Results Of Biochemical Analyses Research Articles

Andrographolide restored production performances and serum biochemical indexes and attenuated organs damage in Mycoplasma gallisepticum-infected broilers

ABSTRACT 1. This study aimed to study the preventive and therapeutic effects of andrographolide (Andro) during Mycoplasma gallisepticum HS strain (MG) infection in ArborAcres (AA) broilers. 2. The minimum inhibitory concentration (MIC) of Andro against MG was measured. Broiler body weight, feed efficiency, morbidity, cure rate and mortality were recorded during the experiment. Air sac lesion scores and immune organ index were calculated. Expression of pMGA1.2 in lung tissue and serum biochemical indices were examined. Histopathological examinations of immune organs, liver, trachea and lung tissue were conducted by Haematoxylin and Eosin stain. 3. MIC was 3.75 μg/mL and Andro significantly inhibited the expression of pMGA1.2 (P ≤ 0.05). Compared with control MG-infected group, Andro low-dose and high-dose prevention reduced the morbidity of chronic respiratory disease in 40.00% and 50.00%, respectively. Mortality of C, D and E group was 16.67%, 10.00% and 6.67%, respectively. Cure rate of E, F, G and H group was 92.00%, 92.86%, 93.33% and 100.0%, respectively. Compared with control MG-infected group, Andro treatment significantly increased average weight gain (AWG), relative weight gain rate (RWG) and feed conversion rate (FCR) at 18 to 24 days (P ≤ 0.05). Compared with control group, Andro alone treatment significantly increased AWG in broilers (P ≤ 0.05). 4. Compared with control MG-infected group, Andro significantly attenuated MG-induced air sac lesion, immune organs, liver, trachea and lung damage in broilers. Andro alone treatment did not induce abnormal morphological changes in these organs in healthy broilers. Serum biochemical analysis results showed, comparing with control MG-infected group, Andro significantly decreased the content of total protein, albumin, globulin, alanine aminotransferase, aspartate aminotransferase, total bilirubin, urea, creatinine, uric acid, total cholesterol, and increased the albumin/globulin ratio and content of alkaline phosphatase, apolipoprotein B and apolipoprotein A-I in a dose-dependent manner (P ≤ 0.05). 5. Andro could act as a potential agent against MG infection in broilers.

Experimental tooth loss affects spatial learning function and blood-brain barrier of mice.

This study aims to investigate how experimental tooth loss affected learning, memory function, and brain pathophysiology in mice. The mice (C57BL/6J, 2-month-old, male) were divided into tooth loss and control groups. The behavioral test battery was performed at 6 and 12months after tooth extraction. The protein levels of the tight junctions in the brains of the mice were analyzed. Hippocampal astrocyte was measured using immunohistochemical staining. The results of behavioral tests and biochemical analysis performed during the 6 months observation period did not show significant differences between the groups. However, the escape latency in the tooth loss group was significantly longer than that in the control group at the 12months after tooth extraction. The level of claudin-5 decreased in the tooth loss group. Additionally, hippocampal astrogliosis was found in the tooth loss group. Experimental tooth loss reduced the level of claudin-5 and caused astrogliosis in the brains of mice, which was accompanied by deterioration of learning functions. This study may provide a new insight about the association between tooth loss and cognitive dysfunction.

The Effect of Oxidized Fish Oil on Blood Parameters and Liver Histology of Nile Tilapia (Oreochromis niloticus)

Fish oil is a virtually unique source of natural LC omega-3 fatty acids EPA, DPA, and DHA, as an important source of lipid in aquafeeds. However during processing and storage, fish oil is prone to oxidation. The oxidized oil has many negative effects on the growth and health of farmed fish, therefore this study was conducted to investigate the effect of oxidized anchovy oil on blood parameters and liver histology of Nile tilapia (Oreochromis niloticus). Nile tilapia fingerlings with an initial average body weight of (15.07±0.58 g) were stocked in nine fiberglass tanks (with a tank capacity of 120 liters) in three groups in triplicates at a density of 50 fish/tank. Fish were fed with three test diets containing 35% crude protein and 8% crude fat (POV: 4.85, 50.10, and 100.80 meq/kg-1) twice a day at a feeding rate of 3% of their body weight per day for 12 weeks. During the experiment fish blood parameters analysis, and liver histology tests were performed on 3 fish taken randomly from each tank at the beginning of the experiment and monthly. The results of blood chemistry analysis showed better results in the group fed with feed that contained fresh oil (A) compared to groups fed with feeds that contained oxidized oils (B and C), but no significant differences (P > 0.05) between feed groups. Whereas the results of serum biochemical analysis showed significant differences (P > 0.05) between feed groups during the experiment period, oxidized fish oil increased serum ALT, AST activity, and TCHO level (P < 0.05), and decreased ALP activity. The liver histology test showed the best results in group A compared to oxidized oil groups (B, C), oxidized fish oil caused damage to the liver structure, and hepatocyte nuclear migration and lipid droplets were observed in liver samples from the oxidized oil groups. In conclusion, the present study demonstrates that oxidized fish oil may have a negative effect on the liver histology and blood parameters of Nile tilapia fingerlings.

