Standardized molecular typing of Candida albicans strains

Summary. A method is presented for the standardization of Candida albicans DNA fingerprinting, which is based on Southern hybridization of Eco RI-digested chromosomal DNA with the moderately repetitive DNA element CARE-2 and the subsequent rehybridization of the blots with a molecular size marker also included in each DNA sample. This method resulted in extremely precise alignment of all strain-specific CARE-2 hybridization patterns, even when analysed on different gels, and will enhance the accuracy of genetic relationship determinations in epidemiological studies including large numbers of strains.
Zusammenfassung. Zur Standardisierung des DNA-Fingerprinting von Candida albicans wurde eine Methode entwickelt, die auf der Southern Hybridisierung Eco RI-gespaltener chromosomaler DNA mit dem mittelrepetitiven DNA-Element CARE-2 und der darauffolgenden Rehybridisierung der Blots mit einem auch in den Proben enthaltenen molekularen Größenmarker beruht. Dies resultierte in einer äußerst präzisen Größen-bestimmung der hybridisierenden Fragmente, so daß alle stammspezifischen CARE-2-Hybridisierungsmuster exakt verglichen werden konnten, auch wenn die Isolate auf verschiedenen Gelen analysiert wurden. Die Methode erhöht die Genauigkeit der Bestimmung genetischer Verwandtschaftsbeziehungen in epidemiologischen Untersuchungen, in denen eine große Anzahl von Stämmen analysiert wird.     

Investigation of automated perimetry in the evaluation of patients for upper lid blepharoplasty.

We investigated the application of automated perimetry in the evaluation of 17 patients for upper lid blepharoplasty. Visual fields were assessed by a 114 point threshold related screening test of the superior visual field on the Humphrey Allergan Model 640 visual field analyzer. Patients enrolled in the study underwent a complete oculoplastic evaluation prior to and at 4-6 weeks after their procedure. Postoperatively, the visual field as measured by the number of points seen, increased by 26.2% (p < 0.000001). Improvement in visual field results was most dramatic in patients whose margin reflex distance (MRD) was < or = 3.5 mm. This effect was related to excision of redundant eyelid tissue rather than a change in MRD after blepharoplasty. Above this MRD level, blepharoplasty did not significantly improve the patients superior visual field. These results suggest that automated perimetry provides valuable information to document visual field changes for medicolegal and insurance purposes.     

THE PHENOTYPE OF MAST CELL IN PRIMARY ADENOID LIVER TUMOR OF RAT AND ITS RELATION TO TUMOR CELL

Mast cells in adenoid liver tumors of 32 rats induced with nitrosomorpholine were observed ultrastructurally, and among them, some were studied immunocytochemically via immunogold techniques. Data indicating that mast cells which located in tumor tissues presented positive expression of rat mast cell protein (RMCP) Ⅰ, Indicating origination from the mucosa mast cells, while those in the connective tissues around tumors were largely stained negatively with either RMCP Ⅰor RMCP Ⅰ antisera, with the exception of only a few cells showing positive RMCP Ⅰ staining. Ultrastructural observation showed that mast cells in tumon contacted closely with the tumor cells. Membranes of the intracytoplasmic granules in these mast cells were fusing together. The content inside the granules were discharged and spread along the intercellular space between the tumor cells. There was not any lesion observed uitrasructrually in the tumor cells contacting with the mast cells. The significance of mucosa mast cells in adenoi     

Virulence determinants of Escherichia coli O6 extraintestinal isolates analysed by Southern hybridizations and DNA long range mapping techniques.

A total of 16 Escherichia coli O6 strains isolated from cases of extraintestinal infections were analysed for the genetic presence and phenotypic expression of fimbrial adhesins (P, S/FIC, type 1), aerobactin and hemolysin. In addition restriction fragment length polymorphisms (RFLPs) of Xbal-cleaved genomic DNA of seven selected strains, separated by orthogonal field alternation gel electrophoresis (OFAGE) were determined and virulence-associated DNA probes were used for Southern hybridization studies of the Xbal-cleaved genomic DNAs. The virulence characteristics and hybridization patterns obtained differed between the various isolates. In three isolates hemolysin genes and P fimbrial determinants were located on the same Xbal fragments. Furthermore, multiple copies of FIC determinants (foc) could be detected in two strains. Our data show that the new technique of pulse field electrophoresis together with Southern hybridization represents a powerful tool for the genetic analysis of pathogenic bacteria.     

