Plasma Enzyme Activities Research Articles

Protective Effects of Phycobiliproteins from Arthrospira maxima (Spirulina) Against Cyclophosphamide-Induced Embryotoxicity and Genotoxicity in Pregnant CD1 Mice.

Background/Objectives: In recent years the global incidence of cancer during pregnancy is rising, occurring in 1 out of every 1000 pregnancies. In this regard, the most used chemotherapy drugs to treat cancer are alkylating agents such as cyclophosphamide (Cp). Despite its great efficacy, has been associated with the production of oxidative stress and DNA damage, leading to embryotoxicity, genotoxicity, and teratogenicity in the developing conceptus. Therefore, this study aimed to investigate the protective role of phycobiliproteins (PBP) derived from Arthrospira maxima (spirulina) in reducing Cp-induced embryotoxicity and genotoxicity in pregnant CD1 mice. Methods: Pregnant CD1 mice were divided into five groups: control, Cp 20 mg/kg, and three doses of PBP (50, 100, and 200 mg/kg) + Cp co-treatment. PBP were administered orally from day 6 to 10.5 dpc, followed by a single intraperitoneal dose of Cp on 10.5 dpc. Embryos were collected at 12.5 dpc to assess morphological development and vascular alterations, while maternal DNA damage was evaluated using micronucleus assays and antioxidant enzyme activity in maternal plasma. Results: PBP exhibited a dose-dependent protective effect against Cp-induced damage. The 200 mg/kg PBP dose significantly reduced developmental abnormalities, micronucleated polychromatic erythrocytes, and oxidative stress, (as evidenced by increased SOD and GPx activity). Conclusions: Phycobiliproteins from Arthrospira maxima (spirulina) effectively reduced Cp-induced morphological and vascular alterations in embryos and genotoxicity in pregnant mice. These findings highlight their potential as a complementary therapy to mitigate teratogenic risks during chemotherapy. Further research is needed to optimize dosing and explore clinical applications.

Influence of Cinnamon Supplementation on Liver Enzymes in Male Rabbits

The liver, a vital organ in the body, plays a central role in managing the metabolism of both internal and external substances. It is crucial for drug processing and detoxification; however, it is vulnerable to damage from foreign chemicals, alcohol intake, poor nutrition, environmental pollutants, illnesses, and various medications. Besides being commonly used as a spice, the inner bark of cinnamon (Cinnamomum zeylanicum L.) has gained popularity for its applications in disease prevention and treatment. Current evidence has indicated potential health benefits associated with cinnamon intake; however, its effects on liver function remain unclear. Therefore, this systematic review and meta-analysis aimed to investigate the effects of cinnamon supplementation on liver enzymes in male rabbits. Relevant studies were identified through a comprehensive search of PubMed/Medline, Scopus, Web of Science, Cochrane Library, and Embase databases up to September 2023. Ten male New Zealand White rabbits were randomly divided into two groups: (1) a control group, and (2) a group treated with cinnamon at a dose of 200 mg/kg body weight. Blood samples and liver homogenates were used to assess liver function markers in both serum and liver tissue. The results showed that cinnamon treatment led to a significant (P<0.05) reduction in the activity of blood plasma and liver enzymes "LDH", "AST", and "ALT", while "ALP" and "AcP" levels significantly (P<0.05) increased compared to the control group. Further high-quality studies are necessary, especially in populations with abnormal liver enzyme levels, to more definitively establish the clinical efficacy of cinnamon on liver function.

Effect of Stocking Density on Selected Blood Physiological, Biochemical and Enzymatical Variables of Broilers Grown to 3 kg under Antibiotic-free Conditions

