Ecological risk assessment of heavy metals contaminated mining sites of eastern india using soil and moss.

The blooming industrialization and urbanization is leading to increased mining operations. These intensified mining activities emit heavy metals into the environment, posing serious threats to ecosystems. Hence, this study focused on assessing heavy metal pollution in mining soil, utilizing mosses as bioindicators. The ecological risk, geo-accumulation factor, and contamination factor have been calculated to know the harmful effect of heavy metals on ecosystem. The study covered three distinct mining sites of eastern India within Odisha: Jajpur's Sukinda Valley (SP1, Cr), Keonjhar's Joda-Barbil (SP2, Fe and Mn), and Sundargarh's Koira-Joda (SP3, Fe). The collection of 48 soil samples through random sampling revealed significant variations in heavy metal concentrations. SP1 recorded Cr concentration of 6572 ± 445 mg/kg and Ni of 8042.47 ± 501.38 mg/kg, surpassing eco-toxicological levels. The storage site in SP2 exhibited the highest Fe concentration at 9872 ± 502 mg/kg, and Mn levels in SP3 were at 7884 ± 432 mg/kg. Storage areas in all three regions held the highest concentrations of heavy metals. Mosses in studied area demonstrated as potential bioindicators for monitoring heavy metal pollution. EF and Igeo assessments showed Cd, Pb, Hg, and other heavy metal contamination compared to earlier investigations. This study indicated higher ecological risks for Pb, As, Cu, Ni, and Zn. The Hyophila involuta accumulates Mn, Cr, Cd, Pb, Fe, and Hg, while Barbula arcuata accumulates Mn, As, and Cu in SP1. Hyophila involuta and Trematodon longicollis accumulate Mn, Cr, Cd, Pb, Fe, Hg, and Zn in SP2. Trematodon ambiguous accumulates Cd, Fe, and Ni, while Fissidens diversifolius accumulates Mn, Cr, Hg, As, Cu, and Zn in SP3. These findings emphasize the necessity of monitoring heavy metal pollution in contaminated zones using moss as a potential bioindicator.

Soil treatment using a biosurfactant producing bacterial consortium in rice fields contaminated with oily sludge- a sustainable approach.

A consortium of two biosurfactant-producing bacteria (Bacillus pumilus KS2 and Bacillus cereus R2) was developed to remediate petroleum hydrocarbon-contaminated paddy soil. Soil samples from a heavily contaminated rice field near Assam's Lakwa oilfield were collected and placed in earthen pots for treatment. After each month of incubation, 50 g of soil from each earthen pot was collected, and the soil TPH (ppm) in each sample was determined. The extracted TPH samples were analysed by Gas chromatography-mass spectrometry (GC-MS) to confirm microbial degradation. The soil samples were examined for changes in pH, conductivity, total organic content (TOC), water holding capacity, and total nitrogen content in addition to TPH degradation. An increasing trend in TPH degradation was observed with each passing month. After six months of treatment, the sample with the lowest initial TPH concentration (1735 ppm) had the highest degradation (91.24%), while the soil with the highest amount of TPH (5780 ppm) had the lowest degradation (74.35%). A wide range of aliphatic hydrocarbons found in soil samples was degraded by the bacterial consortium. The soil samples contained eight different low- and high-molecular-weight PAHs. Some were fully mineralized, while others were significantly reduced. With the decrease in the TPH level in the polluted soil, a significant improvement in the soil's physicochemical qualities (such as pH, electrical conductivity, total organic content, and water-holding capacity) was observed.

Monascin and ankaflavin-Biosynthesis from Monascus purpureus, production methods, pharmacological properties: A review.

Monascus purpureus copiously yields beneficial secondary metabolites , including Monascus pigments, which are broadly used as food additives, as a nitrite substitute in meat products, and as a colorant in the food industry. Monascus yellow pigments (monascin and ankaflavin) have shown potential antidiabetic, antibacterial, anti-inflammatory, antidepressant, antibiotic, anticancer, and antiobesity activities. Cosmetic and textile industries are other areas where it has established its potential as a dye. This paper reviews the production methods of Monascus yellow pigments, biosynthesis of Monascus pigments from M. purpureus, factors affecting yellow pigment production during fermentation, and the pharmacological properties of monascin and ankaflavin.

