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Biomedical subjects

H Schmidt

Publications and source records attributed to H Schmidt.

At least 505 records · Page 28Linked to original sources
PubMed1994

Cloning of human and mouse genes homologous to RAD52, a yeast gene involved in DNA repair and recombination.

The RAD52 gene of Saccharomyces cerevisiae is required for recombinational repair of double-strand breaks. Using degenerate oligonucleotides based on conserved amino acid sequences of RAD52 and rad22, its counterpart from Schizosaccharomyces pombe, RAD52 homologs from man and mouse were cloned by the polymerase chain reaction. DNA sequence analysis revealed an open reading frame of 418 amino acids for the human RAD52 homolog and of 420 amino acid residues for the mouse counterpart. The identity between the two proteins is 69% and the overall similarity 80%. The homology of the mammalian proteins with their counterparts from yeast is primarily concentrated in the N-terminal region. Low amounts of RAD52 RNA were observed in adult mouse tissues. A relatively high level of gene expression was observed in testis and thymus, suggesting that the mammalian RAD52 protein, like its homolog from yeast, plays a role in recombination. The mouse RAD52 gene is located near the tip of chromosome 6 in region G3. The human equivalent maps to region p13.3 of chromosome 12. Until now, this human chromosome has not been implicated in any of the rodent mutants with a defect in the repair of double-strand breaks.

Amino Acid Sequence↗
PubMed1994

Prevalence of attaching and effacing Escherichia coli in stool samples from patients and controls.

Enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli (EHEC) have the ability to cause 'attaching and effacing' (AE) lesions; the genes necessary to cause AE in both of these pathogroups have been identified and termed eae. Using colony hybridization, we screened 237 stool samples from patients with diarrhea, and 237 stool samples from age-matched controls for the presence of E. coli carrying eae. Individual colonies harbouring eae could be recovered from 7 (2.9%) of the patient stools, as well as from 6 (2.5%) of the control stools. All these E. coli isolates were positive in the fluorescence actin staining (FAS) test. In addition, all the samples were also probed for Shiga-like toxin (slt) genes and the EPEC adherence factor (EAF) to evaluate whether testing for eae identified all EHEC and class I EPEC. Of the 7 patient samples harbouring E. coli with eae, 4 had E. coli with eae and slt genes, and 2 had E. coli with eae and EAF sequences. In 2 of the 237 patient stools, E. coli which were eae and EAF negative but slt probe positive could be recovered. These 2 E. coli strains were non-reactive in the FAS test. Of the control samples, none of the E. coli strains, including the 6 samples containing eae positive strains, possessed EAF or slt-sequences. In concrete terms, the similar eae incidence found in both E. coli isolates from patients and controls is currently of limited clinical diagnostic value and more importantly, the eae probe could not identify all slt-harbouring E. coli. On the basis of these results, the use of the eae-probe cannot be recommended in preference to the slt probes for the detection of EHEC.

Adhesins, Bacterial↗
PubMed1994

[Diagnosis of urinary tract infections in puerperium].

Urinary tract infections (UTI) are the most frequent nosocomial infections of women during puerperium. Since, in these women, diagnostic accuracy is affected by lochial secretion, suprapubic urinary bladder puncture (SPB) is recommended. Between December 1989 and January 1993, we subjected 903 women to suprapubic urinary bladder puncture (SpBP) at the 4th or 5th day after delivery. A urine culture of SpBP was done in all cases. Semiquantitative leukocyte counts (n = 891) and nitrite test (n = 830) were done on mid-stream urine (MSU). Side by side with microbiological investigation for urinary tract pathogens via SpPB, MSU was performed in 246 cases immediately after SpPB had been carried out. Leukocyte counts were also estimated in SpBP urine samples. In 370 (41.1%) of 903 SpBP, one or more microorganisms were cultivated. Only 36 (4.0%) of 903 women showed UTI symptoms. Microorganisms were detectable via SpBP in only 26 (72.7%) of these 36 symptomatic patients. Vaginal-operative or secondary caesarean section are related to an increased UTI rate (p < 0.001). UTI were also significantly (p < 0.0001) more frequent in women subjected to catheterisation sub partu (54.5%) compared to no catheterism (24.4%). No significant differences between the number of leukocytes in MSU sediment and the SpBP findings were seen. Semiquantitative leukocyte counts in SpBP offered a significantly (p < 0.001) increased number of leukocytes in cases with microorganism detection in SpBP irrespective of MSU findings. These results justify the designation of uterine tract infection also in the absence of complaints as "infection" and not common "bacteriuria".(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteria↗
PubMed1994

[The determination of the normal values of blood flow velocities in the superior mesenteric artery of premature and term newborns with duplex sonography].

