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Biomedical subjects

E Harms

Publications and source records attributed to E Harms.

At least 91 records · Page 5Linked to original sources
PubMed1988

[Results following anterior levator-plasty].

In this study we report about 172 patients who underwent modified anterior repair in the time from october 1983 up to april 1985 because of vaginal prolapse and/or incontinence. 125 patients could be examined clinically, in the middle 14.2 months after operation. In 64% the modified anterior repair was combined with a colporrhaphia anterior and hysterectomy, and in 20% a colporrhaphia posterior was made additionally. Dehiscence of the anterior vaginal wall occurred in 19.2%, severe pelvic infections in 8.8%. Because of bleeding complications a chirurgical intervention was necessary in 5.6%. A descent of the anterior vaginal wall was seen in 30.4%, 76.8% of the women developed a descent of the posterior vaginal wall, and 8% a stenosis of the vagina. Problems at sexual intercourse were frequent, followed by pain in the lower abdomen. The principle of the operation is presented including the changed anatomy. Many of the adverse effects can be understood and the necessity of prophylaxis of a descensus of posterior vaginal wall was described.

Adult↗
PubMed1988

Translational coupling between the ilvD and ilvA genes of Escherichia coli.

The hypothesis that translation of the ilvD and ilvA genes of Escherichia coli may be linked has been examined in strains in which lacZ-ilvD protein fusions are translated in all three reading frames with respect to ilvD. In these strains, the nucleotide sequence was altered to obtain premature termination of ilvD translation, and in one strain translation termination of ilvD DNA occurred two bases downstream of the ilvA initiation codon. In the wild-type strain, the ilvD translation termination site was located two bases upstream of the ilvA start codon. In each of the mutant strains, expression of ilvA, as determined by the level of threonine deaminase activity, was strikingly lower than in the wild-type strain. The data suggest that expression of ilvD and ilvA is translationally coupled. By inserting a promoterless cat gene downstream of ilvA, it was shown that the differences in enzyme activity were not the result of differences in the amount of ilvA mRNA produced.

DNA Mutational Analysis↗
PubMed1987

Fanconi-Bickel syndrome.

Clinical, biochemical, functional and morphological data are presented in nine infants, children and adults, with Fanconi-Bickel syndrome. Long-term follow-up studies show severe growth retardation, partly compensated for by late onset of puberty. Glomerular filtration rate is normal or slightly decreased. Renal tubular dysfunction is characterized by a specific pattern of impaired proximal tubular transport mechanisms, with marked impairment of glucose transport. The utilization of glucose and galactose is defective, whereas fructose metabolism seems to be normal. Glycogenosis of the liver may be an epiphenomenon. Glycogen accumulation in the kidney is limited to the proximal tubule, with maximal levels in the straight part. The Fanconi-Bickel syndrome is a defined clinical entity which is distinguished from other inherited metabolic diseases by complex defects of renal tubular transport and other forms of glycogenosis.

Adolescent↗
PubMed1987

Prenatal diagnosis of inborn errors of metabolism with renal manifestations.

Prenatal diagnosis has become increasingly important for the prevention of kidney disease due to inborn errors of metabolism. The indications for prenatal diagnosis in a specific disease depend on the degree of renal involvement and the concomitant pathology of other organs, but especially of the central nervous system. The availability of successful postnatal therapy reduces the necessity for prenatal diagnosis. The determination of enzyme activities, the demonstration of storage products in fetal tissues and the analysis of amniotic fluid components are the biochemical methods most frequently used for prenatal detection. Recently, recombinant DNA techniques have enlarged the spectrum of inherited disorders that can be diagnosed before birth.

Female↗
PubMed1987

Rapid analytical and preparative isolation of functional endosomes by free flow electrophoresis.

