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J M Kramer

Publications and source records attributed to J M Kramer.

At least 37 records · Page 2Linked to original sources
PubMed1999

Phage typing of Campylobacter jejuni and Campylobacter coli and its use as an adjunct to serotyping.

Campylobacter is the most commonly reported cause of gastro-intestinal infection in England and Wales, with over 50,000 reported cases in 1997. The majority of human campylobacter isolates in England and Wales are C. jejuni (c. 90%) with most of the remainder being C. coli. We describe the use of phage typing as an extension to serotyping for more detailed characterization within these two species. The scheme was piloted during a study of 2407 C. jejuni and 182 C. coli strains isolated in Wales between April 1996 and March 1997. Fifty-seven C. jejuni phage types were identified, with the ten most prevalent phage types accounting for 60% of isolates tested; 16% of isolates were untypable. The most common phage type was PT 1 which represented c. 20% of isolates. A further 7% of isolates reacted with the phages but did not conform to a designated type (RDNC). Only 12 phage types were identified among C. coli, with the two most common types, PT 2 and PT 7 accounting for 75.2% of isolates. When used in conjunction with serotyping, the ability of phage typing to identify between 6 and 29 subtypes within each of the predominant HS types has enabled a further level of discrimination to be achieved that enhances the epidemiological typing of C. jejuni and C. coli.

Bacteriophage Typing↗
PubMed1998

Negative regulation of the heat shock transcriptional response by HSBP1.

In response to stress, heat shock factor 1 (HSF1) acquires rapid DNA binding and transient transcriptional activity while undergoing conformational transition from an inert non-DNA-binding monomer to active functional trimers. Attenuation of the inducible transcriptional response occurs during heat shock or upon recovery at non-stress conditions and involves dissociation of the HSF1 trimer and loss of activity. We have used the hydrophobic repeats of the HSF1 trimerization domain in the yeast two-hybrid protein interaction assay to identify heat shock factor binding protein 1 (HSBP1), a novel, conserved, 76-amino-acid protein that contains two extended arrays of hydrophobic repeats that interact with the HSF1 heptad repeats. HSBP1 is nuclear-localized and interacts in vivo with the active trimeric state of HSF1 that appears during heat shock. During attenuation of HSF1 to the inert monomer, HSBP1 associates with Hsp70. HSBP1 negatively affects HSF1 DNA-binding activity, and overexpression of HSBP1 in mammalian cells represses the transactivation activity of HSF1. To establish a biological role for HSBP1, the homologous Caenorhabditis elegans protein was overexpressed in body wall muscle cells and was shown to block activation of the heat shock response from a heat shock promoter-reporter construct. Alteration in the level of HSBP1 expression in C. elegans has severe effects on survival of the animals after thermal and chemical stress, consistent with a role for HSBP1 as a negative regulator of the heat shock response.

3T3 Cells↗
PubMed1998

Cuticle chirality and body handedness in Caenorhabditis elegans.

Caenorhabditis elegans adult animals exhibit an inherent chirality of fiber orientation in the basal layer of the cuticle, as well as a naturally invariant but experimentally reversible handedness in the left-right (L-R) asymmetry of the body plan. We have examined the relationship between cuticle chirality and body handedness in normal and L-R reversed animals, using Roller (Rol) mutants and transmission electron microscopy to monitor cuticle properties. Rol phenotypes, several of which have been shown to result from mutations in cuticle collagen genes, are characterized by an invariant, allele-specific handedness in their direction of rolling. We show for several alleles that this direction is not affected by L-R reversal of the body plan. We further show, by electron microscopy, that the chiral orientation of cuticle fibers in animals with normal cuticle is not reversed by L-R body-plan reversal. We conclude that cuticle chirality must be established independently of body-plan handedness. The cues that establish cuticle chirality are still unknown, as are the causes for different rolling directions in different Roller mutants. We discuss the question of how cuticle chirality maintains its independence, and how the orientations of the fiber layers may be determined.

Animals↗
PubMed1998

Leptin does not fully account for the satiety activity of adipose tissue-conditioned medium.

