Search PubMed⌕ Search

Biomedical subjects

J Klein

Publications and source records attributed to J Klein.

At least 361 records · Page 20Linked to original sources
PubMed1994

Organization of Mhc class II B genes in the zebrafish (Brachydanio rerio).

Using three genomic phage libraries, we isolated 26 clones from the zebrafish MHC class II B region. By restriction mapping, the clones could be arranged into six clusters, most clusters consisting of several overlapping clones. The combined clusters cover a total of 161 kb of the zebrafish class II region. Hybridization with specific probes demonstrated the presence in the clusters of two class II A and six class II B genes. Sequencing of the B genes revealed that they represented six different families of class II loci. Only two of the class II B and one of the class II A genes are complete; the others are truncated pseudogenes. Only one of the class II B loci shows extensive restriction fragment length polymorphism. This is also the only locus found to be transcribed in organs with large numbers of lymphoid or myeloid cells. The zebrafish class II genes have promoter regions with sequence elements found previously in mammalian genes and known to be involved in regulation of expression. The exon-intron organization of the zebrafish class II genes is similar to that of the mammalian genes, but the introns are characteristically short, ranging in length from 74 to 362 bp. The distances between A and B genes in a given pair are also short, but the distances between B genes are as long as or longer than those between mammalian class II B genes. All of the zebrafish class II B genes appear to have arisen by duplication and diversification of a single ancestral B gene after the separation of bony fishes from other vertebrate taxa.

Amino Acid Sequence↗
PubMed1994

Physical map of mouse chromosome 17 in the region relevant for positional cloning of the Hybrid sterility 1 gene.

Hybrid sterility 1 (Hst1) is the major gene responsible for sterility of male hybrids between Mus musculus and certain laboratory strains. Thus, Hst1 is of importance in studying both postreproductive isolation of closely related species and male fertility. It has been mapped to mouse chromosome 17 in the region corresponding to the third inversion of the t haplotypes. The aim of the present study was to construct a physical map of the Hst1 region as the first step in an effort to clone the gene. Three yeast artificial chromosome (YAC) libraries (Princeton, Whitehead, and ICRF) were screened with polymerase chain reaction (PCR) oligonucleotide primers and DNA probes specific for loci previously mapped into the region of the third inversion. The isolated YAC clones were restriction mapped and arranged into contigs. Sixteen YAC clones were arranged into a single contig encompassing a region approximately 2000 kb long based on restriction mapping of highly overlapping but independently derived YAC clones. Five new loci in the region of the third inversion were mapped and the order and approximate physical distances of 12 loci established in this contig. The Hst1 gene maps approximately 0.2 cM proximal to the D17Ph1 locus encompassed by the YAC contig. Since the contig extends at least 1200 kb proximal to D17Ph1, it should contain the Hst1 gene.

Animals↗
PubMed1994

Molecular genetics of speciation and human origins.

The major histocompatibility complex (MHC) plays a cardinal role in the defense of vertebrates against parasites and other pathogens. In some genes there are extensive and ancient polymorphisms that have passed from ancestral to descendant species and are shared among contemporary species. The polymorphism at the DRB1 locus, represented by 58 known alleles in humans, has existed for at least 30 million years and is shared by humans, apes, and other primates. The coalescence theory of populations genetics leads to the conclusion that the DRB1 polymorphism requires that the population ancestral to modern humans has maintained a mean effective size of 100,000 individuals over the 30-million-year persistence of this polymorphism. We explore the possibility of occasional population bottlenecks and conclude that the ancestral population could not have at any time consisted of fewer than several thousand individuals. The MHC polymorphisms exclude the theory claiming, on the basis of mitochondrial DNA polymorphisms, that a constriction down to one or few women occurred in Africa, at the transition from archaic to anatomically modern humans, some 200,000 years ago. The data are consistent with, but do not provide specific support for, the claim that human populations throughout the World were at that time replaced by populations migrating from Africa. The MHC and other molecular polymorphisms are consistent with a "multiregional" theory of Pleistocene human evolution that proposes regional continuity of human populations since the time of migrations of Homo erectus to the present, with distinctive regional selective pressures and occasional migrations between populations.

Animals↗
PubMed1994

Intensity of natural selection at the major histocompatibility complex loci.