Background: Reducing stocking densities may play a key role in minimizing difficulties such as physiological welfare when reducing or ending antimicrobials in poultry diets. As the demand for poultry and associated products increases, one of the major concerns centers on the question of whether stocking density influences welfare responses that are characteristic of physiological welfare. This study investigated the effect of stocking density on selected blood physiological variables along with plasma biochemistry variables and enzymes activities of broilers fed antibiotic-free diets grown to 3 kg. Methods: A total of 888 1-d-old Ross ´ Ross 708 chicks were randomly distributed into 24 pens based on stocking density treatments assignment. The treatments consisted of 4 densities (29, 33, 39 and 42 kg/m2) with six replicates. Treatments were blocked within the room to account for any variations in room conditions. Treatment assignments were randomized within each block. Used litter bedding was obtained from commercial farms to mimic commercial conditions and litter microflora. Birds were provided a three phase-feeding program (Starter: 0-14 d, Grower: 15-28 d and Finisher: 29-42 d) antibiotic-free diets. Feed and water provided ad libitum. Blood samples were collected from the brachial wing vein of 3 males and 3 females’ birds per pen on d 28 and 42 and analyzed immediately for blood physiological variables. Blood plasma samples were analyzed for T3, T4, corticosterone, biochemical variables and enzyme activities. Result: The only effect of stocking density was observed on partial pressure of CO2 (pCO2) and uric acid which were within physiological ranges for this species. However, blood glucose and plasma corticosterone concentrations were not affected by stocking density, suggesting no signs of physiological stress. Stocking densities up to 42 kg/m2 with proper environmental management may be suitable for both poultry integrators and contract growers to enhance broilers production efficiency without compromising the welfare of broilers grown to 3 kg body weight.

Preclinical efficacy and safety of adeno-associated virus 5 alpha-galactosidase: A gene therapy for Fabry disease

We developed a novel adeno-associated virus 5 gene therapy (AAV5-GLA) expressing human alpha-galactosidase A (GLA) under the control of a novel, small and strong, liver-restricted promoter. We assessed the preclinical potential of AAV5-GLA for treating Fabry disease, an X-linked hereditary metabolic disorder resulting from mutations in the gene encoding alpha-galactosidase A that lead to accumulation of the substrates globotriaosylceramide and globotriaosylsphingosine, causing heart, kidney, and central nervous system dysfunction. Effects of intravenously administered AAV5-GLA were evaluated in 1) alpha-galactosidase A-knockout mice aged 7–8 weeks (early in disease) and 20 weeks (nociception phenotype manifestation) and 2) cynomolgus macaques during an 8-week period. In both species, AAV5-GLA was observed as safe, generated detectable vector DNA and mRNA levels in liver, and produced stable enzyme activity in liver and plasma. In mice, dose-dependent transgene enzyme activity, cross-correction (substrate reduction) in kidney and heart, and improved nociception lasted over 6 months. Moreover, after delayed administration when animals displayed the nociception phenotype, target organ enzyme activity was present, and accumulated substrates were reduced. Given the strong, durable expression of active GLA with this promoter and favorable profile of adeno-associated virus-based gene therapy in humans, AAV5-GLA warrants further investigation in clinical trials for Fabry disease.

Effects of replacing fish meal with pork meal and supplementing with crystalline amino acids and fish oil on growth performance, body composition, enzyme activity, intestinal morphology, and immune response of Penaeus monodon

IntroductionThe present study aimed to evaluate the effects of substituting fish meal with pork meal in feed on the growth performance, feed utilization, intestinal morphology, and immune function of Penaeus monodon.MethodsA total of 600 uniformly sized juvenile shrimp with an initial weight of 1.20 ± 0.03 g were randomly assigned to four groups, with each group consisting of six replicates of 25 shrimp each. Fish meal protein was replaced with 0%, 5%, 10%, and 15% pork meal, and crystalline amino acids and fish oil were supplemented accordingly. The groups were designated as P0, P5, P10, and P15, and the study duration was 70 days.ResultsResults indicated no significant difference in specific growth rate (SGR) between the P5 and P10 groups compared to the P0 group. However, the SGR in the P15 group was significantly lower than that in the P5 and P10 groups. Feed conversion rates (FCRs) for the P5 and P10 groups were comparable to the P0 group, whereas the P15 group showed a significantly higher FCR than the other groups. There were no significant differences in whole body composition among all the groups. Apart from alanine, the amino acid composition of muscles did not differ significantly among all the groups. Results of enzyme activity in plasma revealed that the replacement of fish meal with pork meal reduced levels of aspartate aminotransferase (AST) and superoxide dismutase (SOD). Specifically, levels of AST in the P5 and P15 groups were significantly lower compared to the P0 and P10 groups. Histological examination of midgut tissues showed that villus height initially increased and then decreased with higher replacement ratios. The P5 group had the highest villus height, while the P15 group had the lowest villus height. Analysis of gene expression related to liver immune functions indicated that genes in TLR signaling pathway were significantly elevated in the P15 group compared to the other groups.DiscussionIn conclusion, even with the supplementation of crystalline amino acids and fish oil, pork meal should not replace more than 10% of fish meal in the feed of P. monodon to maintain optimal growth and feed efficiency.

