Ml Of Medium Research Articles

Xylitol and ethanol co-production from sugarcane bagasse and straw hemicellulosic hydrolysate supplemented with molasses

Co-production systems are a strategy used to diversify production chains and to enable more profitable processes. In addition, the valorization of the sugar-and-alcohol industry through by-product conversion into value-added products is environmentally friendly and enhances their competitiveness to other products available in the market. Thus, the aim of the current study is to evaluate xylitol and ethanol co-production by Candida tropicalis deriving from sugarcane by-products, such as bagasse and straw, supplemented with molasses. Sugarcane bagasse and straw hemicellulosic hydrolysate was obtained through dilute-acid hydrolysis with H2SO4; next, it was concentrated and detoxified with activated charcoal. Different commercial sucrose concentrations (5, 25 and 50 gL−1) were added to the bagasse and straw hemicellulosic hydrolysate in semi-defined medium. In addition, hydrolysate supplementation with sugarcane molasses was evaluated based on sucrose content (10 and 50 gL−1) adjustment. Batch fermentations were carried out in 125-mL Erlenmeyer flasks filled with 50 mL of medium, at 30 °C, 200 rpm, pH 5.5, for 48 h. Molasses supplementation adjusted to 50 gL−1 sucrose in sugarcane bagasse and straw hemicellulosic hydrolysate recorded the maximum xylitol (30.61 gL−1) and ethanol (47.97 gL−1) concentrations observed in the current study. Besides, volumetric xylitol (0.64 gL−1h−1) and ethanol (1.00 gL−1h−1) productivity was observed under the very same condition. Results have shown the likelihood of co-producing xylitol and ethanol through C. tropicalis by using sugarcane by-products to compose the fermentation medium as strategy to intensify and integrate these bioprocesses in sugarcane biorefineries.

Biomass of unconventional plants from Brazilian semiarid as substrate for hydrolytic enzymes production by Aspergillus niger under solid and submerged fermentation

Aspergillus niger KIJH was grown in solid and submerged fermentation using leaves and roots (with and without bark) of plants typically from Brazilian semiarid as substrate to produce a multienzymatic extract, which was characterised for its potential biotechnological applications. Solid-state fermentation (SSF) was applied to select the most promising plants biomass as induction substrates for the production of hydrolytic enzymes by fungus. The best biomasses were used as substrate in submerged fermentation (SmF) assays at two scales. Samples of up scale fermented culture were partially purified by ultrafiltration and activity and pH and temperature stability of CMCase and xylanase were evaluated. A. niger KIJH produced hydrolytic enzymes under SSF containing unconventional plants biomass from Brazilian semiarid. In SmF conditions, maximum CMCase (0.264 U mL-1) and xylanase (1.163 U mL-1) activities were induced by Jacaratia corumbensis. Scaling up the SmF to 500 mL of medium was able to maintain constant the production of CMCase (0.346 U mL-1) and xylanase (1.273 U mL-1) on the fermented culture. Ultrafiltered and concentrated extract presented CMCase activities practically constant in all temperature ranges (30-80°C) and pH (3.0-9.0), while xylanase optimum activity temperature was 50°C and pH in the range of 3.0 to 5.0. CMCase activity remained stable for 24 hours at 50°C and xylanase was reduced in 53% after two hours incubation at the same temperature. CMCase and xylanase obtained by A. niger KIJH cultivated in submerged culture containing J. corumbensis as carbon source may have application in biotechnology processes that require enzymes that remain active under routine extreme conditions.