Long persistence of allelic lineages, prevalence of nonsynonymous over synonymous substitutions in the peptide-binding region (PBR), and deviation from neutrality of the expected gene identity parameter F all indicate indirectly that balancing selection is operating at functional major histocompatibility complex (MHC) loci. Direct demonstrations of the existence of balancing selection at MHC loci are, however, either lacking or not fully convincing. To define the conditions under which balancing selection could be demonstrated, we estimated its intensity from the mean number of nonsynonymous substitutions, KB, at the PBR and the mutation rate mu. We compared the five available methods for estimating KB by computer simulation and chose the most reliable ones for estimation of selection intensity. For the human MHC, the selection coefficients of the HLA-A, -B, -C, -DRB1, -DQB1, -DQA1, and -DPB1 loci are 0.015, 0.042, 0.0026, 0.019, 0.0085, 0.0028, and 0.0007, respectively. This low selection intensity places severe restrictions on the possibility of measuring selection directly in vertebrate populations.

Alleles↗
PubMed1994

The origin of the primate Mhc-DRB genes and allelic lineages as deduced from the study of prosimians.

MHC class II genes of the DRB family were partially sequenced from 10 individuals representing six species of prosimians: Galago senegalensis, G. moholi, Otolemur garnetti, Loris tardigradus, Petterus (Lemur) fulvus, and Lemur catta. Altogether, 41 different genes were discerned, all distinct from genes identified previously. Comparative analysis of the sequences has led to the following conclusions. First, the DRB loci present in human populations diverged from one another before the divergence of prosimian and anthropoid primates. Second, major allelic lineages of the DRB1 locus, such as DRB1*03 (DRB1*13) and DRB1*04, were established more than 85 million years ago. Third, the DRB6 gene was inactivated before the separation of prosimians and anthropoids, and has remained a pseudogene for more than 85 million years. Fourth, the primate DRB region is structurally and functionally unstable. In Lemur catta, for example, all DRB genes have apparently been lost and their function taken over by DOB and/or DPB genes. DRB genes are, however, present in a related species, Petterus (Lemur) fulvus. Fifth, the prosimian DRB3 genes are all inactive; their function seems to have been taken over by new genes. Sixth, several of the prosimian DRB genes and pseudogenes have recently been duplicated. In Otolemur garnetti, for example, one chromosome carries at least three copies of the DRB3 pseudogene.

Alleles↗
PubMed1994

Simultaneous determination of lidocaine, prilocaine and the prilocaine metabolite o-toluidine in plasma by high-performance liquid chromatography.

An HPLC assay is described for the measurement of prilocaine and lidocaine (components of the local anesthetic cream EMLA) as well as the prilocaine metabolite, o-toluidine, in plasma. The method uses UV detection, is simple, sensitive and most important, only a single 200-microliters plasma sample is needed for simultaneous analysis of prilocaine, lidocaine and o-toluidine with a detection limit of 4 ng/ml. The plasma, together with the internal standard (bupivacaine) is extracted with diethyl ether under alkaline conditions, followed by the extraction of the analytes from the organic phase into dilute sulphuric acid. An aliquot of the acid extract is injected onto the HPLC system and the effluent is monitored by a UV detector.

Animals↗
PubMed1994

Hair concentrations of nicotine and cotinine in women and their newborn infants.

