Enzyme Activity In Tissues Research Articles

Drug-Drug Interaction of Aldehyde Oxidase Inhibitor and Xanthine Oxidase Inhibitor with Favipiravir

Aim: Favipiravir is an effective antiviral used in the treatment of COVID-19. It is metabolized by aldehyde oxidase (AO) and xanthine oxidase (XO). This study investigated drug-drug interactions between favipiravir with both AO substrate and XO enzyme inhibitor, allopurinol, and an XO inhibitor, verapamil. 
 Material and Methods: 25 Sprague-Dawley female rats, 250-300 g, were divided into five equal groups. Blood samples were taken from the jugular vein at the end of 0, 15, 30, and 45 minutes, and at the end of the 1st, 2nd, 4th, 6th, and 8th hours after the drugs were administered. The drug-blood concentration was determined in the HPLC-UV device using plasma. The ELISA method measured AO and XO enzyme activities in rat liver tissue.
 Results: Allopurinol prolonged the time taken for favipiravir to reach Cmax (Tmax), decreased maximum serum concentration (Cmax), elimination half-life (T1/2), area under the curve (AUC), and mean residence time (MRT). Allopurinol significantly reduced clearance per unit time (Cl/f) when co-administered with favipiravir. Verapamil accelerated the elimination of favipiravir, significantly reducing T1/2, MRT, and AUC. On the other hand, Favipiravir decreased the absorption of verapamil and slowed its elimination. Cmax, AUC, and Cl values of verapamil decreased. In addition, T1/2, MRT, and volume of distribution (Vd) increased. 
 Conclusion: In conclusion, the concomitant use of favipiravir with other drugs that affect AO and/or XO enzyme activities may cause changes in the pharmacokinetic profiles of drugs and the levels of enzymes that metabolize drugs.

Dietary curcumin supplementation can enhance health and resistance to ammonia stress in the greater amberjack (Seriola dumerili)

We evaluated the effect of dietary curcumin supplementation on the antioxidant capacity of the liver and the resistance of the liver and spleen to ammonia stress in the great amberjack (Seriola dumerili). Three isonitrogenous and isolipidic test diets were prepared by supplementing incremental levels of dietary curcumin at 0 mg/kg (CUR0%, control), 100 mg/kg (CUR0.01%), and 200 mg/kg (CUR0.02%), respectively. Each diet was randomly assigned to triplicate groups of 15 fish per tank. At the end of the feeding experiment, dietary curcumin supplementation positively modulated antioxidant-related genes and enzyme activity in liver tissues. After the ammonia challenge, dietary supplementation with the appropriate level of curcumin alleviated ammonia stress in liver tissue by upregulating the relative expression of GSH-Px and downregulating the relative expression of Keap1 and GR. Meanwhile, ammonia stress in spleen tissue could also be alleviated by upregulating the relative expression of CAT, downregulating the relative expression of GR, and increasing the activity of SOD and GSH. After the recovery, dietary supplementation with curcumin still alleviated ammonia stress in the liver tissue by upregulating the relative expression of CAT, downregulating the relative expression of Keap1 and GR, and increasing the activity of SOD and GSH. On the other hand, ammonia stress in spleen tissue was still alleviated by upregulating the relative expression of Mn-SOD and increasing the activity of SOD and GSH. The histological structure results also showed that liver cells in the curcumin-containing groups exhibited a positive impact on cell boundaries, alignment, and nuclei after the ammonia challenge and recovery. Spleen cells in the curcumin-containing groups exhibited greater aggregation of melano-macrophage centers after the ammonia challenge and recovery. These results suggest that dietary curcumin supplementation at 100 mg/kg can promote the health condition and resistance to ammonia stress of the greater amberjack.

