Enzymes In Tissues Research Articles

Nutritional Components, Biochemical Characteristics, Enzyme Activities, and Growth Differences of Five Freshwater Fish Species?

Common carp (Cyprinus carpio) is an economically important fish worldwide, with many of its species bred for consumption. However, there are few reports on the comprehensive comparative analysis of the muscle nutritional quality and stress resistance characteristics of different common carp species. In this study, after 15 months of feeding in the same environment, the nutritional components, serum biochemical indices, liver antioxidant and intestinal digestive enzyme activities, and muscle growth-related gene expression were determined in Songpu mirror carp (SPM; Cyprinus carpio Songpumirror), Heilongjiang wild carp (HLJ; Cyprinus carpio haematopirus), cold-resistant strain of purse red carp (CPR; Cyprinus carpio ‘Red purse cold-resistant’), Songhe carp (SH; Cyprinus carpio ‘Songhe’), and Songpu carp (SP; Cyprinus carpio Songpu). Muscle nutrient composition showed that HLJ had a significantly lower crude fat content and higher docosahexaenoic acid (DHA) + eicosapentaenoic acid (EPA) proportion than the other four common carp species (p < 0.05). The contents of lysine (Lys) and aspargine (Asp) were significantly higher in the CPR than in other species (p < 0.05). Serum biochemical parameters showed that total protein (TP), total cholesterol (T-CHO), triglycerides (TG), and low-density lipoprotein (LDL) were significantly lower in SPM than in the other species (p < 0.05). The results of tissue enzyme activity showed that the activities of superoxide dismutase (SOD) and catalase from Micrococcus lysodeikticus (CAT) in the liver were significantly higher, while the activities of lipase (LPS), trypsin (TRS), and α-amylase (α-AMS) in the intestine were significantly the lower in HLJ than in the other species (p < 0.05). In addition, the relative expression levels of growth hormone (GH), growth hormone receptor (GHR), insulin-like growth factor 1 (IGF1), insulin-like growth factor receptor (IGF1R), and myoblast determination factor (MyoD) in SP and SH were significantly higher than those in the other species, while the relative expression of myostatin (MSTN) in HLJ was significantly higher (p < 0.05). Therefore, there were significant differences in muscle nutritional quality, serum biochemical indices, liver, and intestinal enzyme activities, and muscle growth potential among the five species of common carp. This study could provide a theoretical basis for the germplasm evaluation and variety improvement of common carp.

Redeployment of Placental Gene Programming: Can Invasive Placentation Molecular Switches Complement the Hallmarks of Cancer?

Placentation arose in mammalian evolution some 150-200 M years ago and integrates in a rather ingenious manner a large number of previously evolved multicellular regulatory pathways which include: angiogenesis, inflammatory cytokines and interleukins, HLA expression, immune peptides, immune regulatory receptors (for NK decidual cells, T and B cells, including Tregs, macrophages, antigen-presenting cells ), endothelial cells and fibroblasts, immune checkpoints (including PD-L1), many paracrine or endocrine hormones and growth factors, tissue enzymes, extracellular vesicles, several different mechanisms for epithelial-mesenchymal transitions, several hypoxia adaptations, morphogenetic changes, and others. Speculating from real genomics and epigenetic data from a single clinical case of a pregnant young woman with breast cancer, it has been recently proposed that mammalian cancer cells do not have to invent “de novo” immune escape mechanisms, including so-called “immune editing”, but to redeploy–probably by epigenetic mechanisms–intrinsic or encrypted gene programmes physiologically used by the process of invasive placentation in mammals. Most of these programmes were not evolved specifically for the placenta–but there were probably some exceptions, such as those still poorly understood key pathways related to foeto-maternal tolerance or perhaps trophoblast differentiation and decidual invasion. In my opinion, invasive placentation molecular switches can complement the hallmarks of cancer, by re-using “placental gene programming” to the competitive advantage of cancer cells.

Oral Administration of Therapeutic Enzyme Capsule for the Management of Inflammatory Bowel Disease.