BACKGROUND: To date, no biological markers have been identified that can predict the extent of fetal exposure to the toxic constituents of cigarette smoke. A variety of xenobiotic agents have been shown to accumulate in growing hair. PATIENTS AND METHODS: We measured maternal and neonatal hair concentrations of nicotine and cotinine in 94 mother-infant pairs. Mothers who were active smokers, nonsmokers, and passive smokers and their infants were included. RESULTS: Mothers who were active smokers (n = 36) had mean (SEM) hair concentrations of 19.2 (4.9) ng/mg for nicotine and 6.3 (4.0) ng/mg for cotinine, significantly higher than concentrations in nonsmokers (n = 35) (1.2 [0.4] ng/mg for nicotine and 0.3 [0.06] ng/mg for cotinine, P < .0001). Infants of smokers had mean hair concentrations of 2.4 (0.9) ng/mg for nicotine (range, 0 to 27.3 ng/mg) and 2.8 (0.8) ng/mg for cotinine (range, 0 to 12.2 ng/mg), significantly higher than concentrations in infants of nonsmokers (0.4 [0.09] ng/mg for nicotine and 0.26 [0.04] ng/mg for cotinine, P < .01). Mothers with passive smoke exposure and their infants (n = 23) had significantly higher hair concentrations of nicotine (3.2 [0.8] ng/mg for mothers and 0.28 [0.05] ng/mg for infants) and cotinine (0.9 [0.3] ng/mg for mothers and 0.6 [0.15] ng/mg for infants) than nonsmoking mothers and their infants (P < .01). There was a significant correlation between maternal and neonatal hair concentrations of nicotine (r = .49, P < .001) or cotinine (r = .85, P = .0001). CONCLUSIONS: This is the first biochemical evidence that infants of passive smokers are at risk of measurable exposure to cigarette smoke. Hair accumulation of cigarette smoke constituents reflects long-term systemic exposure to these toxins and therefore may be well correlated with perinatal risks.

Adult↗
PubMed1994

Phospholipase C and phospholipase D are independently activated in rat hippocampal slices.

In order to investigate a possible G-protein-mediated activation of phospholipase D (PLD) and its relationship to the activation of phosphoinositide-specific phospholipase C (PI-PLC), we measured the effects of aluminium fluoride and carbachol on choline release, the PLD-specific transphosphatidylation reaction (generation of phosphatidylpropanol) and the formation of inositol phosphates in rat hippocampal slices. Aluminium fluoride markedly enhanced the formation of choline and phosphatidylpropanol but failed to increase the formation of inositol phosphates. In contrast, the muscarinic agonist carbachol strongly stimulated PI-PLC but failed to activate PLD. We conclude that PLD in hippocampal slices is activated by a G-protein independently of phosphoinositide hydrolysis.

Aluminum Compounds↗
PubMed1994

Critical comparison of novel and existing methods of compliance assessment during a clinical trial of an oral iron chelator.

The assessment of compliance is critical in the evaluation of the effectiveness of a new therapeutic agent. Fifteen patients with transfusion-dependent beta-thalassemia, many of whom had previously demonstrated erratic compliance with deferoxamine, were enrolled in a clinical trial of a new oral iron chelator, 1,2-dimethyl-3-hydroxypyrid-4-one (L1). Their compliance with this medication was estimated by several existing methods and the novel Medication Event Monitoring System (MEMS). Overall compliance as assessed by the MEMS was 78.5 +/- 13.0% of prescribed doses taken, significantly lower than the corresponding rates calculated by pill counts and diaries (91.5 +/- 9.2% and 94.1 +/- 4.3%, respectively). However, several serious problems were encountered with the MEMS, mostly in the form of incorrect use of the device by the patients. Disclosure of the nature of the MEMS and the compliance monitoring process did not alter the rate of adherence with L1 therapy. Compliance as determined by pill counts did not differ between the 1st and 2nd 6-month periods. Although not reaching statistical significance, a trend towards better L1 compliance occurred in those patients in whom serum ferritin levels decreased. Patients who filled at least 50% of their diaries had significantly better compliance by pill counts than those who completed less than 50% of their diaries (95.9 +/- 4.1% and 86.5 +/- 11.1%, respectively). Steady-state L1 trough concentrations and 24-hour urinary iron excretion did not correlate with L1 compliance.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗
PubMed1994

Cisplatin protein binding in pregnancy and the neonatal period.

The most effective combination regimes for ovarian cancer contain cisplatin, but there is little knowledge about cisplatin use during pregnancy. The high protein binding of cisplatin means that small changes in protein binding result in large changes in the fraction of free drug. It is the free form of cisplatin that crosses the placenta and may contribute to fetal toxicity. The purpose of the present study was to compare protein binding of cisplatin in pregnant women, non-pregnant women, and newborn infants. We found that babies and pregnant women had significantly lower concentrations of both protein and albumin compared to non-pregnant women. Analysis of variance found overall significant differences in protein binding among the three groups over time (P < .05). Babies had statistically less cisplatin protein binding than non-pregnant women at 80 minutes and all time points thereafter (P < .05). In contrast, pregnant women had statistically less cisplatin protein binding than non-pregnant women at 3.3 and 8 hours (P < .05). Of interest, at 75.2 hours, the percentage of free cisplatin was 15% in babies as compared with 9% in non-pregnant women and 10% in pregnant women. This means that the fetus is exposed to 50% higher platinum levels at equal total concentration. Cisplatin protein binding significantly correlated with albumin concentrations at 3.3, 8, and 24 hours (P < .01). Our analysis reveals that pregnancy and fetal changes in cisplatin protein binding are caused in large part by lower albumin levels. The resulting higher levels of free drug in the mother and fetus may increase the risk of toxicity in both.

