Total Alkaline Phosphatase Activity Research Articles

Expression analysis of zinc-metabolizing enzymes in the saliva as a new method of evaluating zinc content in the body: two case reports and a review of the literature

BackgroundThe activity level of alkaline phosphatase, a zinc-requiring enzyme in the serum, is used to indicate zinc nutritional status; however, it does not correlate with serum zinc levels or subjective symptoms of taste disorder in many cases. Hence, this study focused on the total activity of alkaline phosphatase, a zinc-requiring enzyme. The total alkaline phosphatasa activity level in the saliva was measured before and after zinc supplementation, and the results were compared with serum zinc levels.Case presentationThis study included patients with hypozincemia, specifically a patient with zinc-deficient taste disorder (patient 1: a 69-year-old Japanese woman) and a patient with glossodynia with zinc deficiency (patient 2: an 82-year-old Japanese woman). Saliva samples were collected, and blood tests were performed before and after zinc supplementation. Subjective symptoms and serum zinc levels were simultaneously evaluated. Zinc supplementation was performed using zinc acetate hydrate or Polaprezinc.ConclusionsTotal alkaline phosphatase activity levels were found to be associated with serum zinc levels and subjective symptoms. A further study with a higher number of patients is necessary to confirm whether total alkaline phosphatase activity levels more accurately reflect the amounts of zinc in the body than serum zinc levels.

Long-term dietary exposure to the non-steroidal anti-inflammatory drugs diclofenac and ibuprofen can affect the physiology of common carp (Cyprinus carpio) on multiple levels, even at “environmentally relevant” concentrations

The aim of the study was to evaluate the effects of emerging environmental contaminants, the non-steroidal anti-inflammatory drugs (NSAIDs) diclofenac (DCF) and ibuprofen (IBP), on physiological functions in juvenile common carp (Cyprinus carpio). Fish were exposed for 6 weeks, and for the first time, NSAIDs were administered through diet. Either substance was tested at two concentrations, 20 or 2000 μg/kg, resulting in four different treatments (DCF 20, DCF 2000, IBP 20, IBP 2000). The effects on haematological and biochemical profiles, the biomarkers of oxidative stress, and endocrine disruption were studied, and changes in RNA transcription were also monitored to obtain a comprehensive picture of toxicity. Fish exposure to high concentrations of NSAIDs (DCF 2000, IBP 2000) elicited numerous statistically significant changes (p < 0.05) in the endpoints investigated, with DCF being almost always more efficient than IBP. Compared to control fish, a decrease in total leukocyte count attributed to relative lymphopenia was observed. Plasma concentrations of total proteins, ammonia, and thyroxine, and enzyme activities of alanine aminotransferase (ALT), aspartate aminotransferase, and alkaline phosphatase (ALP) were significantly elevated in either group, as were the activities of certain hepatic antioxidant enzymes (superoxide dismutase, glutathione-S-transferase) in the DCF 2000 group. The transcriptomic profile of selected genes in the tissues of exposed fish was affected as well. Significant changes in plasma total proteins, ammonia, ALT, and ALP, as well as in the transcription of genes related to thyroid function and the antioxidant defense of the organism, were found even in fish exposed to the lower DCF concentration (DCF 20). As it was chosen to match DCF concentrations commonly detected in aquatic invertebrates (i.e., the potential feed source of fish), it can be considered “environmentally relevant”. Future research is necessary to shed more light on the dietary NSAID toxicity to fish.

Dietary Selenium Nanoparticles Improved Growth and Health Indices in Asian Seabass (Lates calcarifer) Juveniles Reared in High Saline Water.

A 60-day study was carried out to determine the effect of dietary selenium nanoparticles (SeNP) on growth, digestive enzymes, and health status of Asian seabass (Lates calcarifer, 46.5 ± 0.2 g) juveniles reared in high saline water (48 ppt). Five levels of SeNP were added to a basal diet (45% protein, 15% lipid), including control (0), 0.5 (SeNP0.5), 1.0 (SeNP1), 2 (SeNP2), and 4 (SeNP4) mg SeNP kg-1 diet. Fish were stocked into fifteen 2,000 L tanks (50 fish tank-1) filled with 1,800 L sand-filtered seawater (26.5 ± 1.5°C, 48.0 ± 0.2 ppt) in a flow-through system. Each dietary treatment was performed in three replicates. The growth rate positively increased in both linear and quadratic trends with increasing dietary SeNP level (P < 0.05). The liver Se concentration increased with increasing SeNP in diet (P < 0.05). Gut total protease, trypsin, chymotrypsin, alkaline phosphatase (ALP), lipase, and α-amylase activities were significantly enhanced in the SeNP4 group compared to the other treatments (P < 0.05). Antioxidant capacity improved in fish-fed SeNP2 and SeNP4 diets regarding catalase and superoxide dismutase activities and the liver glutathione content. Serum lysozyme and hemolytic activities and white blood cells' respiratory burst activity in the control were lower than in fish-fed SeNP-supplemented diets. Serum total protein, globulin, and globulin/albumin ratio in fish-fed SeNP1, SeNP2, and SeNP4 diets were higher than the other groups (P < 0.05). The interleukin-10 and granulocyte-macrophage colony-stimulating factor genes' relative transcription levels in the gut of fish-fed SeNP4 were higher than the other groups. Serum cholesterol, triglycerides, ALP, aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase significantly decreased in fish with increasing SeNP content in the diet. In conclusion, supplementing the diet with 4 mg kg-1, SeNP was recommended to improve growth and health indices in L. calcarifer juveniles reared in high saline water (48 ppt).