BackgroundOral administration of proteins/peptides is challenging in clinical application due to their instability and susceptibility in the gastrointestinal tract.Materials and MethodsThe in situ polymerization on the surface of enzymes was used to encapsulate antioxidant enzymes (superoxide dismutase (SOD) and catalase (CAT)) in polymeric shells, and the reactive oxygen species (ROS) scavenging ability was monitored based on DCFH-DA probe using flow cytometry and confocal laser scanning microscopy. The mRNA expression level of pro-inflammatory factors was assessed by real-time qPCR, using lipopolysaccharide-induced RAW264.7 cells as a model. Finally, the enzyme capsules were orally administered for the treatment of inflammatory bowel disease using dextran sodium sulfate (DSS)-induced colitis mice as a model, based on the evaluation of the disease-associated index, ROS level and pro-inflammatory cytokines’ expression.ResultsThe enzyme capsules could effectively scavenge the intracellular reactive oxygen species (ROS) through the cascade catalysis of SOD and CAT, and thus protect the cells from ROS-induced oxidative damage. Meanwhile, the enzyme capsules could inhibit the secretion of pro-inflammatory cytokines from macrophages, thereby achieving favorable anti-inflammation effect. Oral administration of enzyme capsules could facilitate the accumulation of enzymes in the inflamed colon tissues of DSS-induced colitis mice. Moreover, the oral delivery of enzyme capsules could effectively alleviate the symptoms associated with colitis, attributing to the excellent ROS scavenging ability and the inhibition of pro-inflammatory cytokines’ level.ConclusionIn summary, our findings provided a promising approach to construct enzyme-based nano-formulations with favorable therapeutic efficacy and biocompatibility, exhibiting great potential in the treatment of gastrointestinal diseases in an oral administration manner.

Antihepatofibrotic effect of Guizhifuling pill on carbon tetrachloride-induced liver fibrosis in mice.

To evaluate the protective effects and the underlying mechanism of Guizhifuling pill (, GZFL) on carbon tetrachloride (CCl)-induced hepatic fibrosis in mice. Male ICR mice by intraperitoneally administered with 20% CCl (mixed 1∶4 in soybean oil) to induce liver fibrosis. Mice that underwent CCl were orally with GZFL. Using hematoxylin and eosin and Masson staining to examine the pathological changes in liver tissue. Serum biochemical parameters, antioxidant enzyme activity and proinflammatory cytokines was assessed. Nuclear factor-kappaB (NF-κB) pathway and nuclear factor-erythroid 2-related factor 2 (Nrf2) family members were evaluated by Western blotting. Our findings indicated that GZFL could effectively suppress the progression of liver fibrosis in mice, which was determined based on the improvement in liver function and reduction of collagen deposition. GZFL treatment also decreased the level of cytokines and increased the activity of antioxidant enzymes in liver tissue. Moreover, GZFL exerted anti-inflammatory and antioxidant effects through regulating the Nrf2-mediated antioxidant system and inhibiting the NF-κB pathway. GZFL may prevent the progression of liver fibrosis by regulating the Nrf2/ heme oxygenase-1 and NF-κB signaling pathways, thereby highlighting its role in the management of liver fibrosis.

Oxidative Stress Biomarkers Activities and Histological Changes in African Catfish (Clarias gariepinus) Exposed to Carwash Wastewater

The study evaluated the acute toxicity of vehicle wastewater on African catfish (Clarias gariepinus). With a weight range of 250–275.5g and a mean length of 23.5-28.5cm, 150 post-juvenile catfish were purchased from Dalar Kifi Fish Farm BUK New site road, Kano State, and acclimated in a tank for 14 days. Fish were exposed to different vehicle wastewater concentrations of 0.0, 20, 40, 60, and 80% v/v with 5-level exposure concentrations during the acute test (96 hours) in a Completely Randomized Design (CRD) employing renewal bioassay. The acute bioassay showed that the LC50 for the wastewater was 56 percent by volume at 96 hours. Compared to the control, the exposed fish exhibited abnormal swimming, erratic opercular movement, reflex loss, mucus secretion, increased air gulping with an increase in wastewater concentration. Except for water temperature, Biochemical Oxygen Demand (BOD) and pH, the remaining physicochemical parameters were not within the WHO/FAO recommended Standard The order of concentrations of the heavy metal in the wastewater was Cu > Pb > Cd > Cr > Ni. Antioxidant biomarker activities showed that Glutathione reductase (GSH) reduced in the same manner while Catalase (CAT) and Superoxide dismutase (SOD) activities rose considerably (P<0.05) in the liver and gill tissues. Histopathological changes observed in gill tissues were as follows: Hyperplasia, necrosis, primary and secondary lamellae distortion, and epithelial thickening distortion. Hepatic cell breakdown, fatty degeneration, cellular infiltration, localized fibrosis, and cellular necrosis were all visible in the liver tissues. It can be inferred that changes in the histology and activity of the oxidative stress enzymes in fish tissues after exposure to wastewater harmed the experimental fish. As a result of their effects on fish and other non-target aquatic biotas, it is advised that the competent authorities create initiatives to reduce the indiscriminate discharge of untreated vehicle wash wastewater.

