Enzymes In Tissues Research Articles

A study to assess the health effects of an anticancer drug (cyclophosphamide) in zebrafish (Danio rerio): eco-toxicity of emerging contaminants.

Cyclophosphamide (CP) is widely used for treating various kinds of cancer. Because of its high intake, metabolism and excretion, these anticancer medications have been detected in the aquatic environment. There is very limited data on the toxicity and effects of CP on aquatic organisms. The present study aims to assess the toxic effect of CP on certain oxidative stress biomarkers (superoxide dismutase-SOD, catalase-CAT, glutathione peroxidase-GPx, glutathione-GSH, glutathione S-transferases-GST and lipid peroxidation-LPO), protein, glucose, metabolising enzymes (aspartate aminotransferase-AST, alanine aminotransferase-ALT), and ion-regulatory markers (sodium ions-Na+, potassium ions-K+, and chloride ions-Cl-), and histology in the gills and liver of Danio rerio at environmentally relevant concentrations (10, 100 and 1000 ng L-1). Exposure to CP for 42 days led to a significant decrease in SOD, CAT, GST, GPx and GSH levels in the gills and liver tissues of zebrafish. The level of lipid peroxidation in the gills and liver tissues of zebrafish was significantly increased compared to the control group. Chronic exposure significantly changes protein, glucose, AST, ALT, Na+, K+ and Cl- biomarkers. Fish exposed to different levels of CP showed necrosis, inflammation, degeneration and hemorrhage in the gills and hepatic tissues. The observed changes in the studied tissue biomarkers were proportional to both dose and time. In conclusion, CP at environmentally relevant concentrations causes oxidative stress, energy demand, homeostasis disturbances, and enzyme and histological alterations in the vital tissues of zebrafish. These alterations were similar to the toxic effects reported in mammalian models.

The effect of conazoles on reproductive organs structure and function – a review

Conazoles are azole antifungals used in agricultural and pharmaceutical products. Exposure to conazole fungicides leads to several toxic endpoints, including reproductive and endocrine. The results of animal experiments have shown that various conazole fungicides at high doses affect the structure and functions of reproductive organs. In males, adverse effects of conazole fungicides are manifested in the testes, prostate, sperm viability, fertility and sexual behaviour. Reduced testis weight, testis atrophy and reduced or absent sperm production were frequently observed. In female genitalia, structural changes in the ovaries and uterus have been observed. The extent of the changes depends on the dose and duration of treatment. Triazoles affected the expression of multiple genes involved in steroid hormone metabolism and modulate enzyme activity of multiple cytochrome P450 (CYP) and other metabolic enzymes in mammalian liver and other tissues. Conazole fungicides act as endocrine disruptors. Conazoles have been reported to reduce oestradiol and testosterone production and to increase progesterone concentration, indicating the inhibition of enzymes involved in the conversion of progesterone to testosterone. The reproductive effects are consistent with impairment of testosterone homeostasis. The disruption in steroid homeostasis is a common mode of action, leading to abnormal reproductive development and diminished reproductive function. At high doses, azole fungicides affect reproductive organs and fertility in several species.

The effect of ascorbic acid on histopathological, biochemical, pharmacological, and immunological toxicity of chronic lead acetate exposure on the spleen in a rat model

Objective To evaluate the effect of vitamin C on histopathological, biochemical, and immunotoxicity of chronic lead exposure in the spleen of a rat model.Methods The rats were divided into five groups of 10 rats each: group I received normal saline orally as a control group; groups II and III received lead acetate for 4 and 8 weeks, respectively; and groups IV and V received lead acetate and vitamin C for 4 and 8 weeks, respectively. The spleen was excised and processed for light, electron microscopic, histopathological, and biochemical analyses. Quantitative assessments of matrix metalloproteinase-2 (MMP-2), MMP-9, interleukin-2 (IL-2), IL-6, and tumor necrosis factor-alpha gene expressions were performed by real-time PCR.Results The examination of control and vitamin C with lead acetate supplemented groups revealed normal splenic architecture. In contrast, the spleen of lead-intoxicated groups exhibited degenerative changes in the spleen, with a significantly decreased expression of IL-2, glutathione peroxidase, superoxide dismutase, and hemoglobin (P<0.05), with significantly increased proinflammatory cytokine (IL-6 and tumor necrosis factor-alpha) expressions, concomitantly with increased oxidative products (malondialdehyde) and protease enzymes (MMP-2 and MMP-9) in the spleen tissues. The coadministration of vitamin C with lead for 4 weeks markedly resolved these changes.Conclusion This study may specify the efficiency of vitamin C in lead toxicity prevention in the spleen, represented by the reduced splenic harmful changes produced by lead administration.

