Efficiency evaluation of the improved treatment and prevention complex in pregnant women after assisted reproductive technologies

Study objective: to determine the role and effectiveness of the proposed therapeutic and preventive complex and psychoemotional correction of hormonal disorders in the pregnancy dynamics after assisted reproductive technologies (ART) to improve the antenatal observation and prevention of obstetric and perinatal complications.Materials and methods. The study included 299 pregnant women: the main group included 249 women whose pregnancy occurred as an ART result; the control group included 50 pregnant women with spontaneous pregnancy. Therapeutic and prophylactic complex for pregnant women after ART included: micronized progesterone, magnesium oxide, folic acid, L-arginine aspartate, ω3-polyunsaturated fatty acids and long-term psychological correction on the eve of the ART program, at 8–10, 16–18 and 28–30 weeks of pregnancy. Results. There was a significant increase in the β-chorionic gonadotropin (β-hCG) level in women of the study groups in the first trimester of pregnancy against the background of the proposed treatment. Mean β-HCG value at 7–8 weeks of gestation in the subgroup IA exceeded the subgroup IB by 37% (p <0.05), in subgroup IIA it exceeded the subgroup IIB by 33% (p <0.05). The mean β-hCG value in subgroups IIIA and IIIB did not have a significant difference in the dynamics of the first trimester compared with the control group and among themselves (p >0.05).Mean progesterone value at 7–8 weeks of gestation in subgroup IA increased by 38% in comparison with pregnant women who received the conventional treatment complex (p <0.05), in subgroup IIA it was 73% higher than in subgroup IIB (p <0.05). There was no significant difference in the progesterone level in subgroups IIIA and IIIB in the dynamics of the first trimester.The average cortisol value at 23–24 weeks of pregnancy in subgroup IA decreased by 42% (p <0.05), in pregnant women with endocrine infertility against the background of the proposed treatment complex it was 62% less than in subgroup IIB (p <0.05). The average cortisol level in women with a male factor of infertility was 63% lower than in subgroup IIIB against the background of the proposed complex (p <0.05).Conclusion. Advanced therapy with micronized progesterone in combination with magnesium saturation, L-arginine aspartate, folic acid, ω-3 polyunsaturated fatty acids, as well as long-term psychoemotional correction is appropriate and effective compared to conventional therapy for pregnant women.

Lytic bacteriophages facilitate antibiotic sensitization of Enterococcus faecium

Enterococcus faecium, a commensal of the human intestine, has emerged as a hospital-adapted, multi-drug resistant (MDR) pathogen. Bacteriophages (phages), natural predators of bacteria, have regained attention as therapeutics to stem the rise of MDR bacteria. Despite their potential to curtail MDR E. faecium infections, the molecular events governing E. faecium-phage interactions remain largely unknown. Such interactions are important to delineate because phage selective pressure imposed on E. faecium will undoubtedly result in phage resistance phenotypes that could threaten the efficacy of phage therapy. In an effort to understand the emergence of phage resistance in E. faecium, three newly isolated lytic phages were used to demonstrate that E. faecium phage resistance is conferred through an array of cell wall-associated molecules, including secreted antigen A (SagA), enterococcal polysaccharide antigen (Epa), wall teichoic acids, capsule, and an arginine-aspartate-aspartate (RDD) protein of unknown function. We find that capsule and Epa are important for robust phage adsorption and that phage resistance mutations in sagA, epaR, and epaX enhance E. faecium susceptibility to ceftriaxone, an antibiotic normally ineffective due to its low affinity for enterococcal penicillin binding proteins. Consistent with these findings, we provide evidence that phages potently synergize with cell wall (ceftriaxone and ampicillin) and membrane-acting (daptomycin) antimicrobials to slow or completely inhibit the growth of E. faecium. Our work demonstrates that the evolution of phage resistance comes with fitness defects resulting in drug sensitization and that lytic phages could serve as effective antimicrobials for the treatment of E. faecium infections.

Targeted metabolomics as a tool in discriminating endocrine from primary hypertension

Context Identification of patients with endocrine forms of hypertension (EHT) (primary hyperaldosteronism [PA], pheochromocytoma/paraganglioma [PPGL] and Cushing syndrome [CS]) provides the basis to implement individualized therapeutic strategies. Targeted metabolomics (TM) have revealed promising results in profiling cardiovascular diseases and endocrine conditions associated with hypertension. Objective Use TM to identify distinct metabolic patterns between primary hypertension (PHT) and EHT and test its discriminating ability. Design Retrospective analyses of PHT and EHT patients from a European multicentre study (ENSAT-HT). TM was performed on stored blood samples using liquid chromatography mass spectrometry. To identify discriminating metabolites a “classical approach” (CA) (performing a series of univariate and multivariate analyses) and a “machine learning approach” (MLA) (using Random Forest) were used. Patients The study included 282 adult patients (52% female; mean age 49 years) with proven PHT (n=59) and EHT (n=223 with 40 CS, 107 PA and 76 PPGL), respectively. Results From 155 metabolites eligible for statistical analyses, 31 were identified discriminating between PHT and EHT using the CA and 27 using the MLA, of which 15 metabolites (C9, C16, C16:1, C18:1, C18:2, arginine, aspartate, glutamate, ornithine, spermidine, lysoPCaC16:0, lysoPCaC20:4, lysoPCaC24:0, PCaeC42:0, SM C18:1, SM C20:2) were found by both approaches. The ROC curve built on the top 15 metabolites from the CA provided an area under the curve (AUC) of 0.86, which was similar to the performance of the 15 metabolites from MLA (AUC 0.83). Conclusions TM identifies distinct metabolic pattern between PHT and EHT providing promising discriminating performance.

