Maximum Tolerable Concentration Research Articles

Assessment of the Impact of Polycyclic Derivatives of the Frame Series on the Replicative Properties of the SARS-CoV-2 Virus in an <I>in vitro</I> Experiment

The aim of the work was to determine the cytotoxicity and the influence of polycyclic derivatives of the framework series on the replicative properties of the SARS-CoV-2 virus in Vero-E6 cell culture in vitro. Materials and methods. The virus inhibiting effect of 50 adamantane and bicyclo[3.3.1]nonane derivatives with carbocyclic and heterocyclic substituents was investigated. The studies were carried out on Vero-E6 cell culture by assessing the cytopathic effect of the virus. The impact of the compounds on the replicative properties of the SARS-CoV-2 virus was estimated by the decrease in virus titer in the presence of the compounds compared to the control. Based on the virus titer values in the presence of a series of successively decreasing concentrations of the compound, the 50 % effective concentration was calculated.Results and discussion. A study of polycyclic derivatives of the framework series has identified two compounds with antiviral properties against the SARS-CoV-2 virus. Among bicyclo[3.3.1]nonane derivatives containing heterocyclic fragments, compound No. 15144 has showed an inhibitory effect against the SARS-CoV-2 virus. The protective effect of the compound was manifested in maximum tolerable concentration (MTC) (70.0 μg/ml) and ½ MTC (35.0 µg/ml). A decrease in virus titers under the influence of MTC by 0.95 lg TCD50/ml, in ½ MTC (35.0 μg/ml) – by 0.35 lg TCID50/ml has been detected. The effective concentration (EC50) value of the compound No. 15144 was 64.0 μg/ml, the MTC/EC50 ratio was 1.09. Compound No. 14838 (adamantane derivative containing carbocyclic fragments) had less pronounced antiviral activity. As a result of research, it has been established that sample No. 14838 at a dose of MTC (45.0 μg/ml) reduces the infectious titer by 0.78 lg TCD50/ml, in ½ MTC (22.5 μg/ml) by 0.15 lg TCD50/ml compared to the control. The EC50 value of compound No. 14838 was 37.0 μg/ml, the MTC/EC50 ratio was 1.22.

Comparative Study on Phytochemical Composition, Antioxidant, and Anti-HSV-2 Activities of Sambucus nigra L. and Sambucus ebulus L. Extracts

Sambucus nigra (SN) and Sambucus ebulus (SE) are widely used in folk medicine, primarily as antiviral agents for colds and influenza. In the current study, the antiviral activity of extracts of SN and SE fruits, flowers, and leaves were tested against herpes simplex virus type-2 (HSV-2). The HPLC analysis of the investigated extracts revealed the presence of phenolic acids, flavonoids, and anthocyanins. Rutin and chlorogenic acid were the main polyphenol constituents in flower and leaf extracts, whereas anthocyanins were predominant in fruit extracts. The flower extract of SN was characterized by the highest content of rutin and chlorogenic acid—14,232.1 mg/100 g dry weight (DW) and 7086.7 mg/100 g DW, respectively. SN fruit extract revealed the highest antioxidant activity measured using ORAC and HORAC methods—11,443.1 μmol TE/g and 8198.9 μmol GAE/g, respectively. To evaluate cytotoxicity, antiviral, and virucidal activities against HSV-2, the MTT assay and method of Reed and Muench were used. The least toxic extracts were PSNFrE and PSEFrE. The maximum tolerable concentration (MTC) of PSNFrE was 2000 μg/mL and the calculated CC50 value for that extract was 3570 μg/mL. The inhibitory activity against virus replication was established for three of the extracts—PSNFlE, PSNLE, and PSNFrE. PSEFrE showed neither activity against virus replication, nor virucidal activity. The data suggest a significant inactivation of more than 98% after 60 min of contact of HSV-2 virions with the PSNFrE applied in MTC. The current study provides evidence that Sambucus nigra reveals anti-HSV-2 activity; however, the most active parts of the species were fruits. Therefore, SN fruits and their extracts can be used as an attendant therapy for HSV-2 viral infections.