PEMBUATAN BIOETANOL DARI ROTI SISA DENGAN PROSES HIDROLISA ENZIMATIS DAN FERMENTASI

Bread waste contains 57% carbohydrate, that can be used as the raw material of bioethanol production. This research aims was to study the producing bioethanol process from bread waste, and to determine the effect of catalyst concentration of enzyme glucoamylase (Aspergillus Niger). The bioethanol production with bread waste consisted of three stages: hydrolysis (liquification and sacharification), fermentation and distillation. Saccharification performed for 6 hours at 30oC and pH 4.5 using a variable catalyst enzyme glucoamylase (Aspergillus Niger), with 6, 8, 10, and 12 ml/200 ml of media. The glucose level value in sacharification process is obtained of 12.85 gr/ml, 137.236 gr/ml, 154.82 gr/ml, and 198.0327 gr/ml. The optimum value of glucose level occurs in the glucoamylase (Aspergillus Niger) enzyme addition of 12 ml. In separated hydrolysis and fermentation (SHF) process, there is a separation of glucose formation from starch (hydrolysis) and glucose reduction into bioethanol (fermentation). The enzyme catalyst (Aspergillus Niger) with 6, 8, 10, and 12 ml/200 ml media, it is obtained the bioethanol reduction value respective conditions of 14.1261%, 18.4909%, 24.7592%, and 28.6993%. The optimum condition is obtained in the glucoamylase (Aspergillus Niger) enzyme addition of 12 ml.

Development of an optimized and scalable method for isolation of umbilical cord blood-derived small extracellular vesicles for future clinical use.

Extracellular vesicles (EV) are a promising therapeutic tool in regenerative medicine. These particles were shown to accelerate wound healing, through delivery of regenerative mediators, such as microRNAs. Herein we describe an optimized and upscalable process for the isolation of EV smaller than 200 nm (sEV), secreted by umbilical cord blood mononuclear cells (UCB‐MNC) under ischemic conditions and propose quality control thresholds for the isolated vesicles, based on the thorough characterization of their protein, lipid and RNA content. Ultrafiltration and size exclusion chromatography (UF/SEC) optimized methodology proved superior to traditional ultracentrifugation (UC), regarding production time, standardization, scalability, and vesicle yield. Using UF/SEC, we were able to recover approximately 400 times more sEV per mL of media than with UC, and upscaling this process further increases EV yield by about 3‐fold. UF/SEC‐isolated sEV display many of the sEV/exosomes classical markers and are enriched in molecules with anti‐inflammatory and regenerative capacity, such as hemopexin and miR‐150. Accordingly, treatment with sEV promotes angiogenesis and extracellular matrix remodeling, in vitro. In vivo, UCB‐MNC‐sEV significantly accelerate skin regeneration in a mouse model of delayed wound healing. The proposed isolation protocol constitutes a significant improvement compared to UC, the gold‐standard in the field. Isolated sEV maintain their regenerative properties, whereas downstream contaminants are minimized. The use of UF/SEC allows for the standardization and upscalability required for mass production of sEV to be used in a clinical setting.

Assessment of intraocular lens/capsular bag biomechanical interactions following cataract surgery in a human in vitro graded culture capsular bag model

Intraocular lenses (IOLs) are implanted during cataract surgery. For optimum results, stable positioning of the IOL in the capsular bag is important. Wound-healing events following cataract surgery lead to modification of the capsular bag and secondary visual loss due to posterior capsule opacification. At present, it is unclear how these biological events can affect stability of the IOL within the capsular bag. In the present study, a human in vitro graded culture capsular bag model was the experimental system. Capsulorhexis and lens extraction performed on human donor eyes generated suspended capsular bags (5 match-paired experiments). Preparations were secured by pinning the ciliary body to a silicone ring and maintained in 6 mL of medium for 84 days using a graded culture system: days 1–3, 5% human serum and 10 ng/mL transforming growth factor β (TGFβ2); days 4–7, 2% human serum and 1 ng/mL TGFβ2; days 8–14, 1% human serum and 0.1 ng/mL TGFβ2; days 15–84, serum-free Eagle's minimum essential medium (EMEM). A CT LUCIA 611PY IOL was implanted in all preparations. Quantitative measures were determined from whole bag images captured weekly. Images were registered using FIJI and analysed in ImageJ to determine capsular bag area; distortion; angle of contact; haptic stability; capsulorhexis area; and a fusion footprint associated with connection between the anterior and posterior capsules. Cell coverage and light scatter were quantified at end-point. The transdifferentiation marker, α-SMA was assessed by immunocytochemistry. Immediately following surgery, distortion of the capsular bag was evident, such that a long axis is generated between haptics relative to the non-haptic regions (short axis). The angle of contact between the haptics and the peripheral bag appeared inversely correlated to capsular bag area. Growth on the peripheral posterior capsule was observed 1 week after surgery and beneath the IOL within 1 month. As coverage of the posterior capsule progressed this was associated with matrix contraction/wrinkles of both the central posterior capsule and peripheral capsular bag. Cells on the central posterior capsule expressed αSMA. Fusion footprints formed in non-haptic regions of the peripheral bag and progressively increased over the culture period. Within and at the edge of the fusion footprint, refractive structures resembling lens fibre cells and Elschnig's pearls were observed. Cell attachment to the IOL was limited. An impression in the posterior capsule associated with the CT LUCIA 611PY optic edge was evident; cell density was much greater peripheral to this indent. Wound-healing events following surgery reduced capsular bag area. This was associated with the long/short axis ratio and angle of contact increasing with time. In summary, we have developed a human capsular bag model that exhibits features of fibrotic and regenerative PCO. The model permits biomechanical information to be obtained that enables better understanding of IOL characteristics in a clinically relevant biological system. Throughout culture the CT LUCIA 611PY appeared stable in its position and capsular bag modifications did not change this. We propose that the CT LUCIA 611PY optic edge shows an enhanced barrier function, which is likely to provide better PCO management in patients.

