Knockout Pigs as Recipients for Spermatogonial Stem Cell Transplantation.

Spermatogonial stem cell (SSC) transplantation into the testis of a germ cell (GC)-depleted surrogate allows transmission of donor genotype via donor-derived sperm produced by the recipient. Transplantation of gene-edited SSCs provides an approach to propagate gene-edited large animal models. DAZL is a conserved RNA-binding protein important for GC development, and knockout (KO) causes defects in GC commitment and differentiation.

Targeted Gene Editing in Porcine Germ Cells.

As the genetic mutations driving human disease are identified, there is an increasing need for a biomedical model that can accurately represent the disease of interest and provide a platform for potential therapeutic testing. Pigs are a better model for human disease than rodents because of their genetic and physiological similarities to humans. However, current methods to generate porcine models are both technically challenging and expensive.

Targeted Gene Editing in Porcine Spermatogonia.

To study the pathophysiology of human diseases, develop innovative treatments, and refine approaches for regenerative medicine require appropriate preclinical models. Pigs share physiologic and anatomic characteristics with humans and are genetically more similar to humans than are mice. Genetically modified pigs are essential where rodent models do not mimic the human disease phenotype.

ORGANOMETRIC CHANGES IN THYMUS UNDER THE INFLUENCE OF PROPYLENE GLYCOL.

The scientific interest in the influence of xenobiotics on the human body is due to the fact that immune organs are characterized by a pronounced response to the influence of endogenous and exogenous factors. Recently, the immunological impairment, as a manifestation of reactions to ecopathogenic conditions, suggests a major pathogenesis role in the development of cardiovascular, neuropsychiatric diseases, as well as diffuse diseases of connective tissue. Objectives - experiment was designed to elucidate the organometric changes in thymus of male rats due to impact of propylene glycol.

ANTIOXIDANT PROTECTION STATUS AND LIPOPEROXIDATION PROCESSES IN WOMEN WITH PREMATURE BIRTH IN DIFFERENT TERMS OF GESTATION.

The purpose of the study is examination the dynamics of antioxidant defense indicators and lipoperoxidation processes in women with pregnancy termination different periods. 227 pregnant women were examined, 190 of them had clinical signs of premature birth in the gestation period of 23-36 weeks. Formation of clinical groups was carried out depending on the term of pregnancy in the form of premature and timely birth.

TALEN-mediated gene targeting in porcine spermatogonia.

Spermatogonia represent a diploid germ cell population that includes spermatogonial stem cells. In this report, we describe new methods for isolation of highly enriched porcine spermatogonia based on light scatter properties, and for targeted mutagenesis in porcine spermatogonia using nucleofection and TALENs. We optimized a nucleofection protocol to deliver TALENs specifically targeting the DMD locus in porcine spermatogonia.

Phthalate esters affect maturation and function of primate testis tissue ectopically grafted in mice.

Di-n-Butyl (DBP) and Di-(2-EthylHexyl) (DEHP) phthalates can leach from daily-use products resulting in environmental exposure. In male rodents, phthalate exposure results in reproductive effects. To evaluate effects on the immature primate testis, testis fragments from 6-month-old rhesus macaques were grafted subcutaneously to immune-deficient mice, which were exposed to 0, 10, or 500 mg/kg of DBP or DEHP for 14 weeks or 28 weeks (DBP only).

Combined treatment with paclitaxel and suramin prevents the development of metastasis by inhibiting metastatic colonization of circulating tumor cells.

Metastatic disease accounts for most deaths due to breast cancer and thus identification of novel ways to prevent this complication remains a key goal. A frequently employed preclinical model of breast cancer metastasis relies on xenografted human MDA-MB-231 cells, since these reliably produce both soft tissue and osseous metastases when introduced into the arterial circulation of athymic mice. Herein, we explored the ability of suramin (SA), an agent shown to antagonize the effects of various stromal cell-derived growth factors relevant to bone marrow colonization of tumor cells, administered both with and without paclitaxel (PTX), to inhibit the development of MDA-MB-231 metastases.

Goat embryonic stem-like cell derivation and characterization.

Embryonic stem (ES) cells are derived from the inner cell masses of preimplantation embryos. ES cells are pluripotent cells with the capacity for long-term propagation and broad differentiation plasticity. These cells have an exceptional functional feature in that they can differentiate into all tissues and organs, including germ cells.

Viral transduction of male germline stem cells results in transgene transmission after germ cell transplantation in pigs.

Genetic modification of germline stem cells (GSCs) is an alternative approach to generate large transgenic animals where transgenic GSCs are transplanted into a recipient testis to generate donor-derived transgenic sperm. The objective of the present study was to explore the application of viral vectors in delivering an enhanced green fluorescent protein (EGFP) transgene into GSCs for production of transgenic gametes through germ cell transplantation. Both adeno-associated virus (AAV)- and lentivirus (LV)-based vectors were effective in transducing pig GSCs, resulting in the production of transgenic sperm in recipient boars.

Non-viral transfection of goat germline stem cells by nucleofection results in production of transgenic sperm after germ cell transplantation.

Germline stem cells (GSCs) can be used for large animal transgenesis, in which GSCs that are genetically manipulated in vitro are transplanted into a recipient testis to generate donor-derived transgenic sperm. The objectives of this study were to explore a non-viral approach for transgene delivery into goat GSCs and to investigate the efficiency of nucleofection in producing transgenic sperm. Four recipient goats received fractionated irradiation at 8 weeks of age to deplete endogenous GSCs.

