Research on the embryotoxic effect and carcinogenicity of the drug “BTF plus” – a means for normalizing metabolic processes in animals and poul-try

Laboratory studies were conducted to determine the embryotoxic effect and carcinogenicity of the veterinary drug “BTF plus” on white rats and white mice. The drug “BTF plus” is a complex vitamin-mineral drug based on butophosphane, L-carnitine, and cyanocobalamin, which is used to normalize and correct metabolic processes in animals and poultry. The drug is used for various types of animals and poultry as a stimulating, tonic and general strengthening agent for obstetric pathologies (complicated childbirth, postpartum complications, paresis, eclampsia, sexual cycle disorders); metabolic disorders caused by irrational feeding, malnutrition, asthenic syndrome, etc.; anemia with helminthiasis; secondary anemias, as an additional means in the treatment of magnesium and calcium deficiency; to increase muscle activity, with significant loads, overstrain and exhaustion in animals; to increase the body's resistance to various pathogens; to stimulate growth, development and live weight gain in young animals and poultry; as an additional means in the treatment of diseases caused by various factors (infectious and non-infectious origin). The drug “BTF plus”, under the conditions of subcutaneous administration to pregnant female rats in doses (based on the absolute weight of the drug) of 200.0 and 2000.0 mg/kg of body weight, does not cause death and pathological changes in embryos do not have an embryotoxic and teratogenic effect since indicators of total, preimplantation, and postimplantation embryonic lethality in rats of the experimental groups had no significant differences compared to indicators in control and also did not show changes in the weight of the placenta, fetuses, and their cranio-caudal size. The drug “BTF plus”, under conditions of 5-day subcutaneous administration to white mice in doses (based on the absolute weight of the drug) of 200.0 and 2000.0 mg/kg of body weight, does not show a carcinogenic effect (during microscopic studies, the proportion of polychromatophilic erythrocytes was not probable deviations between themselves and was 0.117-0.133%, which is within the normal range of up to 0.2 %). Further studies will be the next stage of pre-registration tests aimed at studying the ecotoxicity of “BTF plus”, which is a mandatory material of the “Safety and residue studies” section of the dossier for this drug

Joint profiling of DNA methylation and chromatin architecture in single cells.

We report a molecular assay, Methyl-HiC, that can simultaneously capture the chromosome conformation and DNA methylome in a cell. Methyl-HiC reveals coordinated DNA methylation status between distal genomic segments that are in spatial proximity in the nucleus, and delineates heterogeneity of both the chromatin architecture and DNA methylome in a mixed population. It enables simultaneous characterization of cell-type-specific chromatin organization and epigenome in complex tissues

Експериментальна оцінка гострої токсичності та подразнювальної дії “БТФ плюс” – ветеринарного лікарського засобу для нормалізації обмінних процесів у тварин і птиці