Anti-cholesterol and Antioxidant Activities of Independent and Combined Microalgae Aqueous Extracts In Vitro

Algae are used as one of important medical sources for its therapeutic properties; and edible algae are acknowledged as complete foods which provide right balance of proteins, carbohydrates, vitamins, mineral salts, carotenoids, and antioxidants. The aim of the current study was to evaluate hot-water extracts of different marine microalgae species (Spirulina platensis, Anabaena oryzae, Scenedesmus obliquus and Chlorella vulgaris) in vitro for their cholesterol-lowering activity, individually or in combinations. The algal extracts (AEs) were prepared from different marine microalgae species and their in vitro anti-cholesterol activities were assessed spectrophotometrically. The algal growth and their bioactive constituents (polyphenol content, antioxidant capacity and tannin content) were quantified using Folin-Ciocalteu, DPPH, vanillin-HCl assays, respectively. The highest value of the biomass concentration (1.367 g) was recorded for S. platensis in 1000 mL of culture media after 21 days. Increasing anti-cholesterol activity by S. platensis extract was observed up to 60 min at a concentration of 15 mg/mL and a highest inhibition was found as 76.01% value (p < 0.05). The combined extracts from S. platensis with C. vulgaris showed maximum cholesterol-lowering effect (77.95%) value (p < 0.05), after the same incubation period. The phytochemical screening indicated the presence of tannins in all algal extracts, with highest tannin contents in S. platensis (4.66 μg TAE/g DW), followed by A. oryzae with (2.99 μg/g) and S. obliquus (2.13 μg/g), A. oryzae extract contained the highest concentration of phenolic compounds (503.67 μg GAE/g). The highest antioxidant activity was obtained by S. platensis extract (92.66%) value (p < 0.05) at 15 mg/mL, while the least one was observed by S. obliquus (16.81%). The extract of S. platensis, used individually or combined with C. vulgaris extract, is recommended as natural bioactive source, with anti-cholesterol and antioxidant potentialities.

From waste to food: utilization of pineapple peels for vinegar production

The present study is aimed at producing vinegar from fermented pineapple byproducts (peels). The vinegar was produced after fermenting the peels of pineapple usig three selected strains of acetic acid bacteria’s. such as propionic bacterium acidipropionici, panteo agglomerans, and pantea dispersa. This article introduces a new type of acetic acid bacteria’s strains for the production of vinegar from pineapple peels. Three fermentation times (24hr, 48hr, and 72hr) and three acetic acid bacterias (propionic bacterium acidipropionici, panteo agglomerans, and pantea dispersa) were considered and arranged in a factorial experimental design. The fermentation was performed in 500 mL Erlenmeyer flasks containing 200 mL of medium at 28℃. Aeration rate, temperature, and carbon source were set as constant factors. pH, total soluble solids, total residual reducing sugar and titratable acidity were evaluated. The results showed that the vinager samples were in the range of 3.5 to 4.31, 1.3 to 2.31 obrix, 0.50 to2.47% and 3.13 to 6.15 mg/100g of pH, total soluble solids, total residual reducing sugar and titratable acidity respectively. There were significant differences (P&lt;0.05) in the values obtained. Therefore bacterial strain and fermentation time were the most important factors affecting vinegar production. From the experimental results and interaction effects, the optimum yield of acetic acid production was found to be 6.15 g/L at fermentation time of 72h by propionic bacterium acidipropionici acetic acid bacterial strain.

