High Respiratory Activity Research Articles

СРАВНИТЕЛЬНАЯ ОЦЕНКА ДЕЙСТВИЯ РАЗЛИЧНЫХ БИОУДОБРЕНИЙ В КОМПЛЕКСЕ С ЦЕОЛИТОМ НА ПРОДУКТИВНОСТЬ И МИКРОБИОЦЕНОЗ ГРЕЧИХИ

The research was conducted to assess the effect of biofertilizers on productivity and microbiocenosis of buckwheat. The experiment was conducted in 2023 in a greenhouse in the Republic of Tatarstan on buckwheat of the Nikolskaya variety. The experimental scheme included the following options: without plants (control 1); without fertilizers (control 2); N60P60K60; zeolite from the Tatar-Shatrashan deposit (particle size 0.04 mm) 1 t/ha; soaking seeds in a suspension of a consortium of microorganisms at the rate of 1 l/t per day. preparation; soaking of seeds in suspension of the biological preparation Azolene (Azotobacter vinelandii IB-4) at the rate of 1 l/t of the preparation; consortium of microorganisms + zeolite; Azolene + zeolite. The consortium of microorganisms included strains of nitrogen-fixing (Azotobacter chroococcum and Pseudomonas brassicacearum) and phosphate-mobilizing (Sphingobacterium multivorum and Achromobacter xylosoxidans) rhizobacteria isolated from the soil of Tatarstan, identified and deposited in a 1:1 ratio by suspension weight (bacterial suspension density up to 8,109 CFU/cm3). Buckwheat yields in the variants with zeolite and mineral fertilizer were 1.3 and 1.8 times higher than in the control, respectively. The maximum weight of fruits in the experiment was noted in variants with a consortium of microorganisms in pure form and together with zeolite - 2.5 and 2.9 times higher than in the control. Biofertilizers, NPK and zeolite can be arranged in the following order according to their effectiveness: consortium of microorganisms > consortium of microorganisms + zeolite > NPK > zeolite. The use of Azolene did not lead to an increase in buckwheat yield compared to the control. During the growing season, in variants with a consortium of microorganisms, not only the best representation of agronomically significant microorganisms (ammonifiers, diazotrophs, phosphate mobilizers, actinobacteria) was noted, but also a high respiratory activity of the soil microbiota. A consortium based on autochthonous nitrogen-fixing and phosphate-mobilizing bacteria can serve as the basis for the creation of an integrated biofertilizer for a wide range of crops.

Abstract B070: Myc synthetic lethality as a therapeutic strategy for resistance: Insights into the targetability of mitochondrial complex I in Myc-high TNBC

Abstract Patients with advanced triple-negative breast cancer (TNBC) frequently develop chemoresistance, emphasizing an urgent need for novel therapeutic strategies able to eradicate treatment resistant tumor cells. Recent studies have highlighted MYC as a common oncogenic alteration and a driver of stem-like TNBC phenotype, which commonly features high mitochondrial respiratory activity instated through OXPHOS system and chemoresistance. Interestingly, emerging evidence suggests that residual tumors from chemo treatment remain susceptible to OXPHOS inhibition through inhibition of respiratory complex I. Our previous studies have revealed a specific MYC-dependent and tumor suppressive synthetic lethal action for combination of metformin and BH3 mimetics in cells, ex vivo and in vivo models of aggressive breast cancer (PMID: 30728358). Metformin is a systemic and local metabolic modulator, which acts as a weak complex I inhibitor and promoter of AMPK activation. Here, we have explored the role of MYC in OXPHOS regulation, to address the question if metformin specifically and lethally targets MYC-modulated OXPHOS in MYC-high TNBC. We found that a selective c-MYC inhibitor (EN4) leads to a significant decrease in complex I activity despite only a modest drop in complex I mass in endogenously MYC-high TNBC cell lines. Conversely, engineered MYC overexpression in these cells upregulated complex I activity. Our current work explores transcriptomic, proteomic, and functional approaches to the mechanisms through which MYC regulates complex I activity, structure, and dynamics, ultimately hoping to define a biological rationale for therapeutic targeting of OXPHOS in Myc-high TNBC. Citation Format: Antti O Hiltunen, Linda J Id, Juha Klefström. Myc synthetic lethality as a therapeutic strategy for resistance: Insights into the targetability of mitochondrial complex I in Myc-high TNBC [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Breast Cancer Research; 2023 Oct 19-22; San Diego, California. Philadelphia (PA): AACR; Cancer Res 2024;84(3 Suppl_1):Abstract nr B070.

Higher impact resistance and lower antibiotic resistance genes proliferation rate in shortcut nitrification-denitrification process under long-term antibiotics selection pressure

Shortcut nitrification–denitrification (SCND) has been widely concerned due to its high nitrogen removal efficiency and low operating cost. However, little information is available on the fate of antibiotic resistance genes (ARGs) in SCND process. This study investigates the dynamics of nitrogen removal and ARGs between SCND and full nitrification–denitrification (FND) under long-term selective pressure conditions of tetracycline and sulfadiazine. The study explores the response patterns of extracellular polymers, microbial activity, and microbial community. Results show that the SCND process exhibits 9.14% higher extracellular polymers and 119.07% higher respiratory activity than FND process, resulting in better TN removal performance (67.16% vs. 49.24%) and stronger impact resistance. Compared to Phase I (0 µg/L) and Phase II (100 µg/L), the relative abundance of total ARGs experienced a sharp increase in Phase III (500 µg/L). The SCND process maintains a lower total ARGs abundance than the FND process in water phase. Specifically, in phase III, the relative abundance of intI1 and ARGs in the FND process are 0.04–2.1 logs and 0.66–1.60 logs higher than that of the SCND process in sludge and water phases, respectively. More denitrifying microbes (>40%) are enriched in the SCND process, while it changes to hydrolytic acidification bacteria (>20%) in the FND process. The Partial Least Squares Path Modeling reveals that microbial community and microbial activity play a crucial role in ARGs transmission, with microbial activity exerting greater influence. Additionally, potential pathogenic bacteria including Clostridium, Gemnobacter, Klebsiella, Enternobacter, Aeromonas, Burkholderia are identified, and SCND over FND process shows better inhibition effect on potential pathogenic bacteria proliferation (0.022% vs 0.080%). This study provides new insights into removing traditional pollutants and ARGs simultaneously from wastewater treatment system.

Recent advances in quality preservation of postharvest golden needle mushroom (Flammulina velutiper).

The golden needle mushroom (Flammulina velutiper) is one of the most productive mushrooms in the world. However, F. velutiper experiences continuous quality degradation in terms of changes in color and textural characteristics, loss of moisture, nutrition and flavor, and increased microbial populations due to its high respiratory activity during the postharvest phase. Postharvest preservation techniques, including physical, chemical and biological methods, play a vital role in maintaining postharvest quality and extending the shelf life of mushrooms. Therefore, in this study, the decay process of F. velutiper and the factors affecting its quality were comprehensively reviewed. Additionally, the preservation methods (e.g., low-temperature storage, packaging, plasma treatment, antimicrobial cleaning and 1-methylcyclopropene treatment) for F. velutiper used for the last 5 years were compared to provide an outlook on future research directions. Overall, this review aims to provide a reference for developing novel, green and safe preservation techniques for F. velutiper. © 2023 Society of Chemical Industry.

Age-related ultrastructural changes in spheroids of the adipose-derived multipotent mesenchymal stromal cells from ovariectomized mice.

Introduction: Adipose-derived multipotent mesenchymal stromal cells (ADSCs) are widely used for cell therapy, in particular for the treatment of diseases of the nervous system. An important issue is to predict the effectiveness and safety of such cell transplants, considering disorders of adipose tissue under age-related dysfunction of sex hormones production. The study aimed to investigate the ultrastructural characteristics of 3D spheroids formed by ADSCs of ovariectomized mice of different ages compared to age-matched controls. Methods: ADSCs were obtained from female CBA/Ca mice randomly divided into four groups: CtrlY-control young (2 months) mice, CtrlO-control old (14 months) mice, OVxY-ovariectomized young mice, and OVxO-ovariectomized old mice of the same age. 3D spheroids were formed by micromass technique for 12-14 days and their ultrastructural characteristics were estimated by transmission electron microscopy. Results and Discussion: The electron microscopy analysis of spheroids from CtrlY animals revealed that ADSCs formed a culture of more or less homogeneous in size multicellular structures. The cytoplasm of these ADSCs had a granular appearance due to being rich in free ribosomes and polysomes, indicating active protein synthesis. Extended electron-dense mitochondria with a regular cristae structure and a predominant condensed matrix were observed in ADSCs from CtrlY group, which could indicate high respiratory activity. At the same time, ADSCs from CtrlO group formed a culture of heterogeneous in size spheroids. In ADSCs from CtrlO group, the mitochondrial population was heterogeneous, a significant part was represented by more round structures. This may indicate an increase in mitochondrial fission and/or an impairment of the fusion. Significantly fewer polysomes were observed in the cytoplasm of ADSCs from CtrlO group, indicating low protein synthetic activity. The cytoplasm of ADSCs in spheroids from old mice had significantly increased amounts of lipid droplets compared to cells obtained from young animals. Also, an increase in the number of lipid droplets in the cytoplasm of ADSCs was observed in both the group of young and old ovariectomized mice compared with control animals of the same age. Together, our data indicate the negative impact of aging on the ultrastructural characteristics of 3D spheroids formed by ADSCs. Our findings are particularly promising in the context of potential therapeutic applications of ADSCs for the treatment of diseases of the nervous system.

Study of the effect of protein synthesis inhibitors on growing <i>Escherichia coli</i> bacteria using electrochemical sensors

Background. The study of the mechanisms of action of antibiotics requires the integrated use of traditional microbiological and physicochemical methods. The aim. To study the response of Escherichia coli bacteria to the action of four antibiotics, inhibitors of protein synthesis, using combined approach. Methods. Bacteria were grown under aerobic conditions on minimal M9 medium with glucose. Tetracycline, kanamycin, streptomycin and chloramphenicol have been tested. The effect of antibiotics on survival (CFU) and growth rate was determined. Respiratory activity, sulfide production, extracellular potassium, as well as pH and Eh of the medium were measured using electrochemical sensors directly in a growing culture in the “real time”, membrane potential was measured using a DiBAC dye and a Leica DM2000 fluorescent microscope. Results. The tested antibiotics were divided into two groups according to their properties. Tetracycline and chloramphenicol showed a pronounced bacteriostatic effect, growth inhibition began immediately after the addition of antibiotics and occurred at a high rate. Both antibiotics inhibited respiration, stimulated sulfide production and an Eh jump. Bacteria treated with tetracycline and chloramphenicol retained the ability to maintain membrane potential and intracellular potassium better. Inhibition of respiration led to a decrease in glucose catabolism, as evidenced by a lower rate of acidification of the medium compared to the control. Growth inhibition with streptomycin and kanamycin was initiated with a 30-minute delay. Both antibiotics showed a bactericidal effect, did not stimulate sulfide production and Eh jump, did not inhibit respiration, but caused a drop in membrane potential and intracellular potassium. High respiratory activity promoted glucose catabolism, as evidenced by the rapid acidification of the medium. Of interest is the detection of kanamycin-induced sulfide production during E. coli growth on MOPS medium. Conclusion. An analysis of the data obtained indicates that the use of electrochemical sensors in combination with traditional methods is a promising approach to studying the mechanisms of action of antibiotics.

2-Deoxyglucose, an Inhibitor of Glycolysis, Enhances the Oncolytic Effect of Coxsackievirus.

Glioblastoma multiforme (GBM) is one of the most common types of brain tumor. Despite intensive research, patients with GBM have a poor prognosis due to a very high rate of relapse and significant side effects of the treatment, with a median survival of 14.6 months. Oncolytic viruses are considered a promising strategy to eliminate GBM and other types of cancer, and several viruses have already been introduced into clinical practice. However, identification of the factors that underly the sensitivity of tumor species to oncolytic viruses or that modulate their clinical efficacy remains an important target. Here, we show that Coxsackievirus B5 (CVB5) demonstrates high oncolytic potential towards GBM primary cell species and cell lines. Moreover, 2-deoxyglucose (2DG), an inhibitor of glycolysis, potentiates the cytopathic effects of CVB5 in most of the cancer cell lines tested. The cells in which the inhibition of glycolysis enhanced oncolysis are characterized by high mitochondrial respiratory activity and glycolytic capacity, as determined by Seahorse analysis. Thus, 2-deoxyglucose and other analogs should be considered as adjuvants for oncolytic therapy of glioblastoma multiforme.

Sulforaphane diminishes moonlighting of pyruvate kinase M2 and interleukin 1β expression in M1 (LPS) macrophages.

Murine macrophages activated by the Toll-like receptor 4 agonist lipopolysaccharide (LPS) polarize to the M1 type by inducing proinflammatory marker proteins and changing their energy metabolism to increased aerobic glycolysis and reduced respiration. We here show that the aliphatic isothiocyanate sulforaphane (Sfn) diminishes M1 marker expression (IL-1β, IL-6, TNF-α, iNOS, NO, and ROS) and leads to highly energetic cells characterized by both high glycolytic and high respiratory activity as assessed by extracellular flux analysis. Focusing on a potential connection between high glycolytic activity and low IL-1β expression in M1 (LPS/Sfn) macrophages, we reveal that Sfn impedes the moonlighting function of pyruvate kinase M2 (PKM2) in M1 macrophages. Sfn limits mono/dimerization and nuclear residence of PKM2 accompanied by reduced HIF-1α levels, Stat3 phosphorylation at tyrosine 705, and IL-1β expression while preserving high levels of cytosolic PKM2 tetramer with high glycolytic enzyme activity. Sfn prevents glutathionylation of PKM2 in LPS-stimulated macrophages which may account for the reduced loss of PKM2 tetramer. Overall, we uncover PKM2 as a novel affected hub within the anti-inflammatory activity profile of Sfn.

Iron availability enhances the cellular energetics of aerobic Escherichia coli cultures while upregulating anaerobic respiratory chains

Aerobic Escherichia coli growth at restricted iron concentrations (≤ 1.75 ± 0.04 μM) is characterized by lower biomass yield, higher acetate accumulation and higher activation of the siderophore iron-acquisition systems. Although iron homeostasis in E. coli has been studied intensively, previous studies focused only on understanding the regulation of the iron import systems and the iron-requiring enzymes. Here, the effect of iron availability on the energy metabolism of E. coli has been investigated. It was established that aerobic cultures growing under limiting iron conditions showed lower ATP yield per glucose, lower growth rate and lower TCA cycle activity and respiration, at the same time as increased glucose consumption, acetate and pyruvate accumulation, practically mimicking microaerobic growth. However, at excess iron, independent of oxygen availability, the cultures showed high cellular energetics (5.8 ATP/mol of glucose) by using pathways requiring iron-rich complex proteins found in the TCA cycle and respiratory chain. In conditions of iron excess, some iron-requiring terminal reductases of the respiratory chain, that were thought to function only under anaerobiosis, were used by the E. coli, when in aerobic conditions, to maintain high respiratory activity. This allowed it to produce more biomass and more reactive oxygen species that were controlled by the higher activity of the antioxidant defenses (SOD, peroxidase and catalase) and the iron-sulfur cluster repair systems.

