Nitroblue Tetrazolium Reduction Research Articles

Pirarucu requires taurine to maximize growth and antioxidant status when fed diets high in plant-based ingredients

Taurine (Tau) is a free conditionally essential amino acid for most fish species that plays a significant nutritional role, mainly in carnivorous fish species. However, limited studies are still available regarding the role of Tau on pirarucu nutrition, an important Amazonian fish species. Therefore, this study used diets high in plant-based ingredients to investigate the effect of taurine (Tau) supplementation on juvenile pirarucu's growth, metabolism, and health (n = 115, Weight = 504.12 ± 19 g). Fish were fed five diets containing increasing levels of Tau: 0.95, 5.1, 9.6, 12.9, and 17.2 g/kg in a completely randomized design with three replicates per treatment. After a 60-day feeding trial, growth, hematology, blood chemistry, Tau content in muscle, carcass, and liver, antioxidant status, and immunological parameters were evaluated. The control group (0.95 g/kg Tau) showed the lowest growth. According to the segmented regression model, the recommended Tau level for maximizing the development of juvenile pirarucu was 4.49 g/kg of feed. Additionally, fish fed the lowest dietary Tau level showed the highest ROS (reactive oxygen species) concentration. Dietary Tau levels did not affect hematological parameters and blood chemistry (p > 0.05). Fish fed 9.6 g/kg of Tau showed increased catalase activity, while the control group had the worst result in NBT (nitroblue tetrazolium reduction test). Fish fed with the highest dietary Tau levels (12.9 and 17.2 g/kg) showed increased total serum immunoglobulin concentration. These results suggest that the optimal inclusion level of Tau in diets for juvenile pirarucu is 4.49 g/kg of feed and that this amino acid acts as a mitigator of oxidative stress.

Role of reactive carbonyls and superoxide radicals in protein damage by cigarette smoke extracts: Comparison of Heat-not-Burn e-cigarettes to conventional cigarettes

Oxidative protein damage involving carbonylation of respiratory tract proteins typically accompanies exposure to tobacco smoke. Such damage can arise via multiple mechanisms, including direct amino acid oxidation by reactive oxygen species or protein adduction by electrophilic aldehydes. This study investigated the relative importance of these pathways during exposure of a model protein to fresh cigarette emission extracts. Briefly, protein carbonyl adducts were estimated in bovine serum albumin following incubation in buffered solutions with whole cigarette emissions extracts prepared from either a single 1R6F research cigarette or a single “Heat-not-Burn” e-cigarette. Although both extracts caused concentration-dependent protein carbonylation, conventional cigarette extracts produced higher adduct yields than e-cigarette extracts. Superoxide radical generation by conventional and e-cigarette emissions was assessed by monitoring nitro blue tetrazolium reduction and was considerably lower in extracts made from “Heat-Not-Burn” e-cigarettes. The superoxide dismutase/catalase mimic EUK-134 strongly suppressed radical production by whole smoke extracts from conventional cigarettes, however, it did not diminish protein carbonyl adduction when incubating smoke extracts with the model protein. In contrast, edaravone, a neuroprotective drug with strong carbonyl-trapping properties, strongly suppressed protein damage without inhibiting superoxide formation. Although these findings require extension to appropriate cell-based and in vivo systems, they suggest reactive aldehydes in tobacco smoke make greater contributions to oxidative protein damage than smoke phase radicals.

Ethanolic extract of Euphorbia Gypsicola induces differentiation and apoptosis in human myeloid leukemia K562 cells

IntroductionThe Euphorbiaceae family, widely distributed and long utilized in traditional medicine, has been recognized for its potential in developing anti-cancer drugs. The current study investigates the capacity of Euphorbia gypsicola extract to induce apoptosis and differentiation in the human myeloid leukemia K562 cell line. MethodsCell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptotic activity was visually evaluated through acridine orange/ethidium bromide (AO/EB) dye staining and annexin V/PI double labeling procedures. Induction of differentiation was examined using nitro blue tetrazolium (NBT) reduction and Wright-Giemsa staining assays. The expression of apoptosis-related proteins including Bax, Bcl2 and Survivin was evaluated by western blotting method. ResultsThe E. gypsicola extract exhibited potent medication properties with IC50 values of 207.59, 48.58, and 7.08 ng/mL at 24, 48, and 72 h, respectively. Fluorescence microscopy, DNA fragmentation, annexin V/PI double staining, and sub-G1 cell cycle arrest confirmed apoptosis occurrence in K562 cells treated with the E. gypsicola extract. Our findings demonstrated that differentiated cells reduced NBT yellow solution after endocytosis, forming dark crystals. Wright Giemsa staining observations indicated a decrease in nuclear volume-to-cytoplasm ratio in K562 cells treated with the crude extract, signifying Induction of differentiation. Furthermore, western blot analysis revealed down-regulation of Survivin protein expression and up-regulation of Bax protein level following treatment with the E. gypsicola extract. However, no significant decrease in Bcl2 protein expression level was observed in K562 cells. ConclusionThese findings indicate the growth inhibitory effects of the E. gypsicola extract on leukemia K562 cells, which supports the further study of this plant to discover novel therapeutic compounds for the treatment of leukemia.

Antioxidant and Anti-Inflammatory Activity of Fractionated Artemia Urmiana Extract in Human Peripheral Blood Neutrophils System

Marine organisms are great resources for discovering bioactive compounds with new antioxidant properties. The aim of this work was to purify and isolate the protein fractions of Artemia urmiana and to evaluate antioxidant activities of them on human peripheral blood neutrophils response. For these purposes, crude extracts protein was extracted from napulii of A. urmiana and sequentially fractionated by ion exchange. The antioxidant activities of the samples were determined by DPPH free radical scavenging, reducing power ability assays in comparison with ascorbic acid. In the second phase, the human peripheral blood neutrophils (HPBN) were separated and cultured. The cells were treated with increasing concentrations of crude and partially purified fractions for 24 hrs. After that, the pellet and supernatant of treated cells were separately collected and assayed for Nitro Blue Tetrazolium (NBT) reduction, the activity of Super Oxide Dismutase (SOD) and Nitric oxide content. The results showed that the IC50 values of ascorbic acid, crude extracts, partially purified protein fractions(F1-F8) were 150, 126, 141, 295, 299, 114, 164 and 132 µg/ml respectively in the DPPH radical scavenging activity. The reducing power activity at the highest concentration (200 µg/ml) based on the optical density of the sample listed, were 2.8, 1.4, 0.13, 0.14, 0.17, 0.14, 0.13, 0.66, 0.18 and 0.35 respectively. In NBT reduction and production of Nitric oxide, treated cells did not show significant difference between different concentrations of protein extract and untreated control cells. The activity of SOD was increased in presence of 50-100 µg/ml of crud extract and fractions 1, 2, 3, 4 and 5 in comparison to untreated control cells (p<0.05).

Highly efficient hydrogen production and selective CO2 reduction by the C3N5 photocatalyst using only visible light.

The production of energy sources by metal-free photocatalysts based on graphitic carbon nitride (g-C3N4) has garnered substantial attention. In this study, nitrogen-rich carbon nitride (C3N5) was successfully synthesized through the thermal polycondensation of 3-amino-1,2,4-triazole. The structural and physical characterization has suggested that a portion of the triazine rings, which constitute the structural framework of g-C3N4, may be substituted with five-membered rings in C3N5. Furthermore, the polymerization of C3N5 proceeded more extensively than that of g-C3N4 from melamine precursors. The increased nitrogen content in C3N5 resulted in a heightened number of π-electrons and a narrowed energy bandgap, with the potential of the valence band maximum being negatively shifted. Additionally, photocatalytic assessments encompassing nitro blue tetrazolium reduction, H2 production from triethanolamine aqueous solution, and CO2 reduction in the liquid phase were performed. All findings demonstrated that C3N5 exhibits significantly superior photocatalytic properties compared to g-C3N4. It is particularly noteworthy that C3N5 selectively generates methanol and H2 from oversaturated CO2 solutions under visible light irradiation, while g-C3N4 selectively generates formaldehyde. These outcomes strongly indicate that C3N5 serves as a metal-free, visible-light-responsive photocatalyst, capable of contributing to both the production of renewable energy sources and the reduction of greenhouse effect gases.

Immunomodulatory effect of Myrtenol on benzo (a) pyrene-induced lung cancer in Swiss albino mice via modulation of tumor markers, cytokines and inhibition of PCNA expression.

Lung cancer is one of the most common cancers in men. Although many diagnostic and treatment regimens have been followed in the treatment for lung cancer, increasing mortality rate due to lung cancer is depressing and hence requires alternative plant based therapeutics with with less side-effects. Myrtenol exhibits anti-inflammatory and antioxidant properties. Hence we intended to study the effect of Myrtenol on B(a)P-induced lung cancer. Our study showed that B(a)P lowered hematological count, decreased phagocyte and avidity indices, nitroblue tetrazolium (NBT) reduction, levels of immunoglubulins, antioxidant levels, whereas Myrtenol treatment restored them back to normal levels. On the other hand, xenobiotic and liver dysfunction marker enzymes and pro-inflammatory cytokines were elevated on B(a)P exposure, which retuned back to normal by Myrtenol. This study thus describes the immunomodulatory and antioxidant effects of Myrtenol on B[a]P-induced immune destruction.

Exploring the Relationship between Neutrophil Activation and Different States of Canine L. infantum Infection: Nitroblue Tetrazolium Test and IFN-γ.

This study aimed to investigate the role of neutrophils in canine leishmaniosis by assessing neutrophil activation and its relationship with different states of L. infantum infection and antibody and IFN-γ production. Dogs were categorized into five groups: healthy-seronegative (n = 25), healthy-seropositive (n = 21), LeishVet-stage I (n = 25), Leishvet-stage II (n = 41), and LeishVet-stage III-IV (n = 16). Results of the nitroblue tetrazolium reduction test (NBT) showed significantly higher neutrophil activation in stage I (median:17.17, range: [7.33-31.50]%) compared to in healthy-seronegative (4.10 [1.20-18.00]%), healthy-seropositive (7.65 [3.98-21.74]%), stage II (6.50 [1.50-28.70]%), and stage III-IV (7.50 [3.00-16.75]%) groups (p < 0.0001). Healthy-seropositive dogs also displayed higher values than all groups except stage I. Stages II and III-IV did not show significant differences compared to healthy-seronegative. Regarding IFN-γ, stage I dogs had higher concentrations (median:127.90, range: [0-3998.00] pg/mL) than healthy-seronegative (0 [0-109.50] pg/mL) (p = 0.0002), stage II (9.00 [0-5086.00] pg/mL) (p = 0.045), and stage III-IV (3.50 [80.00-548.80] pg/mL) (p = 0.02) dogs. Stage II dogs showed increased IFN-γ compared to healthy-seronegative dogs (p = 0.015), while stage III-IV dogs had no significant differences compared to healthy-seronegative dogs (p = 0.12). Healthy-seropositive dogs had elevated IFN-γ concentrations compared to healthy-seronegative dogs (p = 0.001) and dogs in stage III-IV (p = 0.03). In conclusion, neutrophil activation was higher in dogs with mild disease and healthy-seropositive dogs, and a relationship between neutrophil activation and the production of IFN-γ was found.

Effect of storage on the nitro blue tetrazolium reduction test in dog blood samples.