Purification and characterization of Taxol and 10-Deacetyl baccatin III from the bark, needles, and endophytes of Taxus baccata by preparative high-performance liquid chromatography, ultra-high-performance liquid chromatography-mass spectrometry, and nuclear magnetic resonance.

Taxol and 10-Deacetyl baccatin III are major taxanes in the bark, needles, and endophytes of Taxus baccata. The current study aimed to develop a process for their separation from different matrices. Crude taxoid was prepared by extraction of samples with methanol, followed by partitioning with dichloromethane and precipitation with hexane. Analytical high-performance liquid chromatography involved isocratic elution on C18 column (4.6 × 250 mm, 5 µm) with methanol-water (70:30 v/v) at a flow rate of 1 ml/min. Injection volume was 20 µl and detection was carried out at 227 nm. The content of Taxol and 10-Deacetyl baccatin III in bark, needles and endophytic culture broth was 11.19 and 1.75 µg/mg; 11.19 and 1.75 µg/mg; and 2.80 and 0.22 µg/L, respectively. Preparative high-performance liquid chromatography was done on C18 column (10 × 250 mm, 5 µm) at a flow rate of 10 ml/min. About 20 g crude taxoid was processed in < 3 h with a recovery of about 90% for both the analytes. The purity of recovered Taxol and 10-Deacetyl baccatin III determined by ultra-high-performance liquid chromatography-mass spectrometry was found to be 95.78 ± 3.63% and 99.72 ± 0.18%, respectively. The structure of recovered Taxol was confirmed by nuclear magnetic resonance. The method can find use in biotransformation studies.

Nanoprotection from SARS-COV-2: would nanotechnology help in Personal Protection Equipment (PPE) to control the transmission of COVID-19?

The coronavirus disease 2019 (COVID-19) has caused a worldwide outbreak. The severe acute respiratory syndrome coronavirus 2 virus can be transmitted human-to-human through droplets and close contact where personal protective equipment (PPE) is imperative to protect the individuals. The advancement of nanotechnology with significant nanosized properties can confer a higher form of protection. Incorporation of nanotechnology into facemasks can exhibit antiviral properties. Nanocoating on surfaces can achieve self-disinfecting purposes and be applied in highly populated places. Moreover, nano-based hand sanitizers can confer better sterilizing efficacies with low skin irritation as compared to alcohol-based hand sanitizers. The present review discusses the incorporation of nanotechnology into nano-based materials and coatings in facemasks, self-surface disinfectants and hand sanitizers, in the hope to contribute to the current understanding of PPE to combat COVID-19.

Air mapping during COVID-19 and association between air pollutants and physiochemical parameters of the plants using structural equal modeling: a case study.

In urban areas around the world, air pollution introduced by vehicular movement is a key concern. However, restricting vehicular traffic during the COVID-19 shutdown improved air quality to some extent. This study was conducted out in the smart city of Bhubaneswar, which is also the state capital of Odisha, India. The study has tried to map Bhubaneswar by collecting the air quality data before, during, and after the COVID lockdown of six air quality monitoring stations present in Bhubaneswar established under "National Ambient Air Monitoring Program" (NAMP). Furthermore, plants, which are the most vulnerable to air pollution, can show a variety of visible changes depending on their level of sensitivity. Moreover, leaves of Mangifera indica, Monoon longifolium, Azadirachta indica, Millettia pinnata, Aegle marmelos were collected from nearby of six air monitoring stations to assess the "Air Pollution Tolerance Index." M. indica was found to be intermediately tolerant, and all of the other species were found to be sensitive. The structural equation modeling results also revealed a significant relationship between total chlorophyll content, relative water content, ascorbic acid content, leaf extract pH, APTI with species, air quality index, and PM10.