Lentiviral Vector-Mediated Ex Vivo Hematopoietic Stem Cell Gene Therapy for Mucopolysaccharidosis IVA Murine Model.

Mucopolysaccharidosis IVA (MPS IVA) is an autosomal recessive disease caused by a mutation in the N-acetylgalactosamine-6-sulfate-sulfatase (GALNS) gene resulting in progressive systemic skeletal dysplasia. There is currently no effective treatment available for this skeletal condition. Thus, the development of a new therapy stands as an unmet challenge in reversing or alleviating the progression of the disease. Our research, which could be a game-changer, hypothesizes that ex vivo lentiviral (LV) gene therapy (GT) could produce the supraphysiological level of active GALNS enzyme by hematopoietic stem cells (HSCs) transduced with LVs carrying the native GALNS gene under two different promoters (CBh and COL2A1), impacting bone and cartilage abnormalities in MPS IVA. We conditioned newborn knock-out (Galns-/-) MPS IVA mice with busulfan and intravenously transplanted LV-modified HSCs isolated from the bone marrow of Galns-/- donor mice. Transplanted mice were autopsied at 16 weeks, and tissues were collected to assess the therapeutic efficacy of modified HSCs in MPS IVA mice. Although HSC-LV-CBh-hGALNS provided a higher GALNS enzyme activity in plasma, HSC-LV-COL2A1-hGALNS stably corrected heart and bone abnormalities better under a low level of GALNS enzyme. Our findings suggest that ex vivo LV-GT may potentially treat MPS IVA.

Nutritional Assessment and Sub-acute Toxicity Study in Wistar Rats Fed with a Compounded Diet of Roasted Plantain and Fish Sold in Otuoke, Nigeria

The widespread consumption of roasted plantain and fish (Bolle), a common delicacy in Bayelsa State has raised concerns over potential health risks associated with food processing contaminants such as acrylamides, polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs). The roasted plantain and fish are usually prepared by placing the peeled plantain and the dressed fish on a wired gauze and placed over a burning charcoal. The subacute toxicity study of this diet was investigated using twenty adult male rats averagely weighing 117.22 \(\pm\) 4.48 g. The rats were divided into four groups of five rats per group. Group A (Control), was fed with the standard rat chow, Group B; was fed with only the roasted plantain, Group C; was fed with only the roasted fish, and Group D; was fed with a combination of the roasted plantain and fish. The rats were given free access to the feed and portable water ad-libitum throughout the experimental protocol. Body/organ weight data were recorded before and after the completion of the experimental protocol. On day 15th of the experimental protocol, the rats were euthanized and blood specimens were obtained by cardiac puncture for biochemical analysis. The liver, kidney and heart were dissevered instantaneously and weighed. Findings from the study showed that the roasted plantain-fed group had significantly reduced body weight gain, and increased liver and kidney to body weight ratio (p>0.05). The plasma enzyme activity (AST, ALT and ALP) of the plantain-fed group were also significantly increased (p<0.05) while other groups showed non-significant differences (p>0.05) with the control group. The antioxidant status (CAT, SOD and GPx), haematological parameters (RBC, PCV, Hb, MCV, MCH, MCHC and platelet) of the plantain-fed groups were also significantly decreased (p<0.05) while other groups had non-significant changes (p>0.05) when compared to the control group. Malondialdehyde levels in the roasted plantain-fed group were also significantly increased (p<0.05).

Ecological adaptations of amphibians to environmental changes along an altitudinal gradient (Case Study: Bufo gargarizans) from phenotypic and genetic perspectives

BackgroundOrganisms have evolved a range of phenotypic and genetic adaptations to live in different environments along an altitudinal gradient. Herein, we studied the widely distributed Chinese toad, Bufo gargarizans, as a model and used an integrated phenotype-genotype approach to assess adaptations to different altitudinal environments.ResultsComparison of populations from four altitudes (50 m, 1200 m, 2300 m, and 3400 m) showed more effective defenses among high-altitude toads. These included thickened epidermis, more epidermal capillaries and granular glands, greater gland size in skin, and higher antioxidant enzyme activities in plasma. High-altitude toads also showed increased erythrocytes and hematocrit and elevated hemoglobin concentration, potentially improving oxygen delivery. Elevated altitude led to a metabolic shift from aerobic to anaerobic metabolism, and high-altitude populations favored carbohydrates over fatty acids to fuel for energy metabolism. Differentially expressed genes were associated with adaptive phenotypic changes. For instance, expression of genes associated with fatty acid metabolism showed greater suppression at high altitude (3400 m), consistent with decreased flux of β-hydroxybutyric acid and lower free fatty acids levels. Moreover, down-regulation of genes involved in carbon metabolism processes at high altitude (3400 m) were coincident with reduced TCA cycle flux. These results suggest that high-altitude toads adopt a metabolic suppression strategy for survival under harsh environmental conditions. Moreover, the hypoxia-inducible factor signaling cascade was activated at high altitude.ConclusionsCollectively, these results advance our comprehension of adaptation to high-altitude environments by revealing physiological and genetic mechanisms at work in Chinese toads living along altitudinal gradients.