Galactosylceramidase deficiency and pathological abnormalities in cerebral white matter of Krabbe disease

Krabbe Disease (KD) is an autosomal recessive disorder that results from loss-of-function mutations in the GALC gene, which encodes lysosomal enzyme galactosylceramidase (GALC). Functional deficiency of GALC is toxic to myelin-producing cells, which leads to progressive demyelination in both the central and peripheral nervous systems. It is hypothesized that accumulation of psychosine, which can only be degraded by GALC, is a primary initiator of pathologic cascades. Despite the central role of GALC in KD pathomechanism, investigations of GALC deficiency at a protein level are largely absent, due in part, to the lack of sensitive antibodies in the field. Leveraging two custom antibodies that can detect GALC at endogenous levels, we demonstrated that GALC protein is predominantly localized to oligodendrocytes in cerebral white matter of an infant brain, consistent with its functional role in myelination. Mature GALC could also be quantitatively detected as a 26 kDa band by western blotting and correlated to enzyme activity in brain tissues. The p.Ile562Thr polymorphic variant, which is over-represented in the KD population, was associated with reduced mature GALC protein and activity. In three infantile KD cases, homozygous null mutations in GALC lead to deficiency in total GALC protein and activity. Interestingly, although GALC activity was absent, normal levels of total GALC protein were detected by a sandwich ELISA using our custom antibodies in a later-onset KD brain, which suggests that the assay has the potential to differentiate infantile- and later-onset KD cases. Among the infantile KD cases, we quantified a 5-fold increase in psychosine levels, and observed increased levels of acid ceramidase, a key enzyme for psychosine production, and hyperglycosylated lysosomal-associated membrane protein 1, a marker for lysosomal activation, in periventricular white matter, a major pathological brain region, when compared with age-matched normal controls. While near complete demyelination was observed in these cases, we quantified that an early-infantile case (age of death at 10 months) had about 3-fold increases in both globoid cells, a pathological hallmark for KD, and CD8-positive T lymphocytes, a pathological marker for multiple sclerosis, in the white matter when compared with a slower progressing infantile case (age of death at 21 months), which suggests a positive correlation between clinical severity and neuropathology. Taken together, our findings have advanced the understanding of GALC protein biology in the context of normal and KD brain white matter. We also revealed new neuropathological changes that may provide insights to understand KD pathogenesis.

Galactose epimerase deficiency: lessons from the GalNet registry

BackgroundGalactose epimerase (GALE) deficiency is a rare hereditary disorder of galactose metabolism with only a few cases described in the literature. This study aims to present the data of patients with GALE deficiency from different countries included through the Galactosemia Network to further expand the existing knowledge and review the current diagnostic strategy, treatment and follow-up of this not well characterized entity.MethodsObservational study collecting medical data from December 2014 to April 2022 of 22 not previously reported patients from 14 centers in 9 countries. Patients were classified as generalized or non-generalized based on their genotype, enzyme activities in different tissues and/or clinical picture and professional judgment of the treating physician.ResultsIn total 6 patients were classified as generalized and 16 as non-generalized. In the generalized group, acute neonatal illness was reported in 3, cognitive and developmental delays were present in 5 and hearing problems were reported in 3. Four generalized patients were homozygous for the genetic variant NM_001008216.2:c.280G > A (p.Val94Met). In the non-generalized group, no clearly related symptoms were found. Ten novel genetic variants were reported in this study population.ConclusionThe phenotypic spectrum of GALE deficiency ranges from asymptomatic to severe. The generalized patients have a phenotype that is in line with the 9 described cases in the literature and prescribing dietary interventions is the cornerstone for treatment. In the non-generalized group, treatment advice is more difficult. To be able to offer proper counseling, in addition to red blood cell enzyme activity, genetic studies, transferrin glycoform analysis and enzymatic measurements in fibroblasts are recommended. Due to lack of facilities, additional enzymatic testing is not common practice in many centers nor a tailored long-term follow-up is performed.

