THE DIFFICULT PROXIMAL CORONARY ANASTOMOSIS

Two patients are presented in whom severe calcific aortitis made performance of satisfactory proximal coronary anastomosis impossible. Endarterectomy of the ascending aorta after aortic valve replacement, combined with aortocoronary bypass prior to performance of the proximal anastomoses, was successfully employed in one patient. The second patient underwent resection of an anterior segment of aorta, which was replaced by a woven Dacron patch into which the vein grafts were sewn.     

Maximum likelihood estimation by counting methods under polygenic and mixed models in human pedigrees.

For pedigree data, the maximum likelihood estimates of the parameters in polygenic and mixed models are derived analytically although not in closed form but in terms of "counting equations" allowing an iterative solution. Likelihood computations, tests of significance, and tests of goodness of fit are presented. Accelerating the (linear) rate of convergence by a very simple method is demonstrated.     

Cluster-analytic definition of species ecological groups for a submerged barrier reef in Barbados,West Indies

Cluster analytic techniques were employed to define significant ecological groups of benthic species sampled in 2237 0.5 m × 1 m quadrats on a submerged barrier reef located off the Barbados west coast. For measures of species cover, number of colonies, presence-absence, and cover of dead skeletal material, Lance and Williams' flexible sorting procedure defined eight species groups. Only four of these groups could be interpreted with confidence. Resolution of species groups increased with increasing environmental severity and decreased with decreasing species abundance. Highly resolved groups under strong environmental limitation included an Agaricia lamarcki-sponge-antipatharian association on deep outer slopes and a distinctive Acropora cervicornis-Madracis mirabilis-rubble association of ridge-top positions. The next two groups, in order of resolution, differentiated under strong competitive interaction. Associations dominated by Siderastrea siderea-Montastrea cavernosa and Montastrea annularis-Porites astreoides consisted of large colony-forming species as part of a larger complex “mixed” coral association. The other four groups consisted of rare but widely-occurring species. Probable reasons for the low resolution of these groups including severe disturbance, random colonization, niche specialization, and methodological idiosyncracies, were discussed. The efficacy of clustering procedures was discussed in relation to objectivity and efficiency of field recognition.     

A Splicing Mutation in the Novel Mitochondrial Protein DNAJC11 Causes Motor Neuron Pathology Associated with Cristae Disorganization,and Lymphoid Abnormalities in Mice

Mitochondrial structure and function is emerging as a major contributor to neuromuscular disease, highlighting the need for the complete elucidation of the underlying molecular and pathophysiological mechanisms. Following a forward genetics approach with N-ethyl-N-nitrosourea (ENU)-mediated random mutagenesis, we identified a novel mouse model of autosomal recessive neuromuscular disease caused by a splice-site hypomorphic mutation in a novel gene of unknown function, DnaJC11. Recent findings have demonstrated that DNAJC11 protein co-immunoprecipitates with proteins of the mitochondrial contact site (MICOS) complex involved in the formation of mitochondrial cristae and cristae junctions. Homozygous mutant mice developed locomotion defects, muscle weakness, spasticity, limb tremor, leucopenia, thymic and splenic hypoplasia, general wasting and early lethality. Neuropathological analysis showed severe vacuolation of the motor neurons in the spinal cord, originating from dilatations of the endoplasmic reticulum and notably from mitochondria that had lost their proper inner membrane organization. The causal role of the identified mutation in DnaJC11 was verified in rescue experiments by overexpressing the human ortholog. The full length 63 kDa isoform of human DNAJC11 was shown to localize in the periphery of the mitochondrial outer membrane whereas putative additional isoforms displayed differential submitochondrial localization. Moreover, we showed that DNAJC11 is assembled in a high molecular weight complex, similarly to mitofilin and that downregulation of mitofilin or SAM50 affected the levels of DNAJC11 in HeLa cells. Our findings provide the first mouse mutant for a putative MICOS protein and establish a link between DNAJC11 and neuromuscular diseases.     

Basis for receptor specificity of nonecotropic murine leukemia virus surface glycoprotein gp70SU.

Murine leukemia viruses (MuLVs) initiate infection of NIH 3T3 cells by binding of the viral envelope (Env) protein to a cell surface receptor. Interference assays have shown that MuLVs can be divided into four groups, each using a distinct receptor: ecotropic, polytropic, amphotropic, and 10A1. In this study, we have attempted to map the determinants within viral Env proteins by constructing chimeric env genes. Chimeras were made in all six pairwise combinations between Moloney MCF (a polytropic MuLV), amphotropic MuLV, and 10A1, using a conserved EcoRI site in the middle of the Env coding region. The receptor specificity of each chimera was determined by using an interference assay. We found that amphotropic receptor specificity of each chimera was determined by using an interference assay. We found that amphotropic receptor specificity seems to map to the N-terminal portion of surface glycoprotein gp70SU. The difference between amphotropic and 10A1 receptor specificity can be attributed to one or more of only six amino acid differences in this region. Nearly all other cases showed evidence of interaction between Env domains in the generation of receptor specificity. Thus, a chimera composed exclusively of MCF and amphotropic sequences was found to exhibit 10A1 receptor specificity. None of the chimeras were able to infect cells by using the MCF receptor; however, two chimeras containing the C-terminal portion of MCF gp70SU could bind to this receptor, while they were able to infect cells via the amphotropic receptor. This result raises the possibility that receptor binding maps to the C-terminal portion of MCF gp70SU but requires MCF N-terminal sequences for a functional interaction with the MCF receptor.     

Incidence and treatments of postpartum reproductive problems in a dairy herd

Incidence of assisted births, retained fetal membranes (RFM), and metritis were recorded in one hundred dairy cows from parturition through 14 days post-calving. Manual removal of RFM was not attempted. All RFM were excised inside the vulva and observations of natural RFM expulsion were recorded. Fifteen of 100 cows (15%) had assisted births, 27 (27%) had RFM, 8 (8%) had primary metritis not associated with other postpartum reproductive problems, and 26 (26%) had secondary metritis. Uterine swabs for culture were collected during the study from cows with postpartum reproductive problems. E . coli was the most common organism isolated (69.4%). Sensitivities of all isolates to penicillin, tetracycline, and triple sulfa were 44.0%, 59.5%, and 36.9% respectively. One of two antibiotic treatments were administered to cows with these postpartum reproductive problems. Treated animals received either 5 g. tetracycline powder IU on day 1 of treatment plus 10.5 million units procaine penicillin G IM on days 1, 2, and 3; or 4 Sulfaurea boluses IU on day 1. Despite the antibiotic treatments, 26 of 34 cows having either assisted birth and/or RFM developed metritis (76%). Neither treatment regimen was superior to the other. The poor results of antimicrobial therapy suggested the futility of routine administration of therapeutic agents for postpartum reproductive problems. Treatment failure was attributed to ineffective drugs or inadequate dosage regimens.     

Severe hepatocellular disease in mice lacking one or both CaaX prenyltransferases

Protein farnesyltransferase (FTase) and protein geranylgeranyltransferase-I (GGTase-I) add 15- or 20-carbon lipids, respectively, to proteins that terminate with a CaaX motif. These posttranslational modifications of proteins with lipids promote protein interactions with membrane surfaces in cells, but the in vivo importance of the CaaX prenyltransferases and the protein lipidation reactions they catalyze remain incompletely defined. One study concluded that a deficiency of FTase was inconsequential in adult mice and led to little or no tissue pathology. To assess the physiologic importance of the CaaX prenyltransferases, we used conditional knockout alleles and an albumin-Cre transgene to produce mice lacking FTase, GGTase-I, or both enzymes in hepatocytes. The hepatocyte-specific FTase knockout mice survived but exhibited hepatocellular disease and elevated transaminases. Mice lacking GGTase-I not only had elevated transaminases but also had dilated bile cannaliculi, hyperbilirubinemia, hepatosplenomegaly, and reduced survival. Of note, GGTase-I-deficient hepatocytes had a rounded shape and markedly reduced numbers of actin stress fibers. Hepatocyte-specific FTase/GGTase-I double-knockout mice closely resembled mice lacking GGTase-I alone, but the disease was slightly more severe. Our studies refute the notion that FTase is dispensable and demonstrate that GGTase-I is crucial for the vitality of hepatocytes.     

Aspects of cAMP Signaling in Epileptogenesis and Seizures and Its Potential as Drug Target

Neurochemical Research - Epilepsy is one of the most common chronic neurological conditions. Today, close to 30 different medications to prevent epileptic seizures are in use; yet, far from all...     

Prevalence of Alzheimer's disease and vascular dementia: association with education. The Rotterdam study.

OBJECTIVE--To estimate the prevalence of dementia and its subtypes in the general population and examine the relation of the disease to education. DESIGN--Population based cross sectional study. SETTING--Ommoord, a suburb of Rotterdam. SUBJECTS--7528 participants of the Rotterdam study aged 55-106 years. RESULTS--474 cases of dementia were detected, giving an overall prevalence of 6.3%. Prevalence ranged from 0.4% (5/1181 subjects) at age 55-59 years to 43.2% (19/44) at 95 years and over. Alzheimer''s disease was the main subdiagnosis (339 cases; 72%); it was also the main cause of the pronounced increase in dementia with age. The relative proportion of vascular dementia (76 cases; 16%), Parkinson''s disease dementia (30; 6%), and other dementias (24; 5%) decreased with age. A substantially higher prevalence of dementia was found in subjects with a low level of education. The association with education was not due to confounding by cardiovascular disease. CONCLUSIONS--The prevalence of dementia increases exponentially with age. About one third of the population aged 85 and over has dementia. Three quarters of all dementia is due to Alzheimer''s disease. In this study an inverse dose-response relation was found between education and dementia--in particular, Alzheimer''s disease.     

Optical study of DNA surface hybridization reveals DNA surface density as a key parameter for microarray hybridization kinetics

We investigate the kinetics of DNA hybridization reactions on glass substrates, where one 22 mer strand (bound-DNA) is immobilized via phenylene-diisothiocyanate linker molecule on the substrate, the dye-labeled (Cy3) complementary strand (free-DNA) is in solution in a reaction chamber. We use total internal reflection fluorescence for surface detection of hybridization. As a new feature we perform a simultaneous real-time measurement of the change of free-DNA concentration in bulk parallel to the total internal reflection fluorescence measurement. We observe that the free-DNA concentration decreases considerably during hybridization. We show how the standard Langmuir kinetics needs to be extended to take into account the change in bulk concentration and explain our experimental results. Connecting both measurements we can estimate the surface density of accessible, immobilized bound-DNA. We discuss the implications with respect to DNA microarray detection.