The adipokines progranulin and omentin can directly regulate feline ovarian granulosa cell functions.

The aim of the present study was to determine the effects of the adipokines progranulin and omentin on the basic functions of feline ovarian cells. For this purpose, we investigated the effects of the addition of progranulin and omentin (0, 0.1, 1, or 10 ng/ml) on the proliferation (accumulation of PCNA and cyclin B1), apoptosis (accumulation of Bax and caspase 3) and progesterone release of cultured feline ovarian granulosa cells by quantitative immunocytochemistry and enzyme-linked immunosorbent assays (ELISAs). Both progranulin and omentin increased cell proliferation and decreased apoptosis. Both progranulin and omentin promoted progesterone release. The present findings demonstrate that the adipokines progranulin and omentin can directly regulate basic feline ovarian cell functions.

The adipokines progranulin and omentin - novel regulators of basic ovarian cell functions.

The present study aimed to examine the effects of progranulin and omentin on basic ovarian cell functions. For this purpose, we investigated the effects of the addition of progranulin and omentin (0, 0.1, 1, or 10 ng/ml) on the viability, proliferation, apoptosis and steroidogenesis of cultured rabbit ovarian granulosa cells. To determine the importance of the interrelationships between granulosa cells and theca cells, we compared the influence of progranulin and omentin on progesterone and estradiol release in cultured granulosa cells and ovarian fragments containing both granulosa cells and theca cells. Cell viability, proliferation, cytoplasmic apoptosis and release of progesterone and estradiol were measured by Cell Counting Kit-8 (CCK-8), BrdU incorporation, cell death detection, and ELISA. Both progranulin and omentin increased granulosa cell viability and proliferation and decreased apoptosis. Progranulin increased progesterone release by granulosa cells but reduced progesterone output by ovarian fragments. Progranulin decreased estradiol release by granulosa cells but increased it in ovarian fragments. Omentin reduced progesterone release in both models. Omentin reduced estradiol release by granulosa cells but promoted this release in ovarian fragments. The present observations are the first to demonstrate that progranulin and omentin can be direct regulators of basic ovarian cell functions. Furthermore, the differences in the effects of these adipokines on steroidogenesis via granulosa and ovarian fragments indicate that these peptides could target both granulosa and theca cells.

Dose-Dependent Impact of Bee Pollen Supplementation on Macroscopic and Microscopic Structure of Femoral Bone in Rats.

Bee pollen has been successfully used as a feed additive with beneficial impacts on productive, reproductive, and immune conditions of animals. However, its effect on bone structure and bone health remains controversial. Therefore, the purpose of our study was to examine the impact of bee pollen supplementation on macroscopic and microscopic structure of a femoral bone using rats as suitable animal models. Male rats (1 month-old) were assigned into three groups: control (C group) that was fed a standard diet without bee pollen and two bee pollen supplemented groups (P1 and P2 groups) that received an experimental diet including 0.5% and 0.75% of bee pollen, respectively, for 3 months. A number of unfavorable effects of 0.75% bee pollen administration on bone weight, cortical bone thickness, calcium content, alkaline phosphatase activity, sizes of primary osteons' vascular canals, Haversian canals and secondary osteons in the cortical bone have been recorded, whereas these bone parameters were significantly decreased in the P2 group versus the C group. On the contrary, the concentration of 0.5% did not affect any of bone features mentioned above. In conclusion, the impact of bee pollen supplementation on femoral bone structure of rats depends on the dose used.

Maternal Genetic Composition of a Medieval Population from a Hungarian-Slavic Contact Zone in Central Europe.

The genetic composition of the medieval populations of Central Europe has been poorly investigated to date. In particular, the region of modern-day Slovakia is a blank spot in archaeogenetic research. This paper reports the study of mitochondrial DNA (mtDNA) in ancient samples from the 9th-12th centuries originating from the cemeteries discovered in Nitra-Sindolka and Cakajovce, located in western Slovakia (Central Europe). This geographical region is interesting to study because its medieval multi-ethnic population lived in the so-called contact zone of the territory of the Great Moravian and later Hungarian state formations. We described 16 different mtDNA haplotypes in 19 individuals, which belong to the most widespread European mtDNA haplogroups: H, J, T, U and R0. Using comparative statistical and population genetic analyses, we showed the differentiation of the European gene pool in the medieval period. We also demonstrated the heterogeneous genetic characteristics of the investigated population and its affinity to the populations of modern Europe.

The effect of A163G polymorphism in the osteoprotegerin gene on osteoporosis related traits in Slovak postmenopausal women.

Osteoprotegerin (OPG) plays an important role in the osteoclast differentiation as an effective inhibitor of osteoclast maturation and activation. We examined a potential effect of A163G single nucleotide polymorphism in the promoter region of the OPG gene on femoral neck (FN-BMD) and lumbar spine BMD (LS-BMD), as well as circulating alkaline phosphatase, osteocalcin (ALP, OC; formation markers), beta-CrossLaps (CTx; resorption marker) in Slovak postmenopausal women. In addition, fractures of spine, radius and femur were examined.Altogether 284 women (62.28 ± 8.40 years) were selected according to strict inclusion criteria. The polymorphism was detected by PCR-RFLP method. Genotype frequencies were tested using the chi-square test. The differences of quantitative variables between the genotypes were analyzed by covariance analysis (GLM procedure) after correction of the measurements for age and BMI. Fracture incidence in association with OPG genotype was evaluated by Binary Logistic Regression with the genotype, age, and BMI as covariates. The frequencies of genotypes were 76.8 %, 21.1 %, and 2.1 % for AA, AG, and GG, respectively. Statistically significant associations of OPG genotypes with FN-BMD (p < 0.01) and LS-BMD (p < 0.05) were observed. The GG genotype was associated with higher BMD values likewise decreased CTx concentration (p < 0.05) in compared with the other genotypes, which indicates that the allele G has a protective effect on bone. These associations were not followed by the effect of OPG on fracture incidence. Our results suggest that OPG/A163G polymorphism could contribute to the genetic regulation of BMD or bone turnover markers in Slovak population and thus could increase or decreased osteoporosis risk.