Evaluation of changes in serum biochemical parameters in zarudny’s spur-thighed tortoises (Testudo graeca zarudnyi) influenced by gender and season

BackgroundThe Zarudny’s spur-thighed tortoise or Iranian tortoise (Testudo graeca zarudnyi) has just been reported from Iran so far. Therefore, the purpose of this study was to determine the effects of season and gender on serum biochemical parameters of this valuable species in Iran.ResultsThis study was performed on 20 clinically healthy adult Zarudny’s spur-thighed tortoises. Blood samples were collected from the jugular vein and then serum biochemical parameters and body weight were measured in autumn and winter. The following biochemical parameters were measured: total cholesterol, triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, very low-density lipoprotein, total protein, creatinine, urea, glucose, calcium, inorganic phosphorus, total bilirubin, uric acid, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase. Urea concentration increased significantly at emergence from hibernation. Also, in winter, total protein, phosphorous, creatinine, total bilirubin concentrations and alkaline phosphatase activity decreased significantly compared to autumn, but aspartate aminotransferase and lactate dehydrogenase activities were significantly higher than in autumn. There was no significant difference for the parameters mentioned above by gender, except for phosphorous. Phosphorous concentration in females was significantly higher than that of males in autumn. Cholesterol and low-density lipoprotein cholesterol levels in females were significantly higher than males in autumn and winter.ConclusionsThe present study is the first study that monitors the serum biochemical parameters of adult Zarudny’s spur-thighed tortoises based on season and gender. Seasonal reference intervals should be used for biochemical parameters in this valuable species. Also, sex-specific reference intervals for phosphate and cholesterol are necessary.

Effects of fermented feed on growth performance, serum biochemical indexes, antioxidant capacity, and intestinal health of lion-head goslings.

The aim of this study was to evaluate the effects of fermented feed on growth performance, antioxidant indexes and intestinal health in lion-head goslings. 288 male lion-head goslings (one-day-old) were randomly divided into four groups (6 replicates per group, 12 samples per replicate): control group (basal diet) and fermented feed (FF) groups (basal diet supplemented with 2.5, 5.0 and 7.5% FF, respectively). The experimental period lasted 28 days. The results showed that 5.0 and 7.5% FF groups decreased feed conversion rate (FCR) when compared with the control group (p < 0.05). The 5.0% FF group reduced the activity of alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) in serum; while the 7.5% FF group decreased the concentration of total cholesterol (TC), ALP and LDH activity (p < 0.05). Furthermore, the 7.5% FF group significantly increased total antioxidant capacity (T-AOC) in serum (p < 0.05); 2.5% and 5.0% FF groups significantly increased glutathione peroxidase (GSH-Px) in serum (p < 0.05); all FF groups increased the activity of superoxide dismutase (T-SOD) in serum (p < 0.05). For intestinal health, the villous height and villi/crypt ratio in jejunum were increased in all FF groups, but crypt depth was decreased (p < 0.05); The 5.0% FF groups enhanced T-AOC activity in jejunum (p < 0.05); The 2.5% and 5.0% FF groups enhanced GSH-Px activity (p < 0.05) in jejunum; All FF groups reduced malondialdehyde (MDA) level in jejunum (p < 0.05). LEfSe analysis showed that the cecum microbiota was significantly dominant in the 2.5% FF group compared to the control group including Firmicutes, Lactobacillales, Lactobacillus, and Prevotella; the flora that were significantly dominant in the 5.0% FF group compared to the control group included Bacteroidaceae, Bacteroides, Megamonas, and Prevotella; and the groups that were significantly dominant in the 7.5% FF group compared to the control group included Bacteroidota, Bacteroides, Bacteroidaceae, and Ruminococcaceae. In summary, dietary FF supplementation improved growth performance, serum biochemical parameters and antioxidant capacity of lion-head goslings, as well as improved jejunal tissue morphology and optimized intestinal flora structure. In particular, the FF addition at a dose of 7.5% was relatively more effective for lion- head goslings.

Effects of dietary bile acids levels on growth performance, anti-oxidative capacity, immunity and intestinal microbiota of abalone Haliotis discus hannai