Reactions to disaster at a distance. The first week after the earthquake in Soviet Armenia

Armenian-American adolescents from a Los Angeles school were interviewed during the first week after the massive earthquake in Soviet Armenia in December 1988. The authors describe their observations: All the adolescents strongly identified with the victims and felt that the catastrophe personally affected them; they all rushed into hectic activities to escape intolerable grief reactions; there was strong group pressure to accelerate helping efforts; the catastrophe was experienced as fortifying group cohesion; and there was a distinct preference to preserve and strengthen communal values, even over and above helping the earthquake victims. In particular, the authors observed the phenomenon of "participation envy," a special form of survivor guilt involving envious resentment at being excluded from a unique and rallying communal experience.     

Neuroendocrine cell markers for pancreatic islets and tumors.

The authors review the application of a variety of neuroendocrine cell markers to identify pancreatic islet cells and tumors. In the past, several empiric histochemical techniques had been used to demonstrate neuroendocrine cells, particularly the Grimelius argyrophilic stain. The development of immunohistochemistry made it possible to demonstrate specific cell products such as regulatory peptides, thus allowing the classification of pancreatic neuroendocrine tumors with a view to clinical symptoms. However, it is not always possible to visualize regulatory peptides in these tumors. It is therefore important to use broad-spectrum neuroendocrine cell markers to identify the neuroendocrine nature. These markers are proteins localized in the secretory granules (core- or membrane-related), in the cytosol, or in the cellular membrane. The markers most commonly used in routine histopathology are the secretory granule proteins chromogranin A and synaptophysin and the cytosolic enzyme neuronspecific enolase. Other new markers (e.g., synaptic vesicle protein 2) are of general diagnostic value. Region-specific antibodies to chromogranin A can be valuable in differentiating between benign and malignant neuroendocrine tumors. Some markers may be related to the functioning characteristics of pancreatic neuroendocrine tumors, such as prohormone convertases. In addition, markers giving further complementary information have been identified, such as five somatostatin receptor subtypes, the expression of which varies markedly in pancreatic neuroendocrine tumors. Antibodies against all somatostatin receptor subtypes are now commercially available, and immunohistochemical investigation of its expression should be routinely applied when considering treatment with somatostatin analogs.     

Construction and Characterization of an Isogenic slt-ii Deletion Mutant of Enterohemorrhagic Escherichia coli

Enterohemorrhagic Escherichia coli (EHEC) produces Shiga-like toxins (SLT), potent protein synthesis inhibitors. To further dissect the role of SLT-II in the course of disease, we have constructed E. coli TUV86-2, an isogenic SLT-II-negative mutant of EHEC strain 86-24. The slt-ii gene was inactivated by suicide vector mutagenesis. We also isolated derivatives of strain 86-24 that were cured of the phage carrying the toxin genes.     

Contribution of cloned virulence factors from uropathogenic Escherichia coli strains to nephropathogenicity in an experimental rat pyelonephritis model.

Escherichia coli 536 (O6:K15:H31), which was isolated from a case of urinary tract infection, determines high nephropathogenicity in a rat pyelonephritis system as measured by renal bacterial counts 7 days after infection. The loss of S fimbrial adhesin formation (Sfa-) (mannose-resistant hemagglutination [Mrh-] and fimbria production [Fim-]), serum resistance (Sre-), and hemolysin production (Hly-) in the mutant 536-21 led to a dramatic reduction of bacterial counts from almost 10(5) to only 40 cells per g of kidney. The reintroduction of the cloned S fimbrial adhesin determinant (sfa) increases the virulence of the avirulent mutant strain by a factor of 20; almost the same effect was observed after restoration of serum resistance by integration of an sfa+ recombinant cosmid into the chromosome. Additional reintroduction of the Hly+ phenotype by transformation of two hly determinants increased the virulence of the strains. Hemolysin production determined increased renal elimination of leukocytes and erythrocytes. Thus all three determinants investigated, S fimbriae, serum resistance, and hemolysin, contribute to the multifactorial phenomenon of E. coli nephropathogenicity.