To determine whether leptin alone accounts for the satiety activity secreted by native adipose tissue, we prepared culture media conditioned by microdissected adipose tissue from overfed Long-Evans rats, fa/fa rats, or db/db mice (media A, B, and C, respectively). Medium A significantly suppressed food intake following intracerebroventricular delivery to Long-Evans rats (2-h chow intake = 68 +/- 5% of baseline, P < 0.001). Media B and C significantly suppressed food intake following intraperitoneal delivery to ob/ob mice (24-h chow intake = 56 +/- 7% of baseline for medium B, P = 0. 001; 4-day chow intake = 78 +/- 3% of baseline for medium C, P = 0. 004). Using a leptin receptor-based bioassay, we determined that the leptin concentration of medium C was 392 +/- 18 ng/ml. This concentration was 20-fold lower than the concentration of recombinant murine leptin required to produce a similar degree of feeding suppression following 5 days of administration to ob/ob mice. Neither medium conditioned by adipose tissue from ob/ob mice nor medium conditioned by adipose tissue from fa/fa rats and subsequently immunodepleted of leptin had significant satiety activity. We conclude that leptin is necessary but not sufficient to account for the satiety activity of native adipose tissue, perhaps due to the production by adipocytes of a cofactor that augments the ability of leptin to suppress feeding.

Adipose Tissue↗
PubMed1998

Ultrastructural analyses of the Caenorhabditis elegans rol-6 (su1006) mutant, which produces abnormal cuticle collagen.

Roller mutants of Caenorhabditis elegans rotate around their long axis and move in circular paths. Isolation and sequence of the rol-6 gene of C. elegans have shown that it encodes a cuticle collagen. In this paper, we describe the morphological alterations seen in the cuticle of the right roller mutant rol-6 (su1006) at the ultrastructural level. Deep-etched replica analyses showed that the honeycomb elements, fibers organized in a pentagonal fashion above the fishbone fibrous layer, completely fill the intermediate layer, which is observed to be largely empty spaces in the wild-type strain. The honeycomb fibers appear to connect the cortical and basal regions of the mutant cuticle. These fibers are likely to be involved in generating the helical twist of the mutant animals. Deep-etched replicas also revealed a delicate network of filaments on the nematode surface.

Animals↗
PubMed1997

Type IV collagen is detectable in most, but not all, basement membranes of Caenorhabditis elegans and assembles on tissues that do not express it.

Type IV collagen in Caenorhabditis elegans is produced by two essential genes, emb-9 and let-2, which encode alpha1- and alpha2-like chains, respectively. The distribution of EMB-9 and LET-2 chains has been characterized using chain-specific antisera. The chains colocalize, suggesting that they may function in a single heterotrimeric collagen molecule. Type IV collagen is detected in all basement membranes except those on the pseudocoelomic face of body wall muscle and on the regions of the hypodermis between body wall muscle quadrants, indicating that there are major structural differences between some basement membranes in C. elegans. Using lacZ/green fluorescent protein (GFP) reporter constructs, both type IV collagen genes were shown to be expressed in the same cells, primarily body wall muscles, and some somatic cells of the gonad. Although the pharynx and intestine are covered with basement membranes that contain type IV collagen, these tissues do not express either type IV collagen gene. Using an epitope-tagged emb-9 construct, we show that type IV collagen made in body wall muscle cells can assemble into the pharyngeal, intestinal, and gonadal basement membranes. Additionally, we show that expression of functional type IV collagen only in body wall muscle cells is sufficient for C. elegans to complete development and be partially fertile. Since type IV collagen secreted from muscle cells only assembles into some of the basement membranes that it has access to, there must be a mechanism regulating its assembly. We propose that interaction with a cell surface-associated molecule(s) is required to facilitate type IV collagen assembly.

Animals↗
PubMed1997

Characterization of alpha1(IV) collagen mutations in Caenorhabditis elegans and the effects of alpha1 and alpha2(IV) mutations on type IV collagen distribution.