TOXICOLOGICAL EVALUATION OF NATURAL AND SYNTHETIC POLYMER BASED DISSOLVABLE MICRONEEDLE PATCHES HAVING VARIABLE RELEASE PROFILES

Acute toxicity studies of dissolvable microneedle (dMN) patches fabricated from a combination of polymers, i.e. thiolated chitosan (TCS), polyvinyl alcohol (PVA), and polyvinylpyrrolidone (PVP), were performed to determine the safety of polymers on white albino rabbits. The selected albino rabbits were categorized into four groups. Group I was a control group, whereas groups II, III and IV were administered different doses of polymeric dMN patches. The food and water uptake and the weight of the animals were observed before and after administration of the microneedle patch on days 1, 3, 5, 7 and 14. Hematological and serum biochemical changes were observed by taking blood samples of treated and control group rabbits on the 15th day of study. The weight of the main organs was determined and a histopathology study was also performed. Negligible alteration in body weight, meals and water uptake was observed. The control and treated animals displayed similar behavioral pattern. Moreover, the treated groups showed hematological and biochemical analysis results, which were comparable with the results of the control group animals. Animals from both control and treated groups did not present any significant difference in the results of relative organ body weight and urine analysis. A comparison of histopathology results for both treated and control animals’ vital organs confirmed the absence of lesions. The findings of antioxidant effect, dermal and cardiac toxicity studies of chitosan, thiolated chitosan and the TCS/PVA/PVP combination were compared to those of the analysis of circulating oxidative levels, demonstrating that the combination of TCS/PVA/PVP showed improved antioxidant effect, as compared to those of chitosan and thiolated chitosan alone, and showed no significant effect on skin and heart. Thus, the findings of the study established the fact that the fabricated polymeric dMNs are a safe carrier system for different drugs.

Detection of L1014F knockdown resistance mutation in Culex tritaeniorhynchus populations

Culex tritaeniorhynchus is a major Japanese encephalitis virus vector distributed in Southeast Asia and surrounding countries. The aim of the present study is to investigate insecticide resistance status among 10 Cx. tritaeniorhynchus populations of the Mediterranean region of Turkey. Bioassay results indicated that all of the populations were resistant or at least possibly resistant to 1,1′-(2,2,2-Trichloroethane-1,1-diyl) bis (4-chlorobenzene) (DDT) (4%), [(dimethoxyphosphorothioyl) sulfanyl] butanedioate, Diethyl (malathion) (5%), and 2-[(Propan-2-yl) oxy] phenyl methylcarbamate (propoxur) (0,1%). Whereas, some of the populations were still susceptible to 3-Phenoxybenzyl (1RS)-cis, trans-3-(2,2-dichlorovinyl)-2,2-dimethyl cyclopropane carboxylate (permethrin) (0,75%) and (S)-Cyano (3-phenoxy phenyl) methyl (1R,3R)-3-(2,2-dibromoethen-1-yl)-2,2-dimethylcyclopropane-1-carboxylate (deltamethrin) (0,05%). Biochemical analysis results showed altered alpha esterase, beta esterase, para-nitrophenyl acetate (PNPA), and glutathione-s-transferase (GST) levels in some populations while all of the populations had increased oxidase levels except for the Yumurtalık population. Additionally, all of the populations had sensitive acetylcholinesterase (AChE) levels similar to the control group except for the Erzin population. Correlation analysis showed a significant correlation between mortality rates for deltamethrin and alpha esterase, beta esterase, PNPA, and GST levels while mortality rates for permethrin were significantly correlated with GST levels. An allele-specific polymerase chain reaction (AS-PCR) detected high L1014F allele frequency in the populations. Overall results indicate the urgent need for monitoring and mapping of insecticide resistance in Cx. tritaeniorhynchus populations of the study area for effective vector control management.

Alismatis Rhizoma methanolic extract-Effects on metabolic syndrome and mechanisms of triterpenoids using a metabolomic and lipidomic approach.

