The common marmoset, Callithrix jacchus, is an invaluable model in biomedical research. Its use includes genetic engineering applications, which require manipulations of oocytes and production of ...embryos in vitro. To maximize the recovery of oocytes suitable for embryo production and to fulfil the requirements of the 3R principles to the highest degree possible, optimization of ovarian stimulation protocols is crucial. Here, we compared the efficacy of two hormonal ovarian stimulation approaches: 1) stimulation of follicular growth with hFSH followed by triggering of oocyte maturation with hCG (FSH + hCG) and 2) stimulation with hFSH only (FSH-priming).
In total, 14 female marmosets were used as oocyte donors in this study. Each animal underwent up to four surgical interventions, with the first three performed as ovum pick-up (OPU) procedures and the last one being an ovariohysterectomy (OvH). In total, 20 experiments were carried out with FSH + hCG stimulation and 18 with FSH-priming. Efficacy of each stimulation protocol was assessed through in vitro maturation (IVM), in vitro fertilization (IVF) and embryo production rates.
Each study group consisted of two subgroups: the in vivo matured oocytes and the oocytes that underwent IVM. Surprisingly, in the absence of hCG triggering some of the oocytes recovered were at the MII stage, moreover, their number was not significantly lower compared to FSH + hCG stimulation (2.8 vs. 3.9, respectively (ns)). While the IVM and IVF rates did not differ between the two stimulation groups, the IVF rates of in vivo matured oocytes were significantly lower compared to in vitro matured ones in both FSH-priming and FSH + hCG groups. In total, 1.7 eight-cell embryos/experiment (OPU) and 2.1 eight-cell embryos/experiment (OvH) were obtained after FSH + hCG stimulation vs. 1.8 eight-cell embryos/experiment (OPU) and 5.0 eight-cell embryos/experiment (OvH) following FSH-priming. These numbers include embryos obtained from both in vivo and in vitro matured oocytes.
A significantly lower developmental competence of the in vivo matured oocytes renders triggering of the in vivo maturation with hCG as a part of the currently used FSH-stimulation protocol unnecessary. In actual numbers, between 1 and 7 blastocysts were obtained following each FSH-priming. In the absence of further studies, FSH-priming appears superior to FSH + hCG stimulation in the common marmoset under current experimental settings.
Abstract
This paper is aimed at a comparison of Ukrainian and international resource classification systems’ fundamental features. As a result, the most common and different features were outlined. ...The importance of accounting for the social and environmental viability of projects is determined. This study highlights the significance of the harmonization of domestic classification with international standards.
There is an increasing recognition that vitamin D plays important roles in female reproduction. Recent studies demonstrated that 1α,25-dihydroxyvitamin D3 (VD3), the biologically active form of ...vitamin D, improved ovarian follicle survival and growth
. Therefore, we investigated the direct effects of VD3 at the specific preantral and antral stages of follicular development, and tested the hypothesis that vitamin D receptor (VDR) and enzymes critical for vitamin D biosynthesis are expressed in the primate ovary. Fourteen adult rhesus macaques provided ovarian tissue. Secondary and antral follicles were isolated for PCR analysis on VDR, vitamin D3 25-hydroxylase, and 25-hydroxyvitamin D3-1α-hydroxylase. VDR protein localization was determined by immunohistochemistry on ovarian sections. Isolated secondary follicles were cultured under conditions of control and VD3 supplementation during the preantral or antral stage. Follicle survival, growth, steroid and anti-Müllerian hormone (AMH) production, as well as oocyte maturation were evaluated.
- and
-developed follicles were also assessed for genes that are critical for vitamin D biosynthesis and signaling, gonadotropin signaling, steroid and paracrine factor production, and oocyte quality. The mRNA encoding VDR, 25-hydroxylase, and 1α-hydroxylase was detectable in
- and
-developed preantral and antral follicles. The 25-hydroxylase was elevated in cultured follicles relative to
-developed follicles, which further increased following VD3 exposure. VD3 treatment increased 1α-hydroxylase in
-developed antral follicles. The absence of VD3 during culture decreased VDR expression in
-developed antral follicles, which was restored to levels comparable to those of
-developed antral follicles by VD3 supplementation. Positive immunostaining for VDR was detected in the nucleus and cytoplasm of granulosa cells and oocytes. While only survival was improved in preantral follicles treated with VD3, VD3 supplementation promoted both survival and growth of antral follicles with increased estradiol and AMH production, as well as oocyte maturation. Thus, Vitamin D biosynthesis and signaling systems are expressed in primate ovarian follicles. Our findings support a role for VD3 in regulating follicular development in a stage-dependent manner, as well as the intrafollicular vitamin D biosynthesis and signaling, directly in the ovary.