Effects Of Broad-spectrum Antibiotic Administration And Germ-free Status On Biochemical And Endurance Adaptations To Regular Exercise In Mouse Muscle Tissue

PURPOSE: The gut microbiota modulates a variety of physiologic processes. Recent studies from our group have shown that regular endurance exercise alters the composition of the gut microbiome and metabolome in mice and humans, and we hypothesized that the presence of microbes would affect endurance training adaptations in muscular endurance capacity, mitochondrial activity and gene expression in mice. METHODS: To discern differences in adaptations from 6 wks of voluntary wheel running (VWR), we depleted microbes with antibiotics (ABX) and used germ free (GF) mice to compare to control (CON). Male and female C57Bl/6 mice of all groups underwent daily VWR or sedentary (SED) conditions in a 2 x 3 design (VWR/SED, CON/ABX/GF, n = 56). After the intervention, treadmill endurance was assessed and gastrocnemius and soleus tissue were harvested and analyzed for activity of key Krebs cycle enzyme Citrate Synthase (CS) and expression of genes indicating mitochondrial adaptations respectively. RESULTS: Two way ANOVA revealed that VWR increased treadmill endurance, ABX had no effect, and that GF status significantly reduced performance. Additionally, VWR increased Citrate Synthase enzyme activity in gastrocnemius muscle tissue in all groups, and ABX and GF status did not reduce VWR’s effect or have an effect of their own. VWR also tended to increase expression of genes associated with increased mitochondrial activity (PGC-1α, CS, Succinate Dehydrogenase) in soleus tissue, but ABX had no effect on these observations. CONCLUSIONS: We conclude that ABX treatment and GF status do not affect VWR induced adaptations in endurance capacity, gastrocnemius mitochondrial activity or expression of metabolic genes in soleus tissue, but that GF status significantly hinders endurance capacity. This indicates that removing gut microbes does not inhibit muscular endurance training adaptations, but that germ free mice possess hindered endurance exercise capacity.

Ratlarda intravenöz kontrast madde uygulamasının göz içi basıncı, göz yaşı miktarı ve oksidatif stres üzerine etkileri

Although iodinated radiocontrast agents, which are frequently used in radiological procedures such as indirect radiography, angiography, intravenous urography and computed tomography, are generally safe, they can cause serious side effects. In this study, it was aimed to investigate the effect of intravenous ionic high osmolar contrast agent administration on intraocular pressure, tear amount and oxidant and antioxidant parameters of eye tissue. Study groups consisted of 2 groups, Group 1 (Control) and Group 2 (Urographin), and a total of 16 Wistar albino female rats were used. On the first day of the experiment, 6 ml/kg of physiological saline was administered intravenously to the control group, and the same dose of contrast agent was administered to group 2. Intraocular pressure and tear amounts were measured at 1, 6, 12, 24 and 48 hours after intravenous administration. After measurements were made at the 48th hour of the experiment, all rats were euthanized and their eye tissues were removed. In order to reveal the oxidative damage and antioxidant activity in the eye tissue, malondialdehyde and reduced glutathione levels, catalase and glutathione peroxidase enzyme activities were measured spectrophotometrically. Schimer tear test (STT-1) strip was used for tear amount measurements, and rebound tonometer Tonovet® was used as tonometer for intraocular pressure measurements. No statistically significant difference was found between the control and urographin groups in terms of intraocular pressure and tear amounts in the measurements performed at 1,6,12,24 and 48th hours after urographin administration. It was observed that the same application significantly increased the malondialdehyde level (P<0.005) in the eye tissue. There was no significant difference between the groups in terms of reduced glutathione level and catalase and glutathione peroxidase enzyme activities in eye tissue. It was evaluated that intravenous contrast agent administration causes oxidative stress in the eye tissue and this may have a long-term ocular effect

Musculoskeletal stretch injury shows a faster recovery compared to contusion in rats