Alismatis rhizoma is a traditional Chinese medicine. Studies have demonstrated that Alismatis rhizoma also has therapeutic effects on metabolic syndrome. However, the pharmacodynamic material basis and mechanism are still unclear. First, UHPLC/Q-Orbitrap MS was used to detect the chemical components of the Alismatis rhizoma extract, and 31 triterpenoids and 2 sesquiterpenes were preliminarily identified. Then, to investigate the mechanism of the Alismatis rhizoma extract on metabolic syndrome, a mouse model of metabolic syndrome induced by high-fructose drinks was established. The results of serum biochemical analysis showed that the levels of TG, TC, LDL-C, and UA after the Alismatis rhizoma extract treatment were markedly decreased. 1H-NMR was used to conduct non-targeted metabolomics studies. A total of 20 differential metabolites were associated with high-fructose–induced metabolic syndrome, which were mainly correlated with 11 metabolic pathways. Moreover, UHPLC/Q-Orbitrap MS lipidomics analysis found that a total of 53 differential lipids were screened out. The results showed that Alismatis rhizoma extract mainly reduces the synthesis of glycerophospholipid and ceramide and improves the secretion of bile acid. This study shows that the Alismatis rhizoma extract can treat metabolic syndrome mainly by inhibiting energy metabolism, amino acid metabolism, and regulating bile acid to reduce phospholipid content.

Anti-cancer, anti-inflammatory and antioxidant effects of Vit-A/C@SeNPs in mutual diethylnitrosamine and carbon tetrachloride induced hepatocellular damage in albino rats

The purpose of this research was to create a nano-platform containing selenium, vitamin C, and vitamin A (Vit-A/C@SeNPs) as a potential preventive and curative agent against hepatocellular cancer. Transmission electron microscope (TEM), dynamic light scattering (DLS) and UV–visible spectroscopy were used to characterize Vit A/C@SeNPs, and the zeta potential was evaluated to assess their stability. To determine its antioxidant capacity and cytotoxicity; the DPPH (2,2diphenyl-1-picryl-hydrazyl-hydrate) and the sulforhodamine B (SRB) assay were used. For additional in-vivo pharmacokinetic investigations on normal murine, 99mTc-Vit-A/C@SeNPs were radio-synthesized using technetium-99 m. For studying the anti-inflammatory, anti-cancer, and antioxidant effect of this nano-platform in-vivo, Wistar Albino rats suffered hepatocellular damage, induced by N-diethyl nitrosamine (DEN)/carbon tetrachloride (CCl4) administration to produce inflammation and fibrosis in rats, were used. The synthesis provided Vit-A/C@SeNPs with a spherical shape, 70–80 nm average size range, a radical scavenging activity of 60.23% at a concentration of 1 mg/mL, IC50 of 25.6 μg/mL, and −15mV for zetapotential. According to the biochemical analysis results, after administration of Vit-A/C@SeNPs as a liver therapeutic or preventive drug, show distinguishable lowering in liver biochemical marker values such as Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), γ-Glutamyltransferase (GGT), T. bilirubin and Alkaline phosphatase (ALP), as well as remarkable depletion of L-malondialdehyde (L-MDA) in liver, with increase of catalase (CAT) activity & glutathione-S-transferase (GST) conc., and apparent amelioration in the histopathological examination of liver tissue samples. Quantitative real-time polymerase chain reaction (qPCR) for mRNA gene expression levels of inflammatory genes (TGF-β1, NF-κB, PPAR-γ, iNOS & TNFα) and P53 (cellular tumor suppressor gene) revealed a substantial up-regulation. The results were pertinent to damage and inflammation in hepatic cells histopathology, values dropped and tends to normal in both treatment & prevention groups. Interestingly, Vit-A/C@SeNPs usage for treatment or prevention, had showed a significant histological feature improvement in liver tissue.

Finasteride Alleviates High Fat Associated Protein-Overload Nephropathy by Inhibiting Trimethylamine N-Oxide Synthesis and Regulating Gut Microbiota.