Development and Evaluation of an Indirect Capripoxvirus ELISA Based on Truncated P32 Protein Expressed in E. coli.

As notifiable diseases, lumpy skin disease (LSD), sheep pox (SPP), and goat pox (GTP) are associated with a profound effect on cattle, sheep, and goat farming industries. Development of the ELISA method could effectively facilitate serodiagnosis of the infected animals. This study aimed to develop an ELISA system based on the recombinant full-length and truncated P32 protein (Tr.P32) of goat pox virus. The P32 protein was expressed in Rosetta strain of E. coli using pET24a+ vector and evaluated by SDS-PAGE and Western blotting. Then, Tr.P32 was purified by Ni-NTA affinity chromatography under denaturing conditions and used to develop a capripoxvirus-specific ELISA. Checkerboard titration and receiver-operating characteristic (ROC) analysis were used to optimize the ELISA system and determine diagnostic specificity and sensitivity, respectively. The diagnostic potential of the developed ELISA was evaluated using positive and negative control sera collected from goat, sheep, and cattle. Results showed that the expression level of full-length P32 recombinant protein was negligible, while Tr.P32, a ~ 31 kDa recombinant protein, was expressed up to 0.270-0.300 mg/200 mL of culture media. The results of checkerboard titration revealed that 675 ng/well of Tr.P32 antigen and 1:10 dilution of control sera (anti GTPV HIS and healthy goat sera) caused maximum difference in absorbance between positive and negative goat sera. The recombinant Tr.P32 showed good reactions with antibodies against GTP virus (GTPV), SPP virus (SPPV), and LSD virus (LSDV), whereas no cross-reactions with anti-Orf virus antibodies were detected. By comparing with the neutralization index (NI), cut off, diagnostic sensitivity and specificity of the developed indirect-ELISA were estimated, 0.397, 94% and 96.6%, respectively. These findings indicate that the ELISA system based on Tr.P32 protein could potentially be used in sero-surveillance of all capripoxviruses; however, further investigations are required.

In Vitro Germination of Pollen Grains in Pitahaya Species

ABSTRACT The aim of this study was to adjust the basic culture medium for germination of pollen grains for the different species from the genus Hylocereus. Pollen grains from the species Hylocereus polyrhizus Weber and Hylocereus undatus (Haw.) Britton & Rose were withdrawn of anthers from flowers bagged the previous day, before its opening. Then, with a brush, the pollen grains were spread on the surface of Petri dishes containing 20 mL of culture medium. Four experiments were sequentially installed, considering and selecting the best results from the previous experiment to install the next one. Measurements were made on the length of pollen tube under different concentrations of boric acid. Moreover, germination rate readings were performed after 24 h. The pollen viability of species from the genus Hylocereus varies according to species and the pollen germination was influenced by composition and concentration of medium. The culture medium for the germination of pollen grains from H. polyrhizus species must be increased with 100 g L−1 of sucrose, 518 mg L−1 of calcium nitrate and 636 mg L−1 of boric acid, the pH being measured for 5 and the medium solidified with 6 g L−1 of agar. For H. undatus, the medium should be composed of 100 g L−1 of sucrose, 616 mg L−1 of calcium, 619 mg L−1 of boric acid, 6 g L−1 of agar and pH 6. Both species increased the length of pollen tube with the addition of boric acid.