The article proposes to use artificial ecosystems to create comfortable and safe living conditions in the Arctic region. The principles of the organization of artificial ecosystems – bioregenerative ...systems of human life support – are formulated. A brief information about the history of the development of experimental installations and the experiments to find people in isolation from the external environment is given. The scheme of an artificial ecosystem is described, which includes equipment and technologies that make it possible to provide isolation in terms of respiration, water circulation, and human nutrition. The concept of an autonomous non-volatile and environmentally independent module for creating a favourable living environment in the Arctic is described. An example of the implementation of a similar project for the construction of an autonomous module designed to support the livelihoods of the inhabitants of the ecovillage Syuderbin (Gotland Island, Sweden) is given.
There is compelling evidence that a hypercaloric, high-fructose diet can cause metabolic syndrome (MetS) and a whole range of other metabolic changes. In the context of androgen deficiency, MetS in ...boys merits special attention, but the effects of fructose-rich diet in youth on future male reproductive function are still poorly evidenced. The aim of this study was to address this issue and analyse the effects of high-fructose intake starting from weaning to puberty (postnatal day 23 up to 83) on the reproductive function of male rats. For this purpose juvenile male Wistar rats were divided in two groups: control and the group receiving 10 % fructose solution instead of drinking water. Reproductive function was evaluated in terms of fertility, sperm count, testes/epididymis morphology, and serum sex hormones. The fructose-treated group showed a decrease in testosterone and twofold increase in luteinising and follicle-stimulating hormone levels in the serum. This was accompanied with lower testis/epididymis weights, sperm count, and changed testis/epididymis morphology. Their fertility remained unchanged, but the fertility of females mating with these males diminished. In addition, pre-implantation and post-implantation embryonic death rate rose in these females. Our results have confirmed that high fructose consumption from early age until puberty can impair the reproductive function of male rats, and call for further animal and epidemiological investigation.
BACKGROUND This is the first study of the effect of epidermal growth factor (EGF) on marmoset monkey oocytes matured in vitro. METHODS We have evaluated the effects of 10 ng/ml EGF in combination ...with 1 or 10 IU/ml of gonadotrophins (FSH/hCG 1:1 ratio) during in vitro maturation (IVM) of marmoset oocytes. Immature cumulus–oocyte complexes (COCs) were retrieved from ovarian antral follicles of unprimed monkeys. COCs from six animals (n= 268) used in this study were randomly distributed among four experimental groups: (A) 1 FSH +1 hCG; (B) 10 FSH +10 hCG; (C) 1 FSH +1 hCG + EGF; and (D) 10 FSH +10 hCG + EGF (where 1 and 10 are concentrations, IU/ml). After IVM, oocytes were fertilized in vitro and embryos were allowed to progress up to 87–88 h. RESULTS the highest rate of total and radial cumulus expansion was observed in Group A, with the lowest in Group B (P < 0.05). Neither maturation nor fertilization rate were affected by gonadotrophin concentration or presence of EGF. Addition of EGF increased degeneration and decreased first cleavage rate, which was significantly lower in Group C than Group A (P < 0.005). Interestingly, in the EGF groups some embryos cleaved faster than without EGF. CONCLUSIONS The effects of EGF are highly dependent on concentration of gonadotrophins present in IVM medium. EGF has a negative effect on oocytes in the presence of low gonadotrophins, but contrastingly partially protects oocytes from the negative effects of high gonadotrophins. We propose that these observed negative effects of EGF may suggest use of an inappropriate dose of growth factor.
Chronic obstructive pulmonary disease (COPD) is multifactorial disease, which is characterized by airflow limitation and can be provoked by genetic factors, including carriage of the PiZ allele of ...the protease inhibitor (Pi) gene, encoding alpha-1 antitrypsin (A1AT). Both homozygous and heterozygous PiZ allele carriers can develop COPD. It was found recently that normal A1AT regulates cytokine levels, including IL-17, which is involved in COPD progression. The aim of this study was to determine whether homozygous or heterozygous PiZ allele carriage leads to elevated level of IL-17 and other proinflammatory cytokines in COPD patients. Materials and Methods. Serum samples and clinical data were obtained from 44 COPD patients, who included 6 PiZZ, 8 PiMZ, and 30 PiMM A1AT phenotype carriers. Serum concentrations of IL-17, IL-6, IL-8, IFN-γ, and TNF-α were measured by the enzyme-linked immunosorbent assay (ELISA). All A1AT phenotypes were verified by narrow pH range isoelectrofocusing with selective A1AT staining. A turbidimetric method was used for quantitative A1AT measurements. Results. COPD patients with both PiZZ and PiMZ phenotypes demonstrated elevated IL-17 and decreased IFN-γ levels in comparison to patients with the PiMM phenotype of A1AT. Thereafter, the ratio IL-17/IFN-γ in PiZZ and PiMZ groups greatly exceeded the values of the PiMM group. Homozygous PiZ allele carriers also had significantly higher levels of IL-6 and lower levels of IL-8, and IL-6 values correlated negatively with A1AT concentrations. Conclusions. The presence of the PiZ allele in both homozygous and heterozygous states is associated with altered serum cytokine levels, including elevated IL-17, IL-17/IFN-γ ratio, and IL-6 (only PiZZ), but lower IFN-γ and IL-8.