Unhealthy diet especially high-fat diet (HFD) is the major cause of hyperlipidemia leading to deterioration of chronic kidney diseases (CKD) in patients. Trimethylamine N-oxide (TMAO) is a gut-derived uremic toxin. Our previous clinical study demonstrated that the elevation of TMAO was positively correlated with CKD progression. Finasteride, a competitive and specific inhibitor of type II 5a-reductase, has been reported recently to be able to downregulate plasma TMAO level thus preventing the onset of atherosclerosis by our research group. In this study, we established a protein-overload nephropathy CKD mouse model by bovine serum albumin (BSA) injection to investigate whether hyperlipidemia could accelerate CKD progression and the underlying mechanisms. Finasteride was administrated to explore its potential therapeutic effects. The results of biochemical analyses and pathological examination showed that HFD-induced hyperlipidemia led to aggravated protein-overload nephropathy in mice along with an elevated level of circulating TMAO, which can be alleviated by finasteride treatment possibly through inhibition of Fmo3 in liver. The 16 S rRNA sequencing results indicated that HFD feeding altered the composition and distribution of gut microbiota in CKD mice contributing to the enhanced level of TMAO precursor TMA, while finasteride could exert beneficial effects via promoting the abundance of Alistipes_senegalensis and Akkermansia_muciniphila. Immunofluorescence staining (IF) and qRT-PCR results demonstrated the disruption of intestinal barrier by decreased expression of tight junction proteins including Claudin-1 and Zo-1 in HFD-fed CKD mice, which can be rescued by finasteride treatment. Cytokine arrays and redox status analyses revealed an upregulated inflammatory level and oxidative stress after HFD feeding in CKO mice, and finasteride-treatment could alleviate these lesions. To summarize, our study suggested that finasteride could alleviate HFD-associated deterioration of protein-overload nephropathy in mice by inhibition of TMAO synthesis and regulation of gut microbiota.

Therapeutic effect and mechanism of Anemarrhenae Rhizoma on Alzheimer's disease based on multi-platform metabolomics analyses.

Anemarrhenae Rhizoma (AR) has multiple pharmacological activities to prevent and treat Alzheimer’s disease (AD). However, the effect and its molecular mechanism are not elucidated clear. This study aims to evaluate AR’s therapeutic effect and mechanism on AD model rats induced by D-galactose and AlCl3 with serum metabolomics. Behavior study, histopathological observations, and biochemical analyses were applied in the AD model assessment. Gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-QTOF/MS) were combined with multivariate statistical analysis to identify potential biomarkers of AD and evaluate the therapeutic effect of AR on AD from the perspective of metabolomics. A total of 49 biomarkers associated with the AD model were identified by metabolomics, and pathway analysis was performed to obtain the metabolic pathways closely related to the model. With the pre-treatment of AR, 32 metabolites in the serum of AD model rats were significantly affected by AR compared with the AD model group. The regulated metabolites affected by AR were involved in the pathway of arginine biosynthesis, arginine and proline metabolism, ether lipid metabolism, glutathione metabolism, primary bile acid biosynthesis, and steroid biosynthesis. These multi-platform metabolomics analyses were in accord with the results of behavior study, histopathological observations, and biochemical analyses. This study explored the therapeutic mechanism of AR based on multi-platform metabolomics analyses and provided a scientific basis for the application of AR in the prevention and treatment of AD.

Pseudovibrio flavus sp. nov. isolated from the sea sponge Verongula gigantea.

A Gram-negative, motile, rod-shaped marine bacterium, designated RKSG542T, was isolated from the sea sponge Verongula gigantea collected at a depth of 20 m off the west coast of San Salvador, The Bahamas. Phylogenetic analyses based on 16S rRNA gene and genome sequences place RKSG542T in a monophyletic clade with members of the genus Pseudovibrio. Strain RKSG542T shared <96.7 % 16S rRNA gene sequence similarity,<72.2 % average nucleotide identity,<66.7 % average amino acid identity, and <24.8 % digital DNA-DNA hybridization with type strains of the family Stappiaceae. Growth occurred at 22-37 °C (22-30 °C optimum), at pH 7-9 (pH 7 optimum), and with 0.5-5 % (w/v) NaCl (2 % optimum). The predominant fatty acids (>10 %) were summed feature 8 (C18 : 1 ω6c and/or C18 : 1 ω7c), C18 : 0 and C16 : 0, and the respiratory lipoquinone was Q-10. The polar lipid composition comprised phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, three unknown aminolipids, six unknown phospholipids and four unknown lipids. The DNA G+C content of the genome sequence was 52.5 mol%. Based on the results of biochemical, phylogenetic and genomic analyses, RKSG542T (=TSD-76T=LMG 29867T) is presented here as the type strain of a novel species within the genus Pseudovibrio (family Stappiaceae, order Hyphomicrobiales, class Alphaproteobacteria), for which the name Pseudovibrio flavus sp. nov. is proposed.

Brachybacterium kimchii sp. nov. and Brachybacterium halotolerans subsp. kimchii subsp. nov., isolated from the Korean fermented vegetables, kimchi, and description of Brachybacterium halotolerans subsp. halotolerans subsp. nov.

