxidant studies revealed that the plant possesses a highly significant (p < 0.05) concentration of antioxidants. Satisfactory α-amylase inhibitory activity was also observed in in vitro experiments. In vivo studies showed that CE-administered animals had significantly (p < 0.05) lower weight gain and smaller adipocytes than the control group. Moreover, CE resisted any significant (p < 0.05) change in the organ weights. Analogous to findings from its traditional use, the plant extract had a positive modulatory effect on insulin resistance and hyperglycemia. The study also indicated that CE resisted high-fat diet-induced disturbance in lipid profile and countered any pathological changes in liver enzymes caused by fat-infused diet. Furthermore, a study on endogenous antioxidant levels indicated that CE was effective in maintaining catalase and peroxidase levels within the normal range and resisted the effects of lipid peroxidation of thiobarbituric acid reactive substances. Conclusion: In principle, the current study's findings scientifically validate the implication of T. linearis in metabolic syndrome and recommend further studies on molecular insights of the observed therapeutic activity.

The Inhibitory Effect of Polyphenon 60 from Green Tea on Melanin and Tyrosinase in Zebrafish and A375 Human Melanoma Cells.

Polyphenon 60 (PP60) from green tea has long been used as an antioxidant, anticancer, antimicrobial, and antimutagenic. Aim of the Study. To investigate tyrosinase inhibition-related kinetic mechanism and antimelanogenesis potential of PP60. Materials and Methods. The effect of PP60 on melanin and tyrosinase was evaluated in A375 melanoma cells and zebrafish embryos. The melanoma cells were treated with 20, 40, and 60 µg/mL of PP60, and tyrosinase expression was induced by using L-DOPA. The western blot method was used for the evaluation of tyrosinase expression. Cell lysates were prepared from treated and untreated cells for cellular tyrosinase and melanin quantification. Furthermore, zebrafish embryos were treated with 20, 40, and 60 µg/mL of PP60 and reference drug kojic acid for determination of depigmentation and melanin quantification. In vitro assays were also performed to examine the impact of PP60 on mushroom tyrosinase activity. To determine cytotoxicity, MTT was used against melanoma cell line A375. Results. PP60 showed good tyrosinase inhibitory activity with an IC50 value of 0.697 ± 0.021 µg/mL as compared to kojic acid a reference drug with an IC50 value of 2.486 ± 0.085 µg/mL. Kinetic analysis revealed its mixed type of inhibition against mushroom tyrosinase. In addition, western blot analysis showed that at 60 µg/mL dose of PP60 significantly reduced L-DOPA-induced tyrosinase expression in melanoma cells. PP60 significantly inhibits the cellular tyrosinase (p < 0.05) and reduces the melanin (p < 0.05) contents of melanoma cells. Furthermore, PP60 was found to be very potent in significantly reducing the zebrafish embryos' pigmentation (p < 0.05) and melanin (p < 0.05) content at the dose of 60 µg/mL. Conclusions. Our results demonstrate that PP60 has a strong potency to reduce pigmentation. It may be useful for the cosmetic industries to develop skin whitening agents with minimal toxic effects.

Nanoparticles as Alternatives for the Control of Haemonchus contortus: A Systematic Approach to Unveil New Anti-haemonchiasis Agents.