The Effect Of Continuously Dutasteride Monotherapy on The Expression Of Protein Kinase C-Alpha Enzyme In BPH Model Of Wistar Strain Rattus Novergicus Rat.

Benign prostate hyperplasia (BPH) is frequently found in the elderly and significantly impacts the quality of life. One of the risk factors that induce BPH is the androgen hormone. One of the effective medications in reducing the severity of Lower Urinary Tract Symptoms caused by BPH is the α-adrenergic receptor 5α-reductase inhibitor. The study aims to see the effect of long-term dutasteride on the expression of the PKC-α enzyme in prostatic stromal tissue in the BPH Model of Wistar strain Rattus norvegicus rats. This study was an experimental, post-test-only, control group design that used randomization in sample selection. The objective is to measure the expression of PKC-α enzyme from prostate tissue of an adult male Wistar Strain of Rattus Novergicus rat that was given testosterone to induce BPH and given dutasteride in 1,3 and 6 days continuously. Data is shown in mean±SD, and all of the data were analyzed using the software SPSS 21st version with the One Way ANOVA Statistical method after fulfilling the normality test and variant homogeneity test. Data analysis with confidence rate 95% and a=0,05. There was a decrease of PKC-α enzyme and prostate weight in dutasteride monotherapy in 1,3,6 days compared to the positive control, and the lowest value was on the sixth day (SD ± 2876.8). There was a constant decrease of PKC-α enzyme from the first day until the sixth day. In conclusion, long-term dutasteride monotherapy could significantly decrease the level of PKC-α enzyme. There was no upregulation of the PKC-α enzyme in the long term of dutasteride monotherapy.

Moringa oleifera seeds attenuate benzene-induced alterations in lipid peroxidation and antioxidant enzymes in liver and kidney tissues of Wistar rats

Benzene is a notorious toxicant that is responsible for a host of diseases including leukemia. Its concentration in the environment is increasing day-by-day due to excessive automobile use, accelerated industrial activities and cigarette smoke. The awareness on the harmful effects of benzene on health is limited and no antidote has been reported yet. In this study, an attempt has been made to find out a suitable remedy to overcome benzene toxicity in a living organism from a natural source with the seeds of the plant Moringa oleifera (MO). Thirty six Wistar rats were considered for the study and divided into six groups (n=6). While group I remained as control with normal animals, those in groups II – VI received benzene by oral route (800 mg/kg body weight) for 28 consecutive days. On day 29, the benzene-treated animals in groups III – VI received respectively the standard drug ascorbic acid (AA, 25 mg/kg body weight) and MO (50, 100 and 200 mg/kg body weight) for the following 7 days. Group II rats that received only benzene served as negative control without any treatment. On day 36, all the animals were sacrificed and vital organs liver and kidney were removed for studying lipid peroxidation (LPO) and antioxidant markers [Superoxide dismutase (SOD), Total reduced glutathione (TRG), Glutathione peroxidase (GPx) and Catalase (CAT)] in addition to histopathological changes in the tissues. The results of the study revealed that significant changes occurred in the above parameters due to benzene dosing to animals were reverted to near normal values on MO administration in the liver and kidney tissues as compared to untreated animals, suggesting MO’s pro-active role in attenuating benzene toxicity.

Characterization of Orange Peel Extract and Its Potential Protective Effect against Aluminum Chloride-Induced Alzheimer's Disease.