Background: The musculoskeletal repair, in most cases, follows the typical inflammatory process. Although the majority of treatments result in good prognosis, the knowledge concerning basic biochemical principles, regeneration and repair mechanisms of skeletal muscles may help in recovery and accelerate the return to sport, helping to carefully choose the more appropriate therapeutic techniques for the injury management. Objective: This study investigated the effects of contusion and stretch injury on the oxidative damage in the gastrocnemius muscle of rats. Methods: The biochemical analysis was performed on the 1, 3, 5, or 7 days after the injuries and behavior analysis was performed until 48 hours after injuries to evaluate the healing process changing characteristics. Design: Experimental animal study. Results: The contusion and stretch injury increased the oxidative stress markers levels; however, the injuries showed significant differences from each other in the markers of oxidized dichlorofluorescein, superoxide dismutase enzyme activity and N-acetyl-beta-glucosaminidase enzyme activity in skeletal muscle tissue. Furthermore, the injuries showed differences in behavioral analyses in the beam walking test. Conclusion: In conclusion, we could infer that the stretch injury presents an early increase in the activity of the N-acetyl-β-D-glucosaminidase enzyme, indicating a possible macrophage increase in the first hours after injury.

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.

Cross-species comparison of chemical inhibition of human and Xenopus iodotyrosine deiodinase.

The transition to include in vitro-based data in chemical hazard assessment has resulted in the development and implementation of screening assays to cover a diversity of biological pathways, including recently added assays to interrogate chemical disruption of proteins relevant to thyroid signaling pathways. Iodotyrosine deiodinase (IYD), the iodide recycling enzyme, is one such thyroid-relevant endpoint for which a human-based screening assay has recently been developed and used to screen large libraries of chemicals. Presented here is the development of an amphibian IYD inhibition assay and its implementation to conduct a cross-species comparison between chemical inhibition of mammalian and non-mammalian IYD enzyme activity. The successful development of an amphibian IYD inhibition assay was based on demonstration of sufficient IYD enzyme activity in several tissues collected from larval Xenopus laevis. With this new assay, 154 chemicals were tested in concentration-response to provide a basis for comparison of relative chemical potency to results obtained from the human IYD assay. Most chemicals exhibited similar inhibition in both assays, with less than 25% variation in median inhibition for 120 of 154 chemicals and 85% concordance in categorization of "active" (potential IYD inhibitor) versus "inactive". For chemicals that produced 50% or greater inhibition in both assays, rank-order potency was similar, with the majority of the IC50s varying by less than 2-fold (and all within an order of magnitude). Most differences resulted from greater maximum inhibition or higher chemical potency observed with human IYD. This strong cross-species agreement suggests that results from the human-based assay would be conservatively predictive of chemical effects on amphibian IYD.

The Biochemical Assessment of Mitochondrial Respiratory Chain Disorders.

Mitochondrial respiratory chain (MRC) disorders are a complex group of diseases whose diagnosis requires a multidisciplinary approach in which the biochemical investigations play an important role. Initial investigations include metabolite analysis in both blood and urine and the measurement of lactate, pyruvate and amino acid levels, as well as urine organic acids. Recently, hormone-like cytokines, such as fibroblast growth factor-21 (FGF-21), have also been used as a means of assessing evidence of MRC dysfunction, although work is still required to confirm their diagnostic utility and reliability. The assessment of evidence of oxidative stress may also be an important parameter to consider in the diagnosis of MRC function in view of its association with mitochondrial dysfunction. At present, due to the lack of reliable biomarkers available for assessing evidence of MRC dysfunction, the spectrophotometric determination of MRC enzyme activities in skeletal muscle or tissue from the disease-presenting organ is considered the ‘Gold Standard’ biochemical method to provide evidence of MRC dysfunction. The purpose of this review is to outline a number of biochemical methods that may provide diagnostic evidence of MRC dysfunction in patients.

Expression of tyrosinase gene in gingiva: A pilot study.