Haemonchus contortus is an infectious gastrointestinal nematode parasite of small ruminants. This study addresses the in vitro/in vivo anti-haemonchiasis potential, toxicological effects, and mechanism of action of nanoparticles. Online databases were used to search and retrieve the published literature (2000 to 2021). A total of 18 articles were selected and reviewed, out of which, 13 (72.2%) studies reported in vitro, 9 (50.0%) in vivo, and 4 (22.2%) both in vitro/in vivo efficacy of different nanoparticles. Mostly, organic nanoparticles (77.7%) were used including polymeric (85.7%) and lipid nanoparticles (14.3%). The highest efficacy, in vitro, of 100% resulted from using encapsulated bromelain against eggs, larvae, and adult worm mortality at 4, 2, and 1 mg/ml, respectively. While in vivo, encapsulated Eucalyptus staigeriana oil reduced worm burden by 83.75% and encapsulated Cymbopogon citratus nano-emulsion by 83.1%. Encapsulated bromelain, encapsulated Eucalyptus staigeriana oil, and encapsulated Cymbopogon citratus nano-emulsion were safe and non-toxic in vivo. Encapsulated bromelain damaged the cuticle, caused paralysis, and death. Nanoparticles could be a potential source for developing novel anthelmintic drugs to overcome the emerging issue of anthelmintic resistance in H. contortus. Studies on molecular effects, toxicological consequences, and different pharmacological targets of nanoparticles are required in future research.

Exploring threatened traditional knowledge; ethnomedicinal studies of rare endemic flora from Lesser Himalayan region of Pakistan

ABSTRACT Himalayas are one among the world biodiversity hotspots harboring many endemic medicinal plants. Despite augmentation in the documentation of ethnopharmacological knowledge of medicinal plant species, information regarding endemic species is still underway. Current paper highlights the traditional medicinal uses of rare endemic and unexplored group of plants having potential for novel chemical constituents with effective pharmacological activities. In total, 142 informants (91 male and 51 female) including seventeen traditional healers were interviewed using semi-structured questionnaire, personal observations and group discussions. Interviews were taken in field or otherwise photographs were shown for identification. Females were interviewed indirectly through male family members. For data analysis, quantitative analytical approach was adopted using ethnopharmacological indices as Relative frequency of citations and Fidelity Level. In total, 38 endemic plant species belonging to nineteen families were utilized by the local inhabitants. Highest number of endemics was belonging to Ranunculaceae (7), followed by Gentianaceae and Rosaceae (4 each) with respect to number of species. Highest number of endemics was used in fever, wound healing, throat infection and tonic (4 species each). Root was the most widely used part (36.17%) in cure of diseases and the leading mode administered was decoction (25.49%). Highest use reports and RFC values were recorded for Pimpinella stewartii (58 citations, 0.41 RFC), Caltha alba var. alba (52 citations, 0.37 RFC). Endemic plant species considerably contribute toward ethnomedicinal knowledge and despite rarity, the communities prefer their utilization. Conservation of endemics is necessary for future availability to the local communities.

Efficacy of Several Plants Extracts on the Tunneling Activity and Survival of Subterranean Termites (Coptotermes gestroi and Globitermes sulphureus).

This study examined the tunneling activity and the behaviour of two subterranean termites, Globitermes sulphurues and Coptotermes gestroi treated with four plant extracts, namely Pyllanthus niruri, Azadirachta indica, Leucaena leucocephala and Andrographis paniculata. All plants were extracted with three different solvents (methanol, hexane and water) and were diluted into three concentrations (500, 5,000 and 10,000 ppm). A group of 50 workers and 2 soldiers were tested and observed daily during the exposure in order to determine their survivorship and behaviour. Both sections were scored to determine their tunneling activities after seven days. There was a significant difference of tunneling activities of C. gestroi on sand treated with plant extracts (X2(2) = 31.790, p < 0.0001) with a mean rank of 8.50 for methanolic extracts and 32.50 for both hexane and water extracts. Meanwhile, no significant difference was observed on tunneling activity of G. sulphureus on treated sand (X2(2) = 2.200, p = 0.333) with a mean rank of 20.72 for methanolic extracts, 26.31 for water extracts and 26.47 for hexane extracts. Results showed that plants extracted with methanol demonstrated strong repellent properties with 0 tunneling activity on the treated sand and low survivorship of both termites. Moreover, both termites did not survive (0%) when they were treated with methanolic extracts at 10,000 ppm. They also displayed a different behaviour post-exposure such as avoidance, gradually losing the ability to walk and low feeding consumption. These results indicate that there is a strong termiticidal activity of plants extracted with methanol against C. gestroi and G. sulphureus.

