Expression of the Ku70 subunit (XRCC6) and protection from low dose ionizing radiation during zebrafish embryogenesis

The Ku70 protein, a product of the XRCC6 gene, is a component of the nonhomologous end-joining (NHEJ) pathway of DNA repair, which protects cells from the effects of radiation-induced DNA damage. Although the spatial expression of Ku70 during vertebrate embryogenesis has not been described, DNA repair proteins are generally considered to be "housekeeping" genes, which are required for radioprotection in all cells. Here, we report the cloning and characterization of the zebrafish Ku70 ortholog. In situ hybridization and RT-PCR analyses demonstrate that Ku70 mRNA is maternally provided and expressed uniformly among embryonic blastomeres. Later during embryogenesis, zygotically transcribed Ku70 mRNA specifically accumulates in neural tissue, including the retina and proliferative regions of the developing brain. In the absence of genotoxic stress, morpholino-mediated knockdown of Ku70 expression does not affect zebrafish embryogenesis. However, exposure of Ku70 morpholino-injected embryos to low doses of ionizing radiation leads to marked cell death throughout the developing brain, spinal cord, and tail. These results suggest that Ku70 protein plays a crucial role in protecting the developing nervous system from radiation-induced DNA damage during embryogenesis.     

The kinase haspin is required for mitotic histone H3 Thr 3 phosphorylation and normal metaphase chromosome alignment

Post-translational modifications of conserved N-terminal tail residues in histones regulate many aspects of chromosome activity. Thr 3 of histone H3 is highly conserved, but the significance of its phosphorylation is unclear, and the identity of the corresponding kinase unknown. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation of H3 Thr 3 in prophase and its dephosphorylation during anaphase. Furthermore we find that haspin, a member of a distinctive group of protein kinases present in diverse eukaryotes, phosphorylates H3 at Thr 3 in vitro. Importantly, depletion of haspin by RNA interference reveals that this kinase is required for H3 Thr 3 phosphorylation in mitotic cells. In addition to its chromosomal association, haspin is found at the centrosomes and spindle during mitosis. Haspin RNA interference causes misalignment of metaphase chromosomes, and overexpression delays progression through early mitosis. This work reveals a new kinase involved in composing the histone code and adds haspin to the select group of kinases that integrate regulation of chromosome and spindle function during mitosis and meiosis.     

Elevated coronary endothelin-1 but not Nitric Oxide in diabetics during CABG

Background. After coronary artery bypass grafting procedures, a higher incidence of morbidity and mortality has been reported in diabetic patients. We tested whether coronary artery bypass grafting in diabetics affects the endothelin-1 and nitric oxide coronary effluent profile during reperfusion.

Methods. Twenty-one consecutive patients (9 with type II diabetes mellitus, 12 non-diabetics) underwent coronary artery bypass grafting by one surgeon. The two groups did not differ in preoperative ejection fraction, Parsonnet score, number of vessels bypassed, or cross-clamp time. Each patient was treated in the same intraoperative manner with single atrial, aortic, and antegrade and retrograde cardioplegia (CPL) cannulas. Cold CPL arrest was by antegrade and retrograde infusion of modified Buckberg CPL solution. Warm CPL solution was infused before reperfusion. Coronary sinus blood samples were obtained for estimation of endothelin-1 and nitrite plus nitrate before CPL arrest and at 1 and 15 minutes after each of 2 reperfusion periods.

Results. In diabetics, endothelin-1 was significantly increased at all reperfusion times as compared with non-diabetics. Nitrite plus nitrate levels were significantly higher in patients with diabetes than in those without, but did not change with time in either of the groups.

Conclusions. Reperfusion after CPL during coronary artery bypass grafting procedure can trigger the release of endothelin-1 in patients with diabetes mellitus. This may favor increased vascular tone or positive inotropic responses after coronary artery bypass grafting and may contribute to significant cardiovascular consequences in diabetic patients.     

Connections of diffuse bipolar cells in primate retina are biased against S-cones

In mammalian retina, each diffuse bipolar type stratifies in a distinct layer of the inner plexiform layer. Thus, different types of bipolar cells provide output to distinct visual pathways. Here, the question of whether diffuse bipolar cell types differ with respect to their contacts with short wavelength-sensitive (S-) cones was investigated in the retinas of a New World monkey, Callithrix jacchus, and an Old World monkey, Macaca fascicularis. Subpopulations of OFF bipolar cells were labeled with antibodies to the glutamate transporter Glt-1 and ON bipolar cells were labeled with antibodies to the alpha subunit of the Go protein (Goalpha). Two types of diffuse ON bipolar cells, DB4 and DB6, were identified with antibodies to protein kinase Calpha and CD15, respectively. Cone pedicles were labeled either with peanut agglutinin coupled to fluorescein or with antibodies to the ribbon protein, C-terminus binding protein 2. We found that immunoreactivity for Glt-1 (OFF bipolar cells) is reduced at S-cones in comparison to medium/long wavelength-sensitive (M/L-) cones. Immunoreactivity for Goalpha (ON bipolar cells) is comparable at all cone types. Nearly all M/L-cone pedicles contact the diffuse ON bipolar types DB4 and DB6, but only between 60% and 75% of the S-cone pedicles make contact. Furthermore, the number of dendritic tips of DB4 and DB6 cells at S-cone pedicles is lower than that at M/L-cone pedicles. These results suggest that there is a bias in the S-cone connectivity of diffuse bipolar cells.     