Abalone Haliotis discus hannai (initial weight: 38.79 ± 0.70 g) was used as the experimental animal in a 105-day feeding trial to investigate the influence of dietary bile acids levels on the growth, anti-oxidation, immune response and intestinal microbiota. Six isonitrogenous and isolipidic diets were prepared by adding 0 (control group), 15, 30, 60, 120 and 240 mg/kg of bile acids, respectively (named BA0, BA15, BA30, BA60, BA120 and BA240, respectively). It was found that survival of abalone between groups had no significant difference (P > 0.05). Compared to the control, significant improvements in weight gain rate (WGR) were observed in the groups of BA30 and BA60 (P < 0.05). Based on WGR, the broken line regression model analysis showed that the optimum demand for dietary bile acids for abalone was 35.47 mg/kg. Dietary bile acids increased the total anti-oxidative capacity and activities of catalase, superoxide dismutase, lysozyme and alkaline phosphatase, meanwhile decreased the content of malondialdehyde, alanine aminotransferase and aspartate aminotransferase activities in the cell-free hemolymph (P < 0.05). When bile acids were added to the diets, mRNA levels of genes related to pro-inflammatory factors and apoptosis in the digestive gland were down-regulated (P < 0.05). In contrast, the expression of genes related to anti-oxidation was significantly up-regulated (P < 0.05). The Firmicutes, Actinobacteriota and Proteobacteria were the most abundant phyla in intestine. And dietary bile acids significantly decreased the abundance of Actinobacteria and increased the abundance of Firmicutes (P < 0.05). In conclusion, supplementation of dietary bile acids within 120 mg/kg significantly increased the growth of abalone. The 34.62 mg/kg of dietary bile acids significantly increased the anti-oxidative capacity of abalone. Appropriate levels of dietary bile acids (34.62–61.75 mg/kg) promote the immunity of abalone. Application of appropriate levels of bile acids in diets (34.62 mg/kg) changed the intestinal microbiota and promoted the intestinal health of abalone.

Salivary Total Protein and Alkaline Phosphatase Activity as Biomarkers for Skeletal Maturity and Growth Prediction in Healthy Children: An In Vivo Study.

Skeletal maturity assessment involves radiographic analysis and visual inspection of developing bone and their initial appearance or sequential ossification and related changes in size and shape along with the expression of various biomarkers in body fluids. To investigate the correlation of biomarkers such as salivary alkaline phosphatase (S-ALP) and salivary total protein (STP) with skeletal maturity assessment and growth prediction in growing children. A total of 8-15-year-old 150 healthy children were divided into five groups depending upon radiographic stage maturity of the middle phalanx of the left hand's third finger according to the Hagg and Taranger method. Radiographs were taken using intraoral periapical (IOPA) radiographic films. Salivary alkaline phosphatase (S-ALP) activity in the MP3 G group was significantly higher than MP3 F group and MP3 I group. Total protein levels in MP3 F were significantly lower than in MP3 G. The mean value of S-ALP (33541.45 IU/L) and that of STP (2.77 mg/mL) was observed to be highest in the MP3 G group (G3) group. Salivary total protein (STP) and S-ALP may be used as an additional diagnostic tool to assess skeletal maturation and optimize growth prediction during myofunctional orthodontic treatment. Skeletal maturity assessment plays a significant role in orthodontic diagnosis, treatment planning, and stability of orthodontic treatment. Radiographic parameters involve radiographic exposure; hence in this study noninvasive biomarkers such as S-ALP and STP have been evaluated for skeletal maturity assessment and growth prediction. Abhangi KK, Choudhari SR, Butala PB, et al. Salivary Total Protein and Alkaline Phosphatase Activity as Biomarkers for Skeletal Maturity and Growth Prediction in Healthy Children: An In Vivo Study. Int J Clin Pediatr Dent 2023;16(4):603-607.

Dinoflagellate Karenia mikimotoi on the growth performance, antioxidative responses, and physiological activities of the rotifer Brachionus plicatilis.

The harmful dinoflagellate Karenia mikimotoi is responsible for the mortality of aquatic animals. However, the mechanism behind these toxic effects has not been fully determined. Herein, the toxic effects of K. mikimotoi on the growth performance, antioxidative responses, physiological activities, and energetic substance contents of rotifer Brachionus plicatilis were assessed. Rotifers were exposed to Nannochloropsis salina (Eustigmatophyceae), K. mikimotoi, and a mixture of N. salina and K. mikimotoi with biomass ratio proportions of 3:1, 1:1, and 1:3, respectively. Results indicated that K. mikimotoi negatively affected the population growth, survival, and specific growth rates of rotifers within 24 h. The level of reactive oxygen species (ROS), the content of malondialdehyde, and the activity of amylase increased. However, the total antioxidant capacity level, pepsase, cellulase, alkaline phosphatase, xanthine oxidase, and lactate dehydrogenase activities, and glycogen and protein contents decreased with increasing proportions of K. mikimotoi. The mixture of 50% N. salina and 50% K. mikimotoi promoted the increase in glutamic-pyruvic transaminase activity and triglyceride content. These findings underscore ROS-mediated antioxidative responses, physiological responses, and energetic substance content changes in B. plicatilis work together to affect population dynamics inhibition of rotifers by K. mikimotoi.

Chromolaena odorata (L.) R. M. King and H. Robinson Leaves Aqueous Extract Improves the Femoral Head in Ethanol-Induced Osteonecrosis in Rats.

