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Biomedical subjects

J M Kramer

Publications and source records attributed to J M Kramer.

At least 73 records · Page 4Linked to original sources
PubMed1992

Application of pyrolysis mass spectrometry to the investigation of outbreaks of food poisoning and non-gastrointestinal infection associated with Bacillus species and Clostridium perfringens.

Eighteen isolates of Bacillus species and 15 of Clostridium perfringens, all of which had been associated with outbreaks of either food poisoning or non-gastrointestinal infection (NGI), were examined for relatedness by pyrolysis mass spectrometry (PyMS). The PyMS-analysis correctly clustered all the groups of epidemiologically related isolates of both genera, and distinguished all the single, epidemiologically unrelated isolates of the same species. PyMS is a simple, rapid and inexpensive technique which can provide useful and accurate inter-strain comparisons within both the Bacillus and Clostridium genera in complete accord with conventional serological typing results.

Bacillaceae Infections↗
PubMed1992

Media for the detection and enumeration of Bacillus cereus in foods: a review.

Bacillus cereus is an established cause of food poisoning in addition to being a troublesome and persistent contaminant, responsible for a variety of spoilage defects in processed foods and dairy products. A range of diagnostic and selective media has been developed to facilitate the detection and enumeration of B. cereus in routine surveillance situations and food poisoning investigations. These media are reviewed with respect to the selective and diagnostic systems they employ, their ability to recover and differentiate the target organism, and their advantages and limitations in particular applications.

Animals↗
PubMed1992

Bacillus anthracis but not always anthrax.

Gram-positive bacilli isolated during epidemiological investigations which, on the basis of conventional tests, resemble Bacillus anthracis but which fail to produce the capsule or to induce anthrax in test animals have long been dismissed in clinical and veterinary laboratories as B. cereus or simply as unidentified Bacillus spp. and thereupon discarded as inconsequential. In this study, the application of newly available DNA probe, polymerase chain reaction and specific toxin antigen detection technology has revealed that a proportion of such strains are B. anthracis which lack the plasmid carrying the capsule gene (pXO2). While these techniques cannot, of course, be used to confirm the identities of strains resembling B. anthracis but which also lack the plasmid carrying the toxin genes (pXO1), the likelihood that these also are bona fide B. anthracis becomes more acceptable. (As yet no naturally occurring pXO1-/2+ strains have been found.) At this point, the significance of the presence of such avirulent forms of B. anthracis in specimens can only be a subject for speculation, but the possibility that they may be indicators of virulent parents somewhere in the system being examined must be considered.

Animals↗
PubMed1991

Embryonic lethality caused by mutations in basement membrane collagen of C. elegans.

Basement membranes are specialized forms of extracellular matrix with important functions in development. A major structural component of basement membranes is type IV collagen, a heterotrimer of two alpha 1(IV) and one alpha 2(IV) chains, which forms a complex, polygonal network associated with other basement membrane components. Here we report that the alpha 1(IV) collagen chain of Caenorhabditis elegans is encoded by the genetic locus emb-9. Mutations in emb-9 cause temperature-sensitive lethality during late embryogenesis. We have identified single nucleotide alterations that substitute glutamic acid for glycine in the triple-helical Gly-X-Y repeat region of the alpha 1(IV) collagen in three emb-9 mutant strains. These results are direct evidence that defects in basement membranes can disrupt embryonic development and form a basis for the genetic analysis of basement membrane function.

Amino Acid Sequence↗
PubMed1991

Efficient gene transfer in C.elegans: extrachromosomal maintenance and integration of transforming sequences.

We describe a dominant behavioral marker, rol-6(su-1006), and an efficient microinjection procedure which facilitate the recovery of Caenorhabditis elegans transformants. We use these tools to study the mechanism of C.elegans DNA transformation. By injecting mixtures of genetically marked DNA molecules, we show that large extrachromosomal arrays assemble directly from the injected molecules and that homologous recombination drives array assembly. Appropriately placed double-strand breaks stimulated homologous recombination during array formation. Our data indicate that the size of the assembled transgenic structures determines whether or not they will be maintained extrachromosomally or lost. We show that low copy number extrachromosomal transformation can be achieved by adjusting the relative concentration of DNA molecules in the injection mixture. Integration of the injected DNA, though relatively rare, was reproducibly achieved when single-stranded oligonucleotide was co-injected with the double-stranded DNA.

Animals↗
PubMed1990

Tandemly duplicated Caenorhabditis elegans collagen genes differ in their modes of splicing.

Caenorhabditis elegans contains 50 to 150 collagen genes dispersed throughout its genome. We have determined the complete nucleotide sequences of two collagen genes, col-12 and col-13, that are separated by only 1800 bases and are transcribed in the same direction. The 951 nucleotides of their coding regions differ by only five nucleotides (99.5% identity). The amino acid sequences are identical except for two conservative amino acid changes within the putative secretory signal sequences, so the mature forms of the col-12 and col-13 collagens would be identical. The position and sequence of the intron (52 base-pairs) within the coding region of each gene are perfectly conserved. In contrast to the coding regions and the introns, the 5' and 3' flanking regions show little sequence similarity, col-12 and col-13 are expressed at similar levels at the same developmental stages, and appear to utilize conserved TATA boxes and transcription start sites. The major differences between the genes is that, preceding the initiator ATG, col-12 has a cis-spliced intron, while col-13 is transspliced. Thus, col-12 and col-13 are essentially identical in all aspects except that the col-12 mRNA has a 26-nucleotide cis-spliced leader at the same place where the col-13 mRNA has a 22-nucleotide trans-spliced leader. These results suggest that col-12 and col-13 are derived from a gene duplication and that sequence homology in the coding regions, but not in the flanking regions, has been maintained by gene conversion. The fact that the only significant difference between the two genes is in their modes of splicing suggests that cis and trans-splicing can be interchanged during gene evolution.

Amino Acid Sequence↗
PubMed1990

The effect of glucose, starch, and pH on growth, enterotoxin and haemolysin production by strains of Bacillus cereus associated with food poisoning and non-gastrointestinal infection.

Brain-Heart Infusion medium, modified by (a) 1.0% w/v glucose supplement, (b) 1.0% w/v soluble starch supplement, (c) pH adjustment to 8.8 or (d) pH adjustment to 5.0, was used to investigate the influence of glucose, starch and pH on growth, enterotoxin and haemolysin production by Bacillus cereus. The four test strains selected for comparison originated from episodes of emetic-syndrome food poisoning, diarrhoeal-syndrome food poisoning, traumatic wound infection and bovine mastitis, respectively. In the presence of either glucose or starch, growth of all strains was found to be near-optimum and accompanied by enhanced enterotoxin production. However, under the moderately acid and alkaline test conditions B. cereus strains exhibited varying degrees of growth inhibition and partial repression of enterotoxin and haemolysin production. In this respect the food-poisoning isolates proved to have greater resistance to adverse pH environments than did the non-gastrointestinal infection isolates.

Animals↗
PubMed1990

The Caenorhabditis elegans rol-6 gene, which interacts with the sqt-1 collagen gene to determine organismal morphology, encodes a collagen.

The rol-6 gene is one of the more than 40 loci in Caenorhabditis elegans that primarily affect organismal morphology. Certain mutations in the rol-6 gene produce animals that have the right roller phenotype, i.e., they are twisted into a right-handed helix. The rol-6 gene interacts with another gene that affects morphology, sqt-1; a left roller allele of sqt-1 acts as a dominant suppressor of a right roller allele of rol-6. The sqt-1 gene has previously been shown to encode a collagen. We isolated and sequenced the rol-6 gene and found that it also encodes a collagen. The rol-6 gene was identified by physical mapping of overlapping chromosomal deficiencies that cover the gene and by identification of an allele-specific restriction site alteration. The amino acid sequence of the collagen encoded by rol-6 is more similar to that of the sqt-1 collagen than to any of the other ten C. elegans cuticle collagen sequences compared. The locations of cysteine residues flanking the Gly-X-Y repeat regions of rol-6 and sqt-1 are identical, but differ from those in the other collagens. The sequence similarities between rol-6 and sqt-1 indicate that they represent a new collagen subfamily in C. elegans. These findings suggest that these two collagens physically interact, possibly explaining the genetic interaction seen between the rol-6 and sqt-1 genes.

Amino Acid Sequence↗
PubMed1989

The two Caenorhabditis elegans basement membrane (type IV) collagen genes are located on separate chromosomes.

We have identified and characterized the two genes, clb-1 and clb-2, that encode basement membrane collagen (IV) in Caenorhabditis elegans. Both genes encode 5.5-kilobase mRNAs, similar in size to the mammalian and Drosophila type IV collagen gene transcripts but much larger than the cuticle collagen transcripts of C. elegans. The nucleotide sequences of the NC1 regions of both genes were determined. Comparisons of the clb-1 and clb-2 NC1 amino acid sequences with those of mouse and human show that clb-1 shares 72% identity with mammalian alpha 2(IV) and clb-2 shares 63% identity with mammalian alpha 1(IV), suggesting that clb-1 is the alpha 2(IV) homologue and clb-2 is the alpha 1(IV) homologue. The presence of two type IV collagen genes in C. elegans and the mammals, but a single gene in Drosophila, indicates that the primordial type IV collagen gene had already duplicated early in the evolution of the invertebrates and that one of the genes has been subsequently lost from Drosophila. The mouse and human alpha 1(IV) and alpha 2(IV) genes are separated by only about 130 base pairs and are transcribed in opposite directions from overlapping promoters. We show that the C. elegans genes are located on separate chromosomes, clb-1 on X and clb-2 on III, demonstrating that the mammalian arrangement is not a requirement for all type IV collagen genes. We have identified a candidate genetic locus for the clb-2 basement membrane collagen gene, which will allow us to pursue a genetic analysis of basement membrane structure and function in C. elegans.

Amino Acid Sequence↗
PubMed1989

Sequence comparisons of developmentally regulated collagen genes of Caenorhabditis elegans.

Collagen genes col-6, col-7 (partial), col-8, col-14 and col-19 from the nematode Caenorhabditis elegans were sequenced, and compared to the previously sequenced genes col-1 and col-2. The genes are between 1.0 and 1.2 kb in length, and each includes one or two short introns. The presumptive promoter regions contain sequences similar to the eukaryotic TATA promoter element. Two distinct, conserved sequences were found in the presumptive promoter regions of, respectively, the dauer larva-specific genes col-2 and col-6, and the primarily adult-specific genes col-7 and col-19. The domain structures of the collagen polypeptides are similar: each polypeptide contains two triple-helix forming (Gly-X-Y)n domains, one of 30-33 amino acids (aa), and the other of 127-132 aa. The latter domain is interrupted by one to three short (2-8 aa) non-(Gly-X-Y)n segments that occur at relatively conserved locations in each polypeptide. Sets of cysteine residues flank the (Gly-X-Y)n domains in all of the polypeptides. The genes can be placed into three families based upon amino acid sequence similarities. Genes within a family do not always exhibit similar developmental expression programs, suggesting that structural and regulatory regions of the genes have evolved separately. The codon usage in the genes is highly asymmetrical, with adenine appearing in the third position of 85% of the glycine codons, and 93% of the proline codons.

Amino Acid Sequence↗
PubMed1988

The sqt-1 gene of C. elegans encodes a collagen critical for organismal morphogenesis.

Different mutations in the sqt-1 gene of C. elegans can lengthen, shorten, or helically twist the entire animal. We have cloned the sqt-1 gene and have shown that it encodes a collagen. sqt-1 was localized to a 35 kb region of DNA by physical mapping of chromosomal deficiencies. A transposon (Tc1)-induced mutation of sqt-1 was generated and utilized to identify the sqt-1 gene within this 35 kb region. Sequence analysis of the sqt-1 gene shows that it encodes a 32 kd collagen polypeptide that is similar in size and structure to other members of the C. elegans collagen family. The Tc1 insertion mutant has no detectable sqt-1 transcripts, yet it is morphologically normal, indicating that the null phenotype of sqt-1 is wild type. These results demonstrate that collagen mutations can have dramatic effects on organismal morphology.

Alleles↗
PubMed1988

Plasmid, serotypic, and enterotoxin analysis of Bacillus cereus in an outbreak setting.

Bacillus cereus is a recognized agent of food-borne disease. In this report we describe an outbreak of B. cereus gastroenteritis associated with consumption of beef stew among patients and staff at a Rhode Island nursing home. The beef had been improperly stored after preparation. The predominant symptoms of the illness were cramps and diarrhea; it lasted an average of 16 h. No deaths occurred. The organism was recovered from 10 of 23 stools collected from ill patients and 1 of 21 stools collected from controls (P = 0.0044, Fisher's two-tailed exact test). All isolates had the same biotype and serotype, newly designated H.26; all elaborated the diarrheal B. cereus enterotoxin when tested in rabbits by the vascular permeability reaction; and all had identical plasmid profiles, which differed from those of B. cereus strains selected randomly from other outbreaks. Plasmid analysis may prove to be a useful new tool in investigating outbreaks of B. cereus food poisoning.

Adult↗
PubMed1987

Toxigenic Bacillus cereus as a cause of wound infections in the tropics.

A bacteriological survey was conducted on clinically infected traumatic wounds in members of an Operation Raleigh expedition, who were working in the Costa Rican rain forest. Bacillus cereus was isolated from the wounds of 14 of 18 patients, usually in pure and heavy growth. Most of the isolates were strongly toxigenic by in vivo pathogenicity tests. The organism was also isolated from the nose in 15 cases and the throat in five cases. The findings indicate that B. cereus was the principle pathogen in this series of traumatic wounds.

Adolescent↗
PubMed1985

Expression of the Caenorhabditis elegans collagen genes col-1 and col-2 is developmentally regulated.

The total collagen gene expression as well as the specific expression of two sequenced Caenorhabditis elegans collagen genes, col-1 and col-2, has been investigated. Northern blots of RNA isolated from animals at different developmental stages were probed under conditions that allow cross-hybridization of all collagen sequences. The majority of hybridization is to transcripts of 1.1-1.4 kilobases (kb) in length, with weak hybridization to some larger transcripts. Different size patterns, within the 1.1-1.4-kb ranges, are seen in RNAs from different developmental stages. Gene-specific probes were produced from the C. elegans collagen genes col-1 and col-2, and each was shown to hybridize to a single size transcript in the 1.1-1.4-kb region. The col-1 transcript was found in all the developmental stages examined, but its abundance varied between stages. The col-2 transcript was detected only in a single developmental stage, during formation of the dauer larvae. The 5' and the 3' ends of the col-1 and col-2 transcripts were determined by S1 nuclease digestion experiments. Both genes have the common "TATA" and "CAAT" box sequences preceding the 5' end of their transcripts and there is strong sequence homology in their 5' untranslated regions. Multiple copies of an eight-nucleotide repeat sequence were found upstream from both col-1 and col-2.

Animals↗