Clinical efficacy of L-arginin aspartate in complex treatment of patients with essential arterial hypertension with concomitant type 2 diabetes mellitus

Objective. Investigate directly the clinical efficacy of L-arginine aspartate in the treatment of patients with essential arterial hypertension (EAG) and type 2 diabetes mellitus (DM). Materials and methods. The study included 43 patients with EAG II in combination with type 2 DM. The mean age of patients was 55,7±0,6 years, of which 20 (46,5 %) were men and 23 (53,5 %) were women. The duration of the disease on the EAG averaged 8,4±0,4 years, on type 2 DM – 6,5±0,5 years. A comprehensive clinical, laboratory and instrumental examination was carried out. All patients were divided into two groups. Patients in both groups received ramipril 5-10 mg a day in combination with amlodipine 5-10 mg a day and antidiabetic drugs (metformin, gliclazide or a combination thereof) as basic therapy. Patients in group 2 (n=22) were additionally prescribed the drug L-arginine aspartate orally 3 g 3 times a day for 4 weeks. The course of treatment was repeated after 2 months. Results and discussion. In group 2 there was a more pronounced tendency to decrease the average daily and night blood pressure levels, in particular diastolic (3,2 and 2,9 mm Hg; p>0,05) and heart rate (by 17,3 %; p<0,05). The use of L-arginine aspartate significantly improved systolic (ejection fraction increased by 7,1 % vs 4,4 % in group 1; p<0,05) and left ventricular (LV) diastolic function (Em/Am increased by 48,8 % vs 34,7 % in group 1; p<0,05), a decrease in the size of the left atrium (10,2 % vs 8,3 % in group 1; p<0,05) and the reversal of LV hypertrophy (index LV myocardial mass decreased by 20,1 % against 15,9 % in group 1; p<0,05). Additional administration of L-arginine aspartate also led to a decrease in fasting and postprandial plasma glucose (4,9 % and 7,0 %; p<0,05, respectively) than the use of basic therapy alone. At the same time in group 2 there was a decrease in microalbuminuria by 27,6 % (p<0,05) and an increase in glomerular filtration rate by 11,4 % (p>0,05). Conclusions. L-arginine aspartate should be used in patients with EAG in combination with type 2 DM and microalbuminuria to increase the cardio- and nephroprotective efficacy of basic therapy.

Lytic bacteriophages facilitate antibiotic sensitization of Enterococcus faecium

Enterococcus faecium, a commensal of the human intestine, has emerged as a hospital-adapted, multi-drug resistant (MDR) pathogen. Bacteriophages (phages), natural predators of bacteria, have regained attention as therapeutics to stem the rise of MDR bacteria. Despite their potential to curtail MDR E. faecium infections, the molecular events governing E. faecium-phage interactions remain largely unknown. Such interactions are important to delineate because phage selective pressure imposed on E. faecium will undoubtedly result in phage resistance phenotypes that could threaten the efficacy of phage therapy. In an effort to understand the emergence of phage resistance in E. faecium, three newly isolated lytic phages were used to demonstrate that E. faecium phage resistance is conferred through an array of cell wall-associated molecules, including secreted antigen A (SagA), enterococcal polysaccharide antigen (Epa), wall teichoic acids, capsule, and an arginine-arginine-aspartate (RDD) protein of unknown function. We find that capsule and Epa are important for robust phage adsorption and that phage resistance mutations in sagA, epaR, and epaX enhance E. faecium susceptibility to ceftriaxone, an antibiotic normally ineffective due to its low affinity for enterococcal penicillin binding proteins. Consistent with these findings, we provide evidence that phages potently synergize with cell wall (ceftriaxone and ampicillin) and membrane-acting (daptomycin) antimicrobials to slow or completely inhibit the growth of E. faecium. Our work demonstrates that the evolution of phage resistance comes with fitness defects resulting in drug sensitization and that lytic phages could potentially serve as antimicrobial adjuvants in treating E. faecium infections.

Effects of Multi-Strain Probiotics on Immune Responses and Metabolic Balance in Helicobacter pylori-Infected Mice

Chronic inflammation caused by Helicobacter pylori infection increases the risk of developing gastric cancer. Even though the prevalence of H. pylori infection has been decreased in many regions, the development of antibiotic resistance strains has increased the difficulty of eradicating H. pylori. Therefore, exploring alternative approaches to combat H. pylori infection is required. It is well-known that probiotic therapy can improve H. pylori clearance. In this study, H. pylori-infected mice were treated with Lactobacillus fermentum P2 (P2), L. casei L21 (L21), L. rhamnosus JB3 (JB3), or a mixture including the aforementioned three (multi-LAB) for three days. All the lactic acid producing bacteria (LAB) treatments decreased H. pylori loads in the stomach and vacA gene expression, H. pylori specific immunoglobulin (Ig) A, and IgM levels in stomach homogenates, as well as serum levels of interferon-gamma and interleukin-1 beta. The multi-LAB and JB3 treatments further restored the superoxide dismutase and catalase activities suppressed by H. pylori infection. Furthermore, H. pylori infection decreased serum concentrations of 15 kinds of amino acids as well as palmitic acid. The multi-LAB treatment was able to recover the serum levels of alanine, arginine, aspartate, glycine, and tryptophan, which are all important in modulating immune functions. In addition, butyric acid, valeric acid, palmitic acid, palmitoleic acid, stearic acid, and oleic acid levels were increased. In this study, multi-LAB revealed its ability to adjust the composition of metabolites to improve health. To date, the mechanisms underlying how LAB strains crosstalk with the host are not fully understood. Identifying the mechanisms which are regulated by LABs will facilitate the development of effective therapies for infection in the future.

Challenges in the management of an ignored cause of hyperammonemic encephalopathy: pyruvate carboxylase deficiency

Pyruvate carboxylase (PC) deficiency is a rare autosomal recessive disease and provides clinics in three essential phenotypes. Type B PC deficiency is characterized by lactic acidosis and hyperammonemia. We report a Turkish patient who was diagnosed with type B PC deficiency. Despite the application of anaplerotic treatment with biotin, citrate and arginine-aspartate, continuous veno-venous hemodialysis (CVVHD) treatments were applied due to the failure to keep hyperammonemia and lactic acidosis under control. Ammonia values increasing to 860 μmol/L were observed. A homozygous novel variant was detected in PC gene analyses containing a 12-base pair deletion on exon 8. Although the mutation found was not reported previously, it was accepted as a pathogenic variant due to its presence in a functional region of the protein. In type B PC deficiency, although a high level of ammonia is expected, it rarely exceeds 200 μmol/L. As far as we know, the present case has the highest ammonia values in the literature. This paper has been shared to highlight to keep PC deficiency in mind regarding the differential diagnosis of hyperammonemia, particularly in the presence of lactic acidosis, and to serve as a model for the use of different modalities in the management process of PC deficiency.

Genome-Wide Identification, Characterization and Expression Analysis of Non-Arginine Aspartate Receptor like kinase gene family under Colletotrichum truncatum stress conditions in Hot pepper

Receptor Like kinases (RLKs) are conserved upstream signaling molecules that regulate several biological processes, including plant development and stress adaptation. Non arginine aspartate (non-RD) an important class of RLKs plays a vital role in disease resistance and apoptosis in plants. In present investigation, a comprehensive Insilco analysis for non-RD Kinase gene family including identification, sequence similarity, phylogeny, chromosomal localization, gene structures, gene duplication analysis, promoter analysis and transcript expression profiles were elucidated. In this study twenty six genes were observed on nine out of twelve chromosomes. All these genes were clustered into seven subfamilies under large monophyletic group termed as Interleukin-1 Receptor-Associated Kinase (IRAK) family. Structural diversity in genomic structure among non-RD kinase gene family were identified and presence of pathogen induced cis regulatory elements like STRE, MYC, MYB,W box were found. Expression profiles of genes involved in providing resistance to anthracnose pathogen Colletotrichum truncatum in hot pepper were analyzed at different infective stages in both resistant and susceptible genotypes. Among twenty six genes, CaRLK1 gene belonging to LRRXII subfamily was up regulated under severe stress after infection in resistant genotype PBC-80. This integrative approach has helped us to identify candidate genes involved in disease resistance which would be helpful in future crop improvement programs.

Effects of the Chiral Fungicides Metalaxyl and Metalaxyl-M on the Earthworm Eisenia fetida as Determined by 1H-NMR-Based Untargeted Metabolomics

Although metalaxyl and metalaxyl-M are widely used fungicides, very little is known about their subacute and enantiospecific effects on the earthworm metabolome. In this study, Eisenia fetida were exposed to metalaxyl and metalaxyl-M at three concentrations (0.5, 5 and 50 mg/kg) for seven days. 1H nuclear magnetic resonance (1H-NMR)-based untargeted metabolomics showed that metalaxyl and metalaxyl-M exposure disturbed earthworms’ metabolism at all three concentrations. Endogenous metabolites, such as succinate, arginine, aspartate, urea, asparagine, alanine, trimethylamine, taurine, cysteine, serine, threonine, histidine, lysine, glucose, choline, carnitine, citric acid, alpha-ketoisovaleric acid, fumaric acid and so on, were significantly changed. These results indicate that metalaxyl and metalaxyl-M produce different, enantiospecific disturbances in the earthworm metabolism, particularly in the tricarboxylic acid (TCA) and urea cycles. The application of untargeted metabolomics thus provides more information for evaluating the toxic risks of metalaxyl and metalaxyl-M.