Effect of a Total Extract and Saponins from Astragalus glycyphyllos L. on Human Coronavirus Replication In Vitro.

Members of the family Coronaviridae cause diseases in mammals, birds, and wildlife (bats), some of which may be transmissible to humans or specific to humans. In the human population, they can cause a wide range of diseases, mainly affecting the respiratory and digestive systems. In the scientific databases, there are huge numbers of research articles about the antiviral, antifungal, antibacterial, antiviral, and anthelmintic activities of medicinal herbs and crops with different ethnobotanical backgrounds. The subject of our research is the antiviral effect of isolated saponins, a purified saponin mixture, and a methanol extract of Astragalus glycyphyllos L. In the studies conducted for the cytotoxic effect of the substances, CC50 (cytotoxic concentration 50) and MTC (maximum tolerable concentration) were determined by the colorimetric method (MTT assay). The virus was cultured in the MDBK cell line. As a result of the experiments carried out on the influence of substances on viral replication (using MTT-based colorimetric assay for detection of human Coronavirus replication inhibition), it was found that the extract and the purified saponin mixture inhibited 100% viral replication. The calculated selective indices are about 13 and 18, respectively. The obtained results make them promising for a preparation with anti-Coronavirus action.

Research on the antiviral activity of water-soluble melanin from the pharmaceutical chaga mushroom (Inonotus obliquus) against influenza А virus subtypes H5N1, H3N2 and H1N1pdm09 in experiments in vitro

Introduction. Influenza A virus is the cause of epidemics and pandemics that severely affect the health and socioeconomic status of the world's population. The need to develop new methods of etiotropic therapy, and the increasing ability of viruses to counteract the antiviral drugs makes extremely relevant the search for new pharmacologically active substances and the subsequent study of their medicinal properties.
 The aim of the study is to conduct research into the antiviral properties of melanin obtained from the pharmaceutical chaga mushroom in relation to different subtypes of the influenza A virus.
 Materials and methods. A sample of water-soluble melanin from Inonotus obliquus obtained by alkaline hydrolysis and dried at 40C was tested for toxicity and antiviral activity. The commercial anti-influenza drug Tamiflu was used as a reference drug. Statistical processing of the obtained data was carried out according to the Spearman-Kerber method.
 Results. Inonotus obliquus melanin (sample 20-24) toxicity markers, such as a maximum tolerable concentration (MTC) of 237.0 g/mL, and a 50% cytotoxic concentration (CC50) of 153,45 g/mL were established for MDCK cell culture. The assessment of antiviral activity of test sample against three subtypes of the influenza A virus (H5N1, H3N2 and H1N1pdm09) demonstrated a decrease in the infectivity of the influenza virus by 2.53.5 lg with 50% virus-inhibiting concentrations (IC50) of 1.559.52 g/mL. Based on the obtained values of CC50 and IC50, the selectivity indices (SI) of the sample were calculated, characterizing its prospects for further research.
 Conclusions. Melanin obtained from the pharmaceutical chaga mushroom showed the highest activity against the strain of the human pandemic influenza virus A/California/04/2009 (H1N1)pdm09, caused a decrease in its infectivity by 3.5 lg and had an IC50 of 1.6 g/ml. The obtained results indicate the prospects for creating an antiviral drug based on Inonotus obliquus melanins against the influenza virus.

Amoxicillin-Induced Neurotoxicity: Contribution of a Healthcare Data Warehouse to the Determination of a Toxic Concentration Threshold.