The nitro blue tetrazolium (NBT) reduction test (NBTT) has been used for measuring the metabolic activity of phagocytes of mammals. Activated neutrophils transform NBT into formazan in the cytoplasm. The NBTT can detect the activation of neutrophils in peripheral blood and is used to assess neutrophil function in dogs. However, the NBTT is not used frequently in the clinical setting, as samples should be processed after blood collection. The aim of this study was to evaluate the effect of storage on NBTT in dog blood samples. Residual EDTA blood samples from 22 dogs were included of different ages, breeds, and sex. The buffy coat layer was separated from the blood and incubated with 0.1% NBT. The NBTT was performed at 0, 24, 48, and 72 h after the collection of blood. Blood samples were stored at 4°C until the tests were performed. Blood smears were evaluated by light microscopy, and the NBT reduction rate was reported, which represents the percentage of activated neutrophils. The NBT reduction rate was calculated after counting 300 neutrophils in each slide. The means of NBTT in dog blood samples at 0, 24, 48, and 72 h were 8.3%, 8.5%, 8.7%, and 7.8%, respectively. No significant differences were observed between time points. This study showed that the NBTT can be performed up to 72 h after the collection of canine blood if correctly refrigerated at 4°C. This finding supports the performance of the NBTT in the clinical setting.

Oncolytic Newcastle disease virus effects on immune response - a new issue in cancer treatment.

Millions of people are diagnosed with cancer each year, and fighting it puts a heavy financial burden on communities and governments. Numerous advances have been made in the field of cancer; one of the newest methods is using oncolytic viruses. This study aimed to evaluate the effect of oncolytic Newcastle disease virus wild-type strains (NDV-WTS) on the immune system. Forty mice were divided into four groups (10 animals in each group). The control group received phosphate buffered saline, and experimental group 1 (NDV-WTS 1), experimental group 2 (NDV-WTS 2), and experimental group 3 (NDV-WTS 3) received 10-1, 10-2, and 10-3 titers of Newcastle virus on 0, 14th, and 28th days. On the 31st day, 100 µL of Newcastle virus was injected into the left footpads of animals. After 48 hours, delayed-type hypersensitivity (DTH) reactions were measured. On the 33rd day, peritoneal macrophages were isolated. Then proliferation of the cells was measured by the methyl-thiazolyl-tetrazolium (MTT) test. Neutral red uptake and respiratory burst of peritoneal macrophages were also assessed. Data were analyzed using statistical software SPSS, version 19. The results of the DTH test showed that footpad swelling in control, NDV-WTS 1, NDV-WTS 2, and NDV-WTS 3 groups were 23.5%, 23.5%, 23.6% and 23.6%. No significant differences were seen between the groups in this regard (P &gt; 0.05). A negative nitroblue tetrazolium (NBT) reduction test as an indicator of macrophage's respiratory burst, showed no significant difference between the groups (P &gt; 0.05). The neutral red uptake assay and MTT test showed no significant differences between the groups (P &gt; 0.05). The results of this study showed that NDV-WTS in doses of 10-1, 10-2, and 10-3 have no adverse effects on healthy normal cells.

Superoxide Generation by Nicotinamide Coenzymes

Nicotinamide coenzymes can generate superoxide radicalsin alkaline environment. Their formation was registered by reduction of nitroblue tetrazolium (NBT) present in the buffer with formation of diformasan. Inhibition of diformazan formation occurs when superoxide dimutase (SOD) is added to the system, thus confirming generation of O─●. The highest superoxide generating activity was observed with NADPH. In the case of NADPH and NADH, the rate of superoxide generation was significantly lower (by approximately 50%). No O─● was detected when NAD was used under the same conditions and in the same time; however, 4 h later, diformasan was detected in the same sample. The superoxide generating activity decreased in the following order: NADPH &gt; NADH ? NADP &gt; NAD. Other compounds tested (adenosine, ADP and ATP) did not generate superoxide radicals even after prolonged incubation. In a cell, where a local changes in the pH of the environment are possible, nicotinamide coenzymes can be potential sources of O─● and thus participate in cell signaling. A change in pH can initiate this process.