Courtship activity, copulation & insemination success in a mosquito vector fed a herbal aphrodisiac: Implications for sterile insect technology.

BACKGROUND & OBJECTIVES: In sterile insect technology (SIT), mating competitiveness is a pre-condition for the reduction of target pest populations and a crucial parameter for judging efficacy. Still, current SIT trials are being hindered by decreased effectiveness due to reduced sexual performance of released males. Here, we explored the possible role of a herbal aphrodisiac in boosting the mating activity of Aedes aegypti. METHODS: Males were fed one of two diets in this study: experimental extract of Eurycoma longifolia (MSAs) and sugar only (MSOs). Differences in life span, courtship latency, copulation activity and mating success were examined between the two groups. RESULTS: No deaths occurred among MSA and MSO males. Life span of MSOs was similar to that of MSAs. The courtship latency of MSAs was shorter than that of MSOs (P<0.01). MSAs had greater copulation success than MSOs (P<0.001). In all female treatments, MSAs mated more than MSOs, but the differences in rate were significant only in the highest female density (P<0.05). In MSAs, mating success varied significantly with female density (P<0.01), with the 20-female group (P<0.01) having the lowest rate. Single MSA had better mating success at the two lowest female densities. In MSOs, there were no significant differences in mating success rate between the different female densities. INTERPRETATION & CONCLUSIONS: Our results suggested that the herbal aphrodisiac, E. longifolia, stimulated the sexual activity of Ae. aegypti and may be useful for improving the mating competitiveness of sterile males, thus improving SIT programmes.

Anti-inflammatory and anti-oxidant effects of Cardamom (Elettaria repens (Sonn.) Baill) and its phytochemical analysis by 4D GCXGC TOF-MS.

Elleteria repens is a large cardamom used in the culinary preparations. In the present study, we have evaluated in vitro antioxidant and free radical scavenging activities E. repens hexane extract (ERH) exhibited DPPH and metal chelating activity with IC50 values of 464±28.3µg/ml, 199±7.2µg/ml whereas the reducing power and antioxidant activities are found to be 289±14.6 AAE/mg, 468±22.7 GAE/mg. The observed antioxidant activities could be correlated with metabolites such as polyphenol, flavonoid, and terpenoid group of compounds identified in hexane fraction of E. repens by 4D GCXGC TOF-MS. Further ERH was evaluated for its protective properties against macromolecules such as DNA, protein and lipid damage. The extract showed protection against H2O2 induced DNA damage and inhibited AAPH induced protein oxidation and lipid peroxidation. Moreover, ERH administration to rats at 50 and 100mg/kg inhibited carrageenan-induced paw edema, and down-regulated cytokines such as COX-2, IL-6, and TNF-α and inhibited i-NOS mediated NO generation. E. repens also exhibited antioxidant effects by restoring SOD, catalase, GSH levels and inhibited lipid peroxidation in carrageenan challenged rats. Overall, the results suggest that E. repens may be useful in combating inflammation and oxidative stress.

Exposure of a Dengue Vector to Tea and Its Waste: Survival, Developmental Consequences, and Significance for Pest Management.

Dengue mosquitoes are evolving into a broader global public health menace, with relentless outbreaks and the rise in number of Zika virus disease cases as reminders of the continued hazard associated with Aedes vectors. The use of chemical insecticides-the principal strategy against mosquito vectors-has been greatly impeded due to the development of insecticide resistance and the shrinking spectrum of effective agents. Therefore, there is a pressing need for new chemistries for vector control. Tea contains hundreds of chemicals, and its waste, which has become a growing global environmental problem, is almost as rich in toxicants as green leaves. This paper presents the toxic and sublethal effects of different crude extracts of tea on Aedes albopictus. The survival rates of larvae exposed to tea extracts, especially fresh tea extract (FTE), were markedly lower than those in the control treatment group. In addition to this immediate toxicity against different developmental stages, the extracts tested caused a broad range of sublethal effects. The developmental time was clearly longer in containers with tea, especially in those with young larvae (YL) and FTE. Among the survivors, pupation success was reduced in containers with tea, which also produced low adult emergence rates with increasing tea concentration. The production of eggs tended to be reduced in females derived from the tea treatment groups. These indirect effects of tea extracts on Ae. albopictus exhibited different patterns according to the exposed larval stage. Taken together, these findings indicate that tea and its waste affect most key components of Ae. albopictus vectorial capacity and may be useful for dengue control. Reusing tea waste in vector control could also be a practical solution to the problems associated with its pollution.