Amoxicillin (AMX)-induced neurotoxicity is well described and may be associated with AMX overexposure. No neurotoxic concentration threshold has been determined thus far. A better knowledge of maximum tolerable AMX concentrations is of importance to improve the safety of high doses of AMX. We conducted a retrospective study using the local hospital data warehouse EhOP® to generate a specific query related to AMX neurotoxicity symptomatology. All patient medical reports containing a mention of neurotoxicity clinical symptoms coupled with AMX plasma concentration measurements were explored. Patients were classified into two groups according to the imputability of AMX in the onset of their neurotoxicity, on the basis of chronological and semiological criteria. A receiver-operating characteristic curve was performed to identify an AMX neurotoxic steady-state concentration (Css) threshold. The query identified 101 patients among 2054 patients benefiting from AMX TDM. Patients received a median daily dose of 9 g AMX, with a median creatinine clearance of 51 mL/min. A total of 17 of the 101 patients exhibited neurotoxicity attributed to AMX. The mean Css was higher for patients with neurotoxicity attributed to AMX (118 ± 62 mg/L) than those without 74 ± 48 mg/L (p = 0.002). A threshold AMX concentration of 109.7 mg/L predicted the occurrence of neurotoxicity. This study identified, for the first time, an AMX Css threshold of 109.7 mg/L associated with an excess risk of neurotoxicity. This approach needs to be confirmed by a prospective study with systematic neurological evaluation and TDM.

Effect of mercury in the influx and efflux of nutrients in the microalga Desmodesmus armatus

Anthropogenic activities such as mining and the metallurgical industry are the main sources of mercury contamination. Mercury is one of the most serious environmental problems in the world. This study aimed to investigate, using experimental kinetic data, the effect of different inorganic mercury (Hg2+) concentrations on the response of microalga Desmodesmus armatus stress. Cell growth, nutrients uptake and mercury ions from the extracellular medium, and oxygen production were determined. A Compartment Structured Model allowed elucidating the phenomena of transmembrane transport, including influx and efflux of nutrients, metal ions and bioadsorption of metal ions on the cell wall, which are difficult to determine experimentally. This model was able to explain two tolerance mechanisms against mercury, the first one was the adsorption of Hg2+ions onto the cell wall and the second was the efflux of mercury ions. The model predicted a competition between internalization and adsorption with a maximum tolerable concentration of 5.29 mg/L of HgCl2. The kinetic data and the model showed that mercury causes physiological changes in the cell, which allow the microalga to adapt to these new conditions to counteract the toxic effects. For this reason, D. armatus can be considered as a Hg-tolerant microalga. This tolerance capacity is associated with the activation of the efflux as a detoxification mechanism that facilitates the maintenance of the osmotic balance for all the modeled chemical species. Furthermore, the accumulation of mercury in the cell membrane suggests the presence of thiol groups associated with its internalization, leading to the conclusion that metabolically active tolerance mechanisms are dominant over passive ones.

Determination of bioavailable arsenic threshold and validation of modeled permissible total arsenic in paddy soil using machine learning.

Minimizing arsenic intake from food consumption is a key aspect of the public health response in arsenic (As)-contaminated regions. In many of these regions, rice is the predominant staple food. Here, we present a validated maximum allowable concentration of total As in paddy soil and provide the first derivation of a maximum allowable soil concentration for bioavailable As. We have previously used meta-analysis to predict the maximum allowable total As in soil based on decision tree (DT) and logistic regression (LR) models. The models were defined using the maximum tolerable concentration (MTC) of As in rice grains as per the codex recommendation. In the present study, we validated these models using three test data sets derived from purposely collected field data. The DT model performed better than the LR in terms of accuracy and Matthews correlation coefficient (MCC). Therefore, the DT estimated maximum allowable total As in paddy soil of 14mg kg-1 could confidently be used as an appropriate guideline value. We further used the purposely collected field data to predict the concentration of bioavailable As in the paddy soil with the help of random forest (RF), gradient boosting machine (GBM), and LR models. The category of grain As (<MTC and>MTC) was considered as the dependent variable; bioavailable As (BAs), total As (TAs), pH, organic carbon (OC), available phosphorus (AvP), and available iron (AvFe) were the predictor variables. LR performed better than RF and GBM in terms of accuracy, sensitivity, specificity, kappa, precision, log loss, F1score, and MCC. From the better-performing LR model, bioavailable As (BAs), TAs, AvFe, and OC were significant variables for grain As. From the partial dependence plots (PDP) and individual conditional expectation (ICE) of the LR model, 5.70mg kg-1 was estimated to be the limit for BAs in soil.