Two Gram-stain-positive, oxidase-negative, catalase-positive, and coccus-shaped bacterial strains, designated CBA3104T and CBA3105T, were isolated from kimchi. Strain CBA3104T and CBA3105T grew at 10-35°C (optimum, 25°C and 30°C, respectively), at pH 6.0-8.5 (optimum, pH 6.5), and in the presence of 0-15% (w/v) NaCl (optimum, 5%). A phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CBA3104T formed a distinct phylogenetic lineage within the genus Brachybacterium whereas strain CBA3105T was closely positioned with Brachybacterium halotolerans MASK1Z-5T. The 16S rRNA gene sequence similarity between strains CBA3104T and CBA3105T was 99.9%, but ANI and dDDH values between strains CBA3104T and CBA3105T were 93.61% and 51.5%, respectively. Strain CBA3104T showed lower ANI and dDDH values than species delineation against three closely related strains and type species of the genus Brachybacterium, however, strain CBA3105T showed 96.63% ANI value and 69.6% dDDH value with Brachybacterium halotolerans MASK1Z-5T. Among biochemical analysis results, strain CBA3104T could uniquely utilize bromo-succinic acid whereas only strain CBA3105T was positive for alkaline phosphatase and α-fucosidase among two novel strains, closely related strains, and type species of the genus Brachybacterium. Compared with strain CBA3105T and Brachybacterium halotolerans JCM 34339T, strain CBA3105T was differentially positive for acid production of D-arabinose, D-adonitol, and potassium 5-ketogluconate and enzyme activity of β-glucuronidase. Both strains contained menaquinone-7 as the dominant quinone. The cell-wall peptidoglycan of two novel strains contained meso-diaminopimelic acid. The major fatty acids of strains CBA3104T and CBA3105T were anteiso-C15:0, anteiso-C17:0, and iso-C16:0. The major polar lipids of both strains were phosphatidylglycerol and diphosphatidylglycerol. Strain CBA3104T possessed a uniquely higher abundance of tRNA (97 tRNAs) than four Brachybacterium strains used for comparative taxonomic analysis (54-62 tRNAs). Both the CBA3104T and CBA3105T strain harbored various oxidoreductase, transferase, hydrolase, and lyase as strain-specific functional genes compared to closely related strains and Brachybacterium type species. The results of biochemical/physiological, chemotaxonomic, and genomic analyses demonstrated that strains CBA3104T and CBA3105T represent a novel species of the genus Brachybacterium and a novel subspecies of B. halotolerans, respectively, for which the names Brachybacterium kimchii sp. nov. and B. halotolerans subsp. kimchii subsp. nov. are proposed. The type strains of the novel species and the novel subspecies are CBA3104T (= KCCM 43417T = JCM 34759T) and CBA3105T (= KCCM 43418T =JCM 34760T), respectively.

Development of Ultrapure and Potent Tannic Acids as a Pan-coronal Antiviral Therapeutic.

The rampageous transmission of SARS-CoV-2 has been devastatingly impacting human life and public health since late 2019. The waves of pandemic events caused by distinct coronaviruses at present and over the past decades have prompted the need to develop broad-spectrum antiviral drugs against them. In this study, our Pentarlandir ultrapure and potent tannic acids (UPPTA) showed activities against two coronaviral strains, SARS-CoV-2 and HCoV-OC43, the earliest-known coronaviruses. The mode of inhibition of Pentarlandir UPPTA is likely to act on 3-chymotrypsin-like protease (3CLpro) to prevent viral replication, as supported by results of biochemical analysis, a 3CLpro assay, and a “gain-of-function” 3CLpro overexpressed cell-based method. Even in the 3CLpro overexpressed environment, Pentarlandir UPPTA remained its antiviral characteristic. Utilizing cell-based virucidal and cytotoxicity assays, the 50% effective concentrations (EC50) and 50% cytotoxicity concentration (CC50) of Pentarlandir UPPTA were determined to be ∼0.5 and 52.5 μM against SARS-CoV-2, while they were 1.3 and 205.9 μM against HCoV-OC43, respectively. In the pharmacokinetic studies, Pentarlandir UPPTA was distributable at a high level to the lung tissue with no accumulation in the body, although the distribution was affected by the food effect. With further investigation in toxicology, Pentarlandir UPPTA demonstrated an overall safe toxicology profile. Taking these findings together, Pentarlandir UPPTA is considered to be a safe and efficacious pancoronal antiviral drug candidate that has been advanced to clinical development.