Impact of Feeding Artemia franciscana Enriched with Various Oil Resources on Growth, Blood Biochemical and Behavioral Indices, and Survival of Oreochromis Niloticus

Abstract The main objective of the present trial was to examine the efficacy of feeding tilapia fry fish on Artemia franciscana diets supplemented with various oil emulsion resources in terms of performance, behavior indices, survival rate, blood biochemical parameters, and immunological response. Four hundred Nile tilapia fry (weighing 0.15±0.05 g and measuring 2.17±0.08 cm) were randomly allocated into four equal groups (each with five repetitions) and acclimatized for fifteen days. The first group served as the control and received unenriched Artemia franciscana (G0), while the remaining three groups were fed Artemia franciscana diets enriched with different oil resources (0.5 mL oil per liter for 6 hours): soybean oil (G1), sesame oil (G2), and rice bran oil (G3). Behavioral observations were recorded during the 45-day experimental period. At the end of the feeding trial, the chemical composition and fatty acid content of both Artemia and fingerlings were analyzed. Furthermore, the growth performance, survival, and immune response of the fingerlings were evaluated. The results indicated noticeable improvements in behavioral measurements (feeding, foraging and schooling), performance (final length, final weight, net weight gain, feed conversion ratio and specific growth rate), survival, and immune response among fry fish supplemented with enriched Artemia, particularly those enriched with soybean oil. Additionally, the chemical composition and fatty acid content of both Artemia and fish fry were significantly enhanced when oil emulsions are applied, with soybean oil demonstrating the most prominent improvements. Whereas, supplementing fry fish Artemia diets with oil resulted in lower liver enzyme activity and higher protein component levels in plasma in comparison to the control group. In brief, feeding Nile tilapia fry fish Artemia diets enriched with a soybean oil emulsion (0.5 mL/L) is recommended for promoting high performance, immunological activity, and survival throughout the early stage till fingerlings phase.

Study on sperm cryopreservation of hybrid fish derived from Carassius cuvieri (♀) × Carassius auratus red var (♂)

Cryopreservation of sperm is an effective method for conserving germplasm resources in fish genetic breeding. The high-quality hybrid fish (WR) derived from white crucian carp (Carassius cuvieri, WCC, ♀) and red crucian carp (C. auratus red var., RCC, ♂), possesses valuable traits such as high survival rates, strong resistance, and rapid growth, representing an important germplasm resources of crucian carp. This study compared the effects of different antifreeze solutions on sperm viability among three varieties (WR, WCC, and RCC) and examined changes in enzyme activity, fertilization rates, and hatching rates after cryopreservation, aiming to enhance the cryogenic sperm cryopreservation technique in hybrid fish and investigate the mechanisms underlying spermatozoa damage caused by cryopreservation. The results showed that the antifreeze combination of D14 with 15 % dimethyl sulfoxide (DMSO) had the best effect in preserving the sperm of WR and WCC, while D20 with 10 % DMSO was the optimal combination for RCC sperm. After ultra-low temperature preservation, the longevity, fertilization, and hatching rates of frozen sperm were significantly lower (P < 0.05) compared to fresh sperm. The enzyme activities of superoxide dismutase (SOD), lactate dehydrogenase (LDH), and creatine kinase (CK) were significantly decreased (P < 0.05) in spermatozoa, whereas they showed a significant increase (P < 0.05) in sperm plasma. Succinate dehydrogenase (SDH) activity was significantly reduced (P < 0.05) in frozen spermatozoa of RCC compared to fresh spermatozoa, and exhibited lower activity in frozen spermatozoa of WCC and WR. Additionally, SDH activity was significantly elevated (P < 0.05) in frozen sperm plasma, and glutathione reductase (GR) activity was significantly lower (P < 0.05) in both frozen sperm plasma and spermatozoa across all three species. The study demonstrated that cryopreservation had a significant effect on the enzyme activities of spermatozoa and sperm plasma in all three species. These findings provide important technical support for the conservation of high-quality fish germplasm resources, particularly for novel varieties resulting from distant hybridization in fish.