Enhancement of vincristine under in vitro culture of Catharanthus roseus supplemented with Alternaria sesami endophytic fungal extract as a biotic elicitor.

Vincristine, one of the major vinca alkaloid of Catharanthus roseus (L.) G. Don. (Apocynaceae), was enhanced under in vitro callus culture of C. roseus using fungal extract of an endophyte Alternaria sesami isolated from the surface-sterilized root cuttings of C. roseus. Vindoline, a precursor molecule for vincristine production, was detected for the first time in the fungal endophyte A. sesami which was used as a biotic elicitor in this study to enhance vincristine content in the C. roseus callus. It was identified using high-performance liquid chromatography and mass spectroscopy techniques by matching retention time and mass data with reference molecule. Supplementing the heat sterilized A. sesami endophytic fungal culture extract into the callus culture medium of C. roseus resulted in the enhancement of vincristine content in C. roseus callus by 21.717% after 105-day culture.

ß-Carotene-production methods, biosynthesis from Phaffia rhodozyma, factors affecting its production during fermentation, pharmacological properties: A review.

ß-Carotene is the most treasured provitamin A carotenoid molecule exhibiting antioxidant and coloring properties and significant applications in the food, pharmaceutical, and nutraceutical industries. ß-Carotene has many biological functions within the human body; however, it is not synthesized within the human body, so its requirements are fulfilled through food and pharmaceuticals. Its manufacturing via chemical synthesis or extraction from plants offers low yields with excessive manufacturing expenses, which attracted the researchers toward microbial production of ß-carotene. This alternative provides higher yield and low expenses and thus is more economical. Phaffia rhodozyma is a basidiomycetous yeast that is utilized to prevent cardiovascular diseases and cancer and to enhance immunity and antiaging in people. This paper reviews the methods of production of ß-carotene, biosynthesis of ß-carotene fromP. rhodozyma, factors affecting ß-carotene production during fermentation, and pharmacological properties of ß-carotene.

Comparative genomic analyses of Bacillus subtilis strains to study the biochemical and molecular attributes of nattokinases.

The present research work explores the Nattokinase (NK) producing capacity of five Bacillus subtilis strains (MTCC 2616, MTCC 2756, MTCC 2451, MTCC 1427, and MTCC 7164) using soybean varieties as substrate under solid-state fermentation conditions. Subsequently, the biochemical attributes of NKs were analyzed. Soybean variety didn't affect the production of NK to a significant extent; however, the five strains differed substantially for their NK producing capacity. NK produced by MTCC 2451 (R3) showed a low Kmvalue implying its higher specificity for fibrin but this strain (MTCC 2451) didn't produce NK in sufficient quantity. The low Km of MTCC 2451 NK implicates its potential candidature for treating blood clots in cardiovascular patients. The NK produced by MTCC 2616 (R1) was produced in sufficient quantity and showed good fibrin dissolving potential. The aprN of MTCC 2616 substantially varied from the other four strains. The aprN of MTCC 2756 (R2), MTCC 2451 (R3), MTCC 1427 (R4), and MTCC 7164 (R5) shared > 99% sequence identity, but the encoded NKs had significant variations in their Km values. The biochemical-molecular analyses indicate the co-presence of three critical residues (Thr130, Asp140, and Tyr217) as a quintessential attribute in determining the low Km of NK enzymes, and the absence of any one of the three critical residues may affect (highly increase) the Km.

ZnO Nanoparticles of Rubia cordifolia Extract Formulation Developed and Optimized with QbD Application, Considering Ex Vivo Skin Permeation, Antimicrobial and Antioxidant Properties.