Alzheimer's disease (AD) is a devastating neurodegenerative disorder without a cure. Hence, developing an effective treatment or protective agent is crucial for public health. The present study aims to characterize orange peel extract (OPE) through in vitro and in silico studies. Furthermore, it examines the protective effect of OPE against experimentally-induced Alzheimer's disease in rats. The total phenolic and flavonoid content of OPE was 255.86 ± 1.77 and 52.06 ± 1.74 (mg/100 g), respectively. Gallic acid, the common polyphenol in OPE detected by HPLC was 3388.60 μg/100 g. OPE antioxidant IC50 was 67.90 ± 1.05, 60.48 ± 0.91, and 63.70 ± 0.30 by DPPH, ABTS and Hydroxyl radical scavenging activity methods, respectively. In vitro anti-acetylcholinesterase (AChE) IC50 was 0.87 ± 0.025 mg/mL for OPE and 2.45 ± 0.001 mg/mL for gallic acid. Molecular docking analysis for human AChE (4EY7) with donepezil, gallic acid, and acetylcholine showed binding energy ΔG values of -9.47, -3.72, and -5.69 Kcal/mol, respectively. Aluminum chloride injection (70 mg/Kg/day for 6 weeks) induced Alzheimer's-like disease in male rats. OPE (100 and 200 mg/kg/d) and gallic acid (50 mg/kg/d) were administered orally to experimental animals for 6 weeks in addition to aluminum chloride injection (as protective). OPE was found to protect against aluminum chloride-induced neuronal damage by decreasing both gene expression and activity of acetylcholinesterase (AChE) and a decrease in amyloid beta (Aβ42) protein level, thiobarbituric acid-reactive substances (TBARS), and nitric oxide (NO), and increased reduced glutathione (GSH) level and activity of the antioxidant enzymes in the brain tissues. Additionally, gene expressions for amyloid precursor protein (APP) and beta secretase enzyme (BACE1) were downregulated, whereas those for presinilin-2 (PSEN2) and beta cell lymphoma-2 (BCL2) were upregulated. Furthermore, the reverse of mitochondrial alternation and restored brain ultrastructure might underlie neuronal dysfunction in AD. In conclusion, our exploration of the neuroprotective effect of OPE in vivo reveals that OPE may be helpful in ameliorating brain oxidative stress, hence protecting from Alzheimer's disease progression.

Effects of Dietary Supplementation with Iron in Breeding Pigeons on the Blood Iron Status, Tissue Iron Content, and Full Expression of Iron-Containing Enzymes of Squabs.

This study was aimed at investigating the effects of diet iron levels on the blood iron status, tissue iron content, mRNA levels, and the activity of iron-containing enzymes in different tissues of squabs. A total of 120 pairs of healthy Silver Feather King parental pigeons with similar average body weight and egg production were randomly divided into 5 groups with 8 replicates and 3 pairs of pigeons per replicate. The five groups of breeding pigeons were fed an iron-unsupplemented basal diet and basal diet supplemented with 75, 150, 300, and 600mg iron/kg, respectively. The diets were fed in the form of granular feed based on corn, soybean meal, wheat, and sorghum. A broken line model was used for regression analysis. The results showed that plasma iron (PI), serum ferritin, iron contents in crop milk and liver, liver catalase (CAT) activity, and heart succinate dehydrogenase (SDH) activity were affected by iron levels (P < 0.05). And PI, serum ferritin, iron content in crop milk, and heart SDH activity increased quadratically (P < 0.05), but the iron content and CAT activity in the liver decreased quadratically (P < 0.005) as dietary iron level increased. According to the broken-line model of serum ferritin fitting (P < 0.002), the optimal dietary iron level of breeding pigeons was estimated to be 193mg/kg. In conclusion, serum ferritin is a sensitive index to evaluate the iron requirement of the breeding pigeon with two squabs, and the recommended iron supplemental level is 193mg/kg.

Glutathione and Its Metabolic Enzymes in Gliomal Tumor Tissue and the Peritumoral Zone at Different Degrees of Anaplasia.

This research was aimed at investigating the features of free radical activity and the parameters of glutathione metabolism in tumor tissues and the peritumoral zone at different degrees of glial tumor anaplasia. We analyzed postoperative material from 20 patients with gliomas of different degrees of anaplasia. The greatest differences compared to adjacent noncancerous tissues were found in the tumor tissue: an increased amount of glutathione and glutathione-related enzymes at Grades I and II, and a decrease of these parameters at Grades III and IV. For the peritumoral zone of Grades I and II, the indices changed in different directions, while for Grades III and IV, they occurred synchronously with the tumor tissue changes. For Low Grade and High Grade gliomas, opposite trends were revealed regarding changes in the level of glutathione and the enzymes involved in its metabolism and in the free radical activity in the peritumoral zone. The content of glutathione and the enzymes involved in its metabolism decreased with the increasing degree of glioma anaplasia. In contrast, free radical activity increased. The glutathione system is an active participant in the antioxidant defense of the body and can be used to characterize the cell condition of gliomas at different stages of tumor development.

Influence of Effective Microorganisms and Clinoptilolite on Gut Barrier Function, Intestinal Health and Performance of Broiler Chickens during Induced Eimeria tenella Infection

The prohibition of certain coccidiostats in poultry has created a need to seek an alternative to control Eimeria infection. The aim of this study was to evaluate the effects of effective microorganisms (EM) in a multi-strain probiotic (Bokashi®), with clinoptilolite as a feed supplement on the mRNA expression of tight junction proteins and redox enzymes in the caecal tissue of chickens infected with E. tenella. The integrity of the intestinal barrier was tested by determining the concentration of fluorescein isothiocyanate dextran (FITC-d) in the chicken’s serum. A total of 600 1-day-old Ross 308 male chickens received diets with a 0.5% or 0.8% concentration of the probiotic together with clinoptilolite. The experiment used 5 treatment groups, and a control group, each with 5 replicates with 20 birds. The results indicate that the use of the 8 kg/t of feed multi-strain probiotic together with clinoptilolite in the diet of poultry caused a significant reduction in the number of E. tenella oocysts in the faeces and caecum and significantly improved the growth rate of chicken broilers infected with E. tenella. In addition, the probiotic and clinoptilolite enhanced antioxidant processes in the caecal mucosa and reduced oxidative stress induced by E. tenella infection.

Effect of coenzyme Q10 and vitamin E on gentamicin-induced nephrotoxicity in rats

Background/Aim: Gentamicin is an aminoglycoside antibiotic that can cause nephrotoxicity by damaging the kidneys due to high relative blood flow. In this study, the protective and therapeutic effects of CoQ10 and vitamin E on GM-induced nephrotoxicity were investigated. Methods: Fifty Wistar Albino rats were used as live material in this study, which were randomly divided into 5 groups of 10 animals. These groups, and the treatment they received, were as follows: a) control group: Physiological saline (i.p) for 8 days, b) GM group: Gentamycin 80 mg/kg/day/i.p for 8 days, c) GM+CoQ10 group: Gentamycin 80 mg/kg/day/i.p+ CoQ10 10 mg/kg/day/ i.p for 8 days, d) GM+ Vit E group: Gentamycin 80 mg/kg/day/i.p+ Vit E 250 mg/kg /day/ip administrated for 8 days, and e) GM+CoQ10+Vit E group: Gentamycin 80 mg/kg/day/ip + CoQ10 10 mg/kg/day/ip + Vitamin E 250 mg/kg /day/ip for 8 days. Following the test period, urea, creatinine, K, Na, Cl, retinol, vitamin D, and vitamin E levels of the subjects were measured in plasma, while MDA, GSH, AOPP, nitrite, nitrates, CAT, SOD, and GPx activities were measured in kidney tissues. Results: A severe nephrotoxic effect of GM administration in rats were detected as part of this study. The administration of CoQ10 and vitamin E, alone or in combination, was not sufficient to repair the kidney damage at the histopathological level. However, the renal tissue enzyme levels, along with other related to oxidative stress and plasma biochemical parameter analyses, were found to have improved greatly with the administration of vitamin E. Conclusion: CoQ10 and vitamin E can be suggested as supplementary agents to gentamicin treatment to prevent cell degeneration in the kidney and ensure healing

Uniconazole enhances lodging resistance by increasing structural carbohydrate and sclerenchyma cell wall thickness of japonica rice (Oryza sativa L.) under shading stress

Lodging is a major limiting factor for grain yield and quality, and low light enhances lodging risk by increasing stem elongation and decreasing culm thickness. This study evaluated the effect of exogenous uniconazole (UCZ) on the basal stem physiological mechanism, cell structure, carbohydrates, and lignin-related enzymes in culm tissue under shading conditions. The results showed that shading stress significantly decreased grain yield and stem mechanical strength in 2020 and 2021. Compared with normal light (CK), shading rapidly decreased the breaking strength (M) and section modulus (SM). Exogenous UCZ increased culm wall thickness and improved lodging resistance. Uniconazole application also reduced plant height, gravity height, and second basal internode length by inducing a reduction in the cell length, and caused the cells to be densely arranged under both normal light and shading conditions. Scanning electron microscopy (SEM) revealed that the thickness of the sclerenchyma tissue cell wall increased under both conditions under the UCZ treatment, which induced an increase in the cell wall content. Uniconazole application significantly increased non-structural carbohydrate, lignin, cellulose, and hemicellulose contents in culm tissue. Moreover, UCZ application regulated lignin-related enzymes, such as phenylalanine ammonia-lyase (PAL), 4-coumarate: CoA ligase (4CL), cinnamate 4-hydroxylase (C4H), laccase (LAC) and peroxidase (POD). Thus, exogenous UCZ could enlarge sclerenchyma cell wall thickness, alter cell morphology in the stem, and increase stem strength by increasing carbohydrate content, including non-structural carbohydrates, cellulose, lignin, and hemicellulose, to enhance rice (Oryza sativa L.) lodging resistance under shading conditions.

Using Activity-Based Proteomics for the Quantification of Deubiquitinases in Animal Tissue.

Ubiquitination isapost-translational modification, thatregulates essential cellular functions, and the enzymes that control the removal of this modification, deubiquitinases (DUBs), have been well described for the model organisms. However, the information about DUBs is still largely lacking for the non-model organisms, such as agriculturally relevant animals. To understand the expression of these enzymes in animal tissues, we have used chemical proteomics which can be used to identify biologically active DUBs present in tissues based on their reactivity with the activity-based probes (ABPs). Here we describe a sample preparation protocol for ABP-based purification of DUBs from animal tissue using two approaches to homogenize and lyse the animal tissue compatible with ABP labeling of DUBs, including anultrasonication-based tissue processing method and bead-beating method. Both of these methods retain the enzymatic activity of DUBs. In addition, we describe a protocol for ABP labeling of DUBs in tissue lysates and the immunoprecipitation of the probe-reactive DUBs that can be used along with mass spectrometric identification of proteins and the detection of these DUBs by Western blotting.

PSUN84 Chronic Adrenergic Stimulation in Humans Increases Plasma Bile Acids and Expression of Bile Acid Synthesis Enzymes in White Adipose Tissue

Abstract Bile acids (BAs) are classically known as facilitators of fat digestion. Moreover, they are now known to be ligands for receptors involved in glucose and lipoprotein metabolism. BA synthesis was originally thought to be exclusive to hepatic tissue, but emerging evidence in rodents has demonstrated the expression of BA transporters and de novo synthetic enzymes in murine adipose tissue. We previously showed that chronic activation of brown adipose tissue (BAT) and white adipose tissue (WAT) in adult women via the β3-adrenergic receptor (ADRB3) agonist mirabegron increases total plasma BA levels. The largest increases were seen with primary unconjugated BAs. Overall, these data suggest that chronic adrenergic stimulation may increase the synthesis and activity of BA synthesis enzymes in human adipose tissue, which contributes to the increase in total plasma BAs. To confirm this hypothesis, we measured expression levels of BAT and BA synthesis enzymes in liver and adipose tissues collected from healthy patients and those with pheochromocytoma (pheo) who experienced chronic adrenergic activation. As expected, expression of thermogenic UCP1 and ADRB3 was higher in pheo peri-renal BAT than in liver tissue (76,000x, P&amp;lt;0.001; and 6,000x, P=0.02, respectively) or either healthy subcutaneous WAT or pheo subcutaneous WAT (≥1500x, P&amp;lt;0.001 for UCP1). Conversely, expression of the rate-determining enzyme in BA synthesis CYP7A1 was similar in liver and both WAT depots (P&amp;gt;0.05), and liver had slightly higher expression than BAT (3x, P=0.03). Otherwise, most of the principal BA synthesis genes’ expression levels demonstrated the following stepwise pattern: liver tissue &amp;gt; pheo WAT = subcutaneous WAT &amp;gt; BAT (CYP8B1, CYP27A1, AKR1D1, and BAAT). These results suggest that BA production in humans is not limited to the liver. Rather, human WAT in particular may contribute to the BA pool and could serve as a target for producing BA-mediated improvements in glucose and lipoprotein metabolism. Presentation: Sunday, June 12, 2022 12:30 p.m. - 2:30 p.m.

