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[Thermal resistance of the spores of a Bac. stearothermophilus culture used for the preparation of bioindicators].

Thermostability of the spores of Bac. stearothermophilus in ampoules and capillaries in concentrations of 10(9), 10(8) and 10(6) cells per 1 ml of sodium chloride isotonic solution was determined at 119 to 124 degrees C with an interval of 1 degree C and an exposure time of 5, 10, 15, 20, 25, 30, 35 and 40 minutes. The results were used for plotting the survival curves. The time of the microbial death in the ampoules and capillaries at all the temperatures was the same and the ampoules were chosen as the bioindicator vehicle because of their availability and convenience in exploitation. The survival curves may be used for determination of the optimal sterilization conditions. The spore concentration of the thermostable culture in the bioindicator should be equal or exceed the level of the object microbial contamination. In the present study the concentration of the test microbe spores in the bioindicator was 10(6)--10(8) cells/ml.

Culture Media↗

Acute and 8-hour effects of alcohol (0.08% BAC) on younger and older pilots' simulator performance.

This preliminary study examined the acute and 8-hour effects of alcohol at a target peak BAC of 0.08% on pilot performance. Fourteen younger (mean age 27.6 years) and fourteen older (mean age 60.3 years) pilots flew a Frasca 141 simulator in a scenario that included ATC communications and emergencies. Plots were tested during an alcohol and placebo condition at three timepoints: predrink, acute intoxication, and 8 h postdrink. Of eight performance measures, two showed statistically significant effects related to alcohol. First, cockpit monitoring was poorer when pilots were intoxicated, with recovery at 8 h. Second, younger pilots made more communication errors under the influence and there was no significant recovery at 8 h. Older pilots made more communication errors than younger pilots, but possibly because of methodological problems, older pilots' communication performance was not significantly impaired by alcohol. These results provide direction for future research.

Adult↗

[Normal status of Bac. cereus and Cl. perfringens in the body of healthy slaughter animals].

Studied were cattle, pigs, sheep, and young calves intended for slaughter. The experiments were carried out under productional conditions, strictly observing the routinely adopted practice of preslaughter handling. The blood of the animals was sampled prior to slaughter. Samples from the meat (musculature), spleen, kidneys, liver, mesenterial and body lymph nodes as well as feces were taken immediately after slaughter. It was established that Bacillus cereus and Clostridium perfringens were almost lacking in the musculature, mesenterial and body lymph nodes and in some of the parenchymal organs. These organisms were chiefly found in the intestinal tract. Eighty-three up to 100 per cent of the isolated cultures of Cl. perfringens and from 62.5 up to 100 per cent of the isolated cultures of Bac. cereus originated from the feces.

Animals↗

[Viability of a Bac. licheniformis 749/c culture and its formation of penicillinase during storage in the lyophilized state].

The amount of the enzyme produced and the colony morphology of Bac. licheniformis 749/C, the penicillinase-producing organism were studied after storage for 3 years in ampoules at a temperature of 4--10 degrees C, in lyophilized form in sodium glutamate, polyvinylpirrolidone, their mixture and horse serum. The highest rate of the culture growth was observed after storage in lyophlized form in sodium glutamate, though the culture was vialable in all other protective media. Two culture types approximately in an equal ratio were observed in the population platings of the culture lyophilized in various protective media.

Bacillus↗

Some physico-chemical properties micelle--benzalkonium chloride (BAC) adduct on interface in selected eye drops.

In selected eye drops the value of surface tension coefficient delta 25 and critical micelle concentration (cmc) was determined. Calculation of the hydrophilic-lipophilic balance level (HLBcmc) and the solution of the surface state f.Am = K.T form of Gibbs equation enables the estimation of the type of action of micelle with lipid corneal epithelium. Particular attention has been paid to the forms of eye drops which contain benzalkonium chloride (BAC).

Animals↗

Childhood-onset schizophrenia/autistic disorder and t(1;7) reciprocal translocation: identification of a BAC contig spanning the translocation breakpoint at 7q21.

