Reformation of dairy effluent-a phycoremediation approach.

Microalgae are a unique renewable resource utilized since ages, serving as a reservoir for the production of various metabolites. In this study, dairy waste water (DWW) is used as the nutrient media for the cultivation of microalgae. This study focuses on the phycoremediation process of converting rich nutrients in the effluent into biomass and removing contaminants using microalgae. The specific growth rate reached the maximum of 0.55 day-1 in Desmococcus olivaceous, followed by 0.39 day-1 for Scenedesmus dimorphus, 0.23 day-1 in DCS (consortia composing all three strains in equal ratio), and lastly 0.22 day-1 in Chlorella vulgaris. The biomass productivity was 1.44 g L-1 day-1, 1.06 g L-1 day-1, 0.88 g L-1 day-1, and 0.65 g L-1 day-1 in D. olivaceous, S. dimorphus, C. vulgaris, and DCS, respectively. The COD and BOD removal percentage was 82.85% and 45.40% in D. olivaceous, 81.98% and 44.25% in C. vulgaris, 80.73% and 53.45% in S. dimorphus, and 80.10% and 43.10% in DCS, respectively. These results emphasize the promising role of algae in dairy effluent treatment, highlighting the effluent as a suitable medium for microalgae cultivation. It verifies the circular bio-economy concept where the treated wastewater is converted into value-added products.

Antiproliferative effect of Acacia nilotica (L.) leaf extract rich in ethyl gallate against human carcinoma cell line KB.

OBJECTIVES: The objective of this study is to analyze the antiproliferative activity of Acacia nilotica (L.) leaf ethanolic extract against cancer KB cells and to determine the mode of cancer cytotoxicity. MATERIALS AND METHODS: In this study, high-performance liquid chromatography and liquid chromatography-mass spectrometry analysis were done to confirm the presence of ethyl gallate as a major bioactive phenolic in the leaf ethanolic extract of A. nilotica, further dose-dependent (0-120 µg/mL) antiproliferative effect was investigated in human carcinoma cell line KB. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, reactive oxygen species, mitochondrial membrane potential loss, DNA damage, and apoptosis were evaluated. RESULTS: A. nilotica leaf ethanolic extract (ANLEE) showed effective concentration (EC50) of 40 µg/mL. Interference of growth was significantly (P < 0.05) high in KB cells treated with ANLEE when compared to untreated control, but less when compared to the reference drug paclitaxel. In addition, the in vivo acute toxicity study demonstrated the safe limit of administration of 2000 mg/kg body weight ANLEE by the histological analysis in rats. The results from the present study indicate that mitochondria and DNA of KB cells are severely affected leading to apoptosis. CONCLUSIONS: ANLEE is a prospective source for cancer therapy and therefore should be highlighted to explore on its wide range of safety in rats and efficacy against human carcinoma cell line KB.

Development of microsatellite markers for the resin-yielding, non-timber forest product species Boswellia serrata (Burseraceae).

PREMISE OF THE STUDY: Boswellia serrata (Burseraceae) is an economically important aromatic, gum-resin-yielding, non-timber forest tree species. Microsatellite markers were developed for B. serrata for the first time to study genetic diversity and population structure. METHODS AND RESULTS: A magnetic bead enrichment method was used to develop 16 microsatellite markers, of which 11 were polymorphic. The number of alleles per locus in the 60 individuals studied ranged from three to 10, and the levels of observed and expected heterozygosity ranged from 0.50 to 0.90 and 0.666 to 0.861, respectively. The primers successfully amplified in the congeneric species B. ovalifoliolata. CONCLUSIONS: These microsatellite markers can be used to study the genetic variation and population structure of B. serrata and to provide crucial information on population and ecological issues for management and conservation of the species.

Amelioration of methylmercury induced neural damage by essential oil of Selinum vaginatum (Edgew) C. B. Clarke.

Methylmercury (MeHg), an organometallic contaminant is a well spotted cause for a series of disorders, especially in the central nervous system. As there is no proper treatment, Selinum vaginatum (Edgew) C. B. Clarke, a traditional medicinal plant, is taken in the present study for assessing its neuroprotective effect against MeHg induced toxicity using rat brain mitochondrial fractions. The results of 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide dye (MTT) assay indicated that there was a reduction in the mitochondrial viability in MeHg treated sample and IC50 value recorded was 2.5µg/ml. Biochemical analysis showed that there was a significant inhibition of glutathione levels (GSH) and catalase activity and an elevation of thiobarbituric acid reactive substances (TBARS) levels in MeHg treated sample. These changes were prevented by co-incubation with essential oil extracted from Selinum vaginatum. The GSH reduction caused by MeHg is restored by essential oil, endorsing its chelating effect, an important molecular mechanism of defense against oxidative injury. Some of the major compounds are detected in Gas chromatography-mass spectrometry (GC-MS) analysis of essential oil, which could be accountable for its neuroprotection against MeHg.