Efficient co-isolation of microvascular endothelial cells and satellite cell-derived myoblasts from human skeletal muscle.

Vascularization of tissue-engineered constructs remains a key challenge in the field of skeletal muscle tissue engineering. One strategy for vascularizing organoids is in vitro pre-vascularization, relying on de novo assembly of undifferentiated endothelial cells into capillaries, a process termed vasculogenesis. In most endothelial cell research to date, human umbilical vein endothelial cells have been used primarily because of their availability. Nevertheless, this endothelial cell type is naturally not occurring in skeletal muscle tissue. Since endothelial cells display a tissue-specific phenotype, it is of interest to use muscle-specific microvascular endothelial cells to study pre-vascularization in skeletal muscle tissue engineering research. Thus far, tissue biopsies had to be processed in two separate protocols to obtain cells from the myogenic and the endothelial compartment. Here, we describe a novel, detailed protocol for the co-isolation of human skeletal muscle microvascular endothelial cells and satellite cell-derived myoblasts. It incorporates an automated mechanical and enzymatic tissue dissociation followed by magnetically activated cell sorting based on a combination of endothelial and skeletal muscle cell markers. Qualitative, quantitative, and functional characterization of the obtained cells is described and demonstrated by representative results. The simultaneous isolation of both cell types from the same donor is advantageous in terms of time efficiency. In addition, it may be the only possible method to isolate both cell types as the amount of tissue biopsy is often limited. The isolation of the two cell types is crucial for further studies to elucidate cell crosstalk in health and disease. Furthermore, the use of muscle-specific microvascular endothelial cells allows a shift towards engineering more physiologically relevant functional tissue, with downstream applications including drug screening and regenerative medicine.

Short‐term exposure to 5‰ and 15‰ salinity causes the dynamic changes of the NKA gene, enzyme activities and morphological characteristics in fish tissues of Gymnocypris przewalskii

Salinity affects the physiological activities of aquatic animals and determines the distribution of fish. In this study, we found that the naked carp survived under 0–15‰ salinity, but survival rates decreased to varying degrees under 20‰ and 25‰ salinity, which indicates that the safety threshold of salinity tolerance of Gymnocypris przewalskii is 0–15‰. Next, the response rate of NKAα1 in the gills and kidneys was significantly faster than in the livers, and the response intensity of NKAα1 and NKAα3 has certain tissue specificity under different salinity. NKA enzyme activity first increased, then gradually decreased and increased again in the gills and kidneys after 5‰ and 15‰ salinity stress. In contrast, the NKA enzyme activity gradually decreased and then gradually increased in the liver. Salinity stress also could lead to some pathological phenomena in the cytoplasm. These results indicated that changes in salinity can lead to differential expression of the NKA genes and enzyme in tissues. Also, the dynamic changes of the NKA gene, enzyme activities, and morphological characteristics in fish tissues are in response to the salinity changes in the living environment, thus maintaining a dynamic balance of ions inside and outside the cells.

P10.15.B Next generation drug screening platform: generation of patient-derived cells forex vivo drug response studies of gliomas