Melanin is the predominant pigment responsible for the color of skin, hair, iris of eyes, and oral mucosa. Tyrosinase (TYR) is the key enzyme involved in melanin synthesis. Studies in dermatology have shown a positive correlation between TYR enzyme levels and melanin pigmentation of the skin. However, no study has been conducted to assess TYR levels in the gingiva. Hence the present study was conducted to assess TYR levels in gingival melanin hyperpigmentation. To assess the TYR gene expression in gingiva in individuals with moderate to severe gingival melanin hyperpigmentation. Subjects with a chief complaint of blackish appearance of gums with an unesthetic smile were included in the study. Informed consent was obtained. Scaling and root planning were done and subjects were recalled after 2 weeks. The gingival depigmentation procedure was performed using the conventional scalpel technique under adequate local anesthesia. The selected sites underwent conventional gingival depigmentation technique using Bard-Parker handle no: 3 and blade no: 11. The excised layer of epithelium along with a thin layer of underlying connective was sent to the laboratory to assess the TYR gene expression by real-time polymerase chain reaction technique. The levels of the TYR enzyme activity in the gingival tissues from the selected sites were assessed. Table 1 and Graph 1 show the levels of TYR enzyme gene expression in the gingival tissue. TYR gene expression and the degree of gingival melanin hyperpigmentation are positively correlated. Hence the assessment of TYR enzyme activity in gingiva could be of great value in today's cosmetologically conscious individuals.

Silibinin-loaded nanostructured lipid carriers (NLCs) ameliorated cognitive deficits and oxidative damages in aluminum chloride-induced neurotoxicity in male mice

Aluminum chloride (AlCl3) and its accumulation in the brain are associated with neurodegenerative disease. Recent investigations have illustrated that silibinin is known to have neuroprotective properties. The present study investigates the neuroprotective effects of silibinin-loaded nanostructured lipid carriers (Sili-NLCs) against AlCl3-induced neurotoxicity in male mice. Sili-NLCs were prepared using the emulsification-solvent evaporation method and subjected to particle size, zeta potential, and entrapment efficiency (% EE) analysis. Mice were treated with AlCl3 (100 mg/kg/day, p.o.) and with the same concentration of silibinin and Sili-NLCs (50,100, and 200 mg/kg/day, p.o.) for 30 days in different groups. After treating animals, behavioral studies were assessed. Also, the brain tissue samples were collected from all mice to evaluate oxidative damage and histological changes. The particle size, polydispersity index, zeta potential, and entrapment efficiency (% EE) of prepared Sili-NLCs found 239.7 ± 4.04 nm, 0.082 ± 0.003, − 16.33 ± 0.15 mV, and 72.65 ± 2.03 %, respectively. Brain uptake studies showed that Sili-NLCs had a 5.7-fold greater uptake in the mice brain than the free drug. The AlCl3 caused significant cognitive impairment and increased the level of lipid peroxidation accompanied by decreasing antioxidant enzyme activity in the brain tissue. These findings correlated well with the histopathological experiments. Furthermore, treatment with Sili-NLCs significantly improved the AlCl3-induced cognitive impairment, neurochemical anomalies, and histopathological changes. Given these results, silibinin, when delivered using NLCs, is potentially more effective than free silibinin in decreasing AlCl3- induced neurotoxicity.

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.

L-Glutamic Acid Mitigates Carbon Tetrachloride-Induced Acute Tissue Injury by Reducing Oxidative Stress in a Rat Model

Background: Tetrachloromethane (CCl4) is a highly toxic environmental pollutant that causes specific and serious damage to various organs of animals and humans. The study of its effect on physiological and biochemical processes in the mammalian organism is very important, despite the severe restrictions on the use of tetrachloromantane in recent years. The fact is that they can be used as a model for many toxicological studies. Objective: The experimental studies aimed at exploring the ameliorative effects of L-glutamic acid (LGlu) on CCl4 toxicity in the myocardium, lung tissues and blood of male rats. Methods: Rats were exposed to CCl4, and later rats were treated with L-Glu. The GSH level and the activities of antioxidant enzymes were studied. In addition to this, the content of lipid peroxidation products was monitored. Results: The obtained results suggest that CCl4 causes oxidative stress in rat tissues, accompanied by an increase in lipid peroxidation products and a decrease in glutathione peroxidase (GPx), glutathione reductase (GR), glutathione transferase (GST), glucose-6-phosphate dehydrogenase (G6PDH), superoxide dismutase (SOD), catalase (CAT) activities and reduced glutathione (GSH) content. The enzymatic activity in tissues of rats treated with L-Glu was restored. Moreover, the changes, which were observed in the studied parameters, showed to be less significant compared to CCl4 treated group. Conclusion: These results suggest that L-Glu inhibits free radical processes. In summary, this study demonstrates the feasibility of the administration of L-Glu supplementation, which could be used to protect and mitigate the CCl4-induced oxidative stress.