Assessment of physicochemical properties and comparison of the dissolution profile of amoxicillin caplets

Background: Marketed drugs must meet the required standards to guarantee product quality. Amoxicillin is a generic compound marketed under various trademarks as copy drugs. Amoxicillin caplets are an immediate release dosage form of BCS class I. An essential aspect of evaluating copy drugs is to assess the equivalence for their treatment to the innovator drugs to ensure the safety and effectiveness of the circulating copy drugs. Objective: The study aims to evaluate the physicochemical properties and compare the dissolution profile of amoxicillin caplets available in the market. Methods: Five amoxicillin caplet products, four test products, and one reference product were tested for their physicochemical properties and dissolution. The dissolution test was carried out with a type II device at a speed of 75 rpm in 900 mL of media buffered at pH 1.2, 4.5, and 6.8 and at a temperature of 37 +/- 0.5degrees celcius. The dissolution profile was analyzed by comparing it with the similarity factor (f2) parameters. Results: Two of the four amoxicillin caplet products had a similar dissolution profile to the reference products, namely products A and B. Products C and D were dissimilar because f2 was lower than 50 at pH 4.5. The caplets tested had almost the same dimensions, and all caplets met the requirements for uniformity of content, hardness, disintegration time, and dissolution. Conclusion: Not all of the amoxicillin caplets in the market have a similar dissolution profile to the reference products. Keywords: caplets, amoxicillin, dissolution, similarity factor

Cell size influences inorganic carbon acquisition in artificially selected phytoplankton.

Cell size influences the rate at which phytoplankton assimilate dissolved inorganic carbon (DIC), but it is unclear whether volume-specific carbon uptake should be greater in smaller or larger cells. On the one hand, Fick's Law predicts smaller cells to have a superior diffusive CO2 supply. On the other, larger cells may have greater scope to invest metabolic energy to upregulate active transport per unit area through CO2 -concentrating mechanisms (CCMs). Previous studies have focused on among-species comparisons, which complicates disentangling the role of cell size from other covarying traits. In this study, we investigated the DIC assimilation of the green alga Dunaliella tertiolecta after using artificial selection to evolve a 9.3-fold difference in cell volume. We compared CO2 affinity, external carbonic anhydrase (CAext ), isotopic signatures (δ13 C) and growth among size-selected lineages. Evolving cells to larger sizes led to an upregulation of CCMs that improved the DIC uptake of this species, with higher CO2 affinity, higher CAext and higher δ13 C. Larger cells also achieved faster growth and higher maximum biovolume densities. We showed that evolutionary shifts in cell size can alter the efficiency of DIC uptake systems to influence the fitness of a phytoplankton species.

Isolation and evolutionary analyses of gout-associated goose astrovirus causing disease in experimentally infected chickens

Astroviruses are a common cause of gastroenteritis in humans and animals. They are also associated with extraintestinal infections, including hepatitis in ducklings, nephritis in chickens, as well as fatal meningitis and encephalitis in humans and other mammals. Since 2014, outbreaks of disease characterized by visceral gout and swelling of kidneys have been reported in goslings and ducklings in China, with the causative agent revealed to be a novel avian astrovirus designated goose astrovirus (GoAstV). In the present study, this novel gout-associated GoAstV was identified in diseased goslings from 2 farms in Hunan province, China. Three genomes were successfully sequenced and analyzed and were shown to have high identities of 99.7 to 99.8% between each other, with some specific amino acid alterations revealed in open reading frame 2 when compared with other gout-associated GoAstVs. Two strains were further efficiently isolated in the DF-1 chicken fibroblast cell line with high virus titers of 1011 viral genomic copies per mL of culture media. A pilot virus challenge study using GoAstV in chickens demonstrated that this virus can cause clinical visceral gout in chickens, indicating its ability to cross the species barrier. Based on the phylogenetic analyses of capsid sequences, the identified GoAstVs were proposed to be classified into 2 genotypes, GoAstV1 and GoAstV2, and the novel gout-associated GoAstVs were all clustered in GoAstV2. Further Bayesian inference analyses indicated a nucleotide substitution rate of 1.46 × 10−3 substitutions/site/year for avian astrovirus based on open reading frame 2 sequences, and the time to the most recent common ancestor of GoAstVs was estimated to be around 2011. This is the first report to confirm GoAstV can infect chickens while also providing an estimation of the evolutionary rates of Avastroviruses.