In a prospective study blood flow velocity measurements were performed in 110 "healthy" newborn with duplex Doppler sonography in the superior mesenteric artery to obtain standard values. In 49 of these neonates 15, 30, and 45 minutes following feeding examinations were performed. Peak systolic flow velocity, end systolic flow velocity, time average flow velocity and time average maximum flow velocity were determined, the resistance and Pourcelot index as well as the volume blood flow were calculated. The children's gestational age was 27-42 weeks, the postnatal age was 2-68 days and the body weight was 920-4190 g. All measured blood flow velocities showed a synchronous relation to feeding with an increase of blood flow velocity between 15 and 30 minutes and a decrease of blood flow velocity after feeding. The ascertained fasting measurements from 91 newborn (last feeding > 3 h < 12 h) differed significantly from the ascertained basic fasting measurements (last feeding > 12 h) from 15 infants.

Blood Flow Velocity↗
PubMed1994

[The corroborative capacity and value of imaging diagnosis and duplex sonography in newborn infants with necrotizing enterocolitis].

In this prospective study 101 sonographic examinations of superior mesenteric artery blood flow velocities, indices and blood flow volume were consecutively performed in 41 newborn to diagnose necrotising enterocolitis (NEC). Intramural and portal venous gas was also estimated. The artifacts of air in the AMS and the portal system were examined in an experimental study. The results of sonography and abdominal radiography were equivalent in the case of definitely established NEC, portal venous gas being more sensitive to detection by sonography. Abdominal radiography was indicated as the primary examination in case of a clearly identified clinical course. Sonography should be performed so that fewer radiographs will be required in the following course of disease.

Abdomen↗
PubMed1994

The mutator gene swi8 effects specific mutations in the mating-type region of Schizosaccharomyces pombe.

The swi8+ gene of Schizosaccharomyces pombe appears to be involved in the termination step of copy synthesis during mating-type (MT) switching. Mutations in swi8 confer a general mutator phenotype and, in particular, generate specific mutations in the MT region. Sequencing of the MT cassettes of the h90 swi8-137 mutant revealed three altered sites. One is situated at the switching (smt) signal adjacent to the H1 homology box of the expression locus mat1:1. It reduces the rate of MT switching. The alteration at the smt signal arose frequently in other h90 swi8 strains and is probably caused by gene conversion in which the sequence adjacent to the H1 box of mat2:2 is used as template. This change might be generated during the process of MT switching when hybrid DNA formation is anomalously extended into the more heterologous region flanking the H1 homology box. In addition to the gene conversion at mat1:1, two mutations were found in the H3 homology boxes of the silent cassettes mat2:2 and mat3:3.

Base Sequence↗
PubMed1994

Variants of Shiga-like toxin II constitute a major toxin component in Escherichia coli O157 strains from patients with haemolytic uraemic syndrome.

The prevalence and genotype of Shiga-like toxins (SLTs) in Escherichia coli (O)157 strains from patients in Germany with haemolytic uraemic syndrome (HUS) were investigated. This was done by PCR amplification of the B-subunit genes with two primer pairs--one complementary to slt-IB, and the other homologous to both slt-IIB and slt-IIvB sequences. To distinguish between slt-II and slt-IIv, the amplified DNA was digested with restriction endonucleases HaeIII and FokI. Of the 38 strains examined, 17 harboured sequences for slt-IIv; four contained only slt-IIv, three carried both slt-IIv and slt-I, and 10 strains had slt-IIv and slt-II. A further three genotypes (slt-I, slt-II, slt-I/slt-II) were found in the remaining 21 strains resulting in a total of six slt genotypes. To determine whether the slt genes were expressed, and whether genotypes correlated with phenotypes, all strains were subjected to cytotoxicity assays and colony ELISA. All 38 strains displayed cytotoxic activity to Vero cells in similar quantities. The SLT-I-specific monoclonal antibody (MAb)13C4 reacted with all 10 strains in which slt-I sequences were identified. Colony blot ELISA with the SLT-II specific MAb11E10 detected 27 of 28 strains with slt-II sequences, but did not react with any of the seven strains that carried slt-IIv, or slt-I and slt-IIv. The high SLT variability shown here has diagnostic implications and may well have consequences for the host response in infections associated with these pathogens.