Effects of Thiazolidine on AChE Enzyme Activity and Total Protein Level in Zebrafish (Danio rerio) Gill and Liver Tissue

Bu çalışmanın amacı, tiazolidin’in (Tiazolidin-4-karboksilik asit (4S)-2- (4-hidroksi-3-metoksifenil) zebra balığı (Danio rerio) solungaç ve karaciğer dokusunda asetilkolinesteraz enzim (AChE) aktivitesi ve total protein (TP) düzeyleri üzerindeki etkilerinin araştırılmasıdır. Zebra balıkları tiazolidin’in 0,2 ppm, 0,4 ppm ve 0,6 ppm farklı dozlarına 96 saat süreyle maruz bırakılmıştır. AChE enzim aktivitesi karaciğer dokusunda, tiazolidin’in 0,2 ppm, 0,4 ppm ve 0,6 ppm doz gruplarında, kontrol grubuna oranla artmıştır. Solungaç dokusunda ise, tiazolidin’in 0,2 ppm, 0,4 ppm ve 0,6 ppm doz gruplarında, kontrol grubuna oranla AChE enzim aktivitesinin azaldığı görülmüştür. Total protein seviyesi karaciğer dokusunda, tiazolidin’in 0,2 ppm, 0,4 ppm ve 0,6 ppm doz gruplarında, kontrol grubuna oranla azalmıştır. Solungaç dokusunda ise, tiazolidin’in 0,2 ppm, 0,4 ppm ve 0,6 ppm doz gruplarında, kontrol grubuna oranla total protein seviyelerinin önemli sayılabilecek oranda arttığı görülmüştür. Sonuç olarak, bu araştırmada tiazolidinin zebra balığı solungaç ve karaciğer dokuları üzerinde az da olsa zararlı etkilere neden olabileceği görülmüştür.

Anti-aging effect of phlorizin on D-galactose-induced aging in mice through antioxidant and anti-inflammatory activity, prevention of apoptosis, and regulation of the gut microbiota.

Aging is an inevitable and complicated process involving many physiological changes. Screening of natural biologically active anti-aging substances is a current research hotspot. Phlorizin (PZ), an important dihydrochalcone phytoconstituent, has been demonstrated to have antioxidant and anti-tumor effects. In this paper, different doses of PZ (20 and 40mg/kg) were used to research the protective effect on D-galactose (D-gal)-induced aging mice. Following hematoxylin and eosin staining and by observing the hippocampus, we found that PZ alleviated the damage caused by D-gal in neuronal cells, while PZ enhanced the learning and memory abilities of aging mice in a radical eight-arm maze. In order to explain the reasons for these anti-aging effects, we tested the antioxidant enzyme activity and malonic dialdehyde concentration in mouse serum, liver, and brain tissue. The contents of proteins related to anti-inflammation and apoptosis in brain tissue were analyzed, and the gut microbiota was also analyzed. The results indicated that PZ improved antioxidant enzyme activity while significantly reducing the malonic dialdehyde content. Western blotting analysis suggested that PZ effectively alleviated neuro-apoptosis via regulating the expressions of Bax, Bcl-2, and caspase-3. PZ also exerted anti-inflammation effects by regulating the interleukin-1β/inhibitor of nuclear factor kappa B alpha/nuclear factor kappa-light-chain-enhancer of activated B-cells signaling pathways in brain tissues. Importantly, PZ improved the structure and diversity of the gut microbiota, and the microbiota-gut-brain axis may hold a key role in PZ-induced anti-aging effects. In conclusion, PZ can be used as a potential drug candidate to combat aging.

ViphyllinTM, a Standardized Black Pepper Extract Exerts Antihyperglycemic Effect and Improves Sciatic Nerve Conduction in High Fat Diet/Streptozotocin-Induced Diabetic Model Rats.