Accumulation of risk elements in kidney, liver, testis, uterus and bone of free-living wild rodents from a polluted area in Slovakia.

Free-living wild rodents are usually used as indicators of pollution, with elements being determined in either whole body or in specific organs. In the present study, the accumulation of cadmium (Cd), copper (Cu), iron (Fe), and zinc (Zn) in kidney, liver, testis, uterus and bone of yellow-necked mice (Apodemus flavicollis) and bank voles (Myodes glareolus) trapped in a polluted area of Nováky, Slovakia was investigated. Yellow-necked mice and bank voles were collected using standard theriological methods for wood ecosystems. All animals were adults in good physical condition. Concentrations of Cd, Cu, Fe, and Zn in all analyzed organs were determined by atomic absorption spectrophotometry. The highest concentrations of Cd and Zn were found in the bone of both species while Cu and Fe accumulated most in the uterus. Significantly higher concentrations of Cd and Cu were detected in the liver of the bank vole in comparison with the yellow-necked mouse (P<0.05). Similar significantly higher levels of Cd and Zn were found in the bone of the bank vole (P<0.05) than in the yellow-necked mouse, while these rodents had significantly higher Cu and Fe concentrations (P<0.05) in the kidney. Significantly higher levels of Fe and Zn were detected in the testis and uterus of bank voles, respectively. On the other hand, significantly higher concentration of Cu was found in the testis of yellow-necked mice. Results of this study suggest that bank voles are more sensitive heavy metal loaded bioindicators than yellow-necked mice.

Sex determination of early medieval individuals through nested PCR using a new primer set in the SRY gene.

One of the first questions asked about excavated human skeletal remains is the sex. As the morphological sex determination is complicated in cases involving fragmentary bones and in skeletons from infants and children, the development of DNA-based techniques has led to improvements in sex determination. This study is focused on sex determination from ancient DNA obtained from 25 skeletons found in Middle Aged burials in western Slovakia. We performed separate amplifications of DXZ4 repetitive satellite sequences on the X chromosome, and SRY gene - testis determined factor on the Y chromosome, using nested PCR. Our results showed that DXZ4 was amplified in the case of 23 individuals. With newly designed internal and external primer sets for SRY detection with internal PCR products in lengths of 102 bp and 85 bp we succeeded in detecting the SRY locus in 9 samples. Finally, the gender was determined in 23 individuals (14 females and 9 males). In 20 samples, the gender was determined by morphological and molecular methods. Sex determination of 17 samples using nested PCR matched the morphological one, providing evidence of the authenticity and ancient origin of the PCR amplifications. The DXZ4/SRY nested PCR method represents a useful technique in sex determination of medieval human remains and it is a critical addition to anthropological studies.

Histological analysis of ovine compact bone tissue.

Histological sections of femoral diaphysis from adult sheep were investigated in this study. The qualitative characteristics in microstructure of the compact bone were examined in anterior, posterior, medial and lateral views; the quantitative ones were assessed using the software Scion Image. Areas, perimeters, minimum and maximum diameters of the Haversian canals, the Haversian systems and the primary osteons' vascular canals were measured. Our results indicate that ovine diaphysis of the femur is occupied mainly with primary vascular plexiform (laminar, fibrolamellar) tissue. In addition, irregular and dense Haversian bone tissues were observed. All measured variables disposed lower values in comparison with other artiodactyls (cattle and pig).

Differences among species in compact bone tissue microstructure of mammalian skeleton: use of a discriminant function analysis for species identification.

In order to develop an identification key for distinguishing between several mammalian species, bone structure of their compact bone tissue was analyzed using qualitative and quantitative characteristics. Ninety femora of adult male humans, pigs, cows, sheep, rabbits, and rats were studied. The average area, perimeter, minimum, and maximum diameter of 1863 Haversian canals and 1863 secondary osteons were measured using a digital image device. The observed data were first used to evaluate inter- and intraspecies diversity. After that, we applied a discriminant function analysis for differentiation of the species by these variables. Classification functions for investigated species give cross-validated correct classification rates for 76.17% of cases. This percentage value can be increased by integrating conclusions from the qualitative analysis.

Simultaneous detection of malignant hyperthermia and genetic predisposition for improved litter size in pigs by multiplex PCR-RFLP.

The ryanodine receptor gene (RYR1) and the estrogen receptor gene (ESR) are the best commercially used markers for predisposition of stress susceptibility (malignant hyperthermia--MH) and increased litter size, respectively. A simplified method of simultaneous detection of MH and ESR genotypes has been developed. The method is based on simultaneous amplification of fragments of two genes by multiplex PCR and subsequent digestion of the products with two restriction enzymes. The PCR and the digestion could be performed in a single tube and all genotypes could be detected by electrophoretic separation on the same agarose gel. Thus, the development of the method can decrease the cost of the sample analysis and increase the speed and efficiency of the analysis. In our study, frequencies of mutated T allele of the RYR1 gene in Large White (LW), White Meaty (WM) and Landrace (L) were 0.11, 0.13, and 0.15, respectively. Frequencies of the preferred B allele of the ESR gene in the same breeds were 0.35, 0.26, and 0.06, respectively.