ODP057 The Unusual Case of Hypercortisolism in a Patient with Juvenile Onset Hypophosphatasia

Abstract Juvenile onset hypophosphatasia is a rare, genetic disease that is caused by inactivating mutations in the ALPL gene which encodes tissue non-specific alkaline phosphatase (TNSALP). This leads to decreased activity of the TNSALP enzyme in target tissues and accumulation of TNSALP substrates, including inorganic pyrophosphate, an inhibitor of mineralization. This disease is characterized by low levels of alkaline phosphatase in serum and bone, osteomalacia, and periodontal disease. Patients with juvenile onset hypophosphatasia present with premature loss of primary teeth, delayed walking, and a waddling gait. These symptoms may improve spontaneously after puberty and recur later in life. Here we report the case of a 51 year old male who presented initially to endocrinology clinic with a metatarsal stress fracture with subsequent diagnosis of osteoporosis, periodontal infections, and associated myopathy and joint calcifications. The patient also had a prior history of a low alkaline phosphatase level as a child. Genetic testing showed a single variant of undetermined significance found in the ALPA gene. Of note, recent lab work for the patient showed AM cortisol 27.7 mcg/dL (reference range 4. 0-22. 0 mcg/dL) with ACTH 60 pg/mL (reference range 6 - 50 pg/mL). Prior 24 hour urine free cortisol measurements with LC/MS have demonstrated elevated values 159 mcg/24 hr and 256 mcg/24 hr (reference range 4. 0 - 50. 0 mcg/24 hr) . However, most recent testing demonstrated 24 hour urine free cortisol 3.6 mcg/24 hr, and the majority of midnight salivary cortisol tests were within normal. Pituitary MRI demonstrated no abnormalities. The patient has in the past reported findings of a dorsocervical fat pad, abdominal striae, and weight gain. There is no history of diabetes mellitus or hypertension, and the patient at this time reports stable weight without current presence of a fat pad or striae. There are no clinical features of Cushing's syndrome on exam. To our knowledge, this is the second case in the literature detailing hypercortisolism in a patient with hypophosphatasia. The etiology of the hypercortisolism is likely multifactorial, with unknown association with hypophosphatasia. Based on the clinical data it appears that the patient's chronic stress, pain, and anxiety are contributing to the hypercortisolism, and that this is not a true Cushing's or cyclical Cushing's case. Repeated midnight salivary cortisol tests have excluded the possibility of Cushing's as well. Presentation: No date and time listed

Natural history and management of liver dysfunction in lysosomal storage disorders.

Lysosomal storage disorders (LSD) are a rare group of genetic disorders. The major LSDs that cause liver dysfunction are disorders of sphingolipid lipid storage [Gaucher disease (GD) and Niemann-Pick disease] and lysosomal acid lipase deficiency [cholesteryl ester storage disease and Wolman disease (WD)]. These diseases can cause significant liver problems ranging from asymptomatic hepatomegaly to cirrhosis and portal hypertension. Abnormal storage cells initiate hepatic fibrosis in sphingolipid disorders. Dyslipidemia causes micronodular cirrhosis in lipid storage disorders. These disorders must be keenly differentiated from other chronic liver diseases and non-alcoholic steatohepatitis that affect children and young adults. GD, Niemann-Pick type C, and WD also cause neonatal cholestasis and infantile liver failure. Genotype and liver phenotype correlation is variable in these conditions. Patients with LSD may survive up to 4-5 decades except for those with neonatal onset disease. The diagnosis of all LSD is based on enzymatic activity, tissue histology, and genetic testing. Enzyme replacement is possible in GD and Niemann-Pick types A and B though there are major limitations in the outcome. Those that progress invariably require liver transplantation with variable outcomes. The prognosis of Niemann-Pick type C and WD is universally poor. Enzyme replacement therapy has a promising role in cholesteryl ester storage disease. This review attempts to outline the natural history of these disorders from a hepatologist's perspective to increase awareness and facilitate better management of these rare disorders.

Polymethine Molecular Platform for Ratiometric Fluorescent Probes in the Second near-Infrared Window.

Visualizing biomolecules such as enzymes in the deep tissue of living organisms via molecular ratiometric fluorescent probes in the second near-infrared window (NIR-II) with a built-in self-calibration function can provide reliable information about relevant pathophysiological processes directly but so far is not feasible due to the lack of a fluorescence modulation strategy in the NIR-II window. Here we present a molecular platform Py-2 by integrating the rhodamine 6G scaffold and polymethine. The maximal emission wavelength of Py-2 was 1010 nm and blue-shifted to 945 nm when its secondary amine was acylated. Based on Py-2, two molecular ratiometric NIR-II fluorescent probes, nitroreductase-responsive Rap-N and ROS-responsive Rap-R, were constructed and successfully demonstrated in vitro and in vivo. Overall, this report presents a unique approach to developing ratiometric NIR-II molecular probes for in vivo biosensing.

Dietary mixed Bacillus strains promoted the growth indices, enzymatic profile, intestinal immunity, and liver and intestinal histomorphology of Nile tilapia, Oreochromis niloticus

In the current investigation, a 4-week feeding trial was carried out to study the special effects of the dietary probiotic, Sanolife® PRO-F (a combination of Bacillus subtilis 3.25 × 109 CFU/g, Bacillus licheniformis 3.50 × 109 CFU/g, and Bacillus pumilus 3.25 × 109 CFU/g) supplementation on growth performance, serum biochemical immune parameters, intestinal digestive enzymes activities, antioxidants assay, and the expression of pro-inflammatory cytokines and antioxidant-related genes with a specific reference to intestinal histomorphometry of Nile tilapia, Oreochromis niloticus, which fed with Sanolife® PRO-F at 0 (B0), 0.5 (B0.5) and 1 (B1) g kg−1 diet, respectively. At the end of the feeding trial, the achieved results revealed that Bacillus strains probiotic supplementation significantly improved the growth performance including final body weight (FBW), weight gain (WG), specific growth rate (SGR), and feed utilization when matched to the control group (p < 0.05). Concerning serum biochemical parameters, there was a significant reduction in the serum cholesterol and triglyceride levels in fish-fed Bacillus strains (p < 0.01). Serum lysozyme activity showed greater values in the case of the B0.5 and B1 groups (p < 0.01) compared with the control. No changes were estimated in the levels of glucose, hepatic (alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and renal biomarkers (Urea, Creatinine, and Uric acid) in the serum of fish-fed B0.5 and B1 diets. Additionally, the specific activities of protease, amylase, and lipase enzymes were significantly increased upon Bacillus strains՚s incorporation in O. niloticus diets. As well, the antioxidant capacity of catalase (CAT) and superoxide dismutase (SOD) enzymes in the liver and spleen were significantly increased with diminished malondialdehyde (MDA) levels. Correspondingly, the mRNA expression of the antioxidant glutathione peroxidase (GPx) enzyme in the intestinal tissue was notably increased in the control group. The length of intestinal villi, absorptive epithelium, and goblet cell counts were greater in O. niloticus fed on Bacillus strains diets. Given the obtained results, the dietary supplementation of Sanolife® PRO-F to Nile tilapia, O. niloticus at 0.5 g kg 1 and 1 g kg−1 diet can recover the growth, feed utilization, and health condition with the safety of Sanolife® PRO-F probiotic supplementation on liver and kidney function.

Aminopeptidase B can bioconvert L-type amino acid transporter 1 (LAT1)-utilizing amide prodrugs in the brain.