MO053: Are Renal Prognostic Markers Different in Patients with Exostosin 1/2 Positive or Negative Membranous Lupus Nephritis? A Single Center Experience in Turkey

Abstract BACKGROUND AND AIMS Recently, prognostic importance of antibodies in primary membranous nephropathy is well defined. However, knowledge about antibodies related to disease activity in membranous lupus nephritis (MLN) is limited. Exostosin1 and exostosin2 (EXT1/2) are novel antibodies detected in MLN, and further studies are needed for the prognostic significance of them as a marker. The aim of this study is to determine whether renal prognostic markers differ between EXT1/2 positive and negative MLN patients. METHOD This is a single-center, retrospective and observational study. Class 5 MLN patients aged 18 or older were included in the study. Clinical characteristics, results of biochemical analysis and histopathological examination were recorded from 6 to 18 months. The end-point of the study was determined as complete remission, partial remission, need for renal replacement therapy (RRT) and exitus. Group 1: EXT1/2 positive patients. Group 2: EXT1/2 negative patients. Light, immunofluorescent and electron microscopy findings and laboratory parameters were evaluated in EXT1/2 positive and negative patients. RESULTS Group 1: n = 11 patients. Group 2: n = 7 patients. The main demographic, biochemical and histopathological characteristics of the groups were shown in Table 1. EXT1/2 negative patients, a higher number of sclerotic glomeruli, interstitial fibrosis and a higher amount of proteinuria were present than positive patients (P &amp;lt; 0.05). EXT1/EXT2 negative patients had significantly lower C3 and C4 levels than positive patients (P &amp;lt; 0.05). EXT1/EXT2 negative patients had a higher RRT or exitus rate within 18 months than positive patients (P &amp;lt; 0.05). In contrast, EXT1/EXT2 positive patients had a higher rate of achieving complete or partial remission within 18 months than negative patients (P &amp;lt; 0.05). The effect of immunosuppressive drug use other than steroids and glomerulosclerosis remission was statistically significant (P = 0.04 and P = 0.011). CONCLUSION EXT1/2 negative class 5 LMN patients have more poor renal prognostic indicators than positive patients. RRT or exitus rate is higher in EXT1/EXT2 negative patients than positive patients. However, more randomized controlled studies are needed.

The influence of vitamin‐mineral supplementation with prolonged action on cows and on the protein metabolism in calves

It is well established that one of the pathology‘ reason in calves is the disturbances of prenatal development as a result of vitamin and mineral abstinence. The aim of the study was to research the influence of vitamin‐mineral bolus’ supplementation with prolong action to the ration of cows on the calves’ protein metabolism. The studies were carried out in one of the breeding farms of the Leningrad region. Holstein cows were used aged from 3 to 5 years and their calves. 5 groups were formed (n=5 by each). The cows of the 1st experimental group have received two boluses All – mineral plus (the compound: Mg, Zn, Cu, Co, Se, I, Vitamin A, D3, E, Holland Animal Care, Holland), 2d experimental – two boluses Uno Biotin (the compound: Mg, Zn, Cu, Co, Se, I, Vitamin A, D3, E, Holland Animal Care, Holland), 3d experimental group ‐ one Cattle bolus with Iodine (the compound: Cu, Co, Se, I, Telsol Limited, United Kingdom), 4th experimental group – one Cattle Bullet (the compound: Mg, Zn, Cu, Co, Se, I, Vitamin A, D3, E, Holland Animal Care, Holland). The 5th group was control with ordinary ration vitamin‐mineral diet. All cows of experimental groups have got boluses one time which was 60 days before calving. The prolongation of all boluses was 120 days after receiving except bolus All‐mineral plus. The prolongation of their was 180 days. The compound of these boluses includes manganese, zinc, copper, selenium, cobalt, iodine, Vitamin A, D3, E, H. We used the clinical examination of calves and biochemical analysis of calves blood samples on the 3d‐5th days after calving. Serum was analyzed for total protein, albumins and globulins. We were used Student's t‐tests to study differences between groups. Thus, the clinically health animals were nineteen calves of experimental groups and second calves if control group. The body mass of newborn calves was 35‐40 kg. The calves were stood up during first two hours after calving and had a healthy sucking reflex. The temperature of their was fluctuated within 38.5‐39.30°С. The results of biochemical analysis showed that the level of total protein was within the physiological norm in the bloods’samples of 1st, 3d, 4th and 5 th calves groups. The level of total protein was statistically increased in the calves of 1st group in 1.4 times (р&lt;0,05) compared with data of calves in the control group. The value of albumins of calves was decreased in 1.07 times (р&lt;0,05) in 2d group and in 1.4 times in control group (18,19±3,37g/l). We were observed the insignificant increasing trend of albumins’level in calves of 1st,2d,3d and 4th groups compared with data of control group. The difference of globulins’level between calves of 1st, 4th experimental and control group was 1.3 times (р&lt;0,05). The value of globulin in the calves of 2d group was 35,64±5,93 g/l and 3d group ‐ 34,45±7,93 g/l. So, we can assume that the vitamin‐mineral supplementation of cow‐mothers during dry time has a positive influence on the synthesis of the total protein and its fraction ratio.