Adult↗
PubMed1994

Trisomy 8 mosaicism in chorionic villus sampling: case report and counselling issues.

We report an unusual case involving chorionic villus sampling (CVS) and trisomy 8 mosaicism. CVS showed a normal direct preparation while the culture showed mosaicism for trisomy 8. Subsequent amniocentesis revealed only normal chromosomes. A peripheral blood culture after birth revealed low-level trisomy 8 mosaicism. The patient appeared phenotypically and developmentally normal at 30 months of age. We conclude that prenatal counselling for similar cases needs to include the rare but real possibility that chromosome mosaicism detected prenatally may be found postnatally with largely unknown consequences. Secondly, low-level chromosomal mosaicism may be more common than previously recognized. Thirdly, very low-level trisomy 8 mosaicism may be compatible with a normal phenotype but long-term follow-up is required. And lastly, the use of fetal blood sampling is questionable in these cases because the phenotype may not be accurately predicted. Further studies of such cases are needed to address these important and unanswered issues, including the potential implication of mosaicism on academic performance and cognitive functioning.

Adult↗
PubMed1994

Alu elements of the primate major histocompatibility complex.

The chromosomal region constituting the major histocompatibility complex (MHC) has undergone complex evolution that is often difficult to decipher. An important aid in the elucidation of the MHC evolution is the presence of Alu elements (repeats) which serve as markers for tracing chromosomal rearrangements. As the first step toward the establishment of sets of evolutionary markers for the MHC, Alu elements present in selected MHC haplotypes of the human species, the gorilla, and the chimpanzee were identified. Restriction fragments of cosmid clones from the libraries of the three species were hybridized with Alu-specific probes, Alu elements were amplified by the polymerase chain reaction, and the amplification products were sequenced. In some cases, sequences of the regions flanking the Alu elements were also obtained. Altogether, 31 new Alu elements were identified, representing six Alu subfamilies. The average density of Alu elements in the MHC is one element per four kilobases (kb) of sequence. Alu elements have apparently been inserted steadily into the MHC over the last 65 million years (my). On average, one Alu element is inserted into the primate MHC every 4 my. Analysis of the human DR3 haplotype supports its origin by duplication from an ancestral haplotype consisting of DRB1 and DRB2 genes. The sharing of an old Alu element by the DRB1 and DRB2 genes, in turn, supports their divergence from a common ancestor more than 55 my ago.

Animals↗
PubMed1994

Triggers of angina and ST-segment depression in ambulatory patients with coronary artery disease: evidence for an uncoupling of angina and ischemia.

This study evaluated daily-life physical and mental triggers of painful and painless myocardial ischemia and of anginal pain in patients with coronary artery disease (CAD) by using ambulatory electrocardiographic monitoring and a structured diary. Eighty-five percent of ambulant ischemic episodes occurred without chest pain; 66% of anginal pain reports were made in the absence of ischemic ST-segment depression. Chest pain reports in the absence of ischemia could not be attributed to "borderline" ST-segment changes. Compared with silent ischemia, painful ischemia was triggered at higher levels of physical activity (p < 0.05) and at higher levels of self-rated effort and negative emotion (p < 0.05). Painful episodes were associated with greater-magnitude ST depression (p < 0.02), but a substantial percentage of episodes with > or = 2 mm and > or = 3 mm ST depression were silent. These results reveal an uncoupling of anginal symptoms from ambulatory ischemic episodes in patients with CAD during daily life. In addition, specific activities and emotions correlate with the presence or absence of anginal symptoms as much as does the presence of ischemic ST depression. These results may have implications for antiischemic and antianginal treatment strategies.

Activities of Daily Living↗