Laboratory studies were conducted to determine the acute toxicity of the veterinary drug “BTF plus” on white rats and white mice. The preparation “BTF plus” is a complex vitamin and mineral preparation, which is used to normalize and correct metabolic processes in poultry and animals. The drug is used for various types of animals and poultry as a stimulating, tonic and general strengthening agent for: obstetric pathologies (difficult births, postpartum complications, paresis, eclampsia, sexual cycle disorders); metabolic disorders caused by irrational feeding, malnutrition, asthenic syndrome, etc.; anemia with helminthiasis; secondary anemias, as an additional means in the treatment of magnesium and calcium deficiency; to increase muscle activity, with significant loads, overstrain and exhaustion in animals; to increase the body’s resistance to various pathogens; to stimulate growth, development and live weight gain in young animals; as an additional means in the treatment of diseases caused by various factors (infectious and non-infectious origin). Based on the results of determining the parameters of the acute toxicity of the drug “BTF plus” in the case of a single intragastric administration to white female rats, it was not possible to calculate the LD50, since the death of laboratory animals was not detected within 14 days after administration. At the same time, the maximum injected dose (based on the absolute weight of the drug) was 40000.0 mg/kg of body weight, which allows the drug to be classified as VI class of toxicity – the substances are relatively harmless (LD50 > 15000.0 mg/kg of body weight), and according to the degree of danger to IV class – low-hazardous substances (LD50 > 5000.0 mg/kg of body weight). Based on the results of determining the parameters of the acute toxicity of the drug “BTF plus” in the case of a single subcutaneous injection to white female rats and male mice, it was not possible to calculate the LD50, since the death of laboratory animals was not detected within 14 days after administration. At the same time, the maximum injected dose (based on the absolute weight of the drug) was 20000,0 and 40000.0 mg/kg of body weight for both species of animals, respectively, which allows it to be classified as VI class – relatively harmless substances (LD50Subcut > 4500 mg/kg of body weight). Further studies will be the next stage of pre-registration tests aimed at studying the embryotoxic effect of “BTF plus”, which is mandatory material of the “Safety and residue studies” section of the dossier for this drug.Проведені лабораторні дослідження з визначення гострої токсичності ветеринарного препарату “БТФ плюс” на білих щурах, білих мишах. Препарат “БТФ плюс” – комплексний вітамінно-мінеральний препарат, який застосовується для нормалізації та корекції обмінних процесів у птиці та тварин. Препарат застосовують різним видам тварин та птиці як стимулюючий, тонізуючий та загальнозміцнюючий засіб при: акушерських патологіях (складні пологи, післяпологові ускладнення, парези, еклампсії, порушення статевого циклу); порушеннях обміну речовин, що викликані нераціональною годівлею, недоїданням, астенічному синдромі тощо; анеміях при гельмінтозах; вторинних анеміях, як додатковий засіб при лікуванні дефіциту Магнію та Кальцію; для підвищення м’язової активності, при значних навантаженнях, перенапруженнях та виснаженні у тварин; для підвищення резистентності організму до дії різноманітних патогенів; для стимулювання росту, розвитку та приросту живої ваги у молодих тварин; як додатковий засіб при лікуванні захворювань, спричинених різними факторами (інфекційного та неінфекційного походження). За результатами визначення параметрів гострої токсичності препарату “БТФ плюс” – у разі одноразового внутрішньошлункового введення білим щурам-самкам LD50 розрахувати не вдалося, оскільки протягом 14 діб після введення загибелі лабораторних тварин не було виявлено. При цьому максимальна введена доза (за абсолютною масою препарату) становила 40000,0 мг/кг маси тіла, що дозволяє зарахувати препарат до VI класу токсичності – речовини порівняно нешкідливі (LD50 > 15000,0 мг/кг маси тіла), а за ступенем небезпечності до ІV класу – малонебезпечних речовин (LD50 > 5000,0 мг/кг маси тіла). За результатами визначення параметрів гострої токсичності препарату “БТФ плюс” – у разі одноразового підшкірного введення білим щурам-самкам і мишам-самцям LD50 розрахувати не вдалося, оскільки протягом 14 діб після введення загибелі лабораторних тварин не було виявлено. При цьому максимальна введена доза (за абсолютною масою препарату) становила 20000,0 і 40000,0 мг/кг маси тіла для обох видів тварин відповідно, що дозволяє зарахувати його до VІ класу – відносно нешкідливих речовин (LD50Subcut > 4500 мг/кг маси тіла). Подальші дослідження будуть черговим етапом передреєстраційних випробувань, спрямованих на вивчення ембріотоксичної дії “БТФ плюс”, що є обов’язковим матеріалом розділу “Дослідження щодо безпеки і залишків” досьє на даний лікарський засіб

The Drosophila homolog of the mammalian imprint regulator, CTCF, maintains the maternal genomic imprint in Drosophila melanogaster