The objective of the current research is to develop ZnO-Manjistha extract (ZnO-MJE) nanoparticles (NPs) and to investigate their transdermal delivery as well as antimicrobial and antioxidant activity. The optimized formulation was further evaluated based on different parameters. The ZnO-MJE-NPs were prepared by mixing 10 mM ZnSO4·7H2O and 0.8% w/v NaOH in distilled water. To the above, a solution of 10 mL MJE (10 mg) in 50 mL of zinc sulfate was added. Box-Behnken design (Design-Expert software 12.0.1.0) was used for the optimization of ZnO-MJE-NP formulations. The ZnO-MJE-NPs were evaluated for their physicochemical characterization, in vitro release activity, ex vivo permeation across rat skin, antimicrobial activity using sterilized agar media, and antioxidant activity by the DPPH free radical method. The optimized ZnO-MJE-NP formulation (F13) showed a particle size of 257.1 ± 0.76 nm, PDI value of 0.289 ± 0.003, and entrapment efficiency of 79 ± 0.33%. Drug release kinetic models showed that the formulation followed the Korsmeyer-Peppas model with a drug release of 34.50 ± 2.56 at pH 7.4 in 24 h. In ex vivo studies ZnO-MJE-NPs-opt permeation was 63.26%. The antibacterial activity was found to be enhanced in ZnO-MJE-NPs-opt and antioxidant activity was found to be highest (93.14 ± 4.05%) at 100 µg/mL concentrations. The ZnO-MJE-NPs-opt formulation showed prolonged release of the MJE and intensified permeation. Moreover, the formulation was found to show significantly (p < 0.05) better antimicrobial and antioxidant activity as compared to conventional suspension formulations.

Okra (Abelmoschus Esculentus) as a Potential Dietary Medicine with Nutraceutical Importance for Sustainable Health Applications.

Recently, there has been a paradigm shift from conventional therapies to relatively safer phytotherapies. This divergence is crucial for the management of various chronic diseases. Okra (Abelmoschus esculentus L.) is a popular vegetable crop with good nutritional significance, along with certain therapeutic values, which makes it a potential candidate in the use of a variety of nutraceuticals. Different parts of the okra fruit (mucilage, seed, and pods) contain certain important bioactive components, which confer its medicinal properties. The phytochemicals of okra have been studied for their potential therapeutic activities on various chronic diseases, such as type-2 diabetes, cardiovascular, and digestive diseases, as well as the antifatigue effect, liver detoxification, antibacterial, and chemo-preventive activities. Moreover, okra mucilage has been widely used in medicinal applications such as a plasma replacement or blood volume expanders. Overall, okra is considered to be an easily available, low-cost vegetable crop with various nutritional values and potential health benefits. Despite several reports about its therapeutic benefits and potential nutraceutical significance, there is a dearth of research on the pharmacokinetics and bioavailability of okra, which has hampered its widespread use in the nutraceutical industry. This review summarizes the available literature on the bioactive composition of okra and its potential nutraceutical significance. It will also provide a platform for further research on the pharmacokinetics and bioavailability of okra for its possible commercial production as a therapeutic agent against various chronic diseases.

Metabolic differentiation and quantification of gymnemic acid in Gymnema sylvestre (Retz.) R.Br. ex Sm. leaf extract and its fermented products.

INTRODUCTION: Gymnemagenin is the bioactive metabolite found in Gymnema sylvestre leaves and possesses different therapeutic potential. Due to its lower abundance and higher market potential, gymnemagenin was obtained from chemical conversion and bacterial biotransformation. OBJECTIVE: To obtain the probiotic-based fermentative conversion of gymnemic acid-enriched G. sylvestre leaf extract to gymnemagenin-containing nutraceuticals and its metabolites based chromatographic comparison. MATERIAL AND METHODS: Gymnema sylvestre leaves were extracted through soxhalation, and the extract was prepared and characterised. Gymnemic acid was fermented, separately, by Lactobacillus casei, Lactobacillus rhamnosus, Bifidobacterium bifidum, and by their mix co-culture. The fermented materials were analysed for their gymnemagenin content, antioxidant potential, antidiabetic potential, and metabolomics analysis. RESULTS: Extraction yielded about 35% w/w of raw plant material, and 8.5% was found to be as total saponin content. Extract at higher concentration (≥ 5%, w/v) significantly altered the growth behaviour of probiotics. High-performance thin-layer chromatography (HPTLC) based quantification of gymnemagenin revealed that a maximum increase of 95.5% gymnemagenin was found in extract incubated with B. bifidum followed by mix co-culture containing (B. bifidum, L. casei, and L. rhamnosus), L. casei, and L. rhamnosus. However, liquid chromatography mass spectrometry (LC-MS) analysis resulted in the identification of a total of 56 metabolites. CONCLUSION: Chromatographically profiled, and probiotic-based fermented G. sylvestre leaves can be used as a potent nutraceutical for diabetes and other metabolic disorders.

Design of expert guided investigation of native L-asparaginase encapsulated long-acting cross-linker-free poly (lactic-co-glycolic) acid nanoformulation in an Ehrlich ascites tumor model.

Present study explores native L-asparaginase encapsulated long-acting cross-linker-free PLGA-nanoformulation in an Ehrlich ascites tumor model. L-asparaginase-PLGA nanoparticles for tumor were prepared using a double emulsion solvent evaporation technique, optimized and validated by Box-Behnken Design. L-ASN-PNs showed a particle size of 195 nm ± 0.2 nm and a PDI of 0.2. Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) techniques revealed its smooth morphology and elicited an in-vitro release of 80% of the drug, following the Higuchi drug release model. In-vivo studies of L-ASN-PNs on an Ehrlich ascites tumor (EAT) model were completed and compared with the standard medication of 5-fluorouracil (5-FU) treatment. L-ASN-PN treated mice showed a 51.15% decrease in tumor volume and 100% survival rate with no reduction in body weight, no haemotoxicity and no hepatotoxicity, as evident from the hematological parameters, and liver enzyme parameters that were well within the prescribed limits. Chemotherapy has severe side effects and restricted therapeutic success. Henceforth, the purported L-Asparaginase PLGA nanoparticles are a suitable entity for better tumor regression, intra-tumor accumulation and no hematological side-effects.

Attenuation of neurobehavioral and neurochemical abnormalities in animal model of cognitive deficits of Alzheimer's disease by fermented soybean nanonutraceutical.

The present study was performed to evaluate the efficacy of nanonutraceuticals (NN) for attenuation of neurobehavioral and neurochemical abnormalities in Alzheimer's disease. Solid-state fermentation of soybean with Bacillus subtilis was performed to produce different metabolites (nattokinase, daidzin, genistin and glycitin and menaquinone-7). Intoxication of rats with colchicine caused impairment in learning and memory which was demonstrated in neurobehavioral paradigms (Morris water maze and passive avoidance) linked with decreased activity of acetylcholinesterase (AChE). NN treatment led to a significant increase in TLT in the retention trials as compared to acquisition trial TLT suggesting an improved learning and memory in rats. Further, treatment of NN caused an increase in the activity of AChE (42%), accompanied with a reduced activity of glutathione (42%), superoxide dismutase (43%) and catalase (41%). It also decreased the level of lipid peroxidation (28%) and protein carbonyl contents (30%) in hippocampus as compared to those treated with colchicine alone, suggesting a possible neuroprotective efficacy of NN. Interestingly, in silico studies also demonstrated an effective amyloid-ß and BACE-1 inhibition activity. These findings clearly indicated that NN reversed colchicine-induced behavioral and neurochemical alterations through potent antioxidant activity and could possibly impart beneficial effects in cognitive defects associated with Alzheimer's disease.

Lipid Based nanoformulation of lycopene improves oral delivery: formulation optimization, ex vivo assessment and its efficacy against breast cancer.

This study aims at developing an optimised nanostructured lipid carrier (NLC) of lycopene for efficient absorption following oral administration. The optimised formulation showed an average particle size of 121.9 ± 3.66 nm, polydispersity index (PDI) 0.370 ± 0.97 and zeta potential -29.0 ± 0.83 mV. Encapsulation Efficiency (% EE) and drug loading (% DL) was found to be 84.50% ± 4.38 and 9.54% ± 2.65, respectively. In vitro release studies demonstrated the burst release within 4-9 h followed by sustained release over 48 h. The IC50 value of lycopene extract and optimised NLC for ABTS+• were found to be 172.37 µg Trolox equivalent and 184.17 µg Trolox equivalent whereas, for DPPH•, 117.76 µg Trolox equivalent and 143.08 µg Trolox equivalent respectively. Ex vivo studies and MTT assay revealed that the NLC had better permeation and cause sufficiently more cytotoxicity as compared to drug extract due to higher bioavailability and greater penetration.