Vaccinium myrtillus L. extract and its native polyphenol-recombined mixture have anti-proliferative and pro-apoptotic effects on human prostate cancer cell lines.

Vaccinium myrtillus berry extract (VME) and a recombined standard mixture (RSM) of its main native phenolic compounds were investigated for cell growth inhibition and pro-apoptotic activity on hormone-dependent (LNCaP) and hormone-independent (PC3 and DU-145) prostate cancer (PCa) cell lines. Normal prostate epithelial cells (PrEC) were also studied in comparison. VME hindered anchorage-dependent PCa cell proliferation in a dose-dependent manner, that is, at 1/800 (v/v) dilution for LNCaP and PC3, and 1/100 (v/v) dilution for DU-145 (corresponding to 14.15 and 113.2 μg cyanidin-3-O-glucoside equivalents per ml of culture medium), respectively. VME had a growth inhibitory effect towards PrEC at the same dilution of DU-145 cells although the IC50 values indicated that PrEC are more resistant than PCa cell lines. VME also reduced the anchorage-independent growth of PCa cells. The study of the apoptotic profile (i.e., non-apoptotic, early apoptotic, late apoptotic and necrotic cells) evidenced that the apoptotic rate (early+late) was statistically higher in all three cell lines exposed to VME compared to control. Anchorage-dependent and anchorage-independent growth inhibition of RSM was very similar to that displayed by VME. Moreover, RSM exerted its growth inhibitory effect also under hypoxia, the latter representing a biological condition known to sustain PCa proliferation and aggressiveness.

Plant Nitrilase Homologues in Fungi: Phylogenetic and Functional Analysis with Focus on Nitrilases in Trametes versicolor and Agaricus bisporus.

Fungi contain many plant-nitrilase (NLase) homologues according to database searches. In this study, enzymes NitTv1 from Trametes versicolor and NitAb from Agaricus bisporus were purified and characterized as the representatives of this type of fungal NLase. Both enzymes were slightly more similar to NIT4 type than to NIT1/NIT2/NIT3 type of plant NLases in terms of their amino acid sequences. Expression of the synthetic genes in Escherichia coli Origami B (DE3) was induced with 0.02 mM isopropyl β-D-1-thiogalactopyranoside at 20 °C. Purification of NitTv1 and NitAb by cobalt affinity chromatography gave ca. 6.6 mg and 9.6 mg of protein per 100 mL of culture medium, respectively. Their activities were determined with 25 mM of nitriles in 50 mM Tris/HCl buffer, pH 8.0, at 30 °C. NitTv1 and NitAb transformed β-cyano-L-alanine (β-CA) with the highest specific activities (ca. 132 and 40 U mg−1, respectively) similar to plant NLase NIT4. β-CA was transformed into Asn and Asp as in NIT4 but at lower Asn:Asp ratios. The fungal NLases also exhibited significant activities for (aryl)aliphatic nitriles such as 3-phenylpropionitrile, cinnamonitrile and fumaronitrile (substrates of NLase NIT1). NitTv1 was more stable than NitAb (at pH 5–9 vs. pH 5–7). These NLases may participate in plant–fungus interactions by detoxifying plant nitriles and/or producing plant hormones. Their homology models elucidated the molecular interactions with various nitriles in their active sites.

Nutritional medium for differentiation of malignant anthrax

The purpose of the work is to develop a nutrient medium for differentiation of bacillus anthracis from soil aerobic bacilli. In order to achieve the set goal, we used the method of introduction into the environment of cultivation of microorganisms separated from animals and objects of external environment (water, soil, feed, air, scrapes from different surfaces suspected of contamination by their bacillus anthracis) of nutrient substrate - sucrose, used by bacteria of Bacillus genus for synthesis of levan - product of their metabolism, a sign absent in bacillus anthracis. This feature is essential for identification and differentiation of bacillus anthracis from closely related saprophytes. To identify and differentiate the bacillus anthracis, the microbes isolated from the external environment were cultivated in a nutrient medium consisting of meat-and-peptone agar (MPA) and stimulator (inductor) synthesis of levan sucrose in the amount of 10% to 100 ml of melted agar. The proposed nutrient media was prepared as follows. Meat-peptonic agar (500 ml) was melted at 100°C, 10 g of sucrose was added per 100 ml of medium and after the complete dissolution of sucrose, the nutrient medium was poured into Petri dishes and used for sowing the studied material for identification and differentiation of grown crops. The efficiency of the method has been tested in production experiments with positive evaluation. For this purpose, soil samples taken from the territory of old cattle cemeteries were fractionally sown on MPA and thermostatted for 16-18 hours at 37°C and examined crops for the presence of matte and rough (R-form) colonies.