Childhood-onset schizophrenia (COS) is defined by the development of first psychotic symptoms by age 12. While recruiting patients with COS refractory to conventional treatments for a trial of atypical antipsychotic drugs, we discovered a unique case who has a familial t(1;7)(p22;q21) reciprocal translocation and onset of psychosis at age 9. The patient also has symptoms of autistic disorder, which are usually transient before the first psychotic episode among 40-50% of the childhood schizophrenics but has persisted in him even after the remission of psychosis. Cosegregating with the translocation, among the carriers in the family available for the study, are other significant psychopathologies, including alcohol/drug abuse, severe impulsivity, and paranoid personality and language delay. This case may provide a model for understanding the genetic basis of schizophrenia or autism. Here we report the progress toward characterization of genomic organization across the translocation breakpoint at 7q21. The polymorphic markers, D7S630/D7S492 and D7S2410/D7S646, immediately flanking the breakpoint, may be useful for further confirming the genetic linkage for schizophrenia or autism in this region. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:749-753, 2000. Published 2000 Wiley-Liss, Inc.

Autistic Disorder↗

DNA microarrays for comparative genomic hybridization based on DOP-PCR amplification of BAC and PAC clones.

We have designed DOP-PCR primers specifically for the amplification of large insert clones for use in the construction of DNA microarrays. A bioinformatic approach was used to construct primers that were efficient in the general amplification of human DNA but were poor at amplifying E. coli DNA, a common contaminant of DNA preparations from large insert clones. We chose the three most selective primers for use in printing DNA microarrays. DNA combined from the amplification of large insert clones by use of these three primers and spotted onto glass slides showed more than a sixfold increase in the human to E. coli hybridization ratio when compared to the standard DOP-PCR primer, 6MW. The microarrays reproducibly delineated previously characterized gains and deletions in a cancer cell line and identified a small gain not detected by use of conventional CGH. We also describe a method for the bulk testing of the hybridization characteristics of chromosome-specific clones spotted on microarrays by use of DNA amplified from flow-sorted chromosomes. Finally, we describe a set of clones selected from the publicly available Golden Path of the human genome at 1-Mb intervals and a view in the Ensembl genome browser from which data required for the use of these clones in array CGH and other experiments can be downloaded across the Internet.

Animals↗

High resolution tiling-path BAC array deletion mapping suggests commonly involved 3p21-p22 tumor suppressor genes in neuroblastoma and more frequent tumors.

The recurrent loss of 3p segments in neuroblastoma suggests the implication of 1 or more tumor suppressor genes but thus far few efforts have been made to pinpoint their detailed chromosomal position. To achieve this goal, array-based comparative genomic hybridization was performed on a panel of 23 neuroblastoma cell lines and 75 primary tumors using a tiling-path bacterial artificial chromosome array for chromosome 3p. A total of 45 chromosome 3 losses were detected, including whole chromosome losses, large terminal deletions and interstitial deletions. The latter, observed in cell lines as well as a number of distal deletions detected in primary tumors, allowed us to demarcate 3 minimal regions of loss of 3.6 Mb [3p21.31-p21.2, shortest regions of overlap (SRO)1], 1.4 Mb (3p22.3-3p22.2, SRO2) and 3.8 Mb (3p25.3-p25.1, SRO3) in size. The present data significantly extend previous findings and now firmly establish critical regions on 3p implicated in neuroblastoma. Interestingly, the 2 proximal regions coincide with previously defined SROs on 3p21.3 in more frequent tumors including lung and breast cancer. As such, similar tumor suppressor genes may play a critical role in development or progression of a variety of neoplasms, including neuroblastoma.

Cell Line, Tumor↗

Construction of a 780-kb PAC, BAC, and cosmid contig encompassing the minimal critical deletion involved in B cell chronic lymphocytic leukemia at 13q14.3.

A putative tumor suppressor gene involved in B cell chronic lymphocytic leukemia (B-CLL) was mapped to human chromosome 13q14.3 close to the genetic markers D13S25 and D13S319. We constructed a 780-kb-long contig composed of cosmids, bacterial artificial chromosomes, and bacteriophage P1-derived artificial chromosomes that provides essential information and tools for the positional cloning of this gene. The conting contains both flanking markers as well as several additional genetic markers, three ESTs, and one potential CpG island. In addition, using one B-CLL patient, we characterized a small internal deleted region of 550 kb. Comparing this deletion with other recently published deletions narrows the minimally deleted area to less than 100 kb in our physical map. This deletion core region should contain all or part of the disrupted in B cell malignancies tumor suppressor gene.