<p>Abstract</p> <p>Background</p> <p>CTCF is a versatile zinc finger DNA-binding protein that functions as a highly conserved epigenetic transcriptional regulator. CTCF is known to act as a chromosomal insulator, bind promoter regions, and facilitate long-range chromatin interactions. In mammals, CTCF is active in the regulatory regions of some genes that exhibit genomic imprinting, acting as insulator on only one parental allele to facilitate parent-specific expression. In <it>Drosophila</it>, CTCF acts as a chromatin insulator and is thought to be actively involved in the global organization of the genome.</p> <p>Results</p> <p>To determine whether CTCF regulates imprinting in <it>Drosophila</it>, we generated <it>CTCF </it>mutant alleles and assayed gene expression from the imprinted <it>Dp(1;f)LJ9 </it>mini-X chromosome in the presence of reduced <it>CTCF </it>expression. We observed disruption of the maternal imprint when <it>CTCF </it>levels were reduced, but no effect was observed on the paternal imprint. The effect was restricted to maintenance of the imprint and was specific for the <it>Dp(1;f)LJ9 </it>mini-X chromosome.</p> <p>Conclusions</p> <p>CTCF in <it>Drosophila </it>functions in maintaining parent-specific expression from an imprinted domain as it does in mammals. We propose that <it>Drosophila </it>CTCF maintains an insulator boundary on the maternal X chromosome, shielding genes from the imprint-induced silencing that occurs on the paternally inherited X chromosome.</p> <p>See commentary: <url>http://www.biomedcentral.com/1741-7007/8/104</url></p

The Genome of the Stick Insect Medauroidea extradentata Is Strongly Methylated within Genes and Repetitive DNA

BACKGROUND: Cytosine DNA methylation has been detected in many eukaryotic organisms and has been shown to play an important role in development and disease of vertebrates including humans. Molecularly, DNA methylation appears to be involved in the suppression of initiation or of elongation of transcription. Resulting organismal functions are suggested to be the regulation of gene silencing, the suppression of transposon activity and the suppression of initiation of transcription within genes. However, some data concerning the distribution of methylcytosine in insect species appear to contradict such roles. PRINCIPAL FINDINGS: By comparison of MspI and HpaII restriction patterns in genomic DNA of several insects we show that stick insects (Phasmatodea) have highly methylated genomes. We isolated methylated DNA fragments from the Vietnamese Walking Stick Medauroidea extradentata (formerly known as Baculum extradentatum) and demonstrated that most of the corresponding sequences are repetitive. Bisulfite sequencing of one of these fragments and of parts of conserved protein-coding genes revealed a methylcytosine content of 12.6%, mostly found at CpG, but also at CpT and CpA dinucleotides. Corresponding depletions of CpG and enrichments of TpG and CpA dinucleotides in some highly conserved protein-coding genes of Medauroidea reach a similar degree as in vertebrates and show that CpG methylation has occurred in the germline of these insects. CONCLUSIONS: Using four different methods, we demonstrate that the genome of Medauroidea extradentata is strongly methylated. Both repetitive DNA and coding genes appear to contain high levels of methylcytosines. These results argue for similar functions of DNA methylation in stick insects as those already known for vertebrates

The genomes of two key bumblebee species with primitive eusocial organization

Background: The shift from solitary to social behavior is one of the major evolutionary transitions. Primitively eusocial bumblebees are uniquely placed to illuminate the evolution of highly eusocial insect societies. Bumblebees are also invaluable natural and agricultural pollinators, and there is widespread concern over recent population declines in some species. High-quality genomic data will inform key aspects of bumblebee biology, including susceptibility to implicated population viability threats. Results: We report the high quality draft genome sequences of Bombus terrestris and Bombus impatiens, two ecologically dominant bumblebees and widely utilized study species. Comparing these new genomes to those of the highly eusocial honeybee Apis mellifera and other Hymenoptera, we identify deeply conserved similarities, as well as novelties key to the biology of these organisms. Some honeybee genome features thought to underpin advanced eusociality are also present in bumblebees, indicating an earlier evolution in the bee lineage. Xenobiotic detoxification and immune genes are similarly depauperate in bumblebees and honeybees, and multiple categories of genes linked to social organization, including development and behavior, show high conservation. Key differences identified include a bias in bumblebee chemoreception towards gustation from olfaction, and striking differences in microRNAs, potentially responsible for gene regulation underlying social and other traits. Conclusions: These two bumblebee genomes provide a foundation for post-genomic research on these key pollinators and insect societies. Overall, gene repertoires suggest that the route to advanced eusociality in bees was mediated by many small changes in many genes and processes, and not by notable expansion or depauperation