[Essential hypertension as an occupational disease in engine crew members].

The paper raises the question as to whether essential hypertension should be included into a list of occupational diseases (intellectual, sensory, and emotional workloads and working conditions) in engine crew members. In the engine crew members of the South-Eastern Railway, the cases and days lost because of illness and temporary disability are higher than those in the whole railway. 32% of the engine drivers (assistants) are followed up for diagnosed essential hypertension.

Establishment of goat embryonic stem cells from in vivo produced blastocyst-stage embryos.

Embryonic stem (ES) cells with the capacity for germ line transmission have only been verified in mouse and rat. Methods for derivation, propagation, and differentiation of ES cells from domestic animals have not been fully established. Here, we describe derivation of ES cells from goat embryos.

(1)H NMR metabolomics identification of markers of hypoxia-induced metabolic shifts in a breast cancer model system.

Hypoxia can promote invasive behavior in cancer cells and alters the response to therapeutic intervention as a result of changes in the expression many genes, including genes involved in intermediary metabolism. Although metabolomics technologies are capable of simultaneously measuring a wide range of metabolites in an untargeted manner, these methods have been relatively under utilized in the study of cancer cell responses to hypoxia. Thus, (1)H NMR metabolomics was used to examine the effects of hypoxia in the MDA-MB-231 human breast cancer cell line, both in vitro and in vivo.

Lymphoid-specific helicase (HELLS) is essential for meiotic progression in mouse spermatocytes.

Lymphoid-specific helicase (HELLS; also known as LSH) is a member of the SNF2 family of chromatin remodeling proteins. Because Hells-null mice die at birth, a phenotype in male meiosis cannot be studied in these animals. Allografting of testis tissue from Hells(-/-) to wild-type mice was employed to study postnatal germ cell differentiation.

Expression pattern of acetylated alpha-tubulin in porcine spermatogonia.

Mammalian spermatogonial stem cells reside on the basement membrane of the seminiferous tubules. The mechanisms responsible for maintenance of spermatogonia at the basement membrane are unclear. Since acetylated alpha-tubulin (Ac-alpha-Tu) is a component of long-lived, stable microtubules and deacetylation of alpha-tubulin enhances cell motility, we hypothesized that acetylation of alpha-tubulin might be associated with positioning of spermatogonia at the basement membrane.

The lysyl oxidase inhibitor, beta-aminopropionitrile, diminishes the metastatic colonization potential of circulating breast cancer cells.

Lysyl oxidase (LOX), an extracellular matrix remodeling enzyme, appears to have a role in promoting breast cancer cell motility and invasiveness. In addition, increased LOX expression has been correlated with decreases in both metastases-free, and overall survival in breast cancer patients. With this background, we studied the ability of beta-aminopropionitrile (BAPN), an irreversible inhibitor of LOX, to regulate the metastatic colonization potential of the human breast cancer cell line, MDA-MB-231.

Effects of thioredoxin reductase-1 deletion on embryogenesis and transcriptome.

Thioredoxin reductases (Txnrd) maintain intracellular redox homeostasis in most organisms. Metazoan Txnrds also participate in signal transduction. Mouse embryos homozygous for a targeted null mutation of the txnrd1 gene, encoding the cytosolic thioredoxin reductase, were viable at embryonic day 8.

[Clinical features of vocal disorders in population of megapolis].

Vocal problems in population of a megapolis are caused, primarily, by acute inflammation of the upper respiratory tract of viral etiology and exacerbation of chronic pharyngolaryngitis in smokers and vocal professionals. Organic diseases of the vocal folds are rare finding in outpatients (0.2-0.

Reproductive and neurological Quaking(viable) phenotypes in a severe combined immune deficient mouse background.

The quaking(viable) (qkv) mutation, a spontaneous deletion of a multigenic region encompassing roughly 1 Mb at 5.9 cM on the proximal end of mouse chromosome 17, causes severe trembling in all homozygous animals and infertility in all homozygous males. Physiologically, quaking mice exhibit dysmyelination and postmeiotic spermatogenic arrest.

Early vertebrate evolution of the TATA-binding protein, TBP.

TBP functions in transcription initiation in all eukaryotes and in Archaebacteria. Although the 181-amino acid (aa) carboxyl (C-) terminal core of the protein is highly conserved, TBP proteins from different phyla exhibit diverse sequences in their amino (N-) terminal region. In mice, the TBP N-terminus plays a role in protecting the placenta from maternal rejection; however the presence of similar TBP N-termini in nontherian tetrapods suggests that this domain also has more primitive functions.

Fundamental cellular processes do not require vertebrate-specific sequences within the TATA-binding protein.

The 180-amino acid core of the TATA-binding protein (TBPcore) is conserved from Archae bacteria to man. Vertebrate TBPs contain, in addition, a large and highly conserved N-terminal region that is not found in other phyla. We have generated a line of mice in which the tbp allele is replaced with a version, tbp(Delta N), which lacks 111 of 135 N-terminal amino acid residues.

Removing the vertebrate-specific TBP N terminus disrupts placental beta2m-dependent interactions with the maternal immune system.

Mammalian TBP consists of a 180 amino acid core that is common to all eukaryotes, fused to a vertebrate-specific N-terminal domain. We generated mice having a modified tbp allele, tbp(DeltaN), that produces a version of TBP lacking 111 of the 135 vertebrate-specific amino acids. Most tbp(DeltaN/DeltaN) fetuses (>90%) died in midgestation from an apparent defect in the placenta.