Evaluation of ethyl gallate for its antioxidant and anticancer properties against chemical-induced tongue carcinogenesis in mice.

Cancer arising in the oral cavity is one of the major causes of mortality worldwide and demands immediate attention. Regardless of the use of advanced treatment for oral cancer, successful treatment resulting in cancer survival is low. Currently available drugs are ineffective and are toxic. Therefore, successful treatment without toxic effects remains essential. This is quite challenging, leading to the identification of natural bioactive compounds for oral cancer treatment. Thus, a plant extract rich in phenolics is preferred for studying the cellular, biochemical and molecular changes associated with oral carcinogenesis.The present study aims to deal with the above need using Acacia nilotica (L.) leaf extract (AN) and ethyl gallate (EG), a phenolic compound present in AN against oral carcinogenesis. Extension of a tumor cell line to a mouse model was investigated with 4-nitroquinoline 1-oxide (4-NQO) as carcinogen, a surrogate for tobacco. The progression of squamous cell carcinoma (SCC) was achieved through hyperplasia and dysplasia after 4-NQO induction in Swiss albino mice. Administration of AN and EG to animals undergoing dysplasia led to the inhibition of SCC, thereby reducing the tumor burden. The antioxidant capacity of AN and EG was also brought out via biochemical analysis. Further investigation of biomarkers in tongue tissues revealed the involvement of apoptosis in vivo Moreover, no adverse or toxic effect was observed earlier in rats upon oral administration of AN and EG. Thus, AN and EG shows strong hope as drugs against oral cancer progression.

Alleviation of 4-nitroquinoline 1-oxide induced oxidative stress by Oroxylum indicum (L.) leaf extract in albino Wistar rats.

BACKGROUND: 4-nitroquinoline 1-oxide (4-NQO) is a mutagen known to be responsible for causing cancer by generating oxidative stress in humans. Oroxylum indicum (L.) possesses various bioactive compounds with antioxidant properties. In this connection, the present study aims to analyze the alleviation of 4-NQO induced oxidative stress in albino Wistar rats using O. indicum (L.) leaf extract. METHODS: O. indicum (L.) belonging to the family Bignoniaceae, has anticancer and anti-inflammatory properties. In this study, we observed severe oxidative stress in 4-NQO induced albino Wistar rats when compared to untreated control. Alleviation of this condition was seen after the oral administration of O. indicum (L.) leaf extract at 50, 100, and 200 mg/kg body weight. RESULTS: 4-NQO (50 ppm) administration in drinking water resulted in the generation of reactive oxygen species (ROS) leading to cellular damage, lipid peroxidation and imbalance in antioxidant status. Administration of O. indicum (L.) leaf extract has alleviated the level of 4-NQO induced oxidative stress by increasing the antioxidant status and decreasing the elevation of liver markers in serum. CONCLUSIONS: Results clearly suggest that O. indicum (L.) leaf extract when administered orally in a dose dependent manner has the ability to overcome the oxidative stress induced by 4-NQO with hepatoprotective and lipid protective properties.

Neuroprotective effect of Tagara, an Ayurvedic drug against methyl mercury induced oxidative stress using rat brain mitochondrial fractions.

BACKGROUND: Methyl mercury (MeHg), an important environmental toxicant is implicated in neurological disorders such as Hunter-Russell syndrome and Autism. Therefore, the present work is in search of new drugs that can alleviate MeHg toxicity. In this connection, Tagara, an ayurvedic drug is used for assessing its neuro protective effect against MeHg toxicity. METHODS: In the present study, we assessed the phytochemical contents of Tagara by colorimetric and HPLC analyses. The neuroprotective effect of Tagara on MeHg induced neurotoxicity was measured in terms of viability by MTT assay and oxidative stress in terms of catalase activity, glutathione and thiobarbituric acid reactive substance levels. Further, the chelating effect of Tagara towards MeHg was performed to identify the molecular mechanism. Statistical analysis was done by statistical package for social sciences (SPSS) version 16.0. RESULTS: The results demonstrated that Tagara contains significant amounts of phenols and flavonoids. Also, HPLC analysis of Tagara revealed the presence of essential oils such as hydroxyvalerenic and valerenic acids. Our results demonstrated that exposure of rat brain mitochondrial fractions to MeHg resulted in a dose dependent death in MTT assay and IC50 value was found to be 10 µM. However, a 250 µg dose of Tagara effectively prevented MeHg induced mitochondrial damage. The oxidative stress caused by MeHg results in elevated levels of reactive oxygen species as evidenced by elevated TBARS (Thiobarbituric acid-reactive substances) levels and diminished catalase enzyme activity and glutathione content. However, Tagara at 250 µg concentration offsets these alterations caused by MeHg. Further, Tagara also diminished GSH oxidation caused by MeHg, confirming its chelating effect, one of the molecular mechanisms that triggers protection against oxidative damage. CONCLUSION: Our results revealed that MeHg induced toxicity is predominantly mediated through oxidative stress mechanism and the propensity of Tagara to abolish such reactions. Hence, we propose that Tagara with a source of potential neuroprotectants may be a useful approach to alleviate MeHg associated neurotoxicity.