Homotypic regulation of neuronal morphology and connectivity in the mouse retina

The establishment of neuronal circuitry during development relies upon the action of cell-intrinsic mechanisms that specify neuronal form as well as plastic processes that require the transmission of neural activity between afferents and their targets. Here, we examine the role of interactions between neighboring like-type cells within the mouse retina upon neuronal differentiation and circuit formation. Two different genetically modified mouse models were used to modulate the density of homotypic neighbors, the Type 7 cone bipolar cells, without affecting the density of their afferents, the cone photoreceptors. We demonstrate a corresponding plasticity in dendritic field area when the density of Type 7 cone bipolar cells is elevated or reduced. In accord with this variation in dendritic field area across an invariant population of afferents, individual Type 7 cone bipolar cells are also shown to modulate the number of cone pedicles contacted without varying the number of contacts at each cone pedicle. Analysis of developing Type 7 cone bipolar cells reveals that the dendritic tiling present in maturity is achieved secondarily, after an initial stage of dendritic overlap, when the dendritic terminals are stratified at the level of the cone pedicles but are not localized to them. These results demonstrate a conspicuous developmental plasticity in neural circuit formation independent of neural activity, requiring homotypic interactions between neighboring cells that ultimately regulate connectivity within the retina.     

Relevance of neuropeptide Y for the neuroimmune crosstalk

Both cellular and humoral functions of the immune system are modulated by the sympathetic nervous system (SNS). This interaction is mainly mediated by the release of catecholamines (CA) and their receptor-specific action on immune cells. However, neuropeptide Y (NPY), also present in sympathetic nerve terminals, is released upon SNS-stimulation. NPY modulates potent immunological effects in vitro and in vivo, such as differentiation of T helper cells, monocyte mediator release, NK cell activation, and immune cell redistribution. In addition to this direct action within the neuroimmune crosstalk, NPY is also able to modulate the immunomodulatory effects of other neurotransmitters, thereby acting as a neuroimmune co-transmitter. This review will discuss key findings from recent studies, provide implications for the clinical situation, and integrate the pleiotropic functions of NPY in the context of neuroimmune interactions.     

Correlation of renal histopathology with sonographic findings

BACKGROUND: Judgments about irreversible renal disease are frequently based on the sonographic appearance of the kidneys. However, the sensitivity and specificity of sonography in identifying chronic, irreversible disease have never been determined, and the specific pathologic changes that increase renal cortical echogenicity have not been defined. METHODS: We retrospectively compared sonographic parameters (length, quantitative echogenicity, cortical thickness, and parenchymal thickness) to biopsy findings of glomerular sclerosis, tubular atrophy, interstitial fibrosis, and interstitial inflammation in 207 patients. RESULTS: Echogenicity showed the strongest correlation with all 4 histologic parameters (r= 0.28-0.35). Renal size was significantly correlated with glomerular sclerosis (r=-0.26) and tubular atrophy (r= 0.20). Parenchymal thickness, but not cortical thickness, correlated with tubular atrophy (r=-0.23). By multivariate analysis, tubular atrophy and interstitial inflammation, but not interstitial fibrosis, were significant determinants of cortical echogenicity. Severe chronic disease (>50% sclerosed glomeruli or a score of 3 out of 5 or greater for tubular atrophy or interstitial fibrosis) was present in 69% and 47% of patients with combined renal length <20 cm and >20 cm, respectively (P= <0.05). For cortical echogenicity >1.0 (>liver echogenicity) and 1.0. CONCLUSION: Cortical echogenicity is the sonographic parameter that correlates best with renal histopathology. Although size or echogenicity alone are poor predictors of chronic irreversible disease, the likelihood of treatable disease in small kidneys with increased cortical echogenicity is very low.     

Primary prevention of cirrhosis. Public health strategies that can make a difference

Cirrhosis is associated with high morbidity and mortality and often affects persons during the most productive years of life. In the United States, alcoholic liver disease is the leading contributor to the overall prevalence of cirrhosis, followed by infection with hepatitis B virus (HBV) and hepatitis C virus (HCV). In this article, Drs Karsan, Rojter, and Saab examine lifestyle behaviors that can lead to cirrhosis and enumerate public health strategies aimed at primary prevention.