Abstract Background Precision oncology aims to provide individual treatment options for each patient. In this regard, ex vivo drug screening systems have the potential to improve clinical outcomes. Traditionally, cancer drugs are tested on long-term cultured cancer cell line models, but cell lines cannot represent an individual patient from a clinic and are biologically too distinct to be informative for drug screening purposes. Drug screening systems of tumor cells usually rely on viability assays and correlations to genomic alterations. Beside genomic alterations, the cellular metabolism is significantly altered during tumor growth, tumor cell proliferation and tumor cell resistance development. Here we aim to establish a drug screening platform using tumor cells derived directly from the individual patient glial tumor, create patient derived tumor cells (PDCs) and combine the outcomes from standardized viability- and genetic-assays with a new developed metabolomics platform. Material and Methods Fresh native tissue from both low- and high-grade glioma are collected. Tumor tissue is used for NMR-based metabolomic analyses and targeted sequencing based genomic analyses as well as PDC isolation using mechanical and enzymatic tissue dissociation. To preserve the original tumor similarity, tissue is short term cultured for two weeks, and PDCs are seeded and treated with a panel of clinical- and preclinical drugs followed by viability assessment, sequencing and metabolomic profiling. Results Culturing of PDCs is successful in ≥85% of patient cases, provided that at least 2 g of tumor are available. The automatized high throughput ex vivo drug response helps to identify potential drug candidates which might become relevant for therapeutic approaches in future. Further, it is possible to distinguish between IDH1-wild type and IDH1-mutant glial tumors based on the metabolomic profile, which is confirmed by immunohistochemical staining and molecular analysis of IDH1 R132H-mutation. Strong metabolomic variations have been identified, including GABA, lactate, and myo-inositol levels between tumor and healthy tissue. Data retrieved by the systematic evaluation is retrospectively associated with the clinical course of the patients. Conclusion Entangling drug screening and genetic assays with metabolomic profiling of glial tumors enriches the information about cellular drug response and paves the way for future clinical studies and better understanding of underlying drug resistance mechanisms in gliomas. Disclosure Funding: K1 COMET Competence Centre CBmed, funded by the Federal Ministry of Transport, Innovation and Technology; the Federal Ministry of Science, Research and Economy, Land Steiermark (Dep. 12, Business and Innovation), the Styrian Business Promotion Agency (SFG), and the Vienna Business Agency. COMET is executed by the Austrian Research Promotion Agency (FFG).

Role of phenylalanine/tyrosine ammonia lyase and anthocyanidin synthase enzymes for anthocyanin biosynthesis in developing Solanum melongena L. genotypes.

Solanum melongena is a widely consumed vegetable crop comprising health-benefiting phenolic compounds. It has a complex network of biosynthetic enzymes involved in synthesizing nutraceuticals, including anthocyanins. The present study was conducted to investigate the activities of key enzymes involved in biosynthesis and accumulation of anthocyanins in developing genotypes, such as phenylalanine ammonia lyase (PAL), tyrosine ammonia lyase (TAL), and anthocyanidin synthase (ANS). As inadequate information is available in this context, fruit and leaf tissues were analyzed for enzyme activities and anthocyanin accumulation. The study included characterization of extracted anthocyanin followed by expression studies for gateway enzyme (ANS) involved in anthocyanin biosynthesis. Delphinidin was a major anthocyanidin present in fruit tissues (1.46-110.49 mg/100 g) of S. melongena. Anthocyanin accumulation is backed up by the correlation between biochemical analysis and expression studies. The study has shown variation for PAL, TAL and ANS enzymes in different tissues at developmental stages. Enzyme activities had a strong positive correlation with anthocyanin biosynthesis.

The Proteolytic Landscape of Ovarian Cancer: Applications in Nanomedicine.

Ovarian cancer (OvCa) is one of the leading causes of mortality globally with an overall 5-year survival of 47%. The predominant subtype of OvCa is epithelial carcinoma, which can be highly aggressive. This review launches with a summary of the clinical features of OvCa, including staging and current techniques for diagnosis and therapy. Further, the important role of proteases in OvCa progression and dissemination is described. Proteases contribute to tumor angiogenesis, remodeling of extracellular matrix, migration and invasion, major processes in OvCa pathology. Multiple proteases, such as metalloproteinases, trypsin, cathepsin and others, are overexpressed in the tumor tissue. Presence of these catabolic enzymes in OvCa tissue can be exploited for improving early diagnosis and therapeutic options in advanced cases. Nanomedicine, being on the interface of molecular and cellular scales, can be designed to be activated by proteases in the OvCa microenvironment. Various types of protease-enabled nanomedicines are described and the studies that focus on their diagnostic, therapeutic and theranostic potential are reviewed.

Cardiac Protection of a Novel Lupane-Type Triterpenoid from Injuries Induced by Hypoxia-Reperfusion.

Myocardial ischemia–reperfusion injury (MIRI) leads to cardiac remodeling and heart failure associated with acute myocardial infarction, which is one of the leading causes of death worldwide. Betulinic acid (BA), a widely distributed lupane-type triterpenoid, has been reported to possess antioxidative activity and inhibit apoptosis in MIRI. Due to the low bioavailability and water insolubility of BA, a previous study found a series of BA-derivative compounds by microbial transformation. In this study, we observe whether there are anti-MIRI effects of BTA07, a BA derivative, on cardiac injuries induced by hypoxia/reoxygenation (H/R) in adult rat cardiomyocytes in vitro and in Langendorff-perfused hearts ex vivo, and further explore its mechanism of cardioprotection to find more efficient BA derivatives. The hemodynamic parameters of isolated hearts were monitored and recorded by a Lab Chart system. The markers of oxidative stress and apoptosis in isolated hearts and adult rat cardiomyocytes (ARCMs) were evaluated. The expression levels of B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X (Bax), protein kinase B (Akt) and phospho-Akt (pAkt, Ser473) induced by H/R were detected via Western blot. The Langendorff experiments showed that BTA07 improves hemodynamic parameters, reduces myocardium damage and infarct size, inhibits levels of myocardial tissue enzymes lactate dehydrogenase (LDH) and creatine kinase (CK) in the coronary outflow and reduces oxidative stress and the activation of caspase-3 in the myocardium. In vitro, BTA07 reduced cell death and caspase-3 activation and inhibited reactive oxygen species (ROS) generation. Furthermore, the protective effects of BTA07 were attenuated by inhibition of the PI3K/Akt signaling pathway with LY294002 in ARCMs. BTA07 protects ARCMs and isolated hearts from hypoxia–reperfusion partly by inhibiting oxidative stress and cardiomyocyte apoptosis.

Biochar amendment reduces cadmium uptake by stimulating cadmium-resistant PGPR in tomato rhizosphere

Biochar amendment in the soil can exert a positive effect in reducing heavy metal toxicity in plants. However, it remains unclear the extent to which this effect is associated with the modulation of plant growth-promoting rhizobacteria (PGPR). Here, we initially conducted a pot experiment using tomato (Solanum lycopersicum L.) as a model plant grown in soil spiked with cadmium. First, we found biochar amendment to result in reduced cadmium uptake in tomato plants and trackable changes in the tomato rhizosphere microbiome. Then, a rhizosphere transplant experiment validated the importance of this microbiome modulation for cadmium-toxicity amelioration. Sequence-based analyses targeted the isolation of representative isolates of PGPR, including Bacillus and Flavisolibacter spp. that displayed in vitro cadmium tolerance and biosorption capabilities (in addition to abilities to solubilize phosphate and produce indole acetic acid). Last, we performed a soil inoculation experiment and confirmed the effectiveness of these isolates in reducing cadmium toxicity in tomato plants. Besides, we found the inoculation of these taxa as single inoculant and in combination to result in increased activities of specific antioxidant enzymes in tomato tissues. Taken together, this study revealed the ecological and physiological mechanisms by which biochar amendment indirectly alleviate cadmium toxicity in tomato plants, in this case, via the modulation and activity of specific PGPR populations. This study provides new insights into strategies able to promote beneficial PGPR in the rhizosphere with potential application to ameliorate heavy metal toxicity in plants.

Bioaccumulation, Detoxification, and Biological Macromolecular Damage of Benzo[a]pyrene in Exposure in Tissues and Subcellular Fractions of Scallop Chlamys farreri.

Because of the persistence and high toxicity of benzo[a]pyrene (B[a]P), the bioaccumulation and detoxification mechanisms of B[a]P have been studied extensively at the tissue level; but the data at the subcellular level in bivalves have not been reported. The present study was conducted to investigate the effects of B[a]P exposure on bioaccumulation, detoxification, and biomacromolecular damage in gills, digestive glands, and their subcellular fractions of the scallop Chlamys farreri. The subcellular fraction contains cytoplasm, mitochondria, microsome, nucleus, cell membrane, and overall organelle. The results demonstrated that B[a]P accumulation showed a clear time-dose effect. Based on the time-dependent accumulation of B[a]P in subcellular fractions, we speculated that the intracellular migration order of B[a]P was cell membrane, organelle, and nucleus in turn. Considering the difference of B[a]P accumulation may be related to B[a]P metabolism, we have further confirmed that the activities of B[a]P metabolizing enzymes in scallop tissues and subcellular fractions were significantly tempted by B[a]P (p < 0.05), including 7-ethoxyresorufin O-deethylase (increased), glutathione-S-transferase (GST; decreased), and superoxide dismutase (increased). First, GST was detected in bivalve cytoplasm and microsome. Second, B[a]P exposure also caused biomacromolecules damage. The results demonstrated that mitochondria and microsome were more vulnerable to lipid peroxidation than cell membrane and nucleus. Taken together, the present study fills some of the gaps in our knowledge of the bioaccumulation and detoxification mechanisms of C. farreri exposed to B[a]P in subcellular fractions and deeply explores the transportation and the main metabolic and damage sites of polycyclic aromatic hydrocarbons (PAHs) in cells, which helped us to comprehensively understand the toxic mechanism of PAHs on bivalves. Environ Toxicol Chem 2022;41:2353-2364. © 2022 SETAC.

Supplementation with Two New Standardized Tea Extracts Prevents the Development of Hypertension in Mice with Metabolic Syndrome.

Hypertension is considered to be both a cardiovascular disease and a risk factor for other cardiovascular diseases, such as coronary ischemia or stroke. In many cases, hypertension occurs in the context of metabolic syndrome (MetS), a condition in which other circumstances such as abdominal obesity, dyslipidemia, and insulin resistance are also present. The high incidence of MetS makes necessary the search for new strategies, ideally of natural origin and with fewer side effects than conventional pharmacological treatments. Among them, the tea plant is a good candidate, as it contains several bioactive compounds such as caffeine, volatile terpenes, organic acids, and polyphenols with positive biological effects. The aim of this study was to assess whether two new standardized tea extracts, one of white tea (WTE) and the other of black and green tea (CTE), exert beneficial effects on the cardiovascular alterations associated with MetS. For this purpose, male C57/BL6J mice were fed a standard diet (Controls), a diet high in fats and sugars (HFHS), HFHS supplemented with 1.6% WTE, or HFHS supplemented with 1.6% CTE for 20 weeks. The chromatography results showed that CTE is more concentrated on gallic acid, xanthines and flavan-3-ols than WTE. In vivo, supplementation with WTE and CTE prevented the development of MetS-associated hypertension through improved endothelial function. This improvement was associated with a lower expression of proinflammatory and prooxidant markers, and—in the case of CTE supplementation—also with a higher expression of antioxidant enzymes in arterial tissue. In conclusion, supplementation with WTE and CTE prevents the development of hypertension in obese mice; as such, they could be an interesting strategy to prevent the cardiovascular disorders associated with MetS.

Protective effects of small heat shock proteins in Daphnia magna against heavy metal exposure

Daphnia magna is one of the most commonly used model organisms to assess toxicity of heavy metal and other xenobiotics. However, the lack of knowledge about important stress-resistant molecules limits our understanding of the alteration of phenotypic and physiological traits of D. magna upon stress exposures. In this study, we focused on a chaperone family of small heat shock protein (sHSP) that has been found in archaea, bacteria and eukaryotes and plays an important role in stress tolerance. A total of eleven sHSP genes (termed DmsHSP1 - DmsHSP11) were identified from the D. magna genome, whose expression profiles during exposure to heavy metal (Cd2+, Cu2+ and Zn2+) and a few other potential pollutants were evaluated via qRT-PCR and RNA-Seq analysis. The results highlighted the predominant role of DmsHSP1 with the highest basal expression level in adults and robust upregulation upon exposure to heavy metals (Cu2+ > Cd2+ > Zn2+). In vivo, recombinant protein rDmsHSP1–21 and rDmsHSP11–12.8 could not only prevent model substrates agglutination induced by heavy metals or reducer dithiotreitol (DTT), but also protect tissue proteins and enzymes from denaturation and inactivation caused by heavy metals or high temperature. Ectopically expression of DmsHSP1–21 or DmsHSP11–12.8 in E. coli conferred host enhanced resistance against various abiotic stresses including Cd2+, Cu2+ and phenazine methosulfate (PMS). Knockdown of DmsHSP1–21 by RNAi, but not for DmsHSP11–12.8, significantly increased the vulnerability of D. magna to heavy metal exposure. Our work provides systematic information on the evolution and function of sHSPs in D. magna and leads to important insights into the mechanisms by which D. magna survive in adverse environments.

