Gliarin and macrolin, two novel intermediate filament proteins specifically expressed in sets and subsets of glial cells in leech central nervous system.

Using monoclonal antibodies, we have identified two novel intermediate filament (IF) proteins, Gliarin and Macrolin, which are specifically expressed in the central nervous system of an invertebrate. The two proteins both contain the coiled-coil rod domain typical of the superfamily of IF proteins flanked by unique N- and C-terminal domains. Gliarin was found in all glial cells including macro- and microglial cells, whereas Macrolin was expressed in only a single pair of giant connective glial cells. The identification of Macrolin and Gliarin together with the characterization of the strictly neuronal IF protein Filarin in leech central nervous system demonstrate that multiple neuron- and glial-specific IFs are not unique to the vertebrate nervous system but are also found in invertebrates. Interestingly, phylogenetic analysis based on maximum parsimony indicated that the presence of neuron- and glial cell-specific IFs in coelomate protostomes as well as in vertebrates is not of monophyletic origin, but rather represents convergent evolution and appears to have arisen independently.

Amount of calories retained after binge eating and vomiting.

While caloric consumption during binge eating has been measured, it is not known how many of the calories are retained in the gastrointestinal tract after vomiting. In 17 normal weight bulimic patients, there appeared to be a ceiling on the number of calories retained after vomiting. That is, whether or not bulimic patients had small (mean = 1,549 kcal, SD = 505) or large (mean = 3,530 kcal, SD = 438) binges, they retained similar amounts of kilocalories (mean = 1,128, SD = 497, versus mean = 1,209, SD = 574, respectively) after vomiting.

DNA fingerprinting transforms the art of cell authentication.

The increasing diversity of new cell cultures is seriously stretching the capabilities of traditional methods of identification. DNA fingerprinting is set to play an important role in increasing confidence in the authenticity of cultures in research and industry.

DNA fingerprinting--a valuable new technique for the characterisation of cell lines.

DNA fingerprinting is an important new development for the authentication of cell lines. Multilocus methods such as those developed by Alec Jeffreys provide information on a wide range of genetic loci throughout the human genome and thus give a useful genetic "snap-shot" of a cell culture. Our work has shown that Jeffreys multilocus fingerprinting method can be applied to cell lines from a wide range of animals including reptiles, birds, fish and diverse mammals. It can also differentiate very closely related cell lines including those from the same mouse strain. Routine fingerprint analysis has enabled an unprecedented level of confidence in the consistency of cell stocks. Our results demonstrate that this straightforward method represents a powerful and readily interpreted system for cell authentication and exclusion of cross-contamination.

Multilocus DNA fingerprint analysis of cell banks: stability studies and culture identification in human B-lymphoblastoid and mammalian cell lines.

The technique of multilocus DNA fingerprinting has great potential for the authentication of animal cell cultures and in identification of cross-contamination. The Alec Jeffreys probes 33.6 and 33.15 were used as multilocus probes to demonstrate the consistent DNA fingerprint profiles in human peripheral blood and its derivative Epstein-Barr virus (EBV) transformed B-lymphoblastoid cultures maintained by repeated subculture for six months. However, fingerprint analysis of EBV transformed cultures generated from small numbers of cells showed that the majority (seven of eight cultures) had anomalous profiles. Some of these altered profiles shared common features not seen in the peripheral blood pattern. Analysis of seven murine hybridoma clones from a single fusion experiment revealed only two clones which could not be distinguished using probe 33.15. Further studies of master and distribution cell banks for eleven cell lines demonstrated consistent fingerprint profiles in all cases except one (U937). However, this cell line showed only minor differences in the master and distribution bank profiles. These data indicate that, while changes in fingerprint profile may be identified in exceptional instances, the multilocus fingerprinting method using probes 33.6 and 33.15 is a powerful and reliable tool in the quality control of animal cell cultures.

The quality control of cell banks using DNA fingerprinting.

Reproducibility in animal cell culture technology requires careful preparation and characterisation of banks of cell cultures. The two standard techniques used in the quality control of such banks are isoenzyme analysis and cytogenetics which require complex and time-consuming procedures to enable cell line identification. However, DNA fingerprinting is potentially a more powerful method of analysis which can detect mutation and intra-species cross-contamination. At the European Collection of Animal Cell Cultures (ECACC) multilocus fingerprint analysis using probes 33.6 and 33.15 has been assessed in the quality control of cell banks. This method has confirmed consistency between master and working banks, has proven useful over a wide species range and can differentiate closely related cell lines. The key advantage of this method is its ability to detect cross-contamination by cell lines from a wide range of species using a straightforward and economical test. In addition the reproducibility of DNA fingerprints indicates their possible role in cell line authentication procedures which are important for patent and product licence applications.

Carcinosarcoma: a rare tumour of the breast.

This report presents a case of carcinosarcoma, a rare tumour of the breast. The clinical and histological features and management are discussed.

Establishment and characterization of a human CD4 positive cell bank for HIV related studies.

The human CD4 positive cell lines JM, CCRF, CEM, U937, HL60 and THP-1 have been cleared of mycoplasma contamination and defined by DNA fingerprinting and cell surface phenotype marker analysis. These cells have been banked and are now available as a source of standardized cell lines for HIV related research.

Ksp632I, a novel class-IIS restriction endonuclease from Kluyvera sp. strain 632 with the asymmetric hexanucleotide recognition sequence: 5'-CTCTTC(N)1-3' 3'-GAGAAG(N)4-5'.