MEGAMOX MEDIATES ECO-FRIENDLY SYNTHESIS OF SELENIUM NANOPARTICLES AS IMMUNOMODULATORY, ANTIOXIDANT AND ANTIMICROBIAL AGENTS

The research herein includes methodology for eco-friendly preparation of selenium nanoparticles (SeNPs) using the broad-spectrum antibiotic, Megamox (Mega.). Characterization of SeNPs was done using UV-Visible spectroscopy, FTIR, XRD, DLS and TEM imaging. Additionally, the immunomodulatory, antioxidant and antimicrobial activities of both SeNPs and Mega. were checked against different strains of bacteria and fungi. TEM and DLS images showed that SeNPs are polydisperse spheres with mean diameter of 22.4 nm. FTIR analysis indicated that the hydroxyl and nitrogen moieties in Mega. were effective for reduction plus binding manner. According to the results of the nitro blue tetrazolium reduction test, both SeNPs and Mega. presented high intracellular killing activity, which confirmed their immunostimulatory effect. Antioxidant activity of SeNPs and Mega. were 90 and 82 %, respectively. SeNPs presented great activity facing multi-drug resistant bacteria and TB. SeNPs are considered promising cost-effective and eco-friendly anti-bacterial and anti-fungal agents that can represent a new potential nano-platform in both medical and infectious diseases control.

Reprint of: Generation of Superoxide Radical during Autoxidation of Hydroxylamine and an Assay for Superoxide Dismutase

Accompanying the autoxidation of hydroxylamine at pH 10.2, nitroblue tetrazolium was reduced and nitrite was produced in the presence of EDTA. The rate of autoxidation was negligible below pH 8.0, but sharply increased with increasing pH. The reduction of nitroblue tetrazolium was inhibited by superoxide dismutase, indicating the participation of superoxide anion radical in the autoxidation. Hydrogen peroxide stimulated the autoxidation and superoxide dismutase inhibited the hydrogen peroxide-induced oxidation, results which suggest the participation of hydrogen peroxide in autoxidation and in the generation of superoxide radical. An assay for superoxide dismutase using autoxidation of hydroxylamine is described.

Single Cell Assessment of Mitochondrial Function

Cellular mitochondrial function can be assessed using high resolution respirometry that measures O2 consumption rate during various conditions that systematically alter the tricarboxylic acid (TCA) cycle or the electron transport chain (ETC). However, current high resolution respirometry does not measure O2 consumption rate at the single cell level, but actually measures average mitochondrial function across a number of cells (either isolated or in tissues). Thus, respirometry assumes physiological homogeneity across cells. However, in many tissues, mitochondrial function varies across cells and this heterogeneity is physiologically important. Therefore, a direct measurement of cellular mitochondrial function will provide valuable novel information and physiological insight. In the present study, we used a quantitative histochemical technique to measure the activity of succinate dehydrogenase (SDH), a key enzyme located in the inner mitochondrial membrane, and the only enzyme to participate in both the TCA cycle and the ETC as Complex II. SDH mediates the oxidation of succinate to fumarate in the TCA cycle, which is coupled to the reduction of ubiquinone to ubiquinol in the ETC. In this study we determined the maximum velocity of the SDH reaction (SDHmax) in isolated human airway smooth muscle (hASM) cells using 1‐methoxyphenazine methosulphate (mPMS), as an exogenous electron carrier, and azide to inhibit cytochrome oxidase. To measure SDHmax, the cells were exposed to a solution containing 80 mM succinate and 1.5 mM nitroblue tetrazolium (NBT) as the reaction indicator. hASM cells were imaged in 3D (Z optical slice of 0.5 μm) using a Nikon Eclipse A1 laser scanning confocal system with a ×60/1.4 NA oil‐immersion lens. In the quantitative histochemical procedure, changes in cell optical density (OD) due to the progressive reduction of NBT to its diformazan (peak absorbance wavelength of 570 nm) were measured every 15 s over a 10 min period. Linearity of the SDH reaction was confirmed across the 10 min period, and SDHmax was expressed as mM fumarate/liter of tissue/min. Validation of this technique included specific ETC inhibitors including oligomycin (ATP synthase inhibitor), FCCP (proton ionophore), antimycin A (Complex III inhibitor) and rotenone (Complex I inhibitor), similar to those used in high resolution respirometry. We observed that FCCP‐mediated disruption of the mitochondrial proton gradient does not affect SDHmax, while SDHmax is decreased by rotenone and antimycin A. In addition, we used MitoTracker Green to label and image mitochondria in hASM cells and determined mitochondrial volume density. The SDHmax was then normalized to mitochondrial content. Our results confirm that this quantitative technique is rigorous and reproducible, and that measurements of cellular SDHmax can serve as a reliable surrogate for the measurement of maximum mitochondrial respiration in single cells.