EVALUATION OF GROUND WATER QUALITY IN Al-BAIDH FIELD, HODIEDA CITY, YEMEN

Ground water is the main source for drinking water in Hodieda city, Yemen. This study discusses the chemical and physical characteristics of nineteen ground water wells in the Al-baidh field. Thus, the measurement of water quality parameters such as electrical conductivity EC, total dissolved solids TDS, pH, turbidity, color, total alkalinity, total hardness TH , NO3-,SO42-, PO43-, Cl- ,Na+, K+, Ca2+ and Mg2+ in addition to Mn, Cu, Fe and Cr were expressed. The results indicate unacceptable&#x0D; levels of TDS , Cl-, Na+ , Ca2+ and Mg2+comparing with the maximum tolerable concentration given by World Health Organization (WHO) and Yemen standard (YS). Elsewhere, the measurement of SO42- and NO3- exceeded the allowable values given by WHO and Y.S for most the samples which can be attributed to the nature of the geological and chemical structure of soil in Albaidh field. In this study, measurement of the concentration for elements such as Cr, Fe, and Mn were carried out, the results indicate that the values for this elements were within the allowable ranges. In this paper the concepts of star- refinement and strongly star- refinement of covering are extended to fuzzy topological space in the sense of Chang, basic theorem for covering dimension of normal fuzzy topological space is proved . Also, the small inductive dimension function is extended to fuzzy topological space, and some results for this inductive dimension in Chang’s space are obtained.

Enhanced production of high-value polyunsaturated fatty acids (PUFAs) from potential thraustochytrid Aurantiochytrium sp.

Due to growing health concerns, the urban population is utterly inclined towards a healthy lifestyle and incorporated nutritional food supplements to lower common health risks. The ω-3 and ω-6 PUFAs consumption is increasing, hence alternative commercial production is essentially developed. The microbial source is an emerging platform to overcome the global demand for omega PUFAs. Marine oleaginous protist Aurantiochytrium sp. found a potential source to produce substantial DHA and SFA. The objective of the present research was to enhance the PUFA yield by optimizing maximum tolerable glucose concentration with a suitable nitrogen ratio (10:1). The maximum lipid and DHA yield and content were determined 4.30, 1.34 g/L, and 62.4, 33.49 % of total biomass and lipid at 30 g/L glucose respectively, which is one of among highest reported, however relative PUFA was maximum 46.97 % (DHA) in total lipid at 10 g/L glucose. Remaining 42–53.6 % SFA could be used for biodiesel.

Bacterial bio-mobilization and -sequestration of arsenic in contaminated paddy fields of West Bengal, India

One hundred and forty nine bacterial strains were isolated from arsenic contaminated paddy field soil of West Bengal following enrichment upon As(III), As(V) and control plates. The maximum tolerable concentration (MTC) of the bacterial population isolated towards As(V) and As(III) was found to be 600 mM and 10 mM, respectively. Eight isolates with varying MTC values were chosen to study their As ad-/ab-sorption abilities to form a consortium for bioremedial application to As contaminated soil. The As(III) or As(V) ab-/ad-sorption abilities of the chosen isolates positively correlated to their respective As(III) or As(V) MTC values. Phylogenetic analyses of the isolates affiliate them to be the members of genera Bacillus, Paenibacillus, Staphylococcus and Ralstonia. A consortium comprising the eight chosen isolates were incubated under eight microcosm conditions with combinations of flooded or non-flooded, sterile or non-sterile soils with low (6.1 mg/kg) or moderately high (19.5 mg/kg) As concentrations. The consortium revealed an overall appreciable activity of As mobilization and sequestration with best cellular As accumulation under non-flooded low As concentration soil incubation. While the microcosm with highly contaminated soil allowed best microbial As sequestration under flooded condition when incubated solely in sterile soil. Bacterial sequestration and removal of the toxic metalloid may find important implications in As bioremediation in agricultural fields hence, reducing the bioavailability of this toxic element for further translocation into the crop. The study, thereby, provides a novel insight for potential use of the consortia in bioremediating As contaminated soil through microbial biosequestration approach.