Type IV collagen is a major component of basement membranes. We have characterized 11 mutations in emb-9, the alpha1(IV) collagen gene of Caenorhabditis elegans, that result in a spectrum of phenotypes. Five are substitutions of glycines in the Gly-X-Y domain and cause semidominant, temperature-sensitive lethality at the twofold stage of embryogenesis. One is a glycine substitution that causes recessive, non-temperature-sensitive larval lethality. Three putative null alleles, two nonsense mutations and a deletion, all cause recessive, non-temperature-sensitive lethality at the threefold stage of embryogenesis. The less severe null phenotype indicates that glycine substitution containing mutant chains dominantly interfere with the function of other molecules. The emb-9 null mutants do not stain with anti-EMB-9 antisera and show intracellular accumulation of the alpha2(IV) chain, LET-2, indicating that LET-2 assembly and/or secretion requires EMB-9. Glycine substitutions in either EMB-9 or LET-2 cause intracellular accumulation of both chains. The degree of intracellular accumulation differs depending on the allele and temperature and correlates with the severity of the phenotype. Temperature sensitivity appears to result from reduced assembly/secretion of type IV collagen, not defective function in the basement membrane. Because the dominant interference of glycine substitution mutations is maximal when type IV collagen secretion is totally blocked, this interference appears to occur intracellularly, rather than in the basement membrane. We suggest that the nature of dominant interference caused by mutations in type IV collagen is different than that caused by mutations in fibrillar collagens.

Alleles↗
PubMed1997

Fulminant liver failure in association with the emetic toxin of Bacillus cereus.

BACKGROUND: A 17-year-old boy and his father had acute gastroenteritis after eating spaghetti and pesto that had been prepared four days earlier. Within two days, fulminant liver failure and rhabdomyolysis developed in the boy and he died. The father had hyperbilirubinemia and rhabdomyolysis but recovered. We investigated the cause of these illnesses. METHODS: Bacteria were isolated and characterized by conventional methods, and bacterial toxins were quantified by immunoassays and cell-culture techniques. The effect of the isolated toxin on the rates of oxidation of various substrates was analyzed in rat-liver mitochondria. RESULTS: Autopsy of the boy's liver revealed diffuse microvesicular steatosis and midzonal necrosis that suggested impaired beta-oxidation of liver mitochondria due to a mitochondrial toxin. There was no evidence of ingestion of heavy metals, halogenated compounds, hepatotoxic drugs, or staphylococcal enterotoxin. However, high concentrations of Bacillus cereus emetic toxin were found in both the residue from the pan used to reheat the food and the boy's liver and bile. B. cereus was cultured from the intestinal contents and the pan residue. The emetic toxin isolated from the B. cereus cultures was found to be a mitochondrial toxin. CONCLUSIONS: Fulminant liver failure developed after the ingestion of food contaminated with the B. cereus emetic toxin. The toxin inhibits hepatic mitochondrial fatty-acid oxidation, indicating that it caused liver failure in this patient.

Adolescent↗
PubMed1997

The effects of different thermal environments on the physiological and psychological responses of firefighters to a training drill.

Little is known about the impact of thermoregulatory demands on cardiovascular and psychological responses of firefighters during firefighting activities. This study examined selected responses to a training drill in different thermal environments. Male firefighters (n = 16) were randomly assigned to perform a simulated ceiling overhaul task for 16 min in either a neutral (13.7 degrees C) or hot (89.6 degrees C) condition while wearing standard firefighting turnout gear. Physiological and psychological measures were assessed before, after 8 min and 16 min of firefighting activity, and following a 10-min recovery period. The variables assessed included heart rate (HR), tympanic temperature (Ttymp), lactate level (LAC), blood glucose level, ratings of perceived exertion (RPE), perceptions of respiration, thermal sensations (TS) and state anxiety (SA). Significant increases were seen for HR, Ttymp, LAC, RPE and SA, with the increases being much greater following the hot condition. Recovery was significantly slower following work in the hot condition. These findings suggest that the addition of a live fire (a common situation for firefighters) contributes to increased cardiovascular and psychological strain at a standardized workload.

