Studies on preimplantation development of buffalo embryos

A total of 71 lactating and nonlactating buffalo-cows of the Murrah breed and F(1)-F(3) crossbreds of Murrah x Bulgarian buffalo were used for a year as donors of embryos after a preliminary treatment for superovulation induction with pregnant mare serum gonadotrophin (PMSG) or follicle stimulating hormone (FSH) in combination with prostaglandin F-2 alpha analog (PGF-2 alpha) according to general application procedures in cows. From 36 to 72 h following prostaglandin injection, the buffalo-cows were checked with the help of a teaser bull for detection of estrus. The animals in estrus were inseminated twice either naturally or artificially with frozen semen. Nonsurgical flushing of the uterine horns was done in 45 of the buffalo-cows between 108 and 162 h after the onset of estrus. After slaughter the uterine horns and oviducts of the other 26 animals were flushed separately between 74 and 108 h after the beginning of estrus. Seven late morulae and eight hatched blastocysts were recovered between 114 and 116 h from the onset of estrus as a result of nonsurgical flushing. All of the 40 embryos recovered after 117 h were in the hatched blastocyst stage. As a result of flushing the oviducts and the uterine horns of slaughtered donors between 74 and 100 h, eggs were obtained only from the oviducts, while flushing conducted between 102 and 108 yielded eggs from both the oviducts and the uterine horns.     

Standardized Aronia melanocarpa extract regulates redox status in patients receiving hemodialysis with anemia

Molecular and Cellular Biochemistry - The aim of our study was to investigate the effects of one-month consumption of polyphenol-rich standardized Aronia melanocarpa extract (SAE) on redox status...     

Generation of Amyloid-β Is Reduced by the Interaction of Calreticulin with Amyloid Precursor Protein,Presenilin and Nicastrin

Dysregulation of the proteolytic processing of amyloid precursor protein by γ-secretase and the ensuing generation of amyloid-β is associated with the pathogenesis of Alzheimer''s disease. Thus, the identification of amyloid precursor protein binding proteins involved in regulating processing of amyloid precursor protein by the γ-secretase complex is essential for understanding the mechanisms underlying the molecular pathology of the disease. We identified calreticulin as novel amyloid precursor protein interaction partner that binds to the γ-secretase cleavage site within amyloid precursor protein and showed that this Ca²⁺- and N-glycan-independent interaction is mediated by amino acids 330–344 in the C-terminal C-domain of calreticulin. Co-immunoprecipitation confirmed that calreticulin is not only associated with amyloid precursor protein but also with the γ-secretase complex members presenilin and nicastrin. Calreticulin was detected at the cell surface by surface biotinylation of cells overexpressing amyloid precursor protein and was co-localized by immunostaining with amyloid precursor protein and presenilin at the cell surface of hippocampal neurons. The P-domain of calreticulin located between the N-terminal N-domain and the C-domain interacts with presenilin, the catalytic subunit of the γ-secretase complex. The P- and C-domains also interact with nicastrin, another functionally important subunit of this complex. Transfection of amyloid precursor protein overexpressing cells with full-length calreticulin leads to a decrease in amyloid-β₄₂ levels in culture supernatants, while transfection with the P-domain increases amyloid-β₄₀ levels. Similarly, application of the recombinant P- or C-domains and of a synthetic calreticulin peptide comprising amino acid 330–344 to amyloid precursor protein overexpressing cells result in elevated amyloid-β₄₀ and amyloid-β₄₂ levels, respectively. These findings indicate that the interaction of calreticulin with amyloid precursor protein and the γ-secretase complex regulates the proteolytic processing of amyloid precursor protein by the γ-secretase complex, pointing to calreticulin as a potential target for therapy in Alzheimer''s disease.     

Genetic diversity and morphological variability in the Balkan endemic Campanula secundiflora s.l. (Campanulaceae)

We reconstructed the relationships among populations of Campanula secundiflora s.l. and closely related and geographically close populations of C. austroadriatica and C. versicolor. Based on analyses of microsatellite DNA data, the investigated populations have high overall genetic diversity and abundant allelic variation over seven investigated loci. Bayesian model‐based clustering identified four clearly differentiated genetic groups of populations. The genetic differentiation was reflected by morphological differentiation, allowing us to propose a new taxonomic treatment of the constituents of C. secundiflora s.l. The populations distributed in south‐western Serbia and north‐eastern Montenegro were included in C. secundiflora. A new species, C ampanula montenegrina sp. nov. , distributed in the continental part of Montenegro and the northern part of Albania is described. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2016, 180 , 64–88.     

The impact of aerobic and anaerobic training regimes on blood pressure in normotensive and hypertensive rats: focus on redox changes

Molecular and Cellular Biochemistry - Melatonin is a crucial neurohormone synthesized in the pineal gland that influences the physiology of animals. The molecular mechanism of norepinephrine...     

Dosimetric accuracy of three dose calculation algorithms for radiation therapy of in situ non-small cell lung carcinoma

BackgroundStudy determines differences in calculated dose distributions for non-small cell lung carcinoma (NSC LC) patients. NSC LC cases were investigated, being the most common lung cancer treated by radiotherapy in our clinical practice.Materials and methodsA retrospective study of 15 NSCLC patient dose distributions originally calculated using standard superposition (SS) and recalculated using collapsed cone (CC ) and Monte Carlo (MC) based algorithm expressed as dose to medium in medium (MCD_m) and dose to water in medium (MCD_w,) was performed so that prescribed dose covers at least 99% of the gross target volume (GTV). Statistical analysis was performed for differences of conformity index (CI), heterogeneity index (HI), gradient index (GI), dose delivered to 2% of the volume (D_2%), mean dose (D_mean) and percentage of volumes covered by prescribed dose (V_70Gy). For organs at risk (OARs), D_mean and percentage of volume receiving 20 Gy and 5Gy (V_20Gy, V_5Gy) were analysed.ResultsStatistically significant difference for GTVs was observed between MCD_w and SS algorithm in mean dose only. For planning target volumes (PTVs), statistically significant differences were observed in prescribed dose coverage for CC, MCD_m and MCD_w. The differences in mean CI value for the CC algorithm and mean HI value for MCD_m and MCD_w were statistically significant. There is a statistically significant difference in the number of MUs for MCD_m and MCD_w compared to SS.ConclusionAll investigated algorithms succeed in managing the restrictive conditions of the clinical goals. This study shows the drawbacks of the CC algorithm compared to other algorithms used.     

Poly(ADP-ribose) polymerase-1: association with nuclear lamins in rodent liver cells

The distribution of poly(ADP-ribose) polymerase-1 (PARP-1) over different nuclear compartments was studied by nuclear fractionation procedures and Western analysis revealing a prominent role of the nuclear matrix. This structure is operationally defined by the solubility properties of the A- and B-type lamins under defined experimental conditions. We consistently observed that most of the nuclear matrix-associated PARP-1 partitioned, in an active form, with the insoluble, lamin-enriched protein fractions that were prepared by a variety of established biochemical procedures. These PARP-1-protein interactions resisted salt extraction, disulfide reduction, RNase and DNase digestion. An inherent ability of PARP-1 to reassemble with the lamins became evident after a cycle of solubilization/dialysis using either urea or Triton X-100 and disulfide reduction, indicating that these interactions were dominated by hydrophobic forces. Together with in vivo crosslinking and co-immunoprecipitation experiments our results show that the lamins are prominent PARP-1-binding partners which could contribute to the functional sequestration of the enzyme on the nuclear matrix.