Drosophila as a Model for MECP2 Gain of Function in Neurons

Methyl-CpG-binding protein 2 (MECP2) is a multi-functional regulator of gene expression. In humans loss of MECP2 function causes classic Rett syndrome, but gain of MECP2 function also causes mental retardation. Although mouse models provide valuable insight into Mecp2 gain and loss of function, the identification of MECP2 genetic targets and interactors remains time intensive and complicated. This study takes a step toward utilizing Drosophila as a model to identify genetic targets and cellular consequences of MECP2 gain-of function mutations in neurons, the principle cell type affected in patients with Rett-related mental retardation. We show that heterologous expression of human MECP2 in Drosophila motoneurons causes distinct defects in dendritic structure and motor behavior, as reported with MECP2 gain of function in humans and mice. Multiple lines of evidence suggest that these defects arise from specific MECP2 function. First, neurons with MECP2-induced dendrite loss show normal membrane currents. Second, dendritic phenotypes require an intact methyl-CpG-binding domain. Third, dendritic defects are amended by reducing the dose of the chromatin remodeling protein, osa, indicating that MECP2 may act via chromatin remodeling in Drosophila. MECP2-induced motoneuron dendritic defects cause specific motor behavior defects that are easy to score in genetic screening. In sum, our data show that some aspects of MECP2 function can be studied in the Drosophila model, thus expanding the repertoire of genetic reagents that can be used to unravel specific neural functions of MECP2. However, additional genes and signaling pathways identified through such approaches in Drosophila will require careful validation in the mouse model

Immunogenicity and safety of a quadrivalent high-dose inactivated influenza vaccine compared with a standard-dose quadrivalent influenza vaccine in healthy people aged 60 years or older: a randomized Phase III trial

A quadrivalent high-dose inactivated influenza vaccine (IIV4-HD) is licensed for adults 6565&nbsp;y of age based on immunogenicity and efficacy studies. However, IIV4-HD has not been evaluated in adults aged 60\u201364&nbsp;y. This study compared immunogenicity and safety of IIV4-HD with a standard-dose quadrivalent influenza vaccine (IIV4-SD) in adults aged 6560&nbsp;y. This Phase III, randomized, modified double-blind, active-controlled study enrolled 1,528 participants aged 6560&nbsp;y, randomized 1:1 to a single injection of IIV4-HD or IIV4-SD. Hemagglutination inhibition (HAI) geometric mean titers (GMTs) were measured at baseline and D 28 and seroconversion assessed. Safety was described for 180&nbsp;d after vaccination. The primary immunogenicity objective was superiority of IIV4-HD versus IIV4-SD, for all four influenza strains 28&nbsp;d post vaccination in participants aged 60\u201364 and 6565&nbsp;y. IIV4-HD induced a superior immune response versus IIV4-SD in terms of GMTs in participants aged 60\u201364 y and those aged 6565&nbsp;y for all four influenza strains. IIV4-HD induced higher GMTs in those aged 60\u201364 y than those aged 6565 y. Seroconversion rates were higher for IIV4-HD versus IIV4-SD in each age-group for all influenza strains. Both vaccines were well tolerated in participants 6560&nbsp;y of age, with no safety concerns identified. More solicited reactions were reported with IIV4-HD than with IIV4-SD. IIV4-HD provided superior immunogenicity versus IIV4-SD and was well tolerated in adults aged 6560 y. IIV4-HD is assumed to offer improved protection against influenza compared with IIV4-SD in adults aged 6560 y, as was previously assessed for adults aged 6565&nbsp;y