Adult↗
PubMed1997

Caenorhabditis elegans cuticle: a description of new elements of the fibrous layer.

At the ultrastructural level, the Caenorhabdits elegans cuticle shows the presence of well defined layers; 1 of them is the fibrous layer composed by 2 strands of fibers that meet each other at a 60 degrees angle and resembling a fish-bone pattern. In this paper, we describe new elements of the fibrous layer. When thin sections were obtained at a very low angle, i.e., almost tangential, fibers of wavy appearance could be observed. Those elements were 300 nm in length and 20 nm thick and were linked to each other by delicate dots. Deep-etched replicas of C. elegans revealed more details of the arrangement of new elements in the fibrous layer. The wavy fibers were organized in 5-sided, honeycomblike figures. Each pentagonal fiber was 145 nm across and was composed of tightly packed globular structures arranged linearly.

Animals↗
PubMed1996

Medical resources and the internet. Making the connection.

The Internet has provided a new forum through which medical information can be obtained and discussed. We review methods available to take advantage of this resource and provide a glimpse of electronically available information and discussions. These methods are useful for beginning and experienced users of the Internet.

Computer Communication Networks↗
PubMed1996

Evaluation of serotyping, biotyping, plasmid banding pattern analysis, and HEp-2 vacuolation factor assay in the epidemiological investigation of Bacillus cereus emetic-syndrome food poisoning.

To assess the value of the plasmid banding patterns, the vacuolation factor (VF) assay, biotyping, and serological typing as epidemiological markers for strains of Bacillus cereus causing emetic-syndrome illness, 43 isolates from five outbreaks and an additional 76 strains isolated in food-poisoning outbreaks caused by other enteric pathogens were examined by these techniques, and the results were compared. Thirty-eight (88%) of the 43 outbreak strains produced vacuolation responses in HEp-2 cells and were all starch-hydrolysis negative. The other 76 strains associated with outbreaks caused by other food-poisoning bacteria gave all negative VF production results except four strains, and 56 (74%) of these strains produced positive reactions in starch hydrolysis tests. Starch hydrolysis emerged as a convenient screen for VF production, because no starch hydrolysis-positive strains produced VF. With the exception of one isolate, all 38 VF-positive isolates from emtic-syndrome outbreaks were serotype H.1. Isolates from four of the five outbreaks revealed identical plasmid banding patterns in each outbreak, whereas only three of eight serotype H.1 strains from the fifth outbreak exhibited indistinguishable plasmid banding patterns. These results suggest that the plasmid banding pattern analysis may be of value in discriminating between isolates of the same serotype, and establishing if an outbreak arises from a common food source. In conclusion, the vacuolation factor assay combined with the plasmid banding patterns proved to be a valuable tool for the epidemiological investigation of emetic-syndrome outbreaks caused by B. cereus. Moreover, these methods are particularly useful for laboratories that do not have ready access to serotyping facilities.

Bacillus cereus↗
PubMed1996

Physiological, psychophysical, and psychological responses of firefighters to firefighting training drills.

This study was designed to describe the physiological, psychophysical, and psychological responses of firefighters to firefighting drills in a training structure containing live fires. Fifteen male firefighters, wearing standard turnout gear which resulted in full encapsulation, performed two firefighting tasks (advancing fire hose, chopping wood) while inside the training structure. Measurements of heart rate, tympanic membrane temperature, blood lactate, perceptions of respiration, mood, perceived exertion, and thermal sensation were obtained after 8 min of advancing fire hose, and again after 8 min of chopping. Heart rate and temperature increased significantly from baseline and from advancing hose to wood chopping, whereas blood lactate increased initially after advancing the hose and remained elevated at the end of the chopping task. At the completion of the test (both tasks), mean heart rate (182.3 b.min-1), temperature (40.1 degrees C, [104.1 degrees F]), and blood lactate (3.8 mMol) suggested that the firefighting tasks used in this study impose considerable physiological strain on firefighters. Psychophysical and psychological data mirrored the greater physiological strain following firefighting tasks performed in a hot environment while wearing full turnout gear.

Adult↗