Respiratory system as the main determinant of dyspnea in patients with pulmonary hypertension.

Dyspnea on exertion is a devastating symptom, commonly observed in patients with pulmonary hypertension (PH). The pathophysiology of dyspnea in these patients has been mainly attributed to cardiovascular determinants and isolated abnormalities of the respiratory system during exercise, neglecting the contribution of the control of the breathing system. The aim of this review is to provide a novel approach to the interpretation of dyspnea in patients with PH, focused on the impact of the control of the breathing system during exercise. Exercise through multiple mechanisms affects the (1) ventilatory demands, as dictated by respiratory center activity, (2) actual ventilation, and (3) metabolic hyperbola. In patients with PH, exertional dyspnea can be explained by exercise‐induced alterations in these variables. Compared to healthy subjects, at a given CO2 production during exercise, ventilatory demands in patients with PH are higher due to metabolic acidosis (early reaching the anaerobic threshold), hypoxemia, and excessive upward movement of metabolic hyperbola owing to abnormal exercise response of dead space to tidal volume ratio. Simultaneously, dynamic hyperinflation and respiratory muscles weakness decreases the actual ventilation for a given respiratory center activity, creating a dissociation between demands and ventilation. Consequently, a progressive increase in ventilatory demands and respiratory center activity occurs during exercise. The forebrain projection of high respiratory center activity causes exertional dyspnea despite the relatively low ventilation and significant ventilatory reserve. This type of analysis suggests that the respiratory system is the main determinant of exertional dyspnea in patients with PH, with the cardiovascular system being an indirect contributor.

Different dietary protein levels for Podocnemis unifilis subadult farming: hematological and biochemical assessment

ABSTRACT This study evaluated the physiological responses of subadult yellow-spotted Amazon river turtle, Podocnemis unifilis, to incremental crude protein levels (29%, 32%, 35%, and 39% CP) as extruded commercial feeds. The hematological and plasma biochemical parameters of P. unifilis were analyzed at 60, 120, and 180 days of the feeding trial. The increase in total thrombocyte, leukocyte, lymphocyte, heterophil, and eosinophil numbers, besides high respiratory burst activity showed an improvement in animal immune defense response to incremental protein in diets, acting together to maintain the integrity of the tissues against antigens and infectious agents. Elevated subadult P. unifilis plasma total cholesterol and triglyceride levels observed after increasing the feeding time (until 180 days) with incremented protein levels in artificial diets during captivity are a nutritional warning, related to the condition of being less physically active. Based on these results, periodic physiological evaluations are particularly important to ensure the healthy and adequate nutritional conditions of captive-bred animals, such as maintaining lower stocking densities of animals to avoid stress and, if possible, providing them with fresh food in addition to extruded artificial feed.

Synthesis of Silymarin-Gold Nanoparticle Conjugate and Analysis of its Liver-Protecting Activity.

The liver disease problem prompts investigators to search for new methods of liver treatment. Silymarin (Sil) protects the liver by reducing the concentration of free radicals and the extent of damage to the cell membranes. A particularly interesting method to increase the bioavailability of Sil is to use synthesized gold nanoparticles (AuNPs) as reagents. The study considered whether it was possible to use the silymarin-AuNP conjugate as a potential liver-protecting drug. AuNPs were conjugated to Sil and the liver-protecting activity of the conjugate was examined. Experimental hepatitis and hepatocyte cytolysis after carbon tetrachloride action were used as a model system, and the experiments were conducted with laboratory animals. For the first time, silymarin was conjugated to colloidal gold nanoparticles (AuNPs). Electron microscopy showed that the resultant preparations were monodisperse and that the mean conjugate diameter was 18-30 nm ± 0.5 nm (mean diameter of the native nanoparticles, 15 ± 0.5 nm). In experimental hepatitis in mice, conjugate administration interfered with glutathione depletion in hepatocytes in response to carbon tetrachloride. It also was conducive to an increase in energy metabolism and stimulated the monocyte-macrophage function of the liver. The results were confirmed by the high respiratory activity of the hepatocytes in cell culture. We conclude that the silymarin-AuNP conjugate holds promise as a liver-protecting agent in acute liver disease caused by carbon tetrachloride poisoning.