CHANGES IN THE CONTENT OF IMMUNE CELLS CONTAINING THE FcR III RECEPTOR IN THE SLEEP AND BRAIN AFTER EXPERIMENTAL TBI

At TBI there are disturbances in activity of immune system, can complicate regenerative and reparative responses in a brain. The aim of the study was to study the content of CD-16 cells in the spleen and brain parenchyma at different times after TBI in rats and in the correction of disorders in their composition by immunomodulatory drug galalite.Methods. Craniocerebral trauma in animals was modeled by dropping a load weighing 100 g from a height of 120 cm on the head of rats that were anesthetized and were in a state of narcotic sleep, which lasted up to 30 minutes. To correct the resulting disorders used immunomodulatory drug galavit at a dose of 2 mg / kg of animal weight in a volume of0.5 ml, which was administered intramuscularly to animals for 2, 3, 4 days after injury. The spleen and the left and right hemispheres of the brain were homogenized in 5.0 ml of medium 199 and suspensions containing 10.0x106 cells per 1 ml of medium 199 microglia were prepared and infiltrated CNS monocytes were prepared by the method of Sedgwick J. in co-authors. Determination of the level of CD-16 cells in suspensions of spleen and microglia was performed on a flow cytometer using monoclonal antibodies against CD-16 receptor company BD Biosciences according to the instructions for antibodies.Results. In mild TBI in rats, the number of CD- 16 cells containing the FcR III receptor decreases in the spleen for 2 to 5 days, and the number of these cells in the fractions of microglial cells increases. Galavit is a drug with immunomodulatory properties, has virtually no effect on low levels of CD-16 cells in the spleen and stimulates their accumulation in the brain for 10 days after TBI.Conclusions. The results indicate the effect of Galavit on the level of CD-16 cells in the spleen and brain, which indicates the immunomodulatory activity of this drug.

Features of isolation of the anthrax pathogen depending on the type of nutrient medium

The purpose of the work is to develop a nutrient medium for differentiation of bacillus from soil aerobic bacilli. In order to achieve the set goal, we used the method of introduction into the environment of cultivation of microorganisms separated from animals and objects of external environment (water, soil, feed, air, scrapes from different surfaces suspected of contamination by their bacillus ) of nutrient substrate sucrose, used by bacteria of Bacillus genus for synthesis of product of their metabolism, a sign absent in bacillus . This feature is essential for identification and differentiation of bacillus from closely related saprophytes. To identify and differentiate the bacillus , the microbes isolated from the external environment were cultivated in a nutrient medium consisting of agar (MPA) and () synthesis of sucrose in the amount of 10% to 100 ml of melted agar. The proposed nutrient media was prepared as follows. agar (500 ml) was melted at 1000C, 10 g of sucrose was added per 100 ml of medium and after the complete dissolution of sucrose, the nutrient medium was poured into dishes and used for sowing the studied material for identification and differentiation of grown crops. The efficiency of the method has been tested in production experiments with positive evaluation. For this purpose, soil samples taken from the territory of old cattle cemeteries were fractionally sown on MPA and for 16-18 hours at 370C and examined crops for the presence of matte and rough (R-form) colonies.

Application of Cryopreserved Human Intestinal Mucosa and Cryopreserved Human Enterocytes in the Evaluation of Herb-Drug Interactions: Evaluation of CYP3A Inhibitory Potential of Grapefruit Juice and Commercial Formulations of Twenty-Nine Herbal Supplements.