Animals↗
PubMed1994

Polymerase chain reaction for the identification of Porphyromonas gingivalis collagenase genes.

Porphyromonas gingivalis has been shown to exhibit genetic diversity possibly resulting in variation of virulence. In the present study a potential virulence factor was targeted for the detection of P. gingivalis. A 548 bp fragment of the collagenase gene (prtC) from Porphyromonas gingivalis ATCC 33277 was amplified by polymerase chain reaction (PCR) using oligonucleotides derived from the middle portion of prtC. From 16 of 21 clinical P. gingivalis strains, a PCR product of similar size to the prtC could be obtained. These 16 P. gingivalis strains were confirmed as positive for prtC using DNA hybridization with a digoxigenin-labeled prtC PCR product as a probe. In 12 of the 16 prtC positive strains, the restriction analysis of the PCR products revealed fragment patterns identical to the known sequence. In the other 4 prtC positive strains, 4 distinct patterns were found. Of these strains, nucleotide sequence analysis of a 400 bp PCR product stretch revealed 79.1%, 83.0%, 84.8 and 89.5% homology with the known nucleotide sequence for this specific region. Sequence analysis of the PCR products from the ATCC 33277 strain demonstrated 93.7% homology. The limit of detection for the PCR was about 100 organisms. None of the other 48 tested strains of 16 bacterial species derived from oral and extraoral infections yielded a PCR product. The PCR was also used for the detection of prtC sequences in dental plaque. Our data indicate that not all P. gingivalis strains have prtC. Nucleotide heterogeneity exists among P. gingivalis with prtC.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Proteins↗
PubMed1994

Growth and skeletal maturation in children with phenylketonuria.

Growth and skeletal maturation was evaluated in 82 children participating in the German Collaborative Study of Children Treated for Phenylketonuria (PKU). Height, weight, head circumference and bone age were recorded at regular intervals for the first 6 years of life. The mean SD score (SDS) for height was not significantly different from zero at study entry, but decreased mainly during the second year of life to a nadir of -0.78 in boys and -0.54 in girls at 2.5 years. During the subsequent years, a significant trend towards a regain of height SDS was noted in both sexes. Weight-for-height SDS was close to zero in both sexes, with a significant continuous increasing trend throughout the observation period. Head circumference SDS decreased in boys during the first year of life from -0.28 to -0.68, whereas girls showed only a minor change. During the further follow-up period, head circumference SDS remained at approximately -0.3 in boys and 0.0 in girls. While the mean verbal and performance IQ of the total study population at 5 and 6 years of age did not differ from a group of 212 healthy non-PKU children, patients with a head circumference SDS less than the population median at 2 years of age exhibited poorer cognitive abilities at school age than those patients with a relative head size greater than the population median. The children with a head circumference less than the median at 2 years had smaller head sizes already at birth; in addition, the change in relative head size during the first 2 years was correlated significantly with cognitive abilities at school age in boys. Mean bone age was identical to chronological age at each time point of observation. The rate of maturation was one year of bone age per year of chronological age. No correlation between phenylalanine intake or phenylalanine concentrations and the rates of body or head growth or skeletal maturation could be established. We conclude that despite adequate weight gain, moderate growth retardation occurred during the first 2 years of life in this group of children treated for PKU. Growth was more compromised in boys than in girls and tended to be compensated during later follow-up. Early infantile head circumference and growth appear to be predictors of cognitive development.

Age Determination by Skeleton↗
PubMed1994

Development of granulomas and vascular fibrocellular proliferation in the lungs of pigs receiving long-term lipid-based parenteral nutrition.