A prodrug approach is a powerful method to temporarily change the physicochemical and thus, pharmacokinetic properties of drugs. However, in site-selective targeted prodrug delivery, tissue or cell-specific bioconverting enzyme is needed to be utilized to release the active parent drug at a particular location. Unfortunately, ubiquitously expressed enzymes, such as phosphatases and carboxylesterases are well used in phosphate and ester prodrug applications, but less is known about enzymes selectively expressed, e.g., in the brain and enzymes that can hydrolyze more stable prodrug bonds, such as amides and carbamates. In the present study, L-type amino acid transporter 1 (LAT1)-utilizing amide prodrugs bioconverting enzyme was identified by gradually exploring the environment and possible determinants, such as pH and metal ions, that affect amide prodrug hydrolysis. Based on inducement by cobalt ions and slightly elevated pH (8.5) as well as localization in plasma, liver, and particularly in the brain, aminopeptidase B was proposed to be responsible for the bioconversion of the majority of the studied amino acid amide prodrugs. However, this enzyme hydrolyzed only those prodrugs that contained an aromatic promoiety (L-Phe), while leaving the aliphatic promoeities (L-Lys) and the smallest prodrug (with L-Phe promoiety) intact. Moreover, the parent drugs' structure (flexibility and the number of aromatic rings) largely affected the bioconversion rate. It was also noticed in this study, that there were species differences in the bioconversion rate by aminopeptidase B (rodents > human), although the in vitro-in vivo correlation of the studied prodrugs was relatively accurate.

The impact of inflammation on the expression of drug transporters and metabolic enzymes in colonic tissue from ulcerative colitis patients

The intestinal tract forms an important barrier against xenobiotics while allowing nutrients to pass. In ulcerative colitis (UC), a chronic inflammatory bowel disease, this barrier function is impaired leading to an abnormal immune response and inflammation of the colonic mucosa. Transporter proteins and metabolic enzymes are an integral part of the protective barrier in the gut and play an important role in the disposition of nutrients, toxins and oral drugs. In this study, the protein expression of 13 transporters and 13 enzymes was determined in the sigmoid and rectum of UC patients in endoscopic remission and during active inflammation. In inflamed conditions (endoscopic Mayo sub-score 1, 2 or 3), a significant decrease (q < 0.05) was observed in the median expression of the transporters P-gp (0.046 vs 0.529 fmol/µg protein), MRP4 (0.003 vs 0.023 fmol/µg protein) and MCT1 (0.287 vs 1.090 fmol/µg protein), and the enzymes CYP3A5 (0.031 vs 0.046 fmol/µg protein) and UGT2B7 (0.083 vs 0.176 fmol/µg protein). Moreover, during severe inflammation, the decrease was even more pronounced. Expression levels of other proteins were not altered during inflammation (e.g., OATP2B1, CYP3A4, CYP2B6 and UGT2B15). The results suggest a decreased transport and metabolism of xenobiotics in the colon of UC patients during active inflammation potentially altering local drug concentrations and thus treatment outcome.

Anti-urolithiatic Activity of Daidzin in Ethylene Glycol-Induced Urolithiasis in Rats.

Urolithiasis is a common urological disorder, which causes considerable morbidity in both genders at all age groups worldwide. Though treatment options such as diuretics and non-invasive techniques to disintegrate the deposits are available, but often they are found less effective in the clinics. In this work, we planned to investigate the ameliorative effects of daidzin against the ethylene glycol (EG)-induced urolithiasis in rats. The male albino rats were distributed into four groups (n = 6) as control (group I), urolithiasis induced by the administration of 0.75% EG (group II), urolithiasis induced rats treated with 50mg/kg of daidzin (group III), and urolithiasis rats treated with standard drug 750mg/kg of cystone (group IV). The urine volume, pH, and total protein in the urine were assessed. The activities of marker enzymes in both plasma and kidney tissues were analyzed using assay kits. The levels of kidney function markers such as calcium, oxalate, urea, creatinine, uric acid, magnesium, BUN, and phosphorous were estimated using assay kits. The status of antioxidants and inflammatory cytokines were also examined using kits. The renal tissues were examined by histopathological analysis. Our results revealed that the daidzin treatment effectively decreased the urine pH and protein level and increased the urine volume in the urolithiasis rats. Daidzin decreased the calcium, oxalate, uric acid, and urea, creatinine, and BUN levels and also improved the magnesium and phosphorus in the urolithiasis rats. The activities of AST, ALT, ALP, GGT, and LDH were effectively reduced by the daidzin in both serum and renal tissue. Daidzin also reduced the inflammatory marker and increased the antioxidant levels. Histopathology results also proved the therapeutic effects of daidzin. Together, our results displayed that daidzin is effective in the amelioration of EG-induced urolithiasis in rats.