In vitro evaluation of antiproliferative effect of ethyl gallate against human oral squamous carcinoma cell line KB.

Although some polyphenols are known to possess anticancer activity against different cancer cell lines through induction of apoptosis, the mode of antiproliferative effect of ethyl gallate against human oral squamous carcinoma cell line KB was not studied until now. Therefore, the antiproliferative effect of ethyl gallate was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay in comparison with the reference drug paclitaxel. Generation of reactive oxygen species, mitochondrial membrane potential loss, DNA damage and apoptosis were determined using 2,7-diacetyldichlorofluorescein fluorescence, uptake of rhodamine-123 by mitochondria, comet assay and acridine orange/ethidium bromide dual-dye staining method. Both ethyl gallate and paclitaxel exhibited cytotoxicity in a dose-dependent manner. The 50% inhibitory concentration for ethyl gallate was 30 and 20 µg/mL for paclitaxel. A volume of 50 µg/mL of ethyl gallate was found to be significantly effective (P < 0.05) in controlling the cancer cell proliferation leading to acute apoptosis.

Neuroprotective effect of Brahmi, an ayurvedic drug against oxidative stress induced by methyl mercury toxicity in rat brain mitochondrial-enriched fractions.

The present study evaluates the neuroprotective effect of Brahmi against methyl mercury (MeHg) toxicity. The results demonstrated that MeHg-decreased mitochondrial viability in MTT assay and IC50 value was found to be 2.5 µg/mL. However, Brahmi at 250 µg/mL concentration effectively prevented mitochondrial damage caused by MeHg in MTT assay. Our results also demonstrated MeHg significantly inhibited catalase enzyme activity, glutathione content and increased the level of thiobarbituric acid reactive substances in mitochondrial-enriched fractions of rat brain. These alterations were prevented by preincubation with Brahmi. In addition, Brahmi reverted glutathione level to normal that was depleted by MeHg, confirming its chelating effect, one of the molecular mechanisms that underlie protection against oxidative damage. Our study also focused on total phenolic and flavonoid contents of Brahmi, and it was found to contain significant amount of phenols and flavonoids. The presence of saponins was detected by HPLC which might be responsible for neuroprotection against MeHg.

In vitro protection of biological macromolecules against oxidative stress and in vivo toxicity evaluation of Acacia nilotica (L.) and ethyl gallate in rats.

BACKGROUND: Recently, enormous research has been focused on natural bioactive compounds possessing potential antioxidant and anticancer properties using cell lines and animal models. Acacia nilotica (L.) is widely distributed in Asia, Africa, Australia and Kenya. The plant is traditionally used to treat mouth, ear and bone cancer. However, reports on Acacia nilotica (L.) Wild. Ex. Delile subsp. indica (Benth.) Brenan regarding its toxicity profile is limited. Hence in this study, we investigated the antioxidant capacity and acute toxicity of ethyl gallate, a phenolic antioxidant present in the A. nilotica (L.) leaf extract. METHODS: The antioxidant activity of ethyl gallate against Fenton's system (Fe3+/H2O2/ascorbic acid) generated oxidative damage to pBR322 DNA and BSA was investigated. We also studied the interaction of ethyl gallate to CT-DNA by wave scan and FTIR analysis. The amount of ethyl gallate present in the A. nilotica (L.) leaf extract was calculated using HPLC and represented in gram equivalence of ethyl gallate. The acute toxicity profile of ethyl gallate in the A. nilotica (L.) leaf extract was analyzed in albino Wistar rats. Measurement of liver and kidney function markers, total proteins and glucose were determined in the serum. Statistical analysis was done using statistical package for social sciences (SPSS) tool version 16.0. RESULTS: Ethyl gallate was found to be effective at 100 µg/mL concentration by inhibiting the free radical mediated damage to BSA and pBR322 DNA. We also found that the interaction of ethyl gallate and A. nilotica (L.) leaf extract to CT-DNA occurs through intercalation. One gram of A. nilotica (L.) leaf extract was found to be equivalent to 20 mg of ethyl gallate through HPLC analysis. Based on the acute toxicity results, A. nilotica (L.) leaf extract and ethyl gallate as well was found to be non-toxic and safe. CONCLUSIONS: Results revealed no mortality or abnormal biochemical changes in vivo and the protective effect of A. nilotica (L.) leaf extract and ethyl gallate on DNA and protein against oxidative stress in vitro. Hence, A. nilotica (L.) leaf extract or ethyl gallate could be used as potential antioxidants with safe therapeutic application in cancer chemotherapy.