PurposeResearch on plant-based formulations has drawn considerable attention in the management of diabetic neuropathy (DN) for having lesser side effects than the synthetic counterparts. Here, we have investigated for the first time the therapeutic effects of a standardized Piper nigrum L., (black pepper) seed extract, ViphyllinTM in mitigating hyperglycemia and neuropathic pain of type 2 diabetes model rats.MethodsType 2 diabetes was induced in male Wistar rats using high fat diet and a single dose of streptozotocin (60 mg/kg i.p.). The diabetic rats were orally administered with Viphyllin containing not less than 30% β-caryophyllene (BCP), at 25 mg, 50 mg and 100 mg/kg/day doses for 6 weeks. Changes in body weight, fasting blood glucose (FBG), glucose tolerance, and blood biochemical parameters were measured. The nociceptive response to thermal stimulus (tail flick test) and sciatic nerve conduction velocity (NCV) were recorded at the end of study.ResultsViphyllin treatment markedly improved the body weight and glucose tolerance in diabetic rats. Also, the extract could significantly reduce the diabetes-induced elevation in FBG, liver and kidney indices. Further, Viphyllin dose-dependently increased the nociception latency in tail flick test compared to untreated diabetic rats (p<0.05). Viphyllin at 100 mg/kg significantly increased the NCV (44.12±1.91*** m/s vs diabetic control 25.80±1.88 m/s). The antioxidant enzyme activities in sciatic nerve tissue were considerably increased in Viphyllin-treated groups compared to diabetic control. A 6-week treatment with Viphyllin markedly reversed the pathological manifestations of diabetes in vital organs such as liver, kidney and pancreas.ConclusionThe study concludes that Viphyllin exerts antidiabetic effects and improves nerve conduction to mitigate neuropathic pain.

Template-Independent Poly(A)-Tail Decay and RNASEL as Potential Cellular Biomarkers for Prostate Cancer Development.

Simple SummaryThe ultimate need in cancer tissue is to adapt translation machinery to accelerated protein synthesis in a rapidly proliferating environment. Our study was designed with the aim of integrating fundamental and clinical research to find new biomarkers for prostate cancer (PC) with clinical usefulness for the stratification prediction of healthy tissue transition into malignant phenotype. This study revealed: (i) an entirely novel mechanism of the regulatory influence of Poly(A) deadenylase in mRNAs translational activity and the 3′ mRNA untranslated region (3′UTR) length in cancer tissue and its regulation by the poly(A) decay; (ii) the RNASEL interrelationship with the inflammatory pattern of PC and corresponding tumor-adjacent and healthy tissue; and (iii) the sensitivity, specificity, and predictive value of these enzymes. The proposed manuscript is based on the use of specific biochemical and immunoassay methods with the principal research adapted for the use of tissue specimens.The post-transcriptional messenger RNA (mRNA) decay and turnover rate of the template-independent poly(A) tail, localized at the 3′-untranslated region (3′UTR) of mRNA, have been documented among subtle mechanisms of uncontrolled cancer tissue growth. The activity of Poly(A) deadenylase and the expression pattern of RNASEL have been examined. A total of 138 prostate tissue specimens from 46 PC patients (cancer specimens, corresponding adjacent surgically healthy tissues, and in their normal counterparts, at least 2 cm from carcinoma) were used. For the stratification prediction of healthy tissue transition into malignant phenotype, the enzyme activity of tumor-adjacent tissue was considered in relation to the presence of microfocal carcinoma. More than a four-times increase in specific enzyme activity (U/L g.prot) was registered in PC on account of both the dissociation of its inhibitor and genome reprogramming. The obtained ROC curve and Youden index showed that Poly(A) deadenylase identified PC with a sensitivity of 93.5% and a specificity of 94.6%. The RNASEL expression profile was raised significantly in PC, but the sensitivity was 40.5% and specificity was 86.9%. A significantly negative correlation between PC and control tissue counterparts with a higher expression pattern in lymphocyte-infiltrated samples were reported. In conclusion, significantly upregulated Poly(A) deadenylase activity may be a checkpoint for the transition of precancerous lesion to malignancy, while RNASEL may predict chronic inflammation.