Commercial formulations of 29 commonly used herbal supplements (HSs) and grapefruit juice were evaluated for drug interaction potential via quantification of their CYP3A inhibitory potential in two in vitro experimental models of human small intestine, cryopreserved human intestinal mucosa (CHIM), and cryopreserved human enterocytes (CHEs). Two CYP3A substrates were used-in the studies with CHIM, CYP3A activity was quantified via liquid chromatography tandem mass spectrometry quantification of midazolam 1'-hydroxylation, whereas in CHE, luciferin-IPA metabolism to luciferin was quantified by luminescence. Upon treatment of CHIM with the estimated lumen concentration of the HS upon each oral administration (manufacturers' recommended dosage dissolved in 200 ml of culture medium), >80% CYP3A inhibition was observed for green tea extract, St. John's wort, valerian root, horehound, and grapefruit juice. Less than 50% inhibition was observed for fenugreek, aloe vera, guarana, soy isoflavone, maca, echinacea, spirulina, evening primrose, milk thistle, cranberry, red yeast rice, rhodiola, ginkgo biloba, turmeric, curcumin, white kidney bean, garlic, cinnamon, saw palmetto berries, panax ginseng, black elderberry, wheat grass juice, flaxseed oil, black cohosh, and ginger root. The results were confirmed in a a dose-response study with HSs obtained from three suppliers for the four inhibitory HSs (green tea extract, horehound, St. John's wort, valerian root) and three representative noninhibitory HSs (black cohosh, black elderberry, echinacea). Similar results were obtained with the inhibitory HSs in CHE. The results illustrate that CHIM and CHE represent physiologically relevant in vitro experimental models for the evaluation of drug interaction potential of herbal supplements. Based on the results, green tea extract, horehound, St. John's wort, and valerian root may cause drug interactions with orally administered drugs that are CYP3A substrates, as was observed for grapefruit juice. SIGNIFICANCE STATEMENT: In vitro evaluation of 29 popular herbal supplements in cryopreserved human intestinal mucosa identified green tea extract, horehound, St. John's wort, and valerian root to have CYP3A inhibitory potential similar to that for grapefruit juice, suggesting their potential to have clinically significant pharmacokinetic interaction with orally administered drugs that are CYP3A substrates. The results suggest that cryopreserved human intestinal mucosa can be used for in vitro evaluation of drug interactions involving enteric drug metabolism.

GM3 trisaccharide biosynthesis and process optimization using engineered E. coli lysate and whole-cell catalysis

An engineered E. coli strain BL21 (DE3, SiaB, PmST3, △lacZ, △nanZ) that simultaneously encoded CMP-Neu5Ac synthase (SiaB) and α-2-3-sialyltransferase (PmST3) was constructed to synthesize GM3 trisaccharide. The crude E. coli lysate could be readily used for biosynthesis of GM3 trisaccharide with a nearly quantitative yield under optimal conditions. In addition, this engineered E. coli strain as whole-cell catalyst could generate approximately 1 g of GM3 trisaccharide in 40 mL of medium under optimal conditions. The whole-cell catalyst could be recycled and reused for at least 3 rounds without obvious loss of enzymatic activity.

Biological hydrogen production from palm oil mill effluent (POME) by anaerobic consortia and Clostridium beijerinckii

Palm oil mill effluent (POME) was tested as a substrate to produce hydrogen by dark fermentation. Two microbial consortia and a pure culture of Clostridium beijerinckii (ATCC 8260) were cultured anaerobically in raw, diluted and hydrolyzed POME to compare biohydrogen production yields in all three media. Experiments were done in 15 mL Hungate tubes containing 5 mL of medium and 1 mL of inoculum. When Clostridium beijerinckii was cultivated at 30 °C in the hydrolyzed POME (P003), containing 7.5 g/L of sucrose, during 8 days of fermentation and 20 % of the inoculum, the maximum biohydrogen production yield was 4.62 LH2/Lmed. Consortium C3 also showed the best production in hydrolyzed POME while consortium C6 achieved its maximum production in raw POME. This effluent is a potential substrate for biohydrogen production.