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Biomedical subjects

R Herzog

Publications and source records attributed to R Herzog.

At least 19 recordsLinked to original sources

[Problems and complications of leg lengthening with the Wagner apparatus].

Since 1971, we have performed 189 leg lengthening procedures using the Wagner method at our institution. The results obtained in the first 26 cases (1971-1973) showed a high complication rate, which led us to reconsider the indications for this procedure. In the present paper, we analyze the results of 37 leg lengthening procedures carried out in 32 patients during the last 10 years (1981-1990) in the children's unit of the orthopedic department of the University of Basle. We found a complication rate of 78%, and in 46% of cases there was more than one major complication. We did not distinguish between "complications" and "problems", because such distinctions are of little importance to the patient. The average age at the time of surgery was 14.8 years, and the average increase in length was 4.3 (2.2-9.2) cm. For each 1 cm of lengthening, an average of 21 days in hospital and 64 days of reduced weight-bearing were needed. Our conclusion is that the Wagner method makes it possible to attain the goal of leg lengthening, but the second step cannot reduce the length of stay in hospital or the length of time the patient needs the help of crutches. Bone remodeling is disturbed. Our preliminary experience with the Ilizarov method is more encouraging.

Adolescent

Two-dimensional high-performance liquid chromatography at low ng/ml levels of the anti-proliferative agent B859-35 in serum with automated sample clean-up, solid-phase trapping and ultraviolet detection.

An automated non-chiral high-performance liquid chromatographic method is described for the determination of the new anti-proliferative agent B859-35 in serum. This method employs sample clean-up of 1 ml of biofluid by liquid-solid extraction with the AASP (Advanced Automatic Sample Preparation) system. First separation is achieved on a LiChrospher-60-RP-Select-B column. A fraction of this elute is then collected by solid-phase trapping. Thereafter, the final chromatogram is developed on a narrow-bore Hypers1-CPS column and quantified with ultraviolet detection at 230 nm. The limit of quantitation of the assay is 250 ng/ml. Linearity was proven in the range 0.25-100 ng/ml. Typical figures for precision at these concentrations are 7.4 and 3.3%, and for accuracy 8.0 and 1.3%, respectively. An application of this method to the study of pharmacokinetics of B859-35 in serum samples of cancer patients is given.

Antineoplastic Agents

Complete nucleotide sequence of the gene for human heparin cofactor II and mapping to chromosomal band 22q11.

Heparin cofactor II (HCII) is a 66-kDa plasma glycoprotein that inhibits thrombin rapidly in the presence of dermatan sulfate or heparin. Clones comprising the entire HCII gene were isolated from a human leukocyte genomic library in EMBL-3 lambda phage. The sequence of the gene was determined on both strands of DNA (15,849 bp) and included 1749 bp of 5'-flanking sequence, five exons, four introns, and 476 bp of DNA 3' to the polyadenylation site. Ten complete and one partial Alu repeats were identified in the introns and 5'-flanking region. The HCII gene was regionally mapped on chromosome 22 using rodent-human somatic cell hybrids, carrying only parts of human chromosome 22, and the chronic myelogenous leukemia cell line K562. With the cDNA probe HCII7.2, containing the entire coding region of the gene, the HCII gene was shown to be amplified 10-20-fold in K562 cells by Southern analysis and in situ hybridization. From these data, we concluded that the HCII gene is localized on the chromosomal band 22q11 proximal to the breakpoint cluster region (BCR). Analysis by pulsed-field gel electrophoresis indicated that the amplified HCII gene in K562 cells maps at least 2 Mbp proximal to BCR-1. Furthermore, the HCII7.2 cDNA probe detected two frequent restriction fragment length polymorphisms with the restriction enzymes BamHI and HindIII.

Amino Acid Sequence

Large-scale physical mapping within the region 22q12.3-13.1 in meningioma.

The lack of physical mapping data strongly restricts the analysis of the meningioma chromosomal region that was assigned to the bands 22q12.3-qter. Recently, we reported a new marker D22S16 for chromosome 22 that was assigned to the region 22q13-qter by in situ hybridization. Utilizing somatic cell hybrids we now sublocalized the marker D22S16 within the band region 22q12-13.1, thus placing it in the vicinity of the gene for the platelet derived growth factor (PDGFB). A physical map was established for the regions surrounding the PDGFB gene and the D22S16 marker. By means of pulsed-field gel electrophoresis (PFGE) D22S16 and PDGFB were found to be physically linked within 900 kb. We also identified two CpG clusters bordering the PDGFB gene. For the enzyme NotI, a variation of the PDGFB restriction pattern was found between different individuals. PFGE analysis of the two loci (PDFGB and D22S16) failed to identify major rearrangements in meningioma.

Animals

[Failures of knee joint prostheses. An analysis of knee prostheses and component revisions, 1980-1987].

Total knee arthroplasty has become more common in the last 20 years as a result of the continuous improvement in prosthetic design and operative techniques. Therefore, there have also been more failures. To show the reasons for these failures and the difficulties and results of revision surgery we studied the 48 uni- or tricompartmental revisions of total knee arthroplasties we performed in our institute between 1980 and 1987. Twelve to 94 months (mean 40.4) after revision we found 66% good or very good results. The main reason for failure was a poor surgical technique, which we found in 69% in unicompartmental, and in 28% in tricompartmental prostheses. Revision surgery needs a high level of practical knowledge and flexibility. The main problems consist of correction of the alignment and the filling of bone defects. Thus, we reconstructed ten large defects at the tibial plateau, which resulted in one very good, seven good and two moderate results.