Folic acid ameliorates celecoxib cardiotoxicity in a doxorubicin heart failure rat model.

CONTEXT: The cardiotoxic effect of selective cyclo-oxygenase-2 inhibitors is well known. While rofecoxib and valdecoxib have been withdrawn, celecoxib remains on the market. Folic acid, a naturally occurring vitamin, has been shown to reduce myocardial ischemia and post-reperfusion injury in rats. OBJECTIVE: This study examined the cardiac effects of celecoxib and folic acid on doxorubicin-induced cardiomyopathy in rats. MATERIALS AND METHODS: Cardiomyopathy was induced in male Wistar rats with six intraperitoneal injections of 2.5 mg/kg doxorubicin over a period of two weeks. The effect of 28 days of celecoxib (100 mg/kg/day) and its combination with folic acid (10 mg/kg/day) was studied on doxorubicin-induced cardiomyopathy according to serum lactate dehydrogenase (LDH), creatine kinase (CK-MB), troponin-T (Tn-T), tumor necrosis factor alpha (TNF-α), cardiac thiobarbituric acid reactive substance (TBARS), and glutathione (GSH) levels as well as systolic blood pressure (SBP), heart rate (HR) and ultrastructural studies. RESULTS: Celecoxib cardiotoxicity was manifested by significant increases in the LDH, Tn-T, TNF-α, CK-MB, SBP, HR (p < 0.001) and TBARS (p < 0.01) levels and a significant decrease in the GSH (p < 0.05) level when used alone or administered with doxorubicin. However, the combination of folic acid with celecoxib caused a significant reversal of these parameters and reduced the cardiotoxicity of celecoxib that was aggravated by doxorubicin. The ultrastructural study also revealed myocardial protection with this combination. DISCUSSION AND CONCLUSION: Folic acid protects against the cardiotoxic effects of celecoxib, which are aggravated in the presence of doxorubicin. Folic acid may act as a useful adjunct in patients who are taking celecoxib.

In-vitro antimicrobial activity and identification of bioactive components using GC-MS of commercially available essential oils in Saudi Arabia.

This study was designed to evaluate antimicrobial activity and chemical composition of four different plant essential oils i.e. Ginger oil (GiO), Black seed oil (BSO), Oregano oil (OO) and Rose oil (RO) against different bacterial and fungal strains. Anti-microbial activities of selected essential oils were determined by the microbiological technique using Agar well diffusion assay. After in vitro study, most of the essential oils showed antimicrobial activity against all the selected pathogens. Among all the tested oils, GiO showed strong antimicrobial activity. GiO showed highest antimicrobial activity against Shigella (119.79%), Enteococcus hirae (110.61%) and Escherichia coli (106.02%), when compared with the tetracycline (50 µg/mL) activity. However, Antifungal activity of GiO was found to be present against Candida albicans and Aspergilluas flavus, when compared with clotrimazole (50 µg/mL) activity. Among all the selected bacteria, BSO showed maximum antimicrobial activity against the E. coli followed by Citrobacter freundii. Moreover, BSO had highest zone of inhibition against the C. ablicans (33.58%). OO indicated that, Shigella had the highest sensitivity (12.6 ± 0.58, 131.25%), followed by E. hirae (19.1 ± 0.61, 96.46%) and Salmonella typhi (15.2 ± 0.27, 83.06%) when compared with tetracycline activity. OO showed poor sensitivity against all the selected fungal strains. Furthermore, Gas Chromatography analysis revealed that, Gingerol (10.86%) was the chief chemical constituents found in GiO followed by α-Sesquiphellandrene (6.29%), Zingiberene (5.88%). While, BSO, OO and RO had higher percentage of p-Cymene (6.90%), Carvacrol (15.87%) and Citronellol (8.07%) respectively. The results exhibited that the essential oils used for this study was the richest source for antimicrobial activity which indicates the presence of broad spectrum antimicrobial compounds in these essential oils. Hence, essential oils and their components can be recommended for therapeutic purposes as source of an alternative medicine.