Preventive Effect of Limosilactobacillus fermentum SCHY34 on Lead Acetate-Induced Neurological Damage in SD Rats

Lead poisoning caused by lead pollution seriously affects people's health. Lactic acid bacteria has been shown to be useful for biological scavenging of lead. In this experiment, Sprague-Dawley (SD) rats were treated with 200 mg/L of lead acetate solution daily to induce chronic lead poisoning, and oral Limosilactobacillus fermentum (L. fermentum) SCHY34 to study its mitigation effects and mechanisms on rat neurotoxicity. The L. fermentum SCHY34 showed competent results on in vitro survival rate and the lead ion adsorption rate. Animal experiments showed that L. fermentum SCHY34 maintained the morphology of rat liver, kidney, and hippocampi, reduced the accumulation of lead in the blood, liver, kidney, and brain tissue. Further, L. fermentum SCHY34 alleviated the lead-induced decline in spatial memory and response capacity of SD rats, and also regulated the secretion of neurotransmitters and related enzyme activities in the brain tissue of rats, such as glutamate (Glu), monoamine oxidase (MAO), acetylcholinesterase (AchE), cyclic adenosine monophosphate (cAMP), and adenylate cyclase (AC). In addition, the expression of genes related to cognitive capacity, antioxidation, and anti-apoptotic in rat brain tissues were increased L. fermentum SCHY34 treatment, such as brain-derived neurotrophic factor (BDNF), c-fos, c-jun, superoxide dismutase (SOD)1/2, Nuclear factor erythroid 2-related factor 2 (Nrf2), and B-cell lymphoma 2 (Bcl-2), and so on. L. fermentum SCHY34 showed a great biological scavenging and potential effect on alleviating the toxicity of lead ions.

Protective mechanism of traditional Chinese medicine guizhi fuling pills against carbon tetrachloride-induced kidney damage is through inhibiting oxidative stress, inflammation and regulating the intestinal flora

BackgroundChemical or drug-induced kidney damage has been recognized as a critical cause of kidney failure. The oxidative stress, inflammation, and imbalance of intestinal flora caused by carbon tetrachloride (CCl4) play a fundamental role in chronic kidney damage. Guizhi Fuling pills (GZFL) is a traditional formula consisting of five traditional Chinese medicinal herbs, which can promote blood circulation and improve kidney function. The underlying mechanisms of GZFL improving kidney damage are not fully understood yet. AimThe current study aimed to explore the effects of GZFL on CCl4-induced kidney damage and intestinal microbiota in mice. MethodsMale ICR mice were intraperitoneally administered with 20% CCl4 (mixed in a ratio of 1:4 in soybean oil) twice a week, for 4 weeks to induce kidney damage. Creatinine (CRE), urea nitrogen, antioxidant enzymes, and inflammatory cytokines were measured and the histology of the kidney, jejunum, and colon examination to assess kidney and intestinal damage. The expressions of nuclear factor-erythroid 2-related factor 2 (Nrf2) family members, nucleotide-binding domain and leucine-rich repeat protein 3 (NLRP3) inflammasome in kidney tissues, and the tight junction proteins in colonic tissues were detected by Western blot. The gut microbiota was analyzed through 16S rRNA gene sequencing. ResultsGZFL treatment decreased the serum CRE and urea nitrogen levels. Moreover, GZFL reduced the levels of pro-inflammatory cytokines and increased antioxidant enzyme activities in kidney and colonic tissues. GZFL improved the kidney, jejunum, and colon histology. Furthermore, GZFL inhibited the expressions of NLRP3, ASC, and cleaved-Caspase-1, while Nrf2, HO-1, NQO1, GCLM, and tight junction proteins were increased. The dysbiosis of intestinal microbiota improved after GZFL treatment. ConclusionsThis study showed that GZFL could improve kidney damage, which might be mainly via the integrated regulations of the Nrf2 pathway, NLRP3 inflammasome, and composition of intestinal microbiota.