Chronic alcohol consumption damages bone formation and causes bone pathology, including osteonecrosis of the femoral head. The aim of this work was to evaluate the effects of the leaf aqueous extract of Chromolaena odorata (C. odorata) on the femoral head in ethanol-induced osteonecrosis in rats. Animals received alcohol (40°) at 3 g/kg for 12 weeks. A group of animals were sacrificed to attest to the instalment of osteonecrosis by using histopathological analysis. The remaining animals received alcohol concomitantly with the plant extract (150, 300, or 600 mg/kg) or diclofenac (1 mg/kg) for 28 additional days. At the end of the experimental period, biochemical parameters including total cholesterol, triglycerides, calcium, alkaline phosphatase (ALP), reduced glutathione (GSH), malondialdehyde (MDA), nitrite, superoxide dismutase (SOD), and catalase activities were measured. Histopathological and histomorphometry analyses of femurs were also assessed. The administration of alcohol, irrespective of the experimental period, induced a significant increase in total cholesterol (p < 0.05) and triglyceride (p < 0.01) and a decrease in ALP (p < 0.05) and calcium (p < 0.05-p < 0.001) levels. Intoxicated animals showed an alteration in oxidative stress parameters accompanied by a significant drop in bone cortical thickness and density with necrosis and marked bone resorption. The concomitant administration of the plant with ethanol reversed the alcohol-induced bone defect, characterized by the improvement of the lipid profile (p < 0.001), bone calcium concentration (p < 0.05), bone ALP activity (p < 0.001), oxidative stress parameters, improved cortical bone thickness (p < 0.01), and bone density (p < 0.05). These results are supported by the absence of bone resorption with an obvious effect at a dose of 300 mg/kg. The pharmacological effect of the extract on ethanol-induced osteonecrosis of the femoral head is probably due to its osteogenic, hypolipidemic, and antioxidant properties, justifying its use in Cameroonian folk medicine for articulation and bone pain management.

The effects of Cyclina sinensis bioturbation on alkaline phosphatase (APA) and total microbial hydrolytic activities (MBA) in marine clam-shrimp integrated ponds.

This study investigated the influence of Venus clam Cyclina sinensis bioturbation activities on the total benthic microbial and phosphatase activities and selected sediment properties: total phosphorous TP, total organic nitrogen-TON and total organic carbon-TOC. Sediments samples from clam-shrimp integrated-pond and non-clam integrated-pond were sampled for the study in which microbial activity (MBA) and alkaline phosphatase activity (APA), sediment organic contents: TP, TON, TOC, TOM and water quality parameter (dissolved oxygen, temperature, pH and moisture content were analyzed. The p-nitrophenyl phosphate disodium (p-NPP) and Fluorescein diacetate-(FDA) were adopted to measure APA and MBA respectively. Results indicated that microbial activity-MBA and alkaline-phosphatase activity-APA in sediments significantly increased in the pond cultured with clam/shrimp compared to non-clam cultured pond. Phosphate concentration increased significantly and varied among months (p<0.05) implying increased nitrogen concentration in the sediments. The concentrations of total organic nitrogen and total organic carbon didn't differ significantly among months and with the treatments (p>0.05), implying increased TON mineralization. Correlation analyses showed that there was positive correlation with the total microbial activity, alkaline-phosphatase activity, phosphorous concentration and organic matter content within the sediments bioturbated by the Venus clam. The results suggest that, the Venus clam sediment reworking mechanisms in sediment influenced the sediment microbial and APA activities and mineralization thereby affecting the pond alkaline phosphatase enzyme related activities.

Clinical-radiological, hematological and biochemical assessment of bone replacement with germanium-doped calcium-phosphate ceramics for fragment fractures of tubular bones in dogs

Abstract. Among surgical pathology in animals, asignificant share is occupied by various traumatic injuries of the skeleton, the consequence of which are bonefractures. The results of X-ray and biochemical indicators of blood for bone fragment fractures in dogs arepresented.The purpose of the work is to clinically, radiologically, and pathochemically justify bone replacementwith germanium-doped calcium-phosphate ceramicsfor bone fragment fractures in dogs.In the injured animals of the experimental group(n=10), bone defects were replaced with germanium-doped calcium-phosphate ceramics (ГТлГег-700),in the control group (n=10) with unalloyed ceramics(ГТг-700). The activity of total alkaline phosphatase (ALP)and its bone isoenzyme (KiLP), tartrate-resistant acidphosphatase (TrKF) was determined in blood serum.Measurements were made with a Stat Fax 4500 spectrophotometer.On the 60th day, in the animals of the experimentalgroup, the filling of the bone defect with regeneratedmaterial of high X-ray density was noted, with the absence of periosteal reaction, which indicated the complete consolidation of the fracture and was the reasonfor the removal of the means of fixation. In the control,the area of the bone tissue defect that was not filledwith regenerate was visualized.The activity of tartrate-resistant acid phosphatase(TRP) in both groups tended to increase. On the 14th day,the animals of the experimental group had a peak activity of TrKf (33.8±0.31), which was 1.3 times (p&lt;0.001)higher than the level in the control group and 1.5 times(p&lt;0.001) of clinically healthy animals, respectively. Onthe 30th day of research, a 1.2-fold (p&lt;0.001) decreasein the level of TrCF was noted in the experimental groupcompared to the control group, in which the indicatorreached peak values at 34.9±0.25 units/l.The maximum activity of the bone isoenzyme alkaline phosphatase (KiLF) in the experimental groupwas observed on the 14th day of reparative osteogenesis and was 41.7±0.43 units/l, which is 1.2 times(p&lt;0.001) higher than the indicator of the controlgroup. On the other hand, in the last group, the peakactivity of the bone isozyme was noted only on the30th day (40.9±0.48 units/l), while the indicator of theexperimental animals decreased dynamically duringthis period.The use of calcium-phosphate ceramics for fragment fractures of the bones of the tibia in dogs helpsto reduce the intensity of the inflammatory-resorptivestage of reparative osteogenesis and accelerates consolidation by 1.4 times. The dynamics of biochemicalmarkers of bone metabolism testifies to the optimizedcourse of reparative osteogenesis during osteoreplacement of bone defects with germanium-doped calcium-phosphate ceramics. Key words: bioceramics, germanium, total alkaline phosphatase, bone isoenzyme of alkaline phosphatase, tartrate-resistant acid phosphatase, bonetissue.