Residual formaldehyde contents in fresh white cheese in El Salvador: Seasonal changes associated with temperature.

Detection of residual formaldehyde (FA) in dairy products could be explained by direct addition of this preservative to extend the shelf life of raw material or final product at room temperature. FA is not authorized as a preservative by international standards and its addition to dairy products is prohibited due to its potentially harmful effects on consumers. Although the carcinogenicity of FA by oral exposure has not been proven, it is also known it cause histopathological and cytogenic changes in tissues at first contact, so its toxicity by ingestion should not be underestimated. This research determined both residual FA levels in locally produced fresh white cheese and its variation according to the seasons of the year and its association with ambient temperature. None of the FA levels quantified in cheese exceeded the maximum tolerable concentration (2.6mg/kg) and although average FA contents did not vary significantly with seasonal changes (0.093-0.181mg/kg), the number of positive cases did, since the highest prevalence occurred in the dry (60.9 %) and transitional dry-rainy (79.7 %) seasons of 2021, which are characterized by having the highest average ambient temperatures (27.5°C and 28.3°C, respectively). It was also shown that 79.6 % of the variability of FA-positive samples is explained by changes in the average temperature according to the year´s season. The association between these variables and quantified levels of aldehyde in raw milk sampled at the plant could indicate that FA was used to prevent milk and/or the final product from decomposing due to the effect of high ambient temperature. In addition, residual FA contents decreased in both milk and cheese, depending on added preservative levels, and the time elapsed prior to analysis.

Crocetin ameliorates non-alcoholic fatty liver disease by modulating mitochondrial dysfunction in L02 cells and zebrafish model

Ethnopharmacological relevanceTraditional Chinese medicine considers that the etiology and pathogenesis of non-alcoholic fatty liver disease (NAFLD) are related to liver depression and qi stagnation. Saffron and its active ingredient, crocetin (CCT), are used for the treatment of metabolic diseases owing to their “Liver deobstruent” and “Liver tonic” effects. However, the effect of CCT on NAFLD has not been fully elucidated. In the present study, the effect and potential molecular mechanism of CCT were explored in both in vivo and in vitro models of NAFLD. Materials and methodsCCT was isolated from saffron and purity and structure characterization were performed using HPLC, MS, 1H-NMR, and 13C-NMR. The effect of CCT on the viability of L02 cells and its maximum tolerable concentration (MTC) in zebrafish were investigated. Free fatty acids (FFA) and thioacetamide (TAA) were used to induce lipid accumulation in L02 cells and steatosis in zebrafish, respectively. The effects of CCT on indexes related to lipid metabolism, oxidative stress, and mitochondrial function in NAFLD models were explored using biochemical assay kits, Western blot analysis, Reverse Transcription-Polymerase Chain Reaction (RT-PCR), histopathology analysis, and determination of mitochondrial membrane potential (ΔΨm). Morphological analysis of mitochondria was performed using transmission electron microscopy (TEM). ResultsThe levels of triglyceride (TG), total cholesterol (TC), malondialdehyde (MDA), and alanine/aspartate aminotransferases (ALT/AST) activities in FFA treated L02 cells were significantly reduced after CCT treatment. CCT treatment significantly increased ATP concentration, ΔΨm, and activities of superoxide dismutase (SOD), catalase (CAT), and cytochrome c oxidase (COX IV) in FFA treated L02 cells. TEM images showed restoration of mitochondrial morphology. CCT decreased ATP concentration and upregulated expression of B-cell lymphoma-2 (Bcl-2) and COX IV, whereas, CCT downregulated expression of BCL2-Associated X (Bax) and cleaved caspase-3 in TAA treated zebrafish. These findings indicated that mitochondrial dysfunction was alleviated after CCT treatment. Oil Red O staining of L02 cells and zebrafish showed that CCT treatment reversed the accumulation of lipid droplets. ConclusionIn summary, CCT treatment effectively alleviated the symptoms of NAFLD and restored mitochondrial function in L02 cells and zebrafish NAFLD model.

Bacterial Degradation of Bacteriostatic Polyphenols, Tannins

Tannin degradation by bacteria has not been studied much as tannins are commonly known to be bacteriostatic due to enzyme inhibition, substrate deprivation, and the enzyme activity on the bacterial cell wall. However, about a handful of bacteria have been found to tolerate certain concentrations of tannin. This study focuses on isolating and identifying bacteria from decaying portions of tree bark for tannase production and effective catalysis of ester bond hydrolysis in tannins. Different concentrations of commercial tannic acid were used as the sole carbon source on mineral salt medium (MSM) agar plates, to test the maximum tolerable concentrations (MTCs) by the isolates. Tannin degradation was confirmed by a visual reading method and bacterial tannase activity and the biodegradation percentage were determined. One particular isolate was identified to have 50 g/L MTC of tannin, with a tannase activity of 51.61 U/mL that is optimum after 96 hours of incubation. The 16s rRNA sequencing results showed that the isolate belonged to Bacillus genus and the resulting bacterial strain isolate was found to be a new strain of Bacillus subtilis which was submitted to GenBank under the accession number MH330408.

Systematic Evaluation of the Effects of Toxicant Exposure on Survival in Zebrafish Embryos and Larvae.

The use of whole animal models in toxicological studies is essential for understanding the physiological responses caused by chemical exposures. However, such studies can face reproducibility challenges due to unaccounted experimental parameters that can have a marked influence on toxicological outcomes. Zebrafish embryos and larvae are a popular vertebrate animal model for studying cellular, tissue, and organ responses to toxicant exposures. Despite the popularity of this system, standardized protocols that control for the influence of various experimental parameters and culture conditions on the toxicological response in these animals have not been widely adopted, making it difficult to compare findings from different laboratories. Here, we describe a detailed approach for designing and optimizing protocols to assess the impact of chemical exposures on the development and survival of zebrafish embryos and larvae. We first describe our standard procedure to determine two key toxicological thresholds, the maximum tolerable concentration (MTC) and the lethal concentration (LC50 , defined as that in which 50% of larvae die), in response to an exposure that persists from early development through larval maturation. We then describe two protocols to systematically test how key experimental parameters, including genetic background, culture media, animal density, volume, plate material, and developmental stage in which the embryos are exposed, alter the MTC and LC50 . Finally, we provide a step-by-step guide to assess the interaction between two chemicals using this model. These protocols will guide the standardization of toxicological studies using zebrafish and maximize reproducibility. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Zebrafish embryo collection and culture, and establishment of the MTC and LC50 Basic Protocol 2: Evaluation of the impact of culture conditions on toxicant responses of zebrafish embryo and larvae Basic Protocol 3: Identification of the developmental window of sensitivity to toxicant exposure Basic Protocol 4: Testing interaction between multiple toxicants.

Evaluation of heavy metal content in water and removal of metals using native isolated bacterial strains

Abstract. Saleh LIF, Rashed RO, Muhammed SM. 2021. Evaluation of heavy metal content in water and removal of metals using native isolated bacterial strains. Biodiversitas 22: 3163-3174. Natural surface water sources are susceptible to chemical contamination, including heavy metals. The goals of this study were to isolate naturally occurring heavy metal-tolerant bacteria by evaluating their ability to remove heavy metals from Tanjaro River and assessing the river’s water quality. The potency of bacteria to remove heavy metals (Cd, Pb, Cu, Cr, Ni, Zn, Co, and Fe) from the medium was conducted by inductively coupled plasma-optical emission spectrometry. Results of physicochemical parameters of the water samples were as follows: temperature 11.9–31°C, pH 6.1–8.64, total dissolved solids 268–464 mg.l-1, total hardness 232–485 mg.l-1, and alkalinity 122–324.3 mg.l-1. The overall mean concentrations of the heavy metals were in the order of Zn &gt;Cu&gt; Pb&gt; Ni&gt; Co&gt; Fe&gt; Cr&gt; Cd), with values of 0.086, 0.073, 0.71, 0.068, 0.051, 0.056, 0.031, and 0.024ppm, respectively Moreover, the concentrations were generally exceeded the water quality criteria of WHO in some observations. The isolates included both gram-negative (56.5%) and gram-positive (43.5%) bacteria. The isolates displayed different degrees of resistance to heavy metals with maximum tolerable concentrations (MTCs) ranging from (10-250) ppm. Bacillus safensis and Leucobacter chromiiresistens were respectively able to tolerate (80, 90 ppm) Cd, (250, 160 ppm) Pb, (210,100 ppm) Cr, (110, 90 ppm) Ni and (160, 170 ppm) Co, while, Proteus mirabilis could tolerate 90 ppm Cd. The isolates were able to remove 55.4%Pb, 53.1%Cr, 53.7%Ni, 47.7Fe, 61.4%Co, and 55.8% Cu from a medium supplemented with the mixture of these heavy metals. The finding of this study indicated that Tanjaro River was undergoing frequent changes in water quality. Native bacterial isolates possessed considerable tolerance to selected heavy metals, which provides promising candidates for use in bioremediation.

Use of ultraviolet-light mutagenesis to generate a mutant with elevated cadmium resistance, B. agri C15 CdR

Cadmium (Cd) is toxic to living organisms; however, bacteria are adaptable to severe conditions, including Cd contamination. Bacteria can develop in the natural environment, as they generate resistant strains that can be used to remove Cd, but getting such adaptive strains usually takes a long time. In this study, ultraviolet (UV) light mutagenesis was used to accelerate the strain-resistant ability of Brevibacillus agri C15 to generate CdR mutant with high resistance to Cd. Its maximum tolerable concentration (MTC) to Cd was 15.00 mM. The UV light mutagenesis process resulted in the generation of the mutant B. agri C15 CdR (MTC: 20.00 mM Cd). Morphological and biochemical tests showed that there were no major differences between B. agri C15 CdR and B. agri C15. This study provides a basis for developing microbial Cd resistance and facilitating the application of Cd remediation.

The isolation and identification of cadmium-resistant Brevibacillus agri C15

Eight Cd-resistant bacteria were isolated from the soil. Based on the resistance and specific growth rate, one isolate from these was chosen. The 16S rRNA (rrs) gene sequence was used to assert this isolate belongs to the genus of Brevibacillus and closely related to Brevibacillus agri (B.agri) DSM 6348T (AB112716). The isolate was termed B.agri C15. The specific growth yields and the Cd-dependent fall in specific growth yield of B.agri C15 was determined under different concentrations of Cd. The current study provides a basis for isolation Cd resistant bacteria with maximum tolerable concentration (MTC) 15 mM Cd from a contaminated soil 490±50 μmolal Cd that could be suitable for a new Cd-bioremediation process; therefore, further studies are needed to investigate this possibility.

Occurrence of carcinogenic illudane glycosides in drinking water wells

BackgroundPtaquiloside (PTA), caudatoside (CAU) and ptesculentoside (PTE) are carcinogenic illudane glycosides found in bracken ferns (Pteridium spp.) world-wide. The environmentally mobile PTA entails both acute and chronic toxicity. A comparable risk might be associated with the structurally similar CAU and PTE. It is of great health concern if these compounds are present in drinking water, however, it is currently unknown if these compounds can detected in wells in bracken-dominated regions. This study investigates the presence of PTA, CAU, PTE, and their corresponding hydrolysis products pterosins B (PtB), A (PtA) and G (PtG) in water wells in Denmark, Sweden and Spain. Water samples from a total of 77 deep groundwater wells (40–100 m) and shallow water wells (8–40 m) were collected and preserved in the field, pre-concentrated in the laboratory and analysed by liquid chromatography–mass spectrometry (LC–MS).ResultsDeep groundwater wells contained neither illudane glycosides nor their pterosins. However, seven private shallow wells contained at least one of the illudane glycosides and/or pterosins at concentrations up to 0.27 µg L−1 (PTA), 0.75 µg L−1 (CAU), 0.05 µg L−1 (PtB), 0.03 µg L−1 (PtA) and 0.28 µg L−1 (PtG). This is the first finding of illudane glycosides and pterosins in drinking water wells.ConclusionsDetected concentrations of illudane glycosides in some of investigated wells exceeded the suggested maximum tolerable concentrations of PTA, although they were used for drinking water purpose. Contaminated wells were shallow with neutral pH and lower electric conductivity compared to deep groundwater wells with no illudane glycosides nor pterosins.

&lt;i&gt;In Vitro&lt;/i&gt; Wound Healing Effect of Asiaticoside Extracted from &lt;i&gt;Centella asiatica&lt;/i&gt; (‘Pegaga’) on Human Gingival Fibroblast Cell Line

Asiaticoside is a bioactive compound found in the traditional plant Centella asiatica (Asiatic pennywort or ‘Pegaga’) generally used for wound healing applications. Numerous studies have discussed the potential benefits of asiaticoside on different human cells such as keratinocytes and dermal fibroblast cells in healing of wounds. However only very few studies have been conducted to investigate its healing effect on cells originated from human oral cavity. The present study aimed to determine the potential of asiaticoside on human gingival fibroblast cells. Cytotoxic activities of the compounds were assessed by MTT assay. The wound healing was examined by scratch assay. The effect of asiaticoside on Col1A1 gene expression was also analyzed using qRT-PCR. Col1A1 is known to play a crucial role in wound healing. The MTT assay result showed that the maximum tolerable concentration of asiaticoside was 0.25 mg/ml. The scratch assay revealed that asiaticoside significantly accelerated the wound healing compared to the negative control (P&lt;0.05). Moreover, the qRT-PCR demonstrated that asiaticoside markedly increased Col1A1 mRNA expression. These results proved asiaticoside as a potential candidate for wound healing agent in dentistry.

Characterization and bioremediation potential of native heavy-metal tolerant bacteria isolated from rat-hole coal mine environment.

Identification and characterization of endogenous and stress adapted bacterial species, from rat-hole coal mines in Meghalaya, amplify the ambit of bioremediation for eco-restoration. 52 native bacterial isolates, drawn from soil and water samples of these mines, were analysed for bioremediation potential, based on growth and metal tolerance parameters. 12 of these isolates were metal tolerant with Bacillus spp. being the most promising taxon. Three isolates, namely, Serratia marcescens KH-CC, Bacillus altitudinis KH-16F and Bacillus siamensis KH-12A, exhibited high Maximum Tolerable Concentration (MTC) against Fe (500ppm), Mn (830ppm) and Pb (1400ppm). B. siamensis showed highest Fe remediation with 48.34% removal capacity, while maximum removal for Mn and Pb was exhibited by Serratia marcescens at 72.5 and 83%, respectively. The growth profile of the isolates indicated their ability to survive under pH, temperature and salt stress conditions. In vitro growth kinetics studies of the isolates revealed their ability to decrease the acidity of growth media and improve alkalinity from an initial of pH 4.8-5.2 to an alkaline level of pH 8.5-9. These native bacteria, extracted from the stressed coal mine habitat, are potential germane applicants for rehabilitation and eco-restoration of ecologically degraded mine sites.