Endosomes are prelysosomal organelles that serve as an intracellular site for the sorting, distribution, and processing of receptors, ligands, fluid phase components, and membrane proteins internalized by endocytosis. Whereas the overall functions of endosomes are increasingly understood, little is known about endosome structure, composition, or biogenesis. In this paper, we describe a rapid procedure that permits analytical and preparative isolation of endosomes from a variety of tissue culture cells. The procedure relies on a combination of density gradient centrifugation and free flow electrophoresis. It yields a fraction of highly purified, functionally intact organelles. As markers for endosomes in Chinese hamster ovary cells, we used endocytosed horseradish peroxidase, FITC-conjugated dextran, and [35S]methionine-labeled Semliki Forest virus. Total postnuclear supernatants, crude microsomal pellets, or partially purified Golgi fractions were subjected to free flow electrophoresis. Endosomes and lysosomes migrated together as a single anodally deflected peak separated from most other organelles (plasma membrane, mitochondria, endoplasmic reticulum, and Golgi). The endosomes and lysosomes were then resolved by centrifugation in Percoll density gradients. Endosomes prepared in this way were enriched up to 70-fold relative to the initial homogenate and were still capable of ATP-dependent acidification. By electron microscopy, the isolated organelles were found to consist of electron lucent vacuoles and tubules, many of which could be shown to contain an endocytic tracer (e.g., horseradish peroxidase). SDS PAGE analysis of integral and peripheral membrane proteins (separated from each other by condensation in Triton X-114) revealed a unique and restricted subset of proteins when compared with lysosomes, the unshifted free flow electrophoresis peak, and total cell protein. Altogether, the purification procedure takes 5-6 h and yields amounts of endosomes (150-200 micrograms protein) sufficient for biochemical, immunological, and functional analysis.

Animals↗
PubMed1987

Role of codon choice in the leader region of the ilvGMEDA operon of Serratia marcescens.

Leucine participates in multivalent repression of the Serratia marcescens ilvGMEDA operon by attenuation (J.-H. Hsu, E. Harms, and H.E. Umbarger, J. Bacteriol. 164:217-222, 1985), although there is only one single leucine codon that could be involved in this type of control. This leucine codon is the rarely used CUA. The contribution of this leucine codon to the control of transcription by attenuation was examined by replacing it with the commonly used leucine codon CUG and with a nonregulatory proline codon, CCG. These changes left intact the proposed secondary structure of the leader. The effects of the codon changes were assessed by placing the mutant leader regions upstream of the ilvGME structural genes or the cat gene and measuring acetohydroxy acid synthase II, transaminase B, or chloramphenicol acetyltransferase activities in cells grown under limiting and repressing conditions. The presence of the common leucine codon in place of the rare leucine codon reduced derepression by about 70%. Eliminating the leucine codon by converting it to proline abolished leucine control. Furthermore, a possible context effect of the adjacent upstream serine codon on leucine control was examined by changing it into a glycine codon.

Base Sequence↗
PubMed1985

Localization of renin (EC 3.4.23) and converting enzyme (EC 3.4.15.1) in nerve endings of rat brain.

Synaptosomes and lysosomes of rat brain were separated by differential centrifugation and a two-step gradient centrifugation with colloidal silica-gel (Percoll). The organelles were identified by the measurement of established marker-enzymes and by electronmicroscopy. Renin activity, measured by radioimmunoassay for angiotensin I (ANG I), was localized in the synaptosomes and cathepsin D-activity was found in the lysosomal fraction. Converting-enzyme activity was present in the renin-containing synaptosomes. Part of the brain renin activity could be activated by pre-incubation with trypsin. Affinity chromatography of an organelle-enriched brain fraction was carried out using a caseinyl-sepharose column and resulted in the separation of renin from cathepsin D activity; the renin peak was inhibited by antibodies raised against rat kidney renin. We conclude, that the formation of ANG I and its activation to angiotensin II (ANG II) by converting enzyme is possible in synaptosomes. This adds further evidence to an intraneuronal synthesis of ANG I and ANG II in the brain and is in support of previous results demonstrating an intraneuronal localization of the components of the brain renin-angiotensin system.

Animals↗
PubMed1985

[Suprapubic urinary diversion following gynecologic operations].

In a prospective study the results of postoperative urinary discharge by suprapubic catheterisation (SC) are compared with those of transurethral catheterisation (TC) in a randomised collective of gynaecological patients after vaginal hysterectomy with front (resp. front and back) plastic. During November 1979 and September 1980 157 patients were examined, 88 patients by suprapubic catheterisation and 69 by transurethral catheterisation, with random distribution. Additionally, 430 other patients with suprapubic aspiration after vaginal hysterectomy with front and/or back plastic performed between September 1980 and October 1982 were examined retrospectively especially under the aspect of possible complications. The advantages of suprapubic urinary discharge are shown in the significantly reduced rate of primary infections. 20.5 per cent of infections are opposed to 67.1 per cent in the comparative group with TC. Spontaneous miction was possible in the SC group two days earlier than in the group of TC (5.2 days opposed to 7.35 days). Additional drug treatment for restitution of the bladder function was more often necessary for patients with TC, although the results with 43.5 per cent against 29.5 per cent in the SC collective are not significant. The number of renewed catheterisations after removal of the catheter was higher by 27 per cent in the group of patients with SC. Subjective complaints were stated by patients with TC in 66.7 per cent against 18.2 per cent of women with SC. This result is significant. In the TC collective there were significantly more patients with leucocyturia (88.5 per cent against 48.7 per cent). Antibiotic treatment of an urinary tract infection with typical symptoms was necessary in 35.8 per cent for patients with TC and in 14.1 per cent for women with SC.(ABSTRACT TRUNCATED AT 250 WORDS)