Coffee and its waste repel gravid Aedes albopictus females and inhibit the development of their embryos.

BACKGROUND: Dengue is a prevalent arboviral disease and the development of insecticide resistance among its vectors impedes endeavors to control it. Coffee is drunk by millions of people daily worldwide, which is associated with the discarding of large amounts of waste. Coffee and its waste contain large amounts of chemicals many of which are highly toxic and none of which have a history of resistance in mosquitoes. Once in solution, coffee is brownish in colour, resembling leaf infusion, which is highly attractive to gravid mosquitoes. To anticipate the environmental issues related to the increasing popularity of coffee as a drink, and also to combat insecticide resistance, we explored the deterrence potentials of coffee leachates against the ovipositing and embryonic stages of the dengue vector, Aedes albopictus. METHODS: In a series of choice, no-choice, and embryo toxicity bioassays, we examined changes in the ovipositional behaviours and larval eclosion of Ae. albopictus in response to coffee extracts at different concentrations. RESULTS: Oviposition responses were extremely low when ovicups holding highly concentrated extract (HCE) of coffee were the only oviposition sites. Gravid females retained increased numbers of mature eggs until 5 days post-blood feeding. When provided an opportunity to oviposit in cups containing coffee extracts and with water, egg deposition occurred at lower rates in those containing coffee, and HCE cups were far less attractive to females than those containing water only. Females that successfully developed in a coffee environment preferentially oviposited in such cups when in competition with preferred oviposition sites (water cups), but this trait did not continue into the fourth generation. Larval eclosion occurred at lower rates among eggs that matured in a coffee environment, especially among those that were maintained on HCE-moistened substrates. CONCLUSIONS: The observations of the present study indicate a pronounced vulnerability of Ae. albopictus to the presence of coffee in its habitats during the early phases of its life cycle. The observations that coffee repels gravid females and inhibits larval eclosion provide novel possibilities in the search for novel oviposition deterrents and anti-larval eclosion agents against dengue vectors.

Pharmaceutical and biopharmaceutical evaluation of extracts from different plant parts of indigenous origin for their hypoglycemic responses in rabbits.

This study was designed to evaluate the hypoglycemic effects of different plant extracts in single and in combined formulation, in experimentally induced "diabetic rabbits". The extracts were obtained from seeds of Syzygium jambolana, fruits of Momordica charantia and leaves of Azadirachta indica. Treatment of diabetes with plant extracts was started at 8 days after alloxan injection. Rabbits were randomly divided into four groups, each group consisting of six rabbits. Each group of rabbits was given a dose of granules containing 200 mg/kg b.w. concentrated ethanolic extract of a plant while the fourth group was given a dose of granules consisting of combined extract of all three folk plants. Blood samples were drawn at 0, 2, 4, 8, 12, 24, 36, 48, 72 and 96 h. Serum glucose estimation was done by glucose oxidase kit method. Anti-diabetic effect was produced after 72 h in groups 1, 2 and 3 that were administered with a dose of granules of ethanolic extract of single plant but in group 4, treated with 200 mg/kg body weight of combined extract of all three plants, hypoglycemic effect was produced after 96 h. Hypoglycemic effects may be induced in rabbits by administration of extracts of various plant parts. The hypoglycemic effect produced by granules of single plant extract was more pronounced than antidiabetic effect produced by combining three extracts in a single formulation.