Effect of Supplementation with Black Soldier Fly Extract on Intestinal Function in Piglets Infected with Porcine Epidemic Diarrhea Virus.

Porcine epidemic diarrhea virus (PEDV) has developed as a global problem for the pig business, resulting in significant financial losses. Black soldier fly extract (BFE) has been proven to improve intestinal growth in pigs after weaning. Consequently, the goal of the present investigation was to explore the effects of BFE supplementation on intestinal function in PEDV-infected piglets. Eighteen piglets were randomly allocated to three groups: control, PEDV, and BFE + PEDV. The piglets in the BFE + PEDV group received 500 mg/kg BW of BFE orally for seven days from day 4 to 10 of the study. On day 9 of the study, six pigs from each group received either clean saline or PEDV solution at a dosage of 106 TCID50 (50% tissue culture infectious dose) per pig. On day 11, samples of blood and intestine were taken for additional investigation. The results indicated a significant decrease in the average daily gain (ADG) of piglets infected with PEDV (p < 0.05). Additionally, PEDV infection led to an alteration of blood indexes and a reduction in plasma D-xylose concentration and villi height in the small intestine, while it increased plasma diamine oxidase activity and small intestinal crypt depth in piglets (p < 0.05). The PEDV infection significantly reduced antioxidant enzyme activity in plasma and the gut, including total superoxide dismutase and catalase, while increasing contents of oxidation-relevant products such as malondialdehyde and hydrogen peroxide in piglets. Moreover, PEDV infection increased the mRNA expression level of antiviral-related genes (p < 0.05). Nutritional supplementation with BFE improved intestinal histomorphological indicators and reduced oxidative stress produced by PEDV infection in piglets. Interestingly, BFE could significantly promote the mRNA expression level of antiviral-related genes in the ileum (p < 0.05). Overall, the preliminary results suggest that dietary BFE could improve intestinal function in piglets after PEDV infection. Currently, the findings put a spotlight on the role of BFE in the prevention and treatment of PED in piglets.

THE GLA D313Y MUTATION IN FABRY DISEASE: DIAGNOSTIC DILEMMAS AND THERAPEUTIC CONSIDERATIONS

Abstract Fabry disease is a genetic disorder caused by mutations in the GLA gene, which encodes alpha–galactosidase A, leading to enzyme dysfunction and subsequent accumulation of glycosphingolipids. In the classical form, it occurs in childhood or early adolescence with renal, cardiac, and neurological involvement, resulting in a poor prognosis. The GLA D313Y is a missense mutation resulting in a significant reduction of enzyme activity in plasma, while leukocyte enzyme activity remains within normal limits, leading to a "pseudo–deficiency" of enzyme activity with uncertain pathogenicity. In literature, it has been observed that individuals with this mutation may present predominantly with central neurologic manifestations, characterized by multifocal white matter lesions. Historically they do not receive indications for initiating enzyme replacement or molecular chaperone therapies. We present the case of a man affected by arterial hypertension and history of chronic lower limb pain who experienced a lacunar stroke at the age of 35. The GLA D313Y mutation was identified and enzyme activity assay in plasma was within normal limits, precluding the initiation of enzyme replacement therapy. Over the following years, he suffered two additional ischemic strokes, a progressive decline in renal function to stage 3, and acute coronary syndrome. Transthoracic echocardiography revealed hypertrophic non–obstructive cardiomyopathy (SIVd 20 mm) with normal biventricular ejection fraction. Cardiac magnetic resonance showed late gadolinium enhancement of inferior junctional and inferior wall in middle segment (Fig.1). Considering the patient‘s clinical conditions involving multi–organ impairment, an enzymatic activity test was performed on leukocytes, revealing a reduced activity. This prompted the decision to initiate enzyme replacement therapy. This case report supports the hypothesis that the D313Y mutation may be associated with a later presentation and not only neurological involvement. In our experience, it’s mandatory to proceed with the determination of leukocyte enzyme activity in presence of normal enzyme activity in plasma. As an alternative, the detection of glycosphingolipids’ accumulation could be useful, but myocardial and renal biopsy poses a high bleeding risk, especially in anticoagulated or antiaggregated patients. The authors propose considering the initiation of therapy in patients with the D313Y mutation who exhibit significant organ involvement.