Determination of secondary metabolites including curcumin in Rheum ribes L. and surveying of its antioxidant and anticancer activity

Dry stem and root of Rheum ribes are used for the treatment of different diseases. In this study, the organic components in the root of R. ribes were extracted by three extraction methods (soxhlet, ultrasonic and maceration) and the efficiencies of these methods were calculated. To validate the existence of Curcumin, analysis such as UV–Vis (Ultraviolet–Visible), HPLC (High Performance Liquid Chromatography), GC (Gas Chromatography) and GC/Mass were performed and in order to separate Curcuminoids, TLC (Thin Layer Chromatography) technique was implemented on the extract which had the highest yield. Total phenolic, flavonoid and flavonol contents of three kinds of extract were measured. For studying antioxidant activity, DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging was done. Finally, for investigating anticancer activity, MTT (Methyl Thiazolyl Tetrazolium), NBT (Nitro Blue Tetrazolium) and DNA-Binding tests were done. The most yield was belonged to soxhlet extract. The results showed the presence of Curcumin in soxhlet extract has not been reported previously. Furthermore, the total phenolic, flavonoid and flavonol contents were belonged to ultrasonic extract. As well as the maximum antioxidants properties were belonged to ultrasonic and maceration extraction. The MTT assay showed the lowest IC50 in the ultrasonic extract. At NBT test, the highest NBT reduction percentage was belonged to maceration extraction. DNA-binding test revealed that all the extracts have shown some degree of fragmentation. Due to the presence of Curcumin in R. ribes and the notable features that the results showed, it can be a good source for Curcumin extraction and use in the drug industry.

Inactivation of Pathogens via Visible-Light Photolysis of Riboflavin-5'-Phosphate.

Riboflavin-5'-phosphate (or flavin mononucleotide; FMN) is sensitive to visible light. Various compounds, including reactive oxygen species (ROS), can be generated from FMN photolysis upon irradiation with visible light. The ROS generated from FMN photolysis are harmful to microorganisms, including pathogenic bacteria such as Staphylococcus aureus (S. aureus). This article presents a protocol for deactivating S. aureus, as an example, via photochemical reactions involving FMN under visible light irradiation. The superoxide radical anion () generated during the FMN photolysis is evaluated via nitro blue tetrazolium (NBT) reduction. The microbial viability of S. aureus that is attributed to reactive species was used to determine the effectiveness of the process. The bacterial inactivation rate is proportional to FMN concentration. Violet light is more efficient in inactivating S. aureus than blue light irradiation, while the red or green light does not drive FMN photolysis. The present article demonstrates FMN photolysis as a simple and safe method for sanitary processes.