Influence of high-pressure homogenization, ultrasonication, and supercritical fluid on free astaxanthin extraction from ß-glucanase-treated Phaffia rhodozyma cells.

In this study astaxanthin production by Phaffia rhodozyma was enhanced by chemical mutation using ethyl methane sulfonate. The mutant produces a higher amount of astaxanthin than the wild yeast strain. In comparison to supercritical fluid technique, high-pressure homogenization is better for extracting astaxanthin from yeast cells. Ultrasonication of dimethyl sulfoxide, hexane, and acetone-treated cells yielded less astaxanthin than ß-glucanase enzyme-treated cells. The combination of ultrasonication with ß-glucanase enzyme is found to be the most efficient method of extraction among all the tested physical and chemical extraction methods. It gives a maximum yield of 435.71 ± 6.55 µg free astaxanthin per gram of yeast cell mass.

Ultrasound-assisted extraction of gymnemic acids from Gymnema sylvestre leaves and its effect on insulin-producing RINm-5 F ß cell lines.

INTRODUCTION: Gymnema sylvestre is an important anti-diabetic medicinal plant, hence it is necessary to study the effective extraction of its active medicinal components. OBJECTIVE: To develop an efficient ultrasound-assisted extraction method for anti-diabetic gymnemic acids from Gymnema sylvestre leaves and measure their effect on insulin-producing RINm-5 F ß cells. METHODS: Box-Behnken's design and response surface methodology was applied to the ultrasound-assisted extraction of gymnemic acids from Gymnema sylvestre leaves. Analysis of gymnemic acids was carried out by high-performance thin-layer chromatography by converting total gymnemic acids into gymnemagenin by alkali hydrolysis. Effects of extracts on insulin production were tested on cultured, insulin-producing RINm-5 F ß cell lines. RESULTS: The point prediction tool of the design expert software predicted 397.9 mg gymnemic acids per gram of the defatted G. sylvestre leaves using ultrasound-assisted extraction, with ethanol at 60 °C for 30 min. The predicted condition shows 93.34% validity under experimental conditions. The ultrasound-assisted extract caused up to about four times more insulin production from RINm-5 F ß cells than extracts obtained from Soxhlet extraction. CONCLUSIONS: Response surface methodology was successfully used to improve the extraction of gymnemic acids from G. sylvestre leaves. The ultrasound-assisted extraction process may be a better alternative to prepare such herbal extracts because it saves time and may prevent excess degradation of the target analytes.

Effect of sequential bio-processing conditions on the content and composition of vitamin K2 and isoflavones in fermented soy food.

In the present research, effect of sequential addition of Bifidobacterium bifidum, Bacillus subtilis and Rhizopus oligosporus on content and composition of vitamin K2 and isoflavones in fermented soy foods have been investigated. Initially, soybeans were fermented with B. bifidum; then this fermented mass was re-fermented with co-culture of B. subtilis and R. oligosporus. The evolved sequence of microbes inoculation tended towards significantly (p < 0.5) higher enzymes levels (126.16 ± 2.23 IU/mg lipase, 36.52 ± 1.25 IU/mg phytase and 8.52 ± 1.12 IU/mg ß-glucosidase); maximum menaquinone-7 production (9.3 ± 1.27 µg/g); and isoflavone content (84.64 ± 1.97 % daidzein, 99.29 ± 0.86 % genistein, 96.42 ± 1.32 % glycitein) after 72 h of solid-state fermentation. The study showed that co-fermentation of soybean with different microbes in a particular sequence can enhance nutritional value batter than the mono-culture fermentation due to the positive correlation between enzymes (lipase, phytase, ß-glucosidase) levels, menaquinone-7 and soy isoflavones content.