Adaptation of Fish Metabolism to Thermal Pollution of Subarctic Lake Imandra

Kola nuclear power plant, which discharges warm water into one of the bays of sub-arctic Lake Imandra, significantly changes the fish habitat, creating temperature gradient from 2 to 8°C compared to water temperature in the other part of the lake. The permeation, metabolism and bioaccumulation of elements in the stenothermal cold-water species (Coregonus lavaretus L.) in warm water impact zone are characterized. One of the main adaptation mechanisms of ionic regulation is the intense metabolism of Na due to the high respiratory activity of whitefish in warmer water. This feature is associated with elevated accumulation of Rb and loss of Se, Mo, and Si. At elevated temperatures, the main metabolic need is caused by Se deficiency. An extensive metabolism of selenoproteins involves risks of toxic effects and bioaccumulation of Hg, As and Cu with an increase of existing stressor or appearance of new ones.

Fine-Root Traits Reveal Contrasting Ecological Strategies in European Beech and Norway Spruce During Extreme Drought.

Trees adjust multiple structural and functional organ-specific characteristics, “traits”, to cope with diverse soil conditions. Studies on traits are widely used to uncover ecological species adaptability to varying environments. However, fine-root traits are rarely studied for methodological reasons. We analyzed the adaptability of the fine-root systems of European beech and Norway spruce to extreme drought within species-specific tree groups at Kranzberger Forst (Germany), focusing on the seasonality of morphological, physiological, and biochemical key traits in view of carbon (C) and nitrogen dynamics. We hypothesized that fine roots of both species adjust to seasonal drought: with beech representing a “fast” (i.e. with fast C turnover), and spruce a “slow” (i.e. with long-term C retention) ecological strategy. We identified three functional fine-root categories, based on root function (absorptive or transport fine roots), and mycorrhizal status of the absorptive fine-roots (mycorrhizal or non-mycorrhizal). Solely the non-mycorrhizal absorptive roots adjusted in a species-specific manner supporting fine-root ecological strategy hypothesis. During drought, beech produced thin ephemeral (absorptive non-mycorrhizal) fine roots with high specific fine-root area and high respiratory activity, representing fast C turnover and enabling effective resource exploitation. These adjustments reflect a “fast” ecological strategy. Conversely, spruce absorptive fine roots did not respond to the soil moisture deficit by growth but instead increased root suberization. Drastically lowered respiratory activity of this functional category facilitated C retention and structural persistence during drought, indicating a “slow” ecological strategy in spruce. Absorptive mycorrhizal fine roots maintained respiration throughout the drought event in both tree species, but in spruce this was the only fine-root category with high respiration. This suggests, that spruce relies heavily on mycorrhizal associations as a method of drought resistance. Accumulation of non-structural carbohydrates and high C concentrations were observed in the transport fine roots of both species, indicating drought-induced osmotic protection of these roots. Thus, functional classification enabled us to determine that fine-root branches of each species are not tied to one sole ecological strategy. The suggested approach helps to better understand the complex interplay between structure and function belowground.

Influence of Thermal Pollution on the Physiological Conditions and Bioaccumulation of Metals, Metalloids, and Trace Metals in Whitefish (Coregonus lavaretus L.).