Growth Promoter, Immunostimulant and Antioxidant for Rainbow Trout (Oncorhynchus mykiss): Terebinth (Pistacia terebinthus) Extract

In this study, the effects of Pistacia terebinthus (PT) fruit extract supplemented diet on growth performance, haematology, digestive and antioxidant enzyme activities, and non-specific immune responses were evaluated in juvenile rainbow trout (Onchoryhnchus mykiss). The fish were fed diets containing three doses of Pistacia terebinthus extract (0.1, 0.5 and 1% of diet) and a control diet without extract for 63 days. Final weight, weight gain and specific growth rate were significantly improved in all the treated groups. In addition, feed conversion ratio was significantly reduced in all PT diet fed groups. Pepsin and lipase activities were significantly increased in all the treated groups. Trypsin was significantly improved in PT 0.1% and PT 1% groups. Amylase was significantly increased in PT 0.5 and 1% groups. In haematological assays, red blood cell, haemoglobin, haematocrit, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration values were not changed among all experimental groups. Superoxide dismutase, catalase, glutathione peroxidase and glucose-6-phosphate dehydrogenase activities were significantly improved in all the treatment groups. However, catalase activity decreased in PT 0.5% group at the end of 63 days. In addition, hepatic and white muscle lipid peroxidation activities were significant decreased in all the treated groups compared to the control. Non-specific immune parameters, such as nitroblue tetrazolium reduction, myeloperoxidase and lysozyme activities were increased in all the treated fish groups. These results indicated that extract of P. terebinthus can be used to improve fish health in aquaculture.

Bioconversion of chitin waste through Stenotrophomonas maltophilia for production of chitin derivatives as a Seabass enrichment diet

Marine wastes pose a great threat to the ecosystem leading to severe environmental hazards and health issues particularly the shellfish wastes. The shellfish waste which contains half of the amount of chitin can be efficiently transformed into useful products. Various approaches for the hydrolysis of chitin like physical, chemical, and enzymatic processes are there. Still, the use of enzyme chitinase is well documented as an effective and eco-friendly method. The present study summarizes the isolation of chitinase enzyme producing bacteria from different shrimp waste disposal sites in Parangipettai (India), and the possible use of an enzyme hydrolyzate as an immunostimulant to Asian Seabass (Lates calcarifer). The potential chitinase-producing bacteria were identified by 16S rRNA gene sequencing as Stenotrophomonas maltophilia. After purification, the chitinase specific activity was 5.01 (U/ml) and the protein content was 72 mg and the recovery rate was 48.06%. The optimum pH and temperature for the chitinolytic activity were 6.5 and at 35–50 °C, respectively. The animal experiment trial was done with our feed supplements which included 0.0 (control), 0.5%, 1% and 2% of chitin degraded product. All the supplementary feed had an optimal 42% (w/w) of crude protein. The feed protein level was 41–43% on average and gross energy was 13–17 kcal/g and the feed was observed to exhibit a significantly higher (p < 0.05) survival rate, condition factor, specific growth rates, and body weight gain was also found to be promising compared to other fishes fed with control diet only. The red blood cells (RBC) and white blood cell (WBC) counts were found to increase significantly after being challenged with infection in animals fed with chitin derivatives from 1st week to 3rd week when compared to the control. The hematocrit (Hct) values were low on the 2nd and 3rd week in infected fish fed with chitin derivatives. This low level was due to infection lyses of the red blood cells and increased nitro blue tetrazolium reduction. The control diet-fed fish showed 70% mortality but the chitin derivative supplemented fishes showed only 20% mortality post-infection. The results of the study encompass that the use of chitin-derivate enriched feed further is taken into large-scale approaches thereby benefitting the aquaculture sector.

Иммунорегулирующий потенциал омега-3 полиненасыщенных жирных кислот: использование для профилактики частой инфекционной заболеваемости у детей

В последние годы все больше исследований направлено на изучение иммуномодулирующей роли длинноцепочечных полиненасыщенных жирных кислот (ДЦПНЖК), практическое применение которых потенциально способно предотвращать развитие острых инфекционных респираторных заболеваний и аллергических состояний. Из множества ДЦПНЖК наибольший интерес вызывают омега-3-кислоты. В статье приведен обзор некоторых исследований, а также данные собственного исследования о влиянии диетической добавки, содержащей комплекс омега-3-кислот («Омегамi Розумна дитина»), на снижение заболеваемости ОРЗ детей с рекуррентными формами острой респираторной инфекционной патологии. Иммунотропное действие диетической добавки к рациону питания «Омегамi Розумна дитина» у детей характеризовалось усилением функционирования врожденного и адаптивного иммунитета, а иммунорегулирующая направленность проявилась в ограничении провоспалительной настроенности иммунной системы у детей, в виде снижения НСТ-теста, количества CD25 и уменьшения сывороточного содержания ФНО-α при отсутствии сопутствующей иммуносупрессии противомикробного защитного потенциала.