Adeno-associated virus-based gene therapy delivering combinations of two growth-associated genes to MPS IVA mice

Mucopolysaccharidosis type IVA (MPS IVA) is caused by a deficiency of the N-acetylgalactosamine-6-sulfate sulfatase (GALNS) enzyme responsible for the degradation of specific glycosaminoglycans (GAGs). The progressive accumulation of GAGs leads to various skeletal abnormalities (short stature, hypoplasia, tracheal obstruction) and several symptoms in other organs. To date, no treatment is effective for patients with bone abnormalities. To improve bone pathology, we propose a novel combination treatment with the AAV vectors expressing GALNS enzyme and a C-type natriuretic peptide (CNP; NPPC gene) as a growth-promoting agent for MPS IVA. In this study, an MPS IVA mouse model was treated with an AAV vector expressing GALNS combined with another AAV vector expressing NPPC gene, followed for 12 weeks. After the combination therapy, bone growth in mice was induced with increased enzyme activity in tissues (bone, liver, heart, lung) and plasma. Moreover, there were significant changes in bone morphology in CNP-treated mice with increased CNP activity in plasma. Delivering combinations of CNP and GALNS gene therapies enhanced bone growth in MPS IVA mice more than in GALNS gene therapy alone. Enzyme expression therapy alone fails to reach the bone growth region; our results indicate that combining it with CNP offers a potential alternative.

Exploring the dietary and therapeutic potential of licorice (Glycyrrhiza uralensis Fisch.) sprouts.

Ethnopharmacological relevanceThis research substantiates the traditional use of Glycyrrhiza uralensis Fisch. for liver health, with scientific evidence of the non-toxic and lipid-lowering properties of licorice sprout extracts. The sprouts' rich mineral and amino acid content, along with their strong antioxidant activity, reinforce their value in traditional medicine. These findings bridge ancient herbal practices with modern science, highlighting licorice's potential in contemporary therapeutic applications. Aim of the studyThe study aimed to investigate the dietary and medicinal potential of G. uralensis sprouts by assessing their safety, nutritional content, and antioxidant properties using both plant and animal models. Specifically, the study sought to determine the effects of different sizes of licorice sprouts on lipid metabolism in human liver cancer cells and their overall impact on rat health indicators. Materials and methodsThe study examined the effects of aqueous and organic extracts from G. uralensis sprouts of varying lengths on the cytotoxicity, lipid metabolism, and antioxidant activity in HepG2 cells, alongside in vivo impacts on Sprague-Dawley rats, using MTT, ICP, and HPLC. It aimed to assess the potential health benefits of licorice sprouts by analyzing their protective effects against oxidative stress and their nutritional content. ResultsLicorice sprout extracts from G. uralensis demonstrated no cytotoxicity in HepG2 cells, significantly reduced lipid levels, and enhanced antioxidant activities, with the longest sprouts (7 cm) showing higher mineral, sugar, and arginine content as well as increased glycyrrhizin and liquiritigenin. In vivo studies with Sprague-Dawley rats revealed weight gain and improved antioxidant enzyme activities in blood plasma and liver tissues after consuming the extracts, highlighting the sprouts' dietary and therapeutic potential. ConclusionsThis study is the first to demonstrate that G. uralensis sprouts, particularly those 7 cm in length, have no cytotoxic effects, reduce lipids, and have high mineral and antioxidant contents, offering promising dietary and therapeutic benefits.

Effect of Hydrolyzable Tannins of Euphorbia Plants on the Rat Antioxidant System of under Oxidative Stress

Aim: Study of the effect of hydrolyzable tannins isolated from Euphorbia plants on alloxan-induced OS under in vivo conditions.&#x0D; Methodology: Polyphenols were isolated from Euphorbia plants using an extraction method, after which the effect of these polyphenols on malondialdehyde (MDA) and glutathione reductase (GR) activity under oxidative stress in rats was determined in vivo.&#x0D; Results: The data obtained show that the studied hydrolyzable tannins reduce the amount of MDA in organs and tissues to the control level under alloxan-induced OS conditions. It was also found that polyphenols 3GHG, 1GHG and geraniin restore GR activity, and 2DGG had virtually no effect on the restoration of enzyme activity in the blood plasma and pancreas under OS conditions. However, the studied hydrolyzable tannins provide feebly expressed influence on the restoration of GR activity in the liver of rats under OS conditions.&#x0D; Conclusions: Thus, these hydrolyzed tannins exhibit strong antioxidant properties when acting on the amount of MDA, but when the GR activity is restored, their activity is weakly expressed.