A Mouse Model with a Frameshift Mutation in the Nuclear Factor I/X (NFIX) Gene Has Phenotypic Features of Marshall-Smith Syndrome.

The nuclear factor I/X (NFIX) gene encodes a ubiquitously expressed transcription factor whose mutations lead to two allelic disorders characterized by developmental, skeletal, and neural abnormalities, namely, Malan syndrome (MAL) and Marshall-Smith syndrome (MSS). NFIX mutations associated with MAL mainly cluster in exon 2 and are cleared by nonsense-mediated decay (NMD) leading to NFIX haploinsufficiency, whereas NFIX mutations associated with MSS are clustered in exons 6-10 and escape NMD and result in the production of dominant-negative mutant NFIX proteins. Thus, different NFIX mutations have distinct consequences on NFIX expression. To elucidate the in vivo effects of MSS-associated NFIX exon 7 mutations, we used CRISPR-Cas9 to generate mouse models with exon 7 deletions that comprised: a frameshift deletion of two nucleotides (Nfix Del2); in-frame deletion of 24 nucleotides (Nfix Del24); and deletion of 140 nucleotides (Nfix Del140). Nfix +/Del2, Nfix +/Del24, Nfix +/Del140, Nfix Del24/Del24, and Nfix Del140/Del140 mice were viable, normal, and fertile, with no skeletal abnormalities, but Nfix Del2/Del2 mice had significantly reduced viability (p < 0.002) and died at 2-3 weeks of age. Nfix Del2 was not cleared by NMD, and NfixDel2/Del2 mice, when compared to Nfix +/+ and Nfix +/Del2 mice, had: growth retardation; short stature with kyphosis; reduced skull length; marked porosity of the vertebrae with decreased vertebral and femoral bone mineral content; and reduced caudal vertebrae height and femur length. Plasma biochemistry analysis revealed Nfix Del2/Del2 mice to have increased total alkaline phosphatase activity but decreased C-terminal telopeptide and procollagen-type-1-N-terminal propeptide concentrations compared to Nfix +/+ and Nfix +/Del2 mice. Nfix Del2/Del2 mice were also found to have enlarged cerebral cortices and ventricular areas but smaller dentate gyrus compared to Nfix +/+ mice. Thus, Nfix Del2/Del2 mice provide a model for studying the in vivo effects of NFIX mutants that escape NMD and result in developmental abnormalities of the skeletal and neural tissues that are associated with MSS. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

Зміни розрахункових індексів біохімічних показників сироватки крові щурів різного віку після заповнення дефекту в метафізі стегнової кістки алогенними кістковими імплантатами, насиченими мезенхімальними стовбуровими клітинами

The purpose of the study was based on the analysis of calculated biochemical parameters of mineralization in the blood serum of laboratory rats to evaluate the course of metabolic processes in bone tissue after filling a defect of critical size in the metaphysis of the femur with allogeneic bone implants saturated with mesenchymal stem cells. Materials and methods. Indicators of mineralization of bone tissue in the blood serum of white rats were studied (the content of total protein, calcium, alkaline and acid phosphatase activity), the ratio of alkaline to acid phosphatase activity, as well as the degree of mineralization, were calculated. Results and discussion. In 3-month-old rats with alloimplants without mesenchymal stem cells on the 90th day, a decrease in the degree of mineralization was observed, which reflects the final stages of bone tissue remodeling by 1.13 times compared to the level of this in animals of a similar age and conditions of filling the defect on the 14th day (р = 0.008) and 1.12 times compared to the data on the 28th day of the experiment (р = 0.008). In 3-month-old rats with alloimplants saturated with mesenchymal stem cells, a decrease in the degree of mineralization on the 90th day was determined compared to that in experimental rats of a similar age and defect filling conditions on the 14th day by 1.13 times (p = 0.008), and compared with the data of the similar group on the 28th day – 1.14 times (p = 0.008), which reflects the delay in the course of the full stage of bone tissue remodeling due to the use of mesenchymal stem cells. The ratio of serum alkaline to acid phosphatase activity in 3-month-old rats with alloimplants saturated with mesenchymal stem cells was 1.32 times lower on the 14th day compared to the data of a group of the same age with alloimplants without mesenchymal stem cells (p = 0.008), and on the 90th day – by 1.12 times (p = 0.008), which also indicates a delay in the early stages of bone tissue mineralization due to the use of mesenchymal stem cells as part of alloimplants. In 12-month-old rats without the use of mesenchymal stem cells, on the 90th day, there was a decrease in the degree of mineralization by 1.10 times, in accordance with the level of this in a group of rats of a similar age and conditions of defect filling on the 14th day (р = 0.008) and by 1.09 times compared to that on the 28th day (p = 0.008), from which we can conclude about a progressive decrease in the activity of the final stages of bone tissue remodeling. The ratio of the activity of alkaline and acid phosphatase in 12-month-old rats with alloimplants without mesenchymal stem cells on the 90th day was significantly inferior to that in the corresponding 3-month-old animals by 1.12 times (p = 0.008), which indicates a lower activity of the initial stages of bone tissue mineralization in the late stages of defect healing. This indicator progressively decreased during the experiment. Thus, on the 28th day, it was 1.12 times smaller than on the 14th day, and on the 90th day – by 1.53 times less, in accordance with the data on the 14th day (p = 0.008) and by 1.36 times compared to that on the 28th day (p = 0.008). Conclusion. Treatment of experimental rats with a critical size defect in the femoral metaphysis with alloimplants saturated with mesenchymal stem cells, especially in the early stages, leads to a slowing down of bone tissue remodeling processes

