Relationship between Nucleus Swelling and Development Competence of Bovine Cloned Embryos Reconstructed by Enucleated Oocytes with Serum-starved or Serum-fed Fetal Somatic Cells

This study was conducted to examine the occurrence of nuclear remodeling (nucleus swelling) and its effects on the subsequent in vitro development of bovine embryos reconstructed by serum-starved and serum-fed somatic cells. Results from this study demonstrated that all of the reconstructed embryos that received serum-starved and serum-fed somatic cells exhibited condensed-nuclei. More than 90% of the transferred nuclei exhibited nuclear envelope breakdown and premature chromatin condensation which clearly distinct from an intact nucleus. There was no significant difference on the degree of nucleus swelling in SS-NT embryos or SF-NT embryos, indicating that either serum-starved or confluent somatic cell lines could be reprogrammed by the recipient cytoplasm environments in similar pattern. Although the fusion rate was not significantly different among the groups, the proportion of SS-NT embryos which developed to the 2- to 4-cell stage (89.7%) and to the 8- to 16-cell stage (74.7%) was significantly higher than that of SF-NT embryos. Whereas, the proportion of reconstructed embryos that developed to the morula and blastocyst stages were not significantly different among the groups. Results of these studies demonstrate that reconstructed embryos, which received either serum-starved or serum-fed confluent somatic cells, showed similar developmental competence to the blastocyst stage. Keywords: nuclear transplantation technique-somatic cells-nucleus swellin

Study of interleukin-6 in the spread of colorectal cancer: the diagnostic significance of IL-6.

We investigated the diagnostic significance of IL-6 for lymph node metastasis and/or hepatic metastasis from colorectal cancer in 65 patients and evaluated the contributions of 8 factors (IL-6, HGF, IL-1beta, TNF-alpha, TGF-beta1, ELAM-1, ICAM-1, VCAM-1) toward Dukes.s classification of 53 patients. We also examined IL-6 expression in tumor tissue. From the receiver operating characteristic (ROC) curve analysis, an optimal cutoff value of 5.8 pg/ml was determined to classify lymph node and/or hepatic metastasis, and that of 6.3 pg/ml was determined to classify hepatic metastasis. These values indicated sensitivities of 55.0% and 71.4%, and specifi cities of 100% and 88.6%, respectively. IL-6, HGF, and ELAM-1 were very useful for distinguishing among Dukes.s A/B group, C group, and D group. In all cases with high IL-6 values (more than 25.0 pg/ml), immunohistochemical staining was positive for IL-6 in the cytoplasm of cancer cells. IL-6 is strongly suspected to be involved in lymph node and/or hepatic metastasis by promoting it through HGF, and serum IL-6 value (pg/ml) would be useful diagnostically to estimate whether or not there is a high risk of lymph node and/or hepatic metastasis

Baryon superfluidity and neutrino emissivity of neutron stars

For neutron stars with hyperon-mixed core, neutrino emissivity is studied under the equation of state, obtained by introducing repulsive three-body force universal for all baryons so as to assure the maximum mass compatible with the observation. By paying attention to the density-dependence of critical temperatures of baryon superfluids, which reflect the nature of baryon-baryon interaction and control neutron star cooling, we show what neutrino emission processes are efficient in the regions with and without hyperon mixing and remark its implications related to neutron star cooling.Comment: 5pages, 4 figure

Superovulacijski odgovor poludivljeg Dulong goveda (Bos frontalis) i bređost nakon prijenosa njihova embrija u druge vrste.

This study was conducted to examine the superovulatory response in semi-wild Dulong cows (Bos frontalis) and to evaluate whether their embryos can be maintained in pregnancy after interspecies transfer to cross-bred recipients (Bos taurus). Embryos were recovered from five Dulong and nine Red Angus (control) donors after superovulatory treatment. The average numbers of corpora lutea, collected ova and transferable embryos in Dulong donors were similar to those in Red Angus donors. The embryos from Dulong donors were transferred to three cross-bred recipients, two of which became pregnant. However, one embryo was lost between the 40th and 60th day of gestation. The second was lost between the 60th and 90th day of gestation. In Red Angus cows, 44.4% (4/9) of the recipients which received embryos became pregnant, and three delivered calves. We confirmed that Dulong cows respond to superovulatory treatment. Interspecies transfer of Dulong embryos into cross-bred recipients produced pregnancies, but not live offspring.Istraživanje je provedeno da bi se utvrdio superovulacijski odgovor krava poludivljeg Dulong goveda (Bos frontalis) i da bi se procijenilo da li embriji tih krava opstaju tijekom bređosti nakon što su preneseni u primateljice druge vrste (Bos taurus). Embriji su dobiveni nakon superovulacije izazvane kod pet Dulong krava i devet krava pasmine crveni angus (kontrolna skupina). Prosječni broj žutih tijela, jajašaca i prenosivih embrija kod Dulong krava davateljica bio je sličan kao kod davateljica crvene angus pasmine. Embriji Dulong davateljica preneseni su u tri primateljice - križanke od kojih su dvije ostale bređe, no jedan je embrij izgubljen između 40. i 60. dana bređosti. Drugi embrij izgubljen je između 60. i 90. dana bređosti. U slučaju krava pasmine crveni angus, 44,4% (4/9) primateljica ostalo je bređe, od kojih su se otelile njih tri. Može se ustvrditi da Dulong krave reagiraju na postupak superovulacije. Prijenos embrija Dulong krava u primateljice - križanke rezultirao je gravidnošću, ali ne i živooteljenom teladi