Gamma-glutamyl transpeptidase is a promising biological marker of heart failure

Introduction. Currently, the search and study of new biological markers that can help early diagnosis of heart failure, serve as a laboratory tool for assessing the effectiveness of therapy, be a predictive marker of possible adverse clinical outcomes and a significant criterion for risk stratification is very relevant. While cardiospecific markers, including natriuretic peptides, their precursors, and highly sensitive troponins, are widely used in clinical practice, the need to use other markers does not have sufficient evidence. aspect of a biological marker of heart failure.Gamma-glutamyl transpeptidase is an enzyme localized on the outer side of cell membranes and involved in the metabolism of glutathione and cysteine. This enzyme is a dimeric glycoprotein (68 kDa), consisting of 2 subunits – a large and a small (46 and 22 kDa). Gamma-glutamyl transpeptidase is encoded by a multigene family consisting of at least 7 different genes located on chromosome 22; however, only 1 of these genes is involved in the formation of a functional enzyme. Gamma-glutamyl transpeptidase was found in all cells except erythrocytes. There is a significant variability in enzyme activity, which is especially high in tissues with a secretory and absorptive function, such as the kidneys, biliary tract, intestines, and epididymis.Purpose of the review is to present an overview of current publications devoted to the study of γ-glutamyl transpeptidase in the aspect of a biological marker of heart failure.Materials and methods. The analysis of literature sources (foreign and domestic articles) was carried out in the databases: PubMed, RSCI, MedLine, Google Scholar, Science Direct. The search was performed according to the following keywords: biological markers, heart failure, γ-glutamyl transpeptidase, biological markers, heart failure, γ-glutamyl transpeptidase.Results. In addition to its clinical use as a test for liver disease, biliary tract disease, and alcohol abuse, γ-glutamyl transpeptidase is of great interest because of its association with cardiovascular disease, diabetes, metabolic syndrome, and cancer. In the literature available to us, we found a small number of works devoted to the study of γ-glutamyl transpeptidase in patients with heart failure. In the review, we have presented data from experimental and clinical studies indicating a clear link between γ-glutamyl transpeptidase and heart failure. The pathogenetic mechanism of the possible relationship between γ-glutamyl transpeptidase and heart failure is not completely clear. The localization of this enzyme in tissues with a transport function has led to the assumption that it is involved in the transport of amino acids through the γ-glutamyl cycle.Conclusion. Further deeper understanding of the structure and function of the enzyme is needed, as well as future clinical studies to determine the diagnostic, prognostic and possibly therapeutic significance of this biological marker.

A Novel Technique for the Generation of Substantial Numbers of Functional Resident T Cells from Kidney Tissue.

Studying functionality and antigen-specificity of resident kidney T cells derived from a kidney biopsy is hampered by the lack of sufficient numbers of T cells obtained by the standard method of enzymatic tissue dissociation. Enzymatic dissociation of kidney tissue was compared to a novel method of whole kidney tissue culture allowing T cells to migrate into the medium in the presence of exogenous IL-2 and IL-15. T cell numbers were quantified and phenotype of resident T cells (CD69+CD103+/−), TCR Vβ repertoire and functional characteristics were analyzed with multi-parameter flow cytometry. Renal tissue culture for four weeks in the presence of exogenous IL-2 and IL-15 yielded significantly higher numbers of T cells (1.3 × 104/mm3) when compared to cultures without exogenous cytokines (71/mm3) or direct isolation by enzymatic dissociation (662/mm3 T cells, p < 0.05). The proportion of T cells with a resident phenotype did not change in the tissue culture; percentages amounted to 87.2% and 85.1%, respectively. In addition, frequencies of CD4+, CD8+, CD4−CD8−, T cells and MAIT T cells remained similar. For both CD4+ and CD8+, T cells had a more differentiated memory phenotype after tissue culture, but the distribution of TCR Vβ families did not change. In addition, the predominant Th1 cytokine secretion profile and poly-functionality of resident kidney T cell remained intact. T cell proliferation potential was not affected, excluding exhaustion and enrichment of BKV- and CMV-reactive resident T cells was observed. In conclusion, the kidney tissue culture method yields significantly increased numbers of resident T cells without major effects on composition and functionality.

Effects of Soybean Lecithin on Growth Performance, Intestine Morphology, and Liver Tissue Metabolism in Rock Bream (Oplegnathus fasciatus) Larvae