Halomonas antri sp. nov., a carotenoid-producing bacterium isolated from surface seawater.

A Gram-negative, moderately halophilic bacterium, designated as strain Y3S6T, was isolated from a surface seawater sample collected from Dongangyoeng cave, Udo-myeon, Jeju-si, Jeju-do, Repulic of Korea. Cells of strain Y3S6T were aerobic, rod-shaped, non-sporulated, yellow, catalase- negative, oxidase-negative and motile with one polar flagellum. Growth of strain Y3S6T occurred at 15-40 °C (optimum: 25-30 °C), at pH 6.0-9.0 (optimum: pH 7.0) and in the presence of 0-13% NaCl (optimum: 1-6 %, w/v). The novel strain was able to produce carotenoids. Its chemotaxonomic and morphological characteristics were consistent with those of members of the genus Halomonas. Phylogenetic analysis of the 16S rRNA gene sequence revealed that strain Y3S6T formed a clade with Halomonas pellis L5T (98.97 %) and Halomonas saliphila LCB169T(98.90%). The average nucleotide identity and digital DNA-DNA hybridization values of strain Y3S6T with the most closely related strains for which whole genomes are publicly available were 82.3-85.2% and 62.8-66.1 %, respectively. The major fatty acids in strain Y3S6T were C16 : 0, C19 : 0 cyclo ω8c and summed feature 8 (composed of C18 : 1 ω7c and/or C18 : 1 ω6c), and the predominant quinone was Q-9. Its polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unidentified phosphoglycolipid, one unidentified phosphoaminoglycolipid and one unidentified phospholipid. The genomic DNA G+C content based on the draft genome sequence was 64.2 mol%. The results of physiological and biochemical tests and 16S rRNA sequence analysis clearly revealed that strain Y3S6T represents a novel species in the genus Halomonas, for which the name Halomonas antri sp. nov. has been proposed. The type strain is Y3S6T (=KACC 21536T=NBRC 114315=TBRC 15164T).

Hymenobacter armeniacus sp. nov. and Hymenobacter montanus sp. nov., two radiation-resistant bacteria from soil.

Two bacterial strains, designated BT189T and BT664T, were isolated from soil sampled in the Republic of Korea. Phylogenetic analysis based on the 16S rRNA gene sequences showed that strains BT189T and BT664T belonged to the genus Hymenobacter, family Hymenobacteraceae (order Cytophagales). The 16S rRNA genes of the two strains shared a sequence similarity of 93.7 %. The closely related species of strain BT189T were Hymenobacter rubidus DG7BT (97.1 % 16S rRNA similarity) and Hymenobacter terrae DG7AT (96.7 %). The closest related species to strain BT664T were Hymenobacter sedentarius DG5BT (95.3 %) and Hymenobacter terrenus MIMtkLc17T (95.2 %). The genome sizes of strains BT189T and BT664T were 5 285 287 and 5 475 357 bp, respectively. The genomic DNA G+C contents of strains BT189T and BT664T were 63.2 and 59.3 mol%, respectively. The main fatty acids of strain BT189T were iso-C15 : 0, anteiso-C15 : 0 and summed feature 3 (C16 : 1 ω6c/C16 : 1 ω7c), and those of strain BT664T were iso-C15 : 0, C16 : 1 ω5c and summed feature 3 (C16 : 1 ω6c/C16 : 1 ω7c). The main polar lipid in both strains was phosphatidylethanolamine and the predominant respiratory quinone was MK-7, supporting the affiliation of these strains with the genus Hymenobacter. Based on the results of biochemical, chemotaxonomic and phylogenetic analyses, two novel species, Hymenobacter armeniacus BT189T (=KCTC 72341T=NBRC 114843T) and Hymenobacter montanus BT664T (KACC 21967T=NBRC 114856T), are proposed.

Tenebrionibacter intestinalis gen. nov., sp. nov., a member of a novel genus of the family Enterobacteriaceae, isolated from the gut of the plastic-eating mealworm Tenebrio molitor L.

A Gram-negative, white-pigmented, motile and rod-shaped strain, BIT-L3T, was isolated from the gut of plastic-eating mealworm Tenebrio molitor L. Its taxonomic position was determined by using a polyphasic approach. A preliminary analysis based on the 16S rRNA gene sequence (1445 bp) revealed that this strain was closely related to the members within the family Enterobacteriaceae. Phylogenetic trees based on the concatenated partial sequences of seven housekeeping genes (atpD, gyrB, infB, rpoB, pyrG, fusA, leuS) and genome sequences further showed that strain BIT-L3T constituted a separate lineage within the family Enterobacteriaceae. In silico DNA-DNA hybridization values and average nucleotide identity values between strain BIT-L3T and its closest related species within the family Enterobacteriaceae were less than 21.8 and 76.7 %, respectively. The major fatty acids (>5 %) of strain BIT-L3T were C16 : 0, C14 : 0, C17 : 0 cyclo, summed feature 8 (comprising C18 : 1 ω7c and/or C18 : 1 ω6c), summed feature 3 (comprising C16 : 1 ω7c and/or C16 : 1 ω6c and/or iso-C15 : 0 2-OH) and summed feature 2 (comprising iso-C16 : 1 I/C14 : 0 3-OH and/or C12 : 0 aldehyde and/or an unknown fatty acid of equivalent chain length 10.9525). Its genomic DNA G+C content was 53.7 mol%. Based on the results of phylogenetic, physiological and biochemical analyses, strain BIT-L3T is considered to represent a novel species of a novel genus within the family Enterobacteriaceae, for which the name Tenebrionibacter intestinalis gen. nov., sp. nov. is proposed. The type strain is BIT-L3T (=CCTCC AB 2020371T=LMG 32222T=TBRC 14825T).

Optical properties of honey: FTIR spectroscopy and refractometry

115 samples of honey of various botanical types, geographical origin and harvest year (2019– 2021) were analysed using attenuated total reflection infrared spectroscopy of impaired total reflection, refractometry and biochemical analysis. Initial honey samples in liquid and crystallised states were investigated. Crystalline D-glucopyranose (glucose), D-fructopyranose (fructose), their 40% solutions and invert sugar were used as auxiliary substances. Biochemical analysis was used to determine the glucose content in honey samples. Based on the obtained data, a relationship between the results of biochemical analysis and refractometry (refractive index, the content of invert sugars, humidity) was established. We deduced equations that allow the content of glucose and fructose in honey to be evaluated by the refractive index. Studying honey by IR spectroscopy showed that all investigated samples, regardless of the botanical and geographical origins, can be classified into three groups dominated by: I – glucose, II – fructose and III – mixed, with a close content of two monosaccharides. This allowed the bands characteristic of α- and β-pyranose forms of glucose and fructose to be identified, as well as the nature of their changes depending on the ratio of both monosaccharides in honey as a result of their crystallisation to be assessed. It is noted that the ratio of monosaccharides determines not only the stability of the liquid crystal structure and crystallisation rate in honey but also their optical, biochemical and nutritional properties, which are important for the preferred use of honey in medical and pharmacopoeial practice, dietetics and cosmetology.

CASE OF PAROXYSMAL NOCTURNAL HEMOGLOBINURIA. PROBLEMS OF LABORATORY DIAGNOSTICS

The presented case demonstrates the importance of a comprehensive analysis of all the examination data in a patient with paroxysmal nocturnal hemoglobinuria (PNH). A comparison was made of anamnesis data, the clinical picture of the disease, the results of biochemical blood analysis and hematological analyzer data with morphological assessment of cells in a stained blood smear. The interest of this case for hematologists and laboratory specialists is due to the rarity of this disease, difficulties in diagnostics, the presence of 2 types of hemolysis, features of morphological changes in peripheral blood erythrocytes, as well as analysis of the dynamics of manifestations, including against the background of targeted therapy of PNH.

In vivo evaluation of the potential protective effects of prolactin against damage caused by methylmercury.

Non-biodegradable metals such as mercury accumulate in living organisms during life (bioaccumulation) and also within trophic webs (biomagnification) and may reach high concentrations in humans. The contamination of humans by mercury in drinking water and food may be common, in particular in riverside communities that have a diet rich in fish. In vitro studies of human cell lines exposed to the cytotoxic and mutagenic effects of methylmercury have shown that prolactin has potential cytoprotective properties and may act as a co-mitogenic factor and inhibitor of apoptosis. The present in vivo study investigated the protective potential of prolactin against the toxic effects of methylmercury in the mammal Mus musculus. Histological and biochemical analyses, together with biomarker of genotoxicity, were used to verify the protective potential of prolactin in mice exposed to methylmercury. The reduction in kidney and liver tissue damage was not significant. However, results of biochemical and genotoxic analyses were excellent. After prolactin treatment, a significant reduction was observed in biochemical parameters and mutagenic effects of methylmercury. The study results therefore indicated that prolactin has protective effects against the toxicity of methylmercury and allowed us to suggest the continuation of research to propose prolactin in the future, as an alternative to prevent the damage caused by mercury, especially in populations that are more exposed.