The Kola nuclear power plant, which discharges warm water into one of the bays of subarctic Lake Imandra, significantly changes fish habitats. The temperature gradient of the lake is between 2 and 8 °C, which makes it significantly different from the natural temperature of the lake water. The stenothermal cold-water native species (lake whitefish (Coregonus lavaretus L.)), living for more than 40 years under conditions of thermal pollution, has adapted to this stressor. Moreover, this population differs favorably from the population in the natural-temperature environment in terms of its physiological state. Firstly, the hemoglobin concentrations in the fish blood are in the range of the ecological optimum, and secondly, it has a higher somatic growth, as estimated by Fulton’s condition factor. One of its main adaptive mechanisms of ion regulation is an intense metabolism of Na due to the high respiratory activity of the whitefish in warmer water. An increased accumulation of Rb and excretion of Se, Mo, and Si are associated more or less with that feature. Under conditions of an increased water temperature, the main metabolic need is due to a deficiency of Se in fish. The intensive metabolism of selenoproteins may involve risks of toxic effects and the bioaccumulation of Hg, As, and Cu in cases of increased existing stressors or the appearance of new ones.

Oil-addicted biodegradation of macro-alkanes in soils with activator

Three biodegradation were carried out in soils with activator, non-activator and no activator. For oil-addicted biodegradation, macro-alkanes were devoured (soil S1:3313 mg C18-C30/kg, soil S2: 2741 mg C18-C30/kg) in soils with activator, which was 3.8 times of that for non-oil-addicted biodegradation. For oil-addicted biodegradation, high respiratory activity (S1: 0.8 CO2/kg soil, S2: 0.6 mg CO2/kg soil) and rapid growth rate (S1: 48 %, S2: 36 %) of oil-addicted microbe with activator were observed. Moreover, for oil-addicted biodegradation, low consumption of NH4+-N (22 mg/kg) and acetic acid (410 mg/kg) were observed, which were 1/8 and 4/5 for non-oil-addicted biodegradation. Meanwhile, rapid growth and high respiratory activity of the genera Pseudomonas, Diaminobutyricimonas, Dyadobacter and Galbitalea were observed. It indicated that macro-alkanes instead of organic matter such as aromatic protein I and II, fulvic acid-like, soluble microbial metabolites, humic acid-like was utilized rapidly for oil-addicted biodegradation due to less consumption of NH4+-N and acetic acid.

Respiration and Heat Shock Protein After Short-Term Heating/Stretch-Fixing on Smooth Muscle Cells.

A treatment device without a stent is needed for peripheral stenotic artery treatment. We have proposed short-term heating balloon angioplasty, photo-thermo dynamic balloon angioplasty (PTDBA). Though smooth muscle cells (SMCs) after PTDBA are fixed in a stretched formation in a porcine model, influences of this stimulus on SMCs have not been investigated. SMC migration after vascular dilatation would be related to chronic restenosis. The aim of this study was to examine respiratory activity and recovery ability of SMCs after short-term heating/stretch-fixing in vitro for chronic phase treatment effect discussion. SMCs on a stretch chamber were heated for 15s with stretching and fixed in a stretched formation. SMC migration is correlated with the cell respiratory activity. The amount of ATP production was measured using a WST-8 assay for respiratory activity evaluation. The intracellular expression of heat shock protein 70 was measured by an ELISA for recovery ability evaluation. In the case of 60°C heating, SMC respiratory activity after short-term heating/stretch-fixing decreased drastically in all stretching rates. In the case of 50°C heating, SMC respiratory activity decreased and then increased. Alternatively, the recovery ability at 60°C was greater than that at 50°C. SMCs heated at 60°C with stretching would have high recovery ability and low respiratory activity related to SMC migration. These results may be important evidence in determining the treatment condition in PTDBA.

Evaluation the Ozone Treatment To Control the Infection of Saprolegniasis in Cyprinus carpio L.