A new class-IIS restriction endonuclease, Ksp632I, with novel sequence specificity has been discovered in a non-pathogenic species of Kluyvera. The presence of only a single site-specific activity in this Kluyvera sp. strain 632 enables Ksp632I to be isolated in highly purified form free of contaminating nucleases. Ksp632I recognition sites and cleavage positions were deduced using experimental and computer-assisted mapping and sequencing. The cleavage specificity corresponds to the sequence 5'-CTCTTCN decreases NNN-N-3' 3'-GAGAAGN-NNN increases N-5'. The enzyme recognizes an asymmetric hexanucleotide sequence and cleaves in the presence of Mg2+ ions specific phosphodiester bonds in both DNA strands, 1 and 4 nucleotides distal to the recognition sequence. The staggered cuts generate 5'-protruding ends with single-stranded 5'-phosphorylated trinucleotides. Several slow cleavage sites for Ksp632I were observed on lambda cI857Sam7 DNA. Ksp632I may complement other class-IIS enzymes in the universal restriction approach and may serve as a tool for generating defined unidirectional deletions or insertions.

Confirmatory factor analysis of the Interpersonal Support Evaluation List.

Cohen and Hoberman (1983) designed the Interpersonal Support Evaluation List (ISEL) to measure the perceived availability of four relatively independent social support resources and thus to provide tests of stress-buffering hypotheses. The utility of the ISEL for such tests requires evidence that it actually measures distinct functional support dimensions. A confirmatory factor analysis of the ISEL for 133 college students showed that a four-factor model provided a reasonable fit to the data, but the large correlations among the four factors were strongly suggestive of a general, second-order social support factor. However, scoring the ISEL as a unidimensional measure only would result in the loss of unique information contained in the four subscales. Researchers should therefore follow Cohen and Hoberman's procedure of analyzing ISEL subscale scores and the total score.

Role of glutathione in acute N-(3,5-dichlorophenyl) succinimide-induced nephrotoxicity in Sprague-Dawley and Fischer 344 rats.

N-(3,5-Dichlorophenyl)succinimide (NDPS), an experimental agricultural fungicide, has been shown to be a selective nephrotoxin in Sprague-Dawley and Fischer 344 rats. Previous studies have demonstrated that a toxic metabolite contributes to or is responsible for acute NDPS-induced nephrotoxicity. The purpose of this study was to investigate the role of glutathione in NDPS-induced renal effects. In 1 set of experiments, male Sprague-Dawley or Fischer 344 rats received a single intraperitoneal (i.p.) injection of NDPS (0.4 or 1.0 mmol/kg) or sesame oil (2.5 ml/kg). Rats were killed at 1, 3, 6 or 24 h, and reduced (GSH) and oxidized (GSSG) glutathione concentrations determined in liver and renal cortex. In both rat strains NDPS (0.4 or 1.0 mmol/kg) administration produced small decreases in GSH concentrations (1 and 3 h) but moderate increases in GSSG concentrations (1 and 3 h) in liver and kidney. At 24 h both GSH and GSSG concentrations were increased, particularly in kidney. In a second set of experiments, rats were pretreated with the glutathione depletor diethyl maleate (DEM) (0.4 ml/kg, i.p.) 1 h prior to NDPS (0.2, 0.4 or 1.0 mmol/kg, i.p.) or sesame oil (2.5 ml/kg, i.p.) administration, and renal function monitored at 24 and 48 h. DEM pretreatment attenuated the increase in urine volume (24 and 48 h), proteinuria, glucosuria, hematuria and elevated blood urea nitrogen (BUN) concentration produced by NDPS (0.4 or 1.0 mmol/kg) in both Sprague-Dawley and Fischer 344 rats. NDPS-induced increases in kidney weight also were generally prevented by DEM pretreatment. Proximal tubular necrosis produced by NDPS administration was reduced by DEM but not prevented. Pretreatment with the cysteine conjugate beta-lyase inhibitor amino-oxyacetic acid (0.5 mmol/kg, i.p.) 1 h prior to NDPS (0.4 or 1.0 mmol/kg) markedly attenuated all NDPS-induced effects on renal function and morphology. These results suggest that glutathione does not play a protective role against NDPS-induced renal effects and that a glutathione or cysteine conjugate of NDPS might contribute to NDPS-induced nephrotoxicity.

Laser Raman investigation of drug-polymer conjugates: sulfathiazole-povidone coprecipitates.

Laser Raman spectroscopy is used for the investigation of the drug-polymer conjugates, sulfathiazole-povidone. Specifically, Raman spectra, both in the lattice vibration and the intramolecular vibration regions, are used to characterize various polymoprhic forms of sulfathiazole. It is found that sulfathiazole exists in two unsolvated forms, untreated sulfathiazole and another form grown from propanol. The crystals grown from ethanol include varying amounts of ethanol depending on the growth condition. The nature of the povidone-sulfathiazole coprecipitates of various compositions are studied. We find no evidence of any new polymorphic form of sulfathiazole in these coprecipitates. The coprecipitates are found to consist of one of the unsolvated forms of sulfathiazole.

16PF source traits associated with DSM-III symptoms for four diagnostic groups.

Provided evidence of 16PF validity by meta-analysis of source-trait profiles from nine studies for four psychiatric groups (N = 916) (nonparanoid schizophrenics, major depressives, anxiety disorders, alcoholics). All groups had mean sten profiles that deviated markedly from normal personality, and there was considerable convergence between salient traits and diagnostic criteria except for alcoholics.