Calcium homeostasis and certain aspects of its disturbances in juvenile idiopathic arthritis

Background. In the pathogenesis of juvenile idiopathic arthritis (JIA), the main role is played by immunopathological changes in the body with a loss of tolerance to the elements of own tissues; herewith, disorders of calcium and bone metabolism are very important. Such changes occur as a result of autoimmune inflammation, pharmacotherapy, and the influence of a number of other factors that negatively affect calcium homeostasis in the body. Purpose: to study the features of calcium homeostasis and certain aspects of its disorders with an assessment of the structural and functional state of bone tissue, taking into account clinical subtypes and disease activity. Material and methods. Sixty-two children with JIA aged 3.5 to 16 years were examined, of them 11 had systemic and 51 had oligo- and polyarthritis. There were determined serum concentrations of a total calcium using the Lachema test kit (Czech Republic), protein-bound and ultrafiltered fractions, content of inorganic phosphorus (with the generally accepted spectrophotometric method using the Cobas 6000 analyzer and test systems by Roche Diagnostics, Switzerland), the activity of total alkaline phosphatase and its isoenzymes (bone and intestinal) using the Lachema test system (Czech Republic). Ultrasonic osteometry of the calcaneal (trabecular) bone was performed on the Achilles device (Lunar, USA). Results. A significant decrease was found in the average concentration of total calcium, protein-bound calcium in systemic JIA and in high disease activity. The concentration of the ultrafiltered calcium fraction decreased only with high disease activity. The average concentration of inorganic phosphorus in children with oligo- and polyarthritis was within the normal range, while in systemic JIA it decreased. A significant decrease in the serum content of inorganic phosphorus, as well as in the activity of total alkaline phosphatase and its bone isoenzyme was detected in patients with high activity of systemic JIA. In patients with JIA (oligo-, polyarthritis) characterized by a slowly progressive rheumatic process, only the indicator of broadband ultrasound attenuation significantly changed during the first year of the disease, while the speed of ultrasound propagation and the index of bone tissue strength were not changed. In patients with a longer duration of the disease, all densitometric indicators decreased significantly. In the group of patients with systemic JIA and a rapidly progressive course, high activity of the disease that required a glucocorticoid therapy, a significant loss of bone mass was noted by the end of the first year of the disease. Conclusions. In JIA, there are changes in the concentration of total calcium and its protein-bound and ultrafiltered fractions in the blood serum, which indicate the tension of calcium-phosphorus metabolism and possible calcium deficiency in the body already at the early stages of the pathological process. A decrease in the activity of the alkaline phosphatase and its bone isoenzyme is associated with a violation of the structural and functional changes in the bone system of patients with JIA, which progresses with the duration of the disease. Patients with JIA require timely diagnosis and monitoring of calcium-phosphorus metabolism disorders with an assessment of the structural and functional state of the bone system for purposeful correction of comprehensive therapy due to the use of drugs in order to increase bone tissue regeneration, reduce the progression of osteopenia and osteoporosis, and preserve the health of the growing organism.

Novel Finding of Urinary Erythropoietin as an Early Biomarker of Cisplatin-Induced Nephrotoxicity.

Cisplatin is a chemotherapeutic drug used to treat a great variety of solid tumors. Its dose is commonly limited by its nephrotoxicity, manifested as acute kidney injury (AKI). Erythropoietin (Epo) is a glycoprotein hormone that regulates the production of red blood cells. This study was performed to evaluate the presence of endogenous Epo in male Wistar rat urine and to analyse changes in urinary Epo levels in response to cisplatin- induced AKI. Dose-dependent studies and time-dependent experiments were performed to evaluate changes in urea nitrogen and creatinine in plasma as well as Epo, neutrophil gelatinase-associated lipocalin (NGAL), alkaline phosphatase (AP) activity, creatinine and total proteins in urine at 2days post-dosing. Rats received 2, 5 or 10mg/kg b.w., i.p. of cisplatin. At 5mg/kg b.w., i.p. cisplatin, significant increases in urinary Epo were detected. Significant increases in urea nitrogen and creatinine in plasma, NGAL, AP, proteins, and Epo were observed in urine from rats that received 10mg/kg b.w., i.p. of cisplatin. In the time-dependent experiments, rats were injected with a dose of 5mg/kg b.w., i.p. of cisplatin, and sampling occurred 2, 4, and 14 days post-dosing. In these animals, there were significant increases in urea nitrogen and creatinine in plasma and total proteins, AP activity, Epo, and NGAL in urine on day 4. Urinary Epo was also detected on day 2. Taken together, these findings provide weight of evidence for urinary Epo as a promising early biomarker of cisplatin-induced AKI in male rats.

Role of plasma calcium, phosphorus, alkaline phosphatase, and parathyroid ‎hormone in the prediction of vitamin D deficiency‎

Vitamin D deficiency is a highly prevalent medical condition associated with various clinical and biochemical outcomes although these outcomes might be absent in a significant percentage of patients. Altered serum calcium, phosphorus, parathyroid hormone and alkaline phosphatase activity are expected but not proven by many research. The current study aims to investigate the possibility of these biomarkers to be used as indicators of vitamin D deficiency or insufficiency. The study enrolled 150 randomly selected participants who have no acute or chronic medical condition with various levels of serum vitamin D ranging from normal to severely deficient. They were investigated for serum total calcium, phosphorus, parathyroid hormone and alkaline phosphatase activity. The results were statistically compared among the studied groups and correlated with serum vitamin D levels. There was no significant difference in mean serum calcium, phosphorus and alkaline phosphatase activity among the groups with poor correlation with vitamin D. Mean serum parathyroid hormone has shown a significant difference among the studied groups with a strong negative correlation with vitamin D. This difference was apparent even in patients with vitamin D insufficiency. Serum calcium, PO4 and ALP activity seem to have a poor correlation with vitamin D concentration. In addition, serum PTH might be considered a sensitive marker of both vitamin D deficiency and insufficiency.

Взаимосвязь между нарушением иммунобиохимического профиля крови стельных коров и заболеваемостью телят ротавирусной инфекцией

The conducted studies have established that in the group of deep-calving cows with normal and elevated biochemical parameters, an increase in the content of total protein, glucose, concentration of inorganic calcium, an increase in the activity of ALT and alkaline phosphatase was observed. From immunological indicators, the number of T- and B-cells increased. Here, factors that increase the natural resistance of the body of cows, namely, the increase in bactericidal and lysozyme activity of blood serum, are of great importance. In the group of deep-calving cows, where immunobiochemical parameters in most cases were lower than control analogues, the concentration of total protein, glucose, triglycerides, cholesterol, carotene, AST and alkaline phosphatase activity decreased most intensively. Conducting immunological monitoring, a decrease in the number of T- and B-immunocompetent cells was established, bactericidal and lysozyme activity and an indicator of phagocytic activity fell. Draws attention to the weakening of the cellular link of immunity, which indicates the depletion of T-lymphocytes and the development of immunodeficiency states in females. Calves obtained from cow mothers with low immunobiochemical parameters were much more likely to become ill with rotavirus infection, and in half of the diseased calves (26.6 %) a severe form was noted, and in 20 % of the diseased calves a moderate severity of the disease was diagnosed.

Sex differences in the hepatotropic effects of antiulcer drugs and placenta cryoextract in an experimental rat liver injury model

Background/Aim: Sex-related variances in drug metabolism provide a foundation for refining treatment protocols for prevalent conditions based on the patient's sex. Tailoring treatment strategies based on sex is particularly noteworthy among patients with comorbid illnesses due to the potential for drug interactions and the impact of concurrent diseases on clinical outcomes. Aim of this study was to assess the hepatotropic effects of antiulcer drugs (esomeprazole, clarithromycin and metronidazole - E/C/M) and placenta cryoextract (CEP) within a simulated model of tetrachloromethane (CCl4 )-induced hepatitis combined with underlying ethanol-induced liver cirrhosis (EILC), with a focus on the role of subjects' sex. Methods: Using 112 male and female rats, the research explored the effects of different sex hormone levels. Chronic EILC was induced by administering a 50.0 % CCl4 oil solution (8 mL/kg) twice a week, combined with a 5.0 % ethanol solution, over 45 days. Total protein (TP) levels and alkaline phosphatase (AP) activity were measured spectrophotometrically. Results: The research findings indicate that the onset of EILC and the administration of E/C/M resulted in a significantly greater 10.8 % (p = 0.03) reduction in TP levels among females compared to males, without altering hormonal status. Introducing CEP led to a noteworthy (p &lt; 0.001) rise in TP levels, by 30.8 % in males and 33.9 % in females, in the context of EILC and E/C/M administration, while maintaining hormonal status. Among male rats, the most elevated AP activity was observed with excess testosterone propionate administration (5.0 [5.0; 5.9] mmol/L), while the lowest level was recorded in rats after testectomy, measuring 3.8 [2.5; 4.7] mmol/L, exhibiting a significant 20.8 % decrease (p &lt; 0.05) compared to male rats without hormonal status changes. In female rats, the study revealed that against the backdrop of EILC and E/C/M administration, the highest AP level was seen in ovariectomised females, reaching 5.8 [5.1; 6.2] mmol/L, reflecting a substantial 9.4 % increase compared to rats without hormonal status changes. Conclusions: The administration of CEP under similar experimental conditions led to the recovery of the liver's protein-synthesising function in both male and female rats. When female sex hormones were introduced to sham-operated female rats, a significant 20.8 % greater reduction in AP levels was observed. Additionally, gonadectomy led to a more pronounced decrease in this enzyme's levels in male rats compared to female rats, indicating the cytoprotective properties of female sex hormones.

Зміни розрахункових індексів біохімічних показників сироватки крові щурів різного віку після заповнення дефекту в метафізі стегнової кістки алогенними кістковими імплантатами

The purpose of the study was to analyze estimated biochemical parameters of mineralization in the blood serum of laboratory rats, to evaluate the course of metabolic processes in bone tissue after filling the defect in the metaphysis of the femur with allogeneic bone implants. Materials and methods. The work uses a model of creating a transcortical defect in the femur metaphysis of a critical size in white rats. Indicators of mineralization of bone tissue in the blood serum of white rats were studied: the content of total protein, calcium, alkaline and acid phosphatase activity, the ratio of alkaline to acid phosphatase activity, as well as the degree of mineralization, were calculated. Based on the results of a biochemical study of the blood serum of 3- and 12-month-old experimental rats with a critical size defect in the metaphysis of the femur with filling of the defect with bone alloimplants and without filling, it was determined that the introduction of alloimplants into the defect zone of the femoral metaphysis led to an acceleration of the course of mineralization of bone tissue in experimental rats of both age groups. Results and discussion. The introduction of alloimplants into the defect zone of the femoral metaphysis led to an acceleration of bone tissue mineralization in experimental rats of both age groups. Thus, on the 28th day when alloimplants were introduced, the ratio of alkaline and acid phosphatase activity in 3-month-old rats was by 1.26 times higher than the corresponding indicator in experimental animals without defect filling (p=0.008), as well as by 1.34 times – the level indicator on the 14th day. Both in the conditions of using alloimplants to fill the defect, and in the case of an unfilled defect for both age groups, the maximum manifestation of markers of bone tissue formation was recorded on the 28th day of the experiment, and the values of this maximum were higher in the group of animals with alloimplants. If in the case of an unfilled defect on the 90th day there was a sharp decrease in the analyzed indicators, which indicates the actual cessation of the mineralization process, then under the conditions of using alloimplants the decrease was less pronounced, which is a sign of the continuation of these processes. In 3-month-old animals with an unfilled defect, the stages of mineralization processes passed faster and these processes were more developed at the same time. The ratio between the activity of alkaline phosphatase and acid phosphatase in blood serum has proven to be a more informative and sensitive indicator than the degree of mineralization. Conclusion. Treatment of experimental rats with a critical size defect in the metaphysis of the femur with alloimplants leads to the detection of biochemical signs of activation of regenerative processes in them, but this activation quickly decreases in intensity, is insufficient and requires additional strengthening due to certain external influences

Effects of Dietary Eucommia ulmoides Leaf Extract on Growth, Muscle Composition, Hepatopancreas Histology, Immune Responses and Microcystin-LR Resistance of Juvenile Red Claw Crayfish (Cherax quadricarinatus)

The objective of the present study was to investigate the effects of different doses of dietary Eucommia ulmoides leaf extract (ELE) on juvenile red claw crayfish (Cherax quadricarinatus). A total number of 720 red claw crayfish (initial body weight of 0.24 ± 0.01 g) were randomly assigned to six groups and fed diets containing 0 (Diet 1), 0.5 (Diet 2), 1 (Diet 3), 2 (Diet 4), 4 (Diet 5) and 10 (Diet 6) g dry weight (dw) ELE kg (dw)−1 diets for eight weeks and challenged with microcystin-LR stress. The results indicated that dietary supplementation with 1–2 g dw ELE kg (dw)−1 diet could significantly improve the weight gain rate (WGR) and specific growth rate (SGR) of crayfish. Muscle crude protein contents of crayfish fed Diet 2, Diet 3, and Diet 4 were significantly higher than those of the control group. Compared with the control group, dietary ELE could increase total antioxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GPx), acid phosphatase (ACP), alkaline phosphatase (AKP), and phenoloxidase (PO) activities and decrease malondialdehyde (MDA) level of crayfish. Dietary ELE significantly increased the relative expression levels of SOD, thioredoxin 1 (TRX1), GPx, selenium-dependent glutathione peroxidase (Se-GPx), cytochrome P450 (CYP450), anti-lipopolysaccharide factor (ALF) and C-type lysozyme (C-LZM) mRNA of crayfish compared with the control group during the feeding experiment. When subjected to MC-LR stress for 48 h, the mRNA expression levels of SOD, GPx, Se-GPx, glutathione-s-transferase 1 (GST1), ALF, hemocyanin (HEM), and C-LZM in the hepatopancreas could be improved to varying degrees compared with the Diet 1. Supplementation of 1–2 g dw ELE kg (dw)−1 diet could improve the survival rate (SR) of crayfish under MC-LR stress. These results indicated that dietary ELE (1–2 g dw ELE kg (dw)−1 diet) could improve the growth performance, muscle protein, and non-specific immune response and increase the SR of crayfish under MC-LR stress by regulating the mRNA expression levels of the immune- and antioxidant-related genes.