Anti-Bacterial Agents↗
PubMed1985

Comparison of the regulatory regions of ilvGEDA operons from several enteric organisms.

The nucleotide sequence preceding the ilvGEDA operon has been examined and compared in five enteric organisms. The sequence in Escherichia coli B was identical to the earlier-described strain K-12 sequence. The sequences of Salmonella typhimurium and Klebsiella aerogenes were remarkably similar to that of E. coli and identical in that part of the leader region that specified the putative 32-amino-acid peptide. Thus, identical secondary structures could be postulated for the leaders of all three organisms, and regulation of operon expression could be like that postulated earlier for E. coli. Different secondary structures had to be postulated for the leader transcripts of Edwardsiella tarda and Serratia marcescens. Control of attenuation of the operon in these organisms by the level of leucyl tRNA could be explained only if ribosome stalling occurred at a single leucine codon. In both organisms, that single leucine codon is the rarely used CUA rather than the CUG that is in E. coli, S. typhimurium, and K. aerogenes.

Base Sequence↗
PubMed1985

Leucine regulation of the ilvGEDA operon of Serratia marcescens by attenuation is modulated by a single leucine codon.

The effect of leucine limitation and of restricted leucine tRNA charging on the expression of the ilvGEDA operon of Serratia marcescens was examined. In this organism, the ilv leader region specifies a putative peptide containing only a single leucine codon that could be involved in leucine-mediated control by attenuation (E. Harms, J.-H. Hsu, C. S. Subrahmanyam, and H. E. Umbarger, J. Bacteriol. 164:207-216, 1985). A plasmid (pPU134) containing the DNA of the S. marcescens ilv control region and three of the associated structural genes was studied as a single chromosomal copy in an Escherichia coli strain auxotrophic for all three branched-chain amino acids. The S. marcescens ilv genes responded to a multivalent control similar to that found in other enteric organisms. Furthermore, the S. marcescens ilv genes were derepressed when the charging of leucine tRNA was restricted in a leuS derivative of E. coli that had been transformed with pPU134. It was concluded that ribosome stalling leading to deattenuation is not dependent on either tandem or a consecutive series of codons for the regulatory amino acid. However, the fact that the single leucine codon is a less frequently used codon (CUA) may be important. The procedure for obtaining the cloned ilv genes in single chromosomal copy exploited the dependence of ColE1 replicons on the polA gene. The cloning experiments also revealed a branched-chain amino acid-glutamate transaminase in S. marcescens that is different from transaminase B.

Codon↗
PubMed1985

Lysosomal cystine storage in cystinosis and mucolipidosis type II.

Cultured fibroblasts from mucolipidosis II (ML-II) patients demonstrated an elevated cystine content which increased with time in culture compared to fibroblasts from cystinotic patients or normal controls under the same conditions. In both cystinotic and ML-II cells the increased levels of cystine could be derived either from endogenous proteolysis or from in vitro supplementation of the cultured cells with cysteine-glutathione mixed disulfide. Cystine was depleted from both cell types by cysteamine. When cysteamine was replaced with complete medium, the cystine reaccumulated in both cystinotic and ML-II cells within 24 h, although a lag of 4 h was seen with ML-II cells. The intracellular location of the increased cystine in cultured fibroblasts was examined utilizing free-flow electrophoresis and found to be in the purified population of secondary lysosomes of both cystinotic and ML-II cells. White blood cell and hepatic cystine, which was greatly increased in cystinotic patients, was not elevated in ML-II patients. Compared to normal control fibroblasts the efflux of cystine from isolated granular fractions was virtually absent in cystinotic fibroblasts and considerably reduced in ML-II fibroblasts. The examination of such similarities and differences in cystine accumulation and transport in tissues from cystinotic and ML-II patients has provided some insight into the defects in these diseases.

Cells, Cultured↗
PubMed1985

[Prenatally diagnosed fetal hydrothorax].

A predominantly unilateral hydrothorax in an unborn child was diagnosed by sonography during the 34th gestational week. Because of a pathologic cardiotocogram it was necessary to make a primary caesarean section one day later. The premature baby was intubated immediately and the pleural discharge was removed. Later developmental of the child was uneventful. The hydrothorax was interpreted as a partial factor of a hydrops fetalis. Controlling the fetal pleural discharge by ultrasound examination and regular cardiotocogram a caesarean section should be done between the 34th and 36th week of pregnancy.

Cesarean Section↗
PubMed1984

[Malaria relapse in the puerperium].

During the last six years we observed four malaria relapses shortly after delivery. In three cases the relapse occurred subsequent to Caesarean section, and in one case shortly after normal uncomplicated delivery. Case histories revealed that these women had previously stayed in the tropical or subtropical zones. The longest period without relapse was five years, the shortest interval four months. In all cases, microscopic examination revealed a relapse of Malaria tertiana. All the patients were cured within a short time by administration of chloroquine; the clinical signs disappeared. If, in fever of unknown origin, malaria is suspected, diagnosis and therapy will be an easy matter. Good anamnesis will provide a pointer to appropriate therapeutic action.

Adult↗
PubMed1984

Antibodies against lysosomal membranes reveal a 100,000-mol-wt protein that cross-reacts with purified H+,K+ ATPase from gastric mucosa.

Specific antibodies against lysosomal membranes were prepared by using techniques previously described (Louvard, D., H. Reggio, and G. Warren, 1982, J. Cell Biol., 92:92-107) for obtaining organelle-specific antibodies. The purified antibodies stained an acidic vacuolar compartment as shown by double-labeling experiments with acridine orange and indirect immunofluorescence. Characterization of the antibodies by immunoreplica methods revealed one major protein of approximately 100,000 mol wt. The antibodies cross-reacted with purified H+,K+ ATPase from pig gastric mucosa, the enzyme responsible for HCl secretion, but not with ATPases transporting other ions. They may therefore recognize a component of the proton pump involved in the acidification of lysosomes. As was expected, secondary lysosomes contained immunoreactive antigen, as determined by the fine-structural localization of reaction product for peroxidase or immunogold probes in several cell types. The antigen was also found in vacuoles containing phagocytosed bacteria in macrophages so it is present in at least some of the compartments of an endocytic pathway. In liver, the antigen was present in small amounts on the plasma membrane and in large amounts in some coated vesicles (near the sinusoidal surface of hepatocytes), putative endosomes, two cisternae on the cis side of the Golgi complex, adjacent vesicles and vacuoles, and pericanalicular dense bodies. In summary, the antigen seems to be present in those compartments that have recently been demonstrated to be acidified by an ATP-driven pump.

Adenosine Triphosphatases↗
PubMed1983

The intralysosomal pH in cultured human skin fibroblasts in relation to cystine accumulation in patients with cystinosis.

The intralysosomal pH in cultured skin fibroblasts from a patient with cystinosis and from control fibroblasts was estimated by the method of Hollemans et al. (Hollemans, M., Oude Elferink, R.P.J., de Groot, P.G., Strijland, A. and Tager, J.M. (1981) Biochim. Biophys. Acta 643, 140-151). 1. The intralysosomal pH in cystinotic fibroblasts (5.37 +/- 0.04; n = 12) was almost identical to that in the control fibroblasts (5.27 +/- 0.06; n = 10). 2. After depletion of cystine by pretreatment of cystinotic fibroblasts with 1 mM cysteamine, there was no significant change in the intralysosomal pH. Incubation of either cystinotic or control fibroblasts with higher concentrations of cysteamine (10 or 25 mM) led to an increase in intralysosomal pH similar to that seen with other weak bases. 3. The fractional volume of the lysosomes in cystinotic fibroblasts (0.100 +/- 0.012; n = 4) was higher than that in control fibroblasts (0.039 +/- 0.010; n = 4). 4. It is concluded, in contrast to Jonas et al. (Jonas, A.J., Smith, M.L. and Schneider, J.A. (1982) J. Biol. Chem. 257, 13185-13188), that the lysosomal proton-translocating ATPase (Okhuma, S., Moriyama, Y. and Takano, T. (1982) Proc. Natl. Acad. Sci. USA 79, 2758-2762) is not impaired in cystinotic fibroblasts.

Cysteamine↗