Ultrastructural Study of Long-Term Measles Infection in Cultures of Hamster Dorsal-Root Ganglion

The morphogenesis of the Edmonston strain of measles is described in cultures of hamster dorsal-root ganglion maintained for as long as 63 days postinoculation. The patterns observed confirmed those previously reported in both neural and non-neural tissue. However, in the present tissue, the development of viral material could be followed chronologically within different cell types such as neurons and Schwann cells. Active replication was visualized up to 63 days postinoculation. The appearance of cytoplasmic nucleocapsid preceded that of intranuclear nucleocapsid, the latter occurring after 14 days. These intranuclear inclusions were formed after the transformation of the nucleoli into bizarre pleomorphic bodies which eventually segregated into clumps of nucleocapsid. These intranuclear inclusions mimic those seen in subacute sclerosing panencephalitis, now known to be etiologically related to a measles-like virus.     

Herpes Simplex Virus-Host Cell Relationships in Organized Cultures of Mammalian Nerve Tissues

Studies on the replication of herpes simplex virus in organized cultures of rat central nervous system (CNS) and peripheral nervous system (PNS) tissue demonstrated synthesis of intra- and extracellular virus, as determined by plaque assay on HEp-2 cells. Newly synthesized intracellular virus appeared 12 to 14 hr after inoculation of CNS, followed 10 hr later by the appearance of extracellular virus. In PNS cultures, where higher inputs of virus were introduced, intracellular virus appeared 6 to 8 hr after inoculation, followed by extracellular virus 12 hr later. Polykaryocyte formation was observed in CNS and PNS tissue involving neuroglial, meningeal, or Schwann cells. Neuron somas did not participate in polykaryocyte formation, but they underwent progressive morphological changes starting with increased cytoplasmic granularity followed by nucleolar distortions and disintegration, margination of nuclear chromatin, and the appearance of intranuclear inclusions. Finally, all recognizable cellular detail was lost. Immune serum globulin failed to inhibit both the progressive nature of the cytopathic effect and the synthesis of intracellular virus. These findings are discussed in relation to other in vitro systems, as well as to disease processes in man and animals.     

Pseudomonas aeruginosa diaminopimelate decarboxylase: evolutionary relationship with other amino acid decarboxylases

The lysA gene encodes meso-diaminopimelate (DAP) decarboxylase (E.C.4.1.1.20), the last enzyme of the lysine biosynthetic pathway in bacteria. We have determined the nucleotide sequence of the lysA gene from Pseudomonas aeruginosa. Comparison of the deduced amino acid sequence of the lysA gene product revealed extensive similarity with the sequences of the functionally equivalent enzymes from Escherichia coli and Corynebacterium glutamicum. Even though both P. aeruginosa and E. coli are Gram-negative bacteria, sequence comparisons indicate a greater similarity between enzymes of P. aeruginosa and the Gram- positive bacterium C. glutamicum than between those of P. aeruginosa and E. coli enzymes. Comparison of DAP decarboxylase with protein sequences present in data bases revealed that bacterial DAP decarboxylases are homologous to mouse (Mus musculus) ornithine decarboxylase (E.C.4.1.1.17), the key enzyme in polyamine biosynthesis in mammals. On the other hand, no similarity was detected between DAP decarboxylases and other bacterial amino acid decarboxylases.      

Type VIII collagen has a restricted distribution in specialized extracellular matrices

A pepsin-resistant triple helical domain (chain 50,000 Mr) of type VIII collagen was isolated from bovine corneal Descemet's membrane and used as an immunogen for the production of mAbs. An antibody was selected for biochemical and tissue immunofluorescence studies which reacted both with Descemet's membrane and with type VIII collagen 50,000-Mr polypeptides by competition ELISA and immunoblotting. This antibody exhibited no crossreactivity with collagen types I-VI by competition ELISA. The mAb specifically precipitated a high molecular mass component of type VIII collagen (EC2, of chain 125,000 Mr) from the culture medium of subconfluent bovine corneal endothelial cells metabolically labeled for 24 h. In contrast, confluent cells in the presence of FCS and isotope for 7 d secreted a collagenous component of chain 60,000 Mr that did not react with the anti-type VIII collagen IgG. Type VIII collagen therefore appears to be synthesized as a discontinuous triple helical molecule with a predominant chain 125,000 Mr by subconfluent, proliferating cells in culture. Immunofluorescence studies with the mAb showed that type VIII collagen was deposited as fibrils in the extracellular matrix of corneal endothelial cells. In the fetal calf, type VIII collagen was absent from basement membranes and was found in a limited number of tissues. In addition to the linear staining pattern observed in the Descemet's membrane, type VIII collagen was found in highly fibrillar arrays in the ocular sclera, in the meninges surrounding brain, spinal cord, and optic nerve, and in periosteum and perichondrium. Fine fibrils were evident in the white matter of spinal cord, whereas a more generalized staining was apparent in the matrices of cartilage and bone. Despite attempts to unmask the epitope, type VIII collagen was not found in aorta, kidney, lung, liver, skin, and ligament. We conclude that this unusual collagen is a component of certain specialized extracellular matrices, several of which are derived from the neural crest.     

A simplified Finnish diabetes risk score to predict type 2 diabetes risk and disease evolution in a German population.

AIMS: The FINDRISC questionnaire is a screening tool to estimate the risks for type 2 diabetes as well as asymptomatic type 2 diabetes. We aimed to evaluate its performance to predict diabetes in a German population and to compare its predictive and detective ability in the same population. METHODS: A total of 552 subjects with increased risk of type 2 diabetes were investigated. All individuals completed the FINDRISC questionnaires and underwent an oral glucose tolerance test (OGTT). All individuals were followed for 3 years and underwent an OGTT again. The performance of the opportunistic screening was assessed with the area under the receiver operating characteristics curve (AUC). An intervention program was carried out for all diabetic and IFG/IGT patients at baseline. RESULTS: For identification, the asymptomatic type 2 DM was named Condition 1; prediction of type 2 DM risk in the follow-up survey as Condition 2; and diabetes risk predicting in a hypothetical case of survey without intervention program as Condition 3. The ROC-AUC in the three condition were AUC (FINDRISC1)=0.745, AUC (FINDRISC2)=0.789, and AUC (FINDRISC3)=0.775, respectively. A significant association between FINDRISC and evolution of disease was found, but the variation of plasma glucose during the three years follow-up was not associated with FINDRISC. People in the intervention group with an improvement of glucose tolerance had a smaller FINDRISC score than persons with an unchanged or progressive condition of disease. CONCLUSION: FINDRISC was validated in our study as a simple tool with high performance to predict diabetes risk and less efficient to identify asymptomatic type 2 diabetes. People with lower FINDRISC score will benefit easier from preventive intervention.