The lungs of pigs receiving long-term total parenteral nutrition (TPN) have been studied. A total of 20 pigs were tentatively infused with TPN through central venous catheters for 7 weeks. To secure adequate nutrition and gastrointestinal absorbance of nutrients, an additional full oral diet was given to eight of these animals. Fifteen control animals received Ringer solution through central venous catheters in addition to the oral diet. All animals infused with TPN for 7 weeks developed lung granulomas, a finding not observed in control animals. No lung granulomas were seen in three TPN animals sacrificed after 3 to 5 weeks owing to illness. All TPN animals also developed tissue reactions suggesting long-standing lung vascular inflammation. Similar vascular changes were seen in seven control animals that had bacterial infection or endotoxemia. The total amount of neutral fat in lung homogenate tended to increase in animals given TPN, and the linoleic acid content was significantly increased. In conclusion, long-term TPN caused lung granulomas in pigs. Vascular damage seemed to be accelerated by bacterial infection as well as by the TPN. The tissue reactions presumably involved long-term activation of monocytes/macrophages.

Animals↗
PubMed1994

Nucleotide sequence analysis of enteropathogenic Escherichia coli (EPEC) adherence factor probe and development of PCR for rapid detection of EPEC harboring virulence plasmids.

The 1-kb BamHI-SalI fragment from plasmid pMAR2 termed the enteropathogenic Escherichia coli (EPEC) adherence factor (EAF) probe was cloned in pUC19 and pK18. The nucleotide sequence of this fragment was determined, and a set of primers was designed to amplify a 397-bp region associated with pMAR2 by PCR. An analysis of the whole EAF sequence with database libraries indicated no significant homology to any known genes. However, between bases 701 and 787 of the fragment, an 82.8% homology between the EAF and the insertion sequence IS630 of Shigella sonnei exists. The results of PCR with primers of the EAF sequence demonstrated that all of the 151 EAF probe-positive EPEC strains with localized adherence to HEp-2 cells yielded positive EAF PCR results. In contrast, none of the 277 EAF probe-negative strains reacted to the EAF PCR. In addition, the PCR assay was successfully used to generate vector-free digoxigenin-labeled EAF fragments that gave valid results in colony blot hybridization assays. The EAF PCR appears to be a specific and efficient method for the detection of EPEC strains carrying the EAF plasmids.

Adhesins, Escherichia coli↗
PubMed1994

Demonstration of Borrelia burgdorferi DNA in urine samples from healthy humans whose sera contain B. burgdorferi-specific antibodies.

Since the possibility of asymptomatic infection with Borrelia burgdorferi has been suggested by a positive serology found in healthy subjects, we hypothesized that these subjects might excrete borrelial DNA sequences in urine as happens in patients with Lyme borreliosis. We found borrelial sequences by nested PCR in the urine samples from 3 of 13 healthy B. burgdorferi antibody-positive adults but not in urine samples from 79 antibody-negative healthy controls. After therapy with doxycycline, the urine samples were repeatedly negative for B. burgdorferi DNA. We conclude that urinary excretion of borrelial DNA sequences may occur in seropositive healthy subjects during asymptomatic infection. Demonstration of such sequences in urine must be interpreted cautiously and may not necessarily prove a borrelial cause of disease.

Adult↗
PubMed1994

The rad16 gene of Schizosaccharomyces pombe: a homolog of the RAD1 gene of Saccharomyces cerevisiae.