Aged

Isolation and characterization of DNA sequences from flow-sorted human chromosome 22 libraries.

Two flow-sorted chromosome 22 libraries were used to isolate DNA sequences specific for chromosome 22. 45-phage DNAs were probed against human genomic DNA. 12 of them showed unique or low-copy character. Using digested DNA from rodent-human hybrid cell lines, 3 of the 12 recombinants were assigned unique to chromosome 22 and regionally mapped. 1 clone mapped to 22pter-q11, 1 clone to 22q12-qter and 1 clone, for which in situ hybridization was performed, to 22q13.1. 2 low-copy probes, 1 of them displaying polymorphisms in MspI and TaqI digests of individual DNAs, must have similar sequences on 22 and additional chromosomes. Furthermore, a highly repetitive DNA representing a compound locus of some hundred kilobases on chromosome 22 was isolated. These 6 probes may provide useful tools for studying the structure and function of this small chromosome involved in a relatively high number of inherited and acquired diseases.

Chromosomes, Human, Pair 22

Purification and characterization of five variants of phospholipase A2 and complete primary structure of the main phospholipase A2 variant in Heloderma suspectum (Gila monster) venom.

1. Five increasingly anionic variants (Pa1-Pa5) of Ca2+-dependent phospholipase A2 were purified to homogeneity from the venom of the lizard Heloderma suspectum (Gila monster). The purification procedure was based on semi-preparative reverse-phase HPLC followed by anion-exchange HPLC and analytical reverse-phase HPLC. 2. Their Mr were 17,000-18,000, as deduced by SDS/PAGE. Specific activities tested by the capacity to hydrolyze phosphatidylcholines at pH 8.5 decreased as follows: Pa3 greater than Pa5 greater than Pa4 greater than Pa1 greater than Pa2. These activities showed the same optimum pH (9.0), were mainly of the phospholipase A2 type and were lost upon p-bromophenacyl bromide treatment. 3. All five phospholipases efficiently stimulated amylase release from dispersed rat pancreatic acini at pH 7.4, their potency decreasing as follows: Pa2 greater than Pa1 approximately equal to Pa4 greater than Pa3 approximately equal to Pa5. No deleterious effect was apparent based on the lack of lactate dehydrogenase release. 4. The five variants, Pa1-Pa5, differed significantly in amino acid composition and this, together with distinct antigenic properties of Pa2 and Pa5, establishes the subheterogeneity of this new type of phospholipase A2, despite the fact that the N-terminal amino acid sequence (31 residues) of Pa1-Pa5 was exactly the same. 5. The full sequence of the major variant, Pa5, showed that this 142-amino-acid protein exhibited greater similarity to the bee venom enzyme than to any class I or class II secretory phospholipase A2 from snake venom and mammalian pancreas. While Pa5 displayed the highly conserved region between Asp30 and Cys39 (the essential active site of all phospholipases A2), its salient original points included 10 half-cystine residues only, an incomplete N-terminal sequence, large changes in the putative calcium loop, several alterations after the active site and a C-terminal extension never seen in other phospholipases A2, with the only exception being bee venom.

Amino Acid Sequence

[The immunohistochemical growth fraction (Ki-67) of breast cancers: relations to tumor spread, tumor morphology and receptor testing].

245 breast cancer tissues were analysed immunohistochemically, using monoclonal antibody Ki-67, which spezifically reacts with a nuclear antigen of proliferating cells and represents the so-called growth fraction. According to the number of positive cells, three Ki-score categories (I-III) were established. The results were compared with prognostic variables (Histological grading, DNA-cytophotometry), staging parameters, immunocytochemical (ER-ICA, mPRI), and biochemical (DCC) hormone receptor assay. We found a good correlation between Ki-67 and grading (Bloom and Richardson): G 1-tumors had small growth fractions, in contrast to G-III carcinoma, which showed a high percentage of Ki-Score III. Receptor negative cases had a large growth fraction and receptorpositive ones showed an uniform distribution. The comparison between Ki-67 and tumor spread or DNA-Cytophotometry showed no correlation. The majority of 31 recurrent tumors not only had a large number of positive cells, but also a definite dependence was found between Ki-Score and length of the disease-free period. Ki-67 seems to be an important immunohistochemical marker of breast cancer, which contributes to the individual assessment of the disease and can possibly give predictions on tumor development and response to radiation and systemic therapy.

Adult

[Intestinal beta-galactosidase in gnotobiotic animals following monoassociation with various E. coli strains].

Germ-free rats were monoassociated by E. coli germs not utilizing (L-) or utilizing lactose (L+) on endo-medium. There was no influence of germ status on the beta-galactosidase activity in the mucosa of the small and large intestine. With standard food, beta-galactosidase activity were to be measured in the chymus of all intestinal segments and in the faeces of germ-free as well as of monoassociated rats. In the chymus of caecum and colon and in the faeces of E. coli(L+)-animals, only, the short-time (12-16 days) application of lactose containing food resulted in an increase of the enzyme activity. Compared with the E. coli(L-)-animals, the lactose content in the chymus of all intestinal segments of this test group was decreased.

Animals

Usefulness of Ficoll in electric field-mediated cell fusion.

Electrofusion is a technique that enables the production of new cell types with desