Strategy and Tactics in Combinatorial Organic Synthesis. Applications to Drug Discovery

A strategic analysis of various issues which pertain to the enablement of combinatorial organic synthesis to produce libraries of non-polymeric organic molecules is given. Methods and examples of the development of solid-phase organic chemistry and its subsequent application to combinatorial library synthesis for drug discovery is illustrated with successful case studies. The synthetic versatility of resin-bound amino-acid-derived imine intermediates to produces, β-sultams and pyridines is shown. Use of natural products as key components for creation of combinatorial libraries is presented using Rauwolfia alkaloids and the cephalosporin nucleus as examples

Factor analysis of biochemical and clinical indicators of aseptic necrosis of the femoral head

Introduction The study of pathogenesis and improving the diagnosis of aseptic necrosis of the femoral head (ANFH) is one of the challenging problems of regenerative orthopedics. ANFH is a polyetiologic disease characterized by a local increase in the resorption activity of osteoclasts along with inhibition of activity of mesenchymal cells and osteoblasts, which lead to a significant loss of bone tissue. In this connection, it seems to us relevant to study biochemical and clinical markers of bone resorption and remodeling as diagnostic criteria for ANFH. The purpose of the study was to identify the relationship of some biochemical markers of bone tissue metabolism that reflect the balance between resorption and remodeling with clinical ones (gender, age, T- and Z-criteria), evaluate specifically the deficit of vitamin D in patients with ANFH. Materials and methods Clinical and biochemical study included 195 people with a verified diagnosis of aseptic necrosis of the femoral head who had not previously received vitamin D preparations. They were 87 men and 108 women aged 18 to 88 years. Results In the course of the study, the deficiency of vitamin D of varying severity was revealed in 69.7% of the subjects diagnosed with ANFH. Vitamin deficiency in males was higher than in women, 71.2% and 68.5 % respectively (p = 0.02). The average content of vitamin D was 26.5 ng/ml in women and 26.7 ng/ml in men which is lower than the optimal level (> 30 ng/ml). In addition, low values of vitamin D in women before and during the postmenopausal period were almost equal (68% and 68.7 %). In males over 50 years of age, the same tendency was observed: vitamin D deficiency was revealed in 64.6 % of men under 50 years and in 75.9 % over 50 years of age (p = 0.12). The most significant in magnitude and incidence of factorial connections with clinical characteristics were such biochemical parameters as Ca2+, (25OH)D and 1.25 (OH)2D in women and parathyroid hormone, osteocalcin, DPD in men. Discussion Analysis of correlations of biochemical indicators with clinical characteristics established that such indicators as the content of Ca2+, (25OH)D and 1.25 (OH)2D in women, and parathyroid hormone, osteocalcin and DPD in men were the most significant. In women, the most related clinical feature was age. Age correlated with the content of Ca2+, Ca, parathyroid hormone, DPD. It can be assumed that this is due to the biological aging of the female organism (menopause, osteoporosis). Conclusion The factor analysis enabled to determine the main groups of parameters that influence the variation of clinical and biochemical parameters in women and men diagnosed with ANFH, and also to identify the links between biochemical indicators and clinical features. This circumstance makes it possible to conduct a complex and differentiated assessment of metabolic disorders and to justify rational treatment tactics

A standard photomap of ovarian nurse cell chromosomes and inversion polymorphism in Anopheles beklemishevi

Background Anopheles beklemishevi is a member of the Maculipennis group of malaria mosquitoes that has the most northern distribution among other members of the group. Although a cytogenetic map for the larval salivary gland chromosomes of this species has been developed, a high-quality standard cytogenetic photomap that enables genomics and population genetics studies of this mosquito at the adult stage is still lacking. Methods In this study, a cytogenetic map for the polytene chromosomes of An. beklemishevi from ovarian nurse cells was developed using high-resolution digital imaging from field collected mosquitoes. PCR-amplified DNA probes for fluorescence in situ hybridization (FISH) were designed based on the genome of An. atroparvus. The DNA probe obtained by microdissection procedures from the breakpoint region was labelled in a DOP-PCR reaction. Population analysis was performed on 371 specimens collected in 18 locations. Results We report the development of a high-quality standard photomap for the polytene chromosomes from ovarian nurse cells of An. beklemishevi. To confirm the suitability of the map for physical mapping, several PCR-amplified probes were mapped to the chromosomes of An. beklemishevi using FISH. In addition, we identified and mapped DNA probes to flanking regions of the breakpoints of two inversions on chromosome X of this species. Inversion polymorphism was determined in 13 geographically distant populations of An. beklemishevi. Four polymorphic inversions were detected. The positions of common chromosomal inversions were indicated on the map. Conclusions The study constructed a standard photomap for ovarian nurse cell chromosomes of An. beklemishevi and tested its suitability for physical genome mapping and population studies. Cytogenetic analysis determined inversion polymorphism in natural populations of An. beklemishevi related to this species’ adaptatio

Study of polymorphic loci of CALCR, COL1A1, VDR, and LCT genes in patients with aseptic necrosis of the femoral head

Introduction Aseptic necrosis of the femoral head (ANFH) is a multifactorial disease, and genetic predisposition is one of these factors. Considering this circumstance, researchers search for identification of genetic markers of ANFH development. An objective of this research was to study the frequency of alleles and genotypes of polymorphic loci of 1377 C/T gene CALCR; -1997G/T, -1663indelT and +1245 G/T (Sp1) of gene COL1A1; -3731 A/G (Cdx2) и +283 G/A (BsmI) of gene VDR, and -13910 C/T of gene LCT in patients with ANFH and further analyze the association of the molecular-genetic markers under the study with the risk of developing this disease. Material and methods Analysis of association of alleles of genes for studying genetic predisposition to ANFH was carried out. Seven polymorphic markers in genes CALCR, COL1A1, VDR, LCT were detected by pyrosequencing method using the system of genetic analysis PyroMark Q24. Genotyping of 60 DNA samples of individuals with ANFH was conducted, frequencies of alleles and genotypes were determined. Results Genotype A/A of polymorphic locus +283 G/A (BsmI) of gene VDR (OR = 2.92; 95 % CI: 1.16–7.35) was associated with the risk of ANFH development as well as the carriage of allele A of this locus (OR = 1.55; 95 % CI: 1.02-2.37). It was also found that genotype G/G of polymorphic locus -3731 A/G (Cdx2) in gene VDR increased the risk of ANFH development more than twice (OR = 2.09; 95 % CI: 0.51-8.59); the carriage of the allele G of this polymorphic locus is associated with an elevated risk of ANFH (OR = 1.8; 95 % CI: 1.13-2.86). Discussion The results show that the analysis of the polymorphic loci +283 G/A (BsmI) and -3731 A/G (Cdx2) of VDR gene enables an early identification of persons at high risk of ANFH and, consequently, a possibility to prevent this disease. However, the involvement of certain genes in ANFH development requires further study, particularly given the sample sizes and ethnic specificity. Conclusion The risk of developing ANFH increased more than twice in the presence of genotype G/G of the polymorphic locus -3731 A/G (Cdx2) of VDR gene (OR = 2.09; 95 % CI: 0.51–8.59). Association of genotype A/A of locus +283 G/A (BsmI) of the gene of vitamin D receptor VDR with the risk of ANFH was established (OR = 2.92; 95 % CI: 1.16–7.35); it was also found that the A allele carriage was associated with an increased risk of ANFH (OR = 1.55; 95 % CI:1.02–2.37)

Phylogenomics revealed migration routes and adaptive radiation timing of holarctic malaria mosquito species of the Maculipennis group

BackgroundPhylogenetic analyses of closely related species of mosquitoes are important for better understanding the evolution of traits contributing to transmission of vector-borne diseases. Six out of 41 dominant malaria vectors of the genus Anopheles in the world belong to the Maculipennis Group, which is subdivided into two Nearctic subgroups (Freeborni and Quadrimaculatus) and one Palearctic (Maculipennis) subgroup. Although previous studies considered the Nearctic subgroups as ancestral, details about their relationship with the Palearctic subgroup, and their migration times and routes from North America to Eurasia remain controversial. The Palearctic species An. beklemishevi is currently included in the Nearctic Quadrimaculatus subgroup adding to the uncertainties in mosquito systematics.ResultsTo reconstruct historic relationships in the Maculipennis Group, we conducted a phylogenomic analysis of 11 Palearctic and 2 Nearctic species based on sequences of 1271 orthologous genes. The analysis indicated that the Palearctic species An. beklemishevi clusters together with other Eurasian species and represents a basal lineage among them. Also, An. beklemishevi is related more closely to An. freeborni, which inhabits the Western United States, rather than to An. quadrimaculatus, a species from the Eastern United States. The time-calibrated tree suggests a migration of mosquitoes in the Maculipennis Group from North America to Eurasia about 20-25 million years ago through the Bering Land Bridge. A Hybridcheck analysis demonstrated highly significant signatures of introgression events between allopatric species An. labranchiae and An. beklemishevi. The analysis also identified ancestral introgression events between An. sacharovi and its Nearctic relative An. freeborni despite their current geographic isolation. The reconstructed phylogeny suggests that vector competence and the ability to enter complete diapause during winter evolved independently in different lineages of the Maculipennis Group.ConclusionsOur phylogenomic analyses reveal migration routes and adaptive radiation timing of Holarctic malaria vectors and strongly support the inclusion of An. beklemishevi into the Maculipennis Subgroup. Detailed knowledge of the evolutionary history of the Maculipennis Subgroup provides a framework for examining the genomic changes related to ecological adaptation and susceptibility to human pathogens. These genomic variations may inform researchers about similar changes in the future providing insights into the patterns of disease transmission in Eurasia

A nearly complete database on the records and ecology of the rarest boreal tiger moth from 1840s to 2020

Global environmental changes may cause dramatic insect declines but over century-long time series of certain species’ records are rarely available for scientific research. The Menetries’ Tiger Moth (Arctia menetriesii) appears to be the most enigmatic example among boreal insects. Although it occurs throughout the entire Eurasian taiga biome, it is so rare that less than 100 specimens were recorded since its original description in 1846. Here, we present the database, which contains nearly all available information on the species’ records collected from 1840s to 2020. The data on A. menetriesii records (N = 78) through geographic regions, environments, and different timeframes are compiled and unified. The database may serve as the basis for a wide array of future research such as the distribution modeling and predictions of range shifts under climate changes. It represents a unique example of a more than century-long dataset of distributional, ecological, and phenological data designed for an exceptionally rare but widespread boreal insect, which primarily occurs in hard-to-reach, uninhabited areas of Eurasia.Peer reviewe

Molecular identification of Palearctic members of Anopheles maculipennis in northern Iran

BACKGROUND: Members of Anopheles maculipennis complex are effective malaria vectors in Europe and the Caspian Sea region in northern Iran, where malaria has been re-introduced since 1994. The current study has been designed in order to provide further evidence on the status of species composition and to identify more accurately the members of the maculipennis complex in northern Iran. METHODS: The second internal transcribed spacer of ribosomal DNA (rDNA-ITS2) was sequenced in 28 out of 235 specimens that were collected in the five provinces of East Azerbayjan, Ardebil, Guilan, Mazandaran and Khorassan in Iran. RESULTS: The length of the ITS2 ranged from 283 to 302 bp with a GC content of 49.33 – 54.76%. No intra-specific variations were observed. Construction of phylogenetic tree based on the ITS2 sequence revealed that the six Iranian members of the maculipennis complex could be easily clustered into three groups: the An. atroparvus – Anopheles labranchiae group; the paraphyletic group of An. maculipennis, An. messeae, An. persiensis; and An. sacharovi as the third group. CONCLUSION: Detection of three species of the An. maculipennis complex including An. atroparvus, An. messae and An. labranchiae, as shown as new records in northern Iran, is somehow alarming. A better understanding of the epidemiology of malaria on both sides of the Caspian Sea may be provided by applying the molecular techniques to the correct identification of species complexes, to the detection of Plasmodium composition in Anopheles vectors and to the status of insecticide resistance by looking to related genes

Effect of aliskiren on post-discharge outcomes among diabetic and non-diabetic patients hospitalized for heart failure: insights from the ASTRONAUT trial

Aims The objective of the Aliskiren Trial on Acute Heart Failure Outcomes (ASTRONAUT) was to determine whether aliskiren, a direct renin inhibitor, would improve post-discharge outcomes in patients with hospitalization for heart failure (HHF) with reduced ejection fraction. Pre-specified subgroup analyses suggested potential heterogeneity in post-discharge outcomes with aliskiren in patients with and without baseline diabetes mellitus (DM). Methods and results ASTRONAUT included 953 patients without DM (aliskiren 489; placebo 464) and 662 patients with DM (aliskiren 319; placebo 343) (as reported by study investigators). Study endpoints included the first occurrence of cardiovascular death or HHF within 6 and 12 months, all-cause death within 6 and 12 months, and change from baseline in N-terminal pro-B-type natriuretic peptide (NT-proBNP) at 1, 6, and 12 months. Data regarding risk of hyperkalaemia, renal impairment, and hypotension, and changes in additional serum biomarkers were collected. The effect of aliskiren on cardiovascular death or HHF within 6 months (primary endpoint) did not significantly differ by baseline DM status (P = 0.08 for interaction), but reached statistical significance at 12 months (non-DM: HR: 0.80, 95% CI: 0.64-0.99; DM: HR: 1.16, 95% CI: 0.91-1.47; P = 0.03 for interaction). Risk of 12-month all-cause death with aliskiren significantly differed by the presence of baseline DM (non-DM: HR: 0.69, 95% CI: 0.50-0.94; DM: HR: 1.64, 95% CI: 1.15-2.33; P < 0.01 for interaction). Among non-diabetics, aliskiren significantly reduced NT-proBNP through 6 months and plasma troponin I and aldosterone through 12 months, as compared to placebo. Among diabetic patients, aliskiren reduced plasma troponin I and aldosterone relative to placebo through 1 month only. There was a trend towards differing risk of post-baseline potassium ≥6 mmol/L with aliskiren by underlying DM status (non-DM: HR: 1.17, 95% CI: 0.71-1.93; DM: HR: 2.39, 95% CI: 1.30-4.42; P = 0.07 for interaction). Conclusion This pre-specified subgroup analysis from the ASTRONAUT trial generates the hypothesis that the addition of aliskiren to standard HHF therapy in non-diabetic patients is generally well-tolerated and improves post-discharge outcomes and biomarker profiles. In contrast, diabetic patients receiving aliskiren appear to have worse post-discharge outcomes. Future prospective investigations are needed to confirm potential benefits of renin inhibition in a large cohort of HHF patients without D

Design and baseline characteristics of the finerenone in reducing cardiovascular mortality and morbidity in diabetic kidney disease trial

Background: Among people with diabetes, those with kidney disease have exceptionally high rates of cardiovascular (CV) morbidity and mortality and progression of their underlying kidney disease. Finerenone is a novel, nonsteroidal, selective mineralocorticoid receptor antagonist that has shown to reduce albuminuria in type 2 diabetes (T2D) patients with chronic kidney disease (CKD) while revealing only a low risk of hyperkalemia. However, the effect of finerenone on CV and renal outcomes has not yet been investigated in long-term trials. Patients and Methods: The Finerenone in Reducing CV Mortality and Morbidity in Diabetic Kidney Disease (FIGARO-DKD) trial aims to assess the efficacy and safety of finerenone compared to placebo at reducing clinically important CV and renal outcomes in T2D patients with CKD. FIGARO-DKD is a randomized, double-blind, placebo-controlled, parallel-group, event-driven trial running in 47 countries with an expected duration of approximately 6 years. FIGARO-DKD randomized 7,437 patients with an estimated glomerular filtration rate >= 25 mL/min/1.73 m(2) and albuminuria (urinary albumin-to-creatinine ratio >= 30 to <= 5,000 mg/g). The study has at least 90% power to detect a 20% reduction in the risk of the primary outcome (overall two-sided significance level alpha = 0.05), the composite of time to first occurrence of CV death, nonfatal myocardial infarction, nonfatal stroke, or hospitalization for heart failure. Conclusions: FIGARO-DKD will determine whether an optimally treated cohort of T2D patients with CKD at high risk of CV and renal events will experience cardiorenal benefits with the addition of finerenone to their treatment regimen. Trial Registration: EudraCT number: 2015-000950-39; ClinicalTrials.gov identifier: NCT02545049