Investigations have demonstrated a strong and positive association between dietary intact phospholipid (PL) inclusion and aquatic larval growth, nevertheless, the precise molecular mechanism underlying PL inclusion on growth performance has not been well elucidated. This study aimed to investigate the effects of dietary soybean lecithin (SL) inclusion on growth performance, liver metabolism, resistance to hypoxia stress, and potential molecular mechanisms in rock bream (Oplegnathus fasciatus) larvae. Four types of equal-protein and equal-lipid content microdiets (MDs) were formulated with graded levels of SL to achieve phospholipid levels of (PLs, dry matter) 3.84% (SL0), 6.71% (SL4), 9.38% (SL8), and 12.21% (SL12). Rock bream larvae (25 days post-hatching) were fed the respective MDs for 30 days with three replicates. We found that dietary SL inclusion promoted growth performance, survival rate, and stress resistance to hypoxia stress. The increased dietary SL inclusion improved intestinal structure, as shown by the increased perimeter ratio, muscular thickness, and mucosal fold height of the mid-intestinal tissue. Moreover, a high SL inclusion diet (SL12) increased the activity of the key lipolysis-related enzyme (lipase [LP]) in liver tissue but decreased the activity of amino acid catabolism-related enzymes (aspartate aminotransferase [AST] and alanine aminotransferase [ALT]). RNA sequencing results in liver tissue revealed that the SL12 diet increased the transcriptional level of fatty acid activation-related genes (acsl6 and acsbg2), phospholipid catabolism-related genes (acat2, lpin2, and crls), and amino acid synthesis-related genes (gs, csb, aldh18a1, and oct), but decreased the expression of amino acid catabolism-related gene gprt2. Notably, the SL12 diet significantly increased the expression of ribosome biogenesis-related genes (pes1, nop56, nop58, and rpf2) in liver tissue. The ribosome protein-related pathways were the most enriched pathways mapped in the GO database. Collectively, this study demonstrated the necessity of dietary SL for survival, growth performance, promotion of mid-intestinal morphology, and hypoxia stress during the rock bream larval stage. The SL-induced growth performance promotion was likely attributed to increasing nutrient acquisition by intestinal morphology improvement and to increasing SL catabolism and thereby sparing amino acids for protein synthesis.

Controlled Release of Therapeutics From Enzyme-Responsive Biomaterials

Numerous promising drug leads are regularly abandoned due to having poor pharmacokinetic profiles. Biomaterials are often used as drug delivery systems to improve the pharmacokinetics of these otherwise promising drug candidates. Hydrogels are a subset of biomaterials that offer porous matrices, permeable to endogenous nutrients in aqueous in vivo environments. Environmentally sensitive hydrogels have become of interest to further tailor these materials to only allow therapeutic release in response to specific environmental cues instead of simple encapsulation and subsequent diffusion. Enzyme-responsive materials allow for the exploitation of endogenous tissue enzyme expression levels and/or altered expression levels during pathological states. The simplest and most common method for stimulus-dependant release is through the destruction of the matrix to release encapsulated therapeutics that would otherwise be trapped indefinitely. A second approach is to covalently attach therapeutics to the hydrogel scaffold and include enzymatically sensitive cross linkages throughout the scaffold backbone. The third, and least common approach, is to use labile linkers between the therapeutic and the scaffold which affords controlled, precise release of the therapeutic with a known molecular structure. These linkers can also be tailored to specific enzymes that are elevated in certain disease states. This review will; 1) briefly describe matrix degradation; 2) present the cleavage of covalently attached therapeutics and; 3) highlight the few examples of targeted cleavage of therapeutics from specific matrix locations and the potential use of these systems in biomedicine.

Characterizing Tissue Enzyme Activities in the Marine Toad (Rhinella marina)

Abstract Plasma biochemistry tests are routine diagnostic tools in veterinary medicine that require an understanding of the enzyme activity profiles of individual organs for accurate interpretation. Tissue enzyme activity has been found to vary among and within taxa and has not been described in the marine toad (Rhinella marina). This study measured enzyme activities in the plasma, liver, heart, skeletal muscle, gastrointestinal system, kidney, and pancreas of five marine toads, to evaluate the tissue specificity of analytes commonly included on plasma biochemical analysis. In the marine toad, alanine transaminase (ALT) and aspartate transaminase activities were not specific to one organ, with minimal ALT activity measured in any organ tissue. Alkaline phosphatase activity was significantly higher in heart tissues and gamma-glutamyltransferase (GGT) activity was significantly higher in pancreatic tissues than in all other organ tissues, and plasma. GGT activity in the kidney was also significantly higher than that found in plasma, but not all other tissues. Amylase and lipase were highest in the plasma. Creatine phosphokinase and lactate dehydrogenase (LDH) were specific to skeletal muscle, with significant LDH activity also found in cardiac tissue. This information will be vital to the interpretation of plasma biochemistry analysis in the marine toad and shows similarities to tissue enzyme activity in the Cuban treefrog (Osteopilus septentrionalis), suggesting extrapolation to other anurans is possible.