The objective of present study is to set up an alternative method of controlling of saprolegniasis, in Cyprinus carpio L. using ozone. For this purpose, six different treatments were applied of 120 fish (20 fish/treatment), of which three infected with Saprolegnia spp. (2×104 zoospors/l) and were treated with ozone (O3) in concentrations of 0.25, 0.50 and 0.75 mg l−1 (1hour), one reference treatment with formalin (0.15 ml l−1, 30 min), two treatments one as positive control (infected with Saprolegnia spp. without disinfectants) and one as negative control without any disinfectants. After 14 days of treatment with ozone and formalin, blood samples were collected from fish for hematological, biochemical and Immunological tests. Survival rate was also studied. Considerable changes have been recorded in the mean values of blood indices. RBCs count and Hemoglobin content, were found significantly increased (p<0.05) in all ozone treatments compared to C- group. Total protein and globulin were significantly increased (p<0.05) in all ozone and formalin treatments in comparison to positive and negative groups (C- and C+). Albumin showed a significant decrease (p<0.05) in all ozone and formalin treatments relative to C+ and C-group. The results of respiratory burst activity (measured by Nitro blue tetrazolium reduction assay) showed significantly different (p<0.05) in all ozone and formalin treatments respectively compared to control groups (C+ and C-); Highest respiratory activity was observed in ozone treatment at 0.50 mg/l. Among Ozone treatments 0.50 mg l−1 showed the highest survival rate (90%) followed by formalin treatment (80%). Survival rate of the control group (without disinfectant) was 20% and there was a significant difference (P<0.05) with other treatments. It was obvious that the obtained results were highly suggestive for the efficiency of ozone as an efficient antifungal disinfectant for fish.

A constitutive active allele of the transcription factor Msn2 mimicking low PKA activity dictates metabolic remodeling in yeast.

In yeast, protein kinase A (PKA) adjusts transcriptional profiles, metabolic rates, and cell growth in accord with carbon source availability. PKA affects gene expression mostly via the transcription factors Msn2 and Msn4, two key regulators of the environmental stress response. Here we analyze the role of the PKA-Msn2 signaling module using an Msn2 allele that harbors serine-to-alanine substitutions at six functionally important PKA motifs (Msn2A6). Expression of Msn2A6 mimics low PKA activity, entails a transcription profile similar to that of respiring cells, and prevents formation of colonies on glucose-containing medium. Furthermore, Msn2A6 leads to high oxygen consumption and hence high respiratory activity. Substantially increased intracellular concentrations of several carbon metabolites, such as trehalose, point to a metabolic adjustment similar to diauxic shift. This partial metabolic switch is the likely cause for the slow-growth phenotype in the presence of glucose. Consistently, Msn2A6 expression does not interfere with growth on ethanol and tolerated is to a limited degree in deletion mutant strains with a gene expression signature corresponding to nonfermentative growth. We propose that the lethality observed in mutants with hampered PKA activity resides in metabolic reprogramming that is initiated by Msn2 hyperactivity.

Intergeneric Hybridization between Streptomyces albulus and Bacillus subtilis Facilitates Production of ε-Poly-L-lysine from Corn Starch Residues

Intergeneric hybridization between S.albulus and B. subtilis to produce e-poly-L-lysine (e-PL) from corn starch residues (CSR) was investigated in this study. One hybrid, designated S. albulus LS-84, which incorporated the protease gene from B. subtilis, could effectively utilize the protein in CSR as a nitrogen source. In fed-batch fermentation, LS-84 produced 32.6 g/L e-PL in the presence of 20 g/L CSR. This was an increase of 256.1% compared to that of the parent strain S. albulus LS-01. The rapid hydrolysis of CSR by protease caused rapid growth for LS-84, which allowed higher respiratory activity. As a result, activities of several key enzymes in LS-84 were higher than those in LS-01; additionally, the content of several intracellular amino acids, such as Asp, Glu, and Arg, was also much higher in LS-84. Therefore, intergeneric hybridization between S. albulus and B. subtilis to produce e-PL from CSR is an economical method for effective utilization of waste resources.