The rad10, rad16, rad20, and swi9 mutants of the fission yeast Schizosaccharomyces pombe, isolated by their radiation sensitivity or abnormal mating-type switching, have been shown previously to be allelic. We have cloned DNA correcting the UV sensitivity or mating-type switching phenotype of these mutants and shown that the correcting DNA is encompassed in a single open reading frame. The gene, which we will refer to as rad16, is approximately 3 kb in length, contains seven introns, and encodes a protein of 892 amino acids. It is not essential for viability of S. pombe. The predicted protein is the homolog of the Saccharomyces cerevisiae RAD1 protein, which is involved in an early step in excision-repair of UV damage from DNA. The approximately 30% sequence identity between the predicted proteins from the two yeasts is distributed throughout the protein. Two-hybrid experiments indicate a strong protein-protein interaction between the products of the rad16 and swi10 genes of S. pombe, which mirrors that reported for RAD1 and RAD10 in S. cerevisiae. We have identified the mutations in the four alleles of rad16. They mapped to the N-terminal (rad10), central (rad20), and C-terminal (rad16 and swi9) regions. The rad10 and rad20 mutations are in the splice donor sequences of introns 2 and 4, respectively. The plasmid correcting the UV sensitivity of the rad20 mutation was missing the sequence corresponding to the 335 N-terminal amino acids of the predicted protein. Neither smaller nor larger truncations were, however, able to correct its UV sensitivity.

Amino Acid Sequence↗
PubMed1994

Markedly accelerated catabolism of apolipoprotein A-II (ApoA-II) and high density lipoproteins containing ApoA-II in classic lecithin: cholesterol acyltransferase deficiency and fish-eye disease.

Classic (complete) lecithin:cholesterol acyltransferase (LCAT) deficiency and Fish-eye disease (partial LCAT deficiency) are genetic syndromes associated with markedly decreased plasma levels of high density lipoprotein (HDL) cholesterol but not with an increased risk of atherosclerotic cardiovascular disease. We investigated the metabolism of the HDL apolipoproteins (apo) apoA-I and apoA-II in a total of five patients with LCAT deficiency, one with classic LCAT deficiency and four with Fish-eye disease. Plasma levels of apoA-II were decreased to a proportionately greater extent (23% of normal) than apoA-I (30% of normal). In addition, plasma concentrations of HDL particles containing both apoA-I and apoA-II (LpA-I:A-II) were much lower (18% of normal) than those of particles containing only apoA-I (LpA-I) (51% of normal). The metabolic basis for the low levels of apoA-II and LpA-I:A-II was investigated in all five patients using both exogenous radiotracer and endogenous stable isotope labeling techniques. The mean plasma residence time of apoA-I was decreased at 2.08 +/- 0.27 d (controls 4.74 +/- 0.65 days); however, the residence time of apoA-II was even shorter at 1.66 +/- 0.24 d (controls 5.25 +/- 0.61 d). In addition, the catabolism of apoA-I in LpA-I:A-II was substantially faster than that of apoA-I in LpA-I. In summary, genetic syndromes of either complete or partial LCAT deficiency result in low levels of HDL through preferential hypercatabolism of apoA-II and HDL particles containing apoA-II. Because LpA-I has been proposed to be more protective than LpA-I:A-II against atherosclerosis, this selective effect on the metabolism of LpA-I:A-II may provide a potential explanation why patients with classic LCAT deficiency and Fish-eye disease are not at increased risk for premature atherosclerosis despite markedly decreased levels of HDL cholesterol and apoA-I.

Adult↗
PubMed1994

A wet smear criterion for bacterial vaginosis.

OBJECTIVE: To evaluate a simpler criterion for bacterial vaginosis than the recommended criterion of Amsel. DESIGN: Cross-sectional study comparing diagnostic data with a recommended standard. SETTING: Danish general practice. PARTICIPANTS: 595 non-pregnant women aged 15-49 years who were gynaecologically examined, were divided into two groups in accordance with a complaint of vaginal discharge. MAIN OUTCOME MEASURE: The associations of pH > 4.5, homogeneity, amine odour, clue cells, and wet smear assessment of predominance of lactobacilli, Gardnerella vaginalis, motile rods with the criterion of Amsel. RESULTS: The four components of Amsel showed a considerable variation of association. Predominance of GV was more highly associated than several of the components. The association of a wet smear criterion composed of the presence of clue cells in combination with predominance of Gardnerella vaginalis and absence of lactobacilli was superior to other combinations. In the group without a complaint of vaginal discharge, preceding screening with use of a pH-measurement seemed advantageous. CONCLUSION: The wet smear criterion seems suitable for use in general practice. But before a definitive establishment, calculation of the reproducibility of interpretation of this criterion, preferably with a more explicit setting of the components, has to be carried out.

Adolescent↗