DNA methylation dynamics in mouse preimplantation embryos revealed by mass spectrometry

Following fertilization in mammals, paternal genomic 5-methyl-2′-deoxycytidine (5 mC) content is thought to decrease via oxidation to 5-hydroxymethyl-2′-deoxycytidine (5 hmC). This reciprocal model of demethylation and hydroxymethylation is inferred from indirect, non-quantitative methods. We here report direct quantification of genomic 5 mC and 5 hmC in mouse embryos by small scale liquid chromatographic tandem mass spectrometry (SMM). Profiles of absolute 5 mC levels in embryos produced by in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) were almost identical. By 10 h after fertilization, 5 mC levels had declined by ∼40%, consistent with active genomic DNA demethylation. Levels of 5 mC in androgenotes (containing only a paternal genome) and parthenogenotes (containing only a maternal genome) underwent active 5 mC loss in the first 6 h, showing that both parental genomes can undergo demethylation independently. We found no evidence for net loss of 5 mC 10-48 h after fertilization, implying that any passive â€'demethylation' following DNA replication was balanced by active 5 mC maintenance methylation. However, levels of 5 mC declined during development after 48 h, to 1% (measured as a fraction of G-residues) in blastocysts (∼96 h). 5 hmC levels were consistently low (<0. 2% of G-residues) throughout development in normal diploid embryos. This work directly quantifies the dynamics of global genomic DNA modification in mouse preimplantation embryos, suggesting that SMM will be applicable to other biomedical situations with limiting sample sizes

A novel missense PTEN mutation identified in a patient with macrocephaly and developmental delay

Phosphatase and tensin homolog (PTEN) plays an important role in tumor suppression. A germline mutation in the PTEN gene induces not only PTEN hamartoma tumor syndrome, including Cowden syndrome, but also macrocephaly/autism syndrome. Here, we describe a boy with macrocephaly/ autism syndrome harboring a novel missense heterozygous PTEN mutation, c.959T>C (p.Leu320Ser). Interestingly, a previously reported nonsense mutation resulting in p.Leu320X was found in Cowden syndrome patients. Our case may be suggestive of a genotype-phenotype correlation

A case of autism spectrum disorder with cleft lip and palate carrying a mutation in exon 8 of AUTS2

We report a patient with autism and cleft lip and palate carrying a de novo heterozygous AUTS2 mutation, c.1464_1467del ACTC (p.Tyr488*). Although the causal relationship between cleft lip and palate and this mutation is unclear, this case report may expand the clinical phenotype of AUTS2 syndrome

Effects of calmodulin and protein kinase C modulators on transient Ca2+ increase and capacitative Ca2+ entry in human platelets : relevant to pathophysiology of bipolar disorder

Disturbed intracellular calcium (Ca2+) homeostasis has been implicated in bipolar disorder, which mechanisms may be involved in the dysregulation of protein kinase C (PKC) and calmodulin systems. In this study, we investigated a transient intracellular Ca2+ increase induced by thapsigargin, a inhibitor of sarco/endoplasmic reticulum Ca2+-ATPase pump (SERCA), and a capacitative Ca2+ entry followed by addition of extracellular Ca2+, in the presence or absence of PKC/calmodulin modulators in the platelets of healthy subjects in order to elucidate the role of SERCA in Ca2+ homeostasis and to assess how both PKC and calmodulin systems regulate the two Ca2+ responses. Moreover, we also examined the thapsigargin-elicited transient Ca2+ increase and capacitative Ca2+ entry in patients with mood disorders. PKC and calmodulin systems have opposite regulatory effects on the transient Ca2+ increase and capacitative Ca2+ entry in the platelets of normal subjects. The inhibitory effect of PKC activation on capacitative Ca2+ entry is significantly increased and the stimulatory effect of PKC inhibition is significantly decreased in bipolar disorder compared to major depressive disorder and normal controls. These results suggest the possibility that increased PKC activity may activate the inhibitory effect of capacitative Ca2+ entry in bipolar disorder. However, this is a preliminary study using a small sample, thus further studies are needed to examine the PKC and calmodulin modulators on the capacitative Ca2+ entry in a larger sample