Study of in-vitro anti-inflammatory and immunomodulatory effect of Ayurvedic plants – Murva

Background and Aims: Toxic chemicals, pathogens and damaged cells trigger the inflammatory process and cause disease in the kidney, heart, brain, liver, lung and reproductive system. The present study evaluates the anti-inflammatory and immunomodulatory effect of Ayurvedic plants under the Murva category. Methods: Marsdenia tenacissima is an acceptable source of Murva. Sansevieria roxburghiana is considered as Murva in west Bengal. Methanol extracts of both plants were used to evaluate anti-inflammatory and immunomodulatory effects by certain in vitro methods. The in vitro anti-inflammatory effects of methanol extract Marsdenia tenacissima (MEMT) and methanol extract of Sansevieria roxburghiana (MESR) were assessed by heat and hypotonic induced haemolysis on the red blood cells of rats. Nitro blue tetrazolium (NBT) assay and inhibition of TNF-α release in DAL cell lines were conducted to assess the immunomodulatory potential of extracts. Results: MEMT, MESR and a combination of MEMT&amp;MESR showed significant (p&lt;0.001) inhibition of haemolysis on heat and hypotonic induced methods compared with the standard Diclofenac Sodium. In NBT reduction test, MEMT showed a more significant (p&lt;0.001) result than MESR. Similarly, the inhibition of TNF-α release was also significantly enhanced by 400 µg/ mL of MEMT. Conclusion: The present research results revealed that Murva is a safe Ayurveda drug for the treatment of cancer and cardiac disease with the protection of systemic immunity and set free from an inflammatory condition.

Comparative Study of Antioxidant and Pro-Oxidant Properties of Homoleptic and Heteroleptic Copper Complexes with Amino Acids, Dipeptides and 1,10-Phenanthroline: The Quest for Antitumor Compounds.

In a search for new antitumoral agents, a series of homoleptic copper(II) complexes with amino acids and dipeptides, as well as heteroleptic complexes containing both dipeptides and 1,10-phenanthroline, were studied. Furthermore, a single-crystal structure containing alanyl-leucinato ([Cu3(AlaLeu)3(H2O)3(CO3)]·PF6·H2O), which is the first homotrinuclear carbonato-bridged copper(II) complex with a dipeptide moiety, is presented. To assess possible antitumor action mechanisms, we focused on the comparative analysis of pro- and antioxidant behaviors. Pro-oxidant activity, in which the reactive oxygen species (ROS) formed by the reaction of the complexes with H2O2 produce oxidative damage to 2-deoxy-d-ribose, was evaluated using the TBARS method. Additionally, the antioxidant action was quantified through the superoxide dismutase (SOD)-like activity, using a protocol based on the inhibitory effect of SOD on the reduction of nitrobluetetrazolium (NBT) by the superoxide anion generated by the xanthine/xanthine oxidase system. Our findings show that Cu–amino acid complexes are strong ROS producers and moderate SOD mimics. Conversely, Cu–dipeptide–phen complexes are good SOD mimics but poor ROS producers. The activity of Cu–dipeptide complexes was strongly dependent on the dipeptide. A DFT computational analysis revealed that complexes with high SOD-like activity tend to display a large dipole moment and condensed-to-copper charge, softness and LUMO contribution. Moreover, good ROS producers have higher global hardness and copper electrophilicity, lower copper softness and flexible and freely accessible coordination polyhedra.