The study focuses on the problem of severe tetanus cases, which continue to be reported worldwide. The population of countries with low vaccination rates, including Ukraine, is particularly at risk. ...Despite the increase in the level of DPT vaccination in our country among adults and children, it remains insufficient, which leads to the registration of new cases of tetanus.
The aim of the study was to analyze the main directions in the treatment of tetanus. The authors searched for information in electronic databases such as MEDLINE/Pub Med and Google Scholar for the last 20 years. The search was performed on such terms as tetanus, treatment, benzodiazepines, tetanus immunoglobulin (medical topics). The main components of modern treatment are analyzed in the paper, which include removal of the pathogen by careful surgical treatment of wounds, rational antibiotic therapy. Drugs of choice for the eradication of vegetative forms of the pathogen are benzylpenicillin and metronidazole, but can also be used antibacterial drugs of other groups (macrolides, tetracyclines, cephalosporins). Specific immunotherapy with tetanus immunoglobulin can neutralize the toxin that freely circulates in the blood. Those who do not have vaccination data are also indicated for the introduction of tetanus toxoid. Intrathecal administration of anti-tetanus immunoglobulin remains a debatable issue. In the treatment of convulsions, drugs of the benzodiazepine group are preferred. Infusions of high doses of magnesium, dexmetomedin, baclofen, propofol or their combination can also be used. In the case of severe muscular rigidity, muscle relaxants of nondepolarizing action are used. If the spasms are prolonged, botulinum toxin A may be recommended as an injection into peripheral muscles. B-blockers (esmolol), dexmetomedin, clonidine are used to overcome autonomic dysfunction of the nervous system. Supportive therapy is important. It is aimed at providing the patient with complete nutrition. Adequate ventilation with early tracheostomy helps to avoid complications of prolonged intubation.
The above methods can significantly reduce mortality from tetanus from 100% (in the absence of treatment) to 10%–20%.
Keywords: tetanus, management, benzodiazepines, magnesium sulfate, tetanus immunoglobin, tracheostomy.
Representatives of the new heterocyclic system, pyrimido5’,4’:3,4pyrrolo1,2-
f
phenanthridine, were obtained
via
the intramolecular C–H-arylation of 6-aryl-5-(2-bromophenyl) pyrrolo3,4-
d
...pyrimidine-2,4-diones catalyzed by palladium complexes with
N
-heterocyclic carbene ligands (Pd/NHC).
The article is devoted to research of preconditions and urgency of the reform of decentralization and change of state administrative-territorial system. The main reasons for the necessity of ...realization of the reform in Ukraine are determined: excessive fragmentation of territorial communities, subsidization of local budgets, allocation of a significant part of financial resources to the maintenance of the control apparatus, degradation of rural territories, lack of initiatives from local self-government bodies. The article reflects the results of sociological research on Ukrainians' awareness of the decentralization reform, their expectations and the evaluation of the effectiveness of its implementation on the territory of Ukraine. Іt is determined that among the main problems / obstacles in the implementation of the decentralization reform, the respondents distinguish: financial imbalances; inadequate legal framework; lack of clear division of powers between district administration, district councils and UTC; the need to comply with the declared data; lack of awareness of decentralization; the lack of agreement within communities; political processes. The stages of administrative reform implementation in Ukraine are investigated, in accordance with the legislation of Ukraine. The first stage involves the transfer of the state powers to the lowest administrative-territorial level to the newly created united territorial communities (UTC). The second stage of the reform involves the creation of administrative-territorial units of the regional level that is prefectures (counties or districts), empowered with management authorities. The third stage of the reform involves the formation of a third regional level of the ATU of the country.The advantages, intermediate results of the reform in the context of the problem-oriented orientation are shown; the first consequences of the reform were identified, first of all, in the growth of the financial potential of the territory and the strengthening of the resource base of local budgets. Areas of use of financial resources by the united territorial communities are investigated. The emphasis is on changing the course from the consumer position (the expectation of subsidies from district and provincial budgets), to attempts to create an effective administrative apparatus and to direct resources to the communities development, to carry out analysis of budget spending and to prevent cases of inefficient or irrational use.
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