The therapeutic effect and metabolic mechanism analysis of Guilingji on idiopathic oligo-asthenoteratozoospermia

IntroductionGuilingji, a famous traditional Chinese medicine (TCM) formula, has been used to combat aging and male sexual dysfunction in China for centuries. To date, there has been little evidence-based clinical research on the use of Guilingji to treat idiopathic oligo-asthenoteratozoospermia (OAT), and the therapeutic mechanism from a metabolic perspective needs to be investigated further. MethodsThis was a multicenter, double-blind, randomized controlled clinical study of 240 patients with idiopathic OAT recruited from four hospitals between January 2020 and January 2022. Patients were randomly assigned in a 1꞉1 ratio to receive oral Guilingji capsules or placebo for 12 weeks. The total progressive motile sperm count (TPMSC) was considered the primary outcome, and the other sperm parameters, seminal plasma parameters and serum hormones were considered the secondary outcome. A nontargeted metabolomics analysis of serum from OAT patients before and after Guilingji administration was performed by HPLC–MS to identify key metabolites. Furthermore, we used a rat model to show spermatogenesis phenotypes to validate the effect of the key metabolites screened from the patients. ResultsAt weeks 4, 8 and 12, TPMSC and other sperm parameters were significantly improved in the Guilingji group compared with the placebo group (P < 0.05 for all comparisons). At week 4, superoxide dismutase (SOD) and acrosomal enzyme activity of seminal plasma were significantly elevated in the Guilingji group compared with the placebo group, while reactive oxygen species (ROS) levels were significantly reduced (P < 0.05). Lactate dehydrogenase-X (LDHX) levels appeared to be significantly increased after 12 weeks continuous medication compared with Placebo group (P = 0.032). The metabolomics analysis of serum from OAT patients before and after Guilingji administration showed that the glucose-6-phosphate (G6P) concentration in patients' serum was significantly elevated after Guilingji treatment. Compared to the control, when Kidney-Yang deficiency model rats were treated with Guilingji or its key intermediate metabolite G6P, their sperm concentration and spermatozoic activity were improved similarly, and their structural damage of rat's testicular and epididymal tissues were recovered. ConclusionThis study provided valuable clinical evidence for the utility of Guilingji as a treatment for OAT. These findings thus demonstrate that G6P is involved in the therapeutic mechanism of Guilingji in OAT treatment based on clinical and rat intervention studies.

Changes in plasma metabolite concentrations and enzyme activities in aging riding horses.

In older horses, basal metabolic rate decreases, and plasma metabolite and hormone concentrations related to energy metabolism change. The occurrence of age-related diseases, which increases in old animals, may enhance inflammatory reactivity (inflammaging). Finding the appropriate treatment for inflammaging at an early stage may prevent various age-related diseases. Changes in metabolite and hormone concentrations and enzyme activities involved in energy metabolism in the plasma of clinically healthy riding horses of various ages were measured to identify biomarkers of inflammaging (persistent low-grade inflammation that occurs with aging). All horses were clinically healthy, and their body condition scores (BCSs) were 4 or 5 (9-point scale). Plasma triglyceride (TG), total cholesterol (T-Cho), blood urea nitrogen (BUN), insulin concentrations, malondialdehyde (MDA), and serum amyloid A (SAA) concentrations generally increased with age. Adiponectin concentrations, plasma superoxide dismutase (SOD), and leukocyte AMP-activated protein kinase (AMPK) activities decreased, while plasma aspartate aminotransferase (AST), alanine aminotransferase (ALT), and glutathione peroxidase (GPx) remained unchanged as horses aged. Although riding horses that partake in continuous exercise seems to be less likely to develop inflammaging, horses over 17 years of age tend to show proinflammatory signs with disordered lipid metabolism. In riding horses, SAA, in combination with other markers, may be a useful biomarker for inflammaging and dysregulated lipid metabolism in aging horses.

Study of age-related changes in plasma metabolites and enzyme activity of healthy small dogs that underwent medical checkups.

In Japan, the importance of medical checkups for pet dogs is increasing. In this study, we retrospectively explored the effects of age on plasma biomarkers in healthy small dogs that underwent medical checkups. Based on the modified American Animal Hospital Association Canine Life Stage Guidelines, 52 healthy small dogs were divided into 3 groups according to their life stage: young adult (1-4 years old), mature adult (5-11 years old), senior (12-15 years old). None of the dogs were obese. Plasma was collected from animals that underwent medical checkups at Muromi Animal Hospital (Fukuoka, Japan). Plasma glucose, triglyceride (TG), total protein, blood urea nitrogen (BUN), creatinine, total cholesterol, and albumin concentrations; alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase (ALP) activities; c-reactive protein (CRP), non-esterified fatty acid, malondialdehyde (MDA), serum amyloid A (SAA), insulin, and adiponectin (ADN) concentrations; glutathione peroxidase (GPx), superoxide dismutase (SOD), malate dehydrogenase (MDH), and lactate dehydrogenase (LDH) activities; and M/L ratio (MDH/LDH) were examined. Changes in the abovementioned plasma biomarker levels were compared between canines in different life stages. Plasma ADN concentrations and GPx, SOD, and MDH activities significantly decreased with age, whereas plasma ALP, BUN, TG, and MDA concentrations gradually increased. Plasma SAA levels measured by the latex agglutination method in 51 of the 52 small dogs that underwent medical checkups were below the detection limit. Plasma ADN concentrations, GPx, SOD activity, and BUN levels may be important biomarkers for clarifying the effect of age in healthy dogs that undergo medical checkups. However, plasma SAA values obtained by the latex agglutination method were not considered an age-related inflammation marker for healthy dogs.

Sunitinib induces cardiotoxicity through modulating oxidative stress and Nrf2-dependent ferroptosis in vitro and in vivo

SUN, a multi-targeted tyrosine kinase inhibitor, exerts cardiotoxicity which hinders its clinical use. It is necessary to elucidate molecular mechanism of SUN-induced cardiotoxicity. To elucidate molecular mechanism of SUN-induced cardiotoxicity and whether it is related to Nrf2-dependent ferroptosis, in vitro model with H9c2 cells derived from rat heart tissue and in vivo model (C57BL/6J male mouse) were used in the present study. In vivo model was established by oral treatment of SUN at dose of 10, 20, 40 mg/kg for 14 days. Body weight, ECG, plasma enzyme activities, histology staining were performed to evaluate heart function. Western-blot was performed to analyze the level of ferroptosis-related proteins. In vitro results indicated that SUN markedly induced ferroptosis embodied as collapsed MMP, accumulated iron and elevated ROS. In vivo results showed that SUN significantly impaired cardiac function. Abnormal electrocardiogram, increased serum CK and lactate LDH levels were significantly observed in SUN groups. Histology staining showed that SUN caused structural injuries and fibrosis deposition. Moreover, SUN increased the level of MDA and Fe2+ content, decreased the level of GSH. Both in vitro and in vivo experiments indicated that SUN reduced the expression of Nrf2, HO-1, NQO1, GPX4 and FTH1, enhanced the TfR expression. This study suggested that oxidative stress and Nrf2-dependent ferroptosis played a vital role in SUN-induced cardiotoxicity.

Evaluation of the relationship among gene expressions and enzyme activities with antioxidant role and presenilin 1 expression in Alzheimer's disease.

It is known that oxidative stress originating from reactive oxygen species plays a role in the pathogenesis of Alzheimer's disease. In this study, the role of antioxidant status associated with oxidative stress in Alzheimer's disease was investigated. Peripheral blood samples were obtained from 28 healthy individuals (as control) and 28 Alzheimer's patients who met the National Institute of Neurological and Communicative Diseases and Stroke/Alzheimer's Disease and Related Disorders Association criteria. Catalase, glutathione S-transferase and paraoxonase 1 enzyme activities in blood plasma and glutathione S-transferase enzyme activities in erythrocytes were determined by spectrophotometer. Catalase, glutathione S-transferase and presenilin 1 gene expressions in leukocytes were determined using qRT-PCR. Data were analysed with SPSS one-way anova, a LSD post hoc test at p < 0.05. The activity of each enzyme was significantly reduced in Alzheimer's patients compared to control. The catalase gene expression level did not change compared to the control. Glutathione S-transferase and presenilin 1 gene expression levels were increased compared to the control.