Bacteriophages↗

Mutant rescue by BAC clone injection in zebrafish.

Genes essential for vertebrate body plan specification, organ development, and organ function are likely to be shared between mammals and zebrafish, but only in zebrafish have large-scale, genome-wide mutagenesis screens been conducted to isolate embryonic lethal mutations. Discovering the roles played by these disrupted genes requires their molecular characterization, which would be facilitated by assaying large cloned genomic DNAs for their potential to rescue mutant phenotypes. Here we demonstrate that bacterial artificial chromosomes can rescue the phenotype of floating head (flh) mutants. Homozygous flh embryos lack a differentiated notochord and have a reduced, discontinuous floor plate. Mutant embryos injected with genomic clones containing the flh+ gene often had stretches of several to many notochord cells overlaid by a row of floor-plate cells. In contrast, control mutant embryos injected with artificial chromosomes lacking the flh+ gene failed to form notochord. We conclude that the injection of large-insert genomic clones will speed the isolation of zebrafish genes disrupted by mutation and hence the identification of gene functions necessary for development of vertebrate embryos.

Animals↗

A 500-kb YAC and BAC contig encompassing the high-growth deletion in mouse chromosome 10 and identification of the murine Raidd/Cradd gene in the candidate region.

The mouse high growth (hg) gene was identified in a selection experiment for rapid growth. It produces a 30-50% increase in weight gain of homozygous individuals without resulting in obesity. Recently, hg was mapped to a deletion around marker D10Mit69. Here we report a map of yeast artificial chromosome and bacterial artificial chromosome clones spanning the entire region deleted in high-growth mice. The size of the deletion estimated from the clone lengths is approximately 500 kb. Using exon trapping, the murine Raidd/Cradd gene was identified in the hg region, and its cDNA sequence and expression pattern were determined. The human RAIDD/CRADD was mapped using radiation hybrid mapping to human chromosome 12, 2.9 cR distal to marker AFMB311WC5. The identified Raidd/Cradd gene, which is deleted in high-growth mice, presents a potential candidate for hg.

Adaptor Proteins, Signal Transducing↗

An integrated somatic cell hybrid, YAC, and BAC map of the Rmc1 region of mouse chromosome 1.

Rmc1, the cellular receptor for the polytropic class of murine retroviruses, determines the tissue tropism of the virus and therefore plays a critical role in the pathogenesis of polytropic virus-induced leukemia. Previously we reported the physical mapping of this gene to a 5-cM region of mouse chromosome 1 and the construction of a yeast artificial chromosome (YAC) contig across this region. In this report we describe the refinement of the Rmc1 candidate region to approximately 600 kb and the generation of an integrated somatic cell hybrid, YAC, and bacterial artificial chromosome contig spanning the region. A number of genes and loci were physically ordered along the chromosome, including a recently identified candidate for Rmc1.

Animals↗

A transcript map of a 2-Mb BAC contig in the proximal portion of the mouse X chromosome and regional mapping of the scurfy mutation.

A physical clone contig has been constructed, spanning 2 Mb on the proximal mouse X chromosome containing the mouse scurfy (sf) and tattered (Td) mutations. Extensive transcript mapping in this interval has identified 37 potential transcription units, including a number of novel genes, and 4 pseudogenes. These genes have been ordered by STS content and restriction mapping. Comparison of the transcript map to the corresponding region in human Xp11.23-p11.22 shows extensive homology, with complete conservation of gene order for loci in common between the two maps. Further, using a novel method to identify simple sequence length polymorphisms, we have developed a number of genetic markers, which has enabled the region containing the sf mutation to be narrowed to <300 kb. This contig has already allowed the cloning of the Td gene using a candidate gene approach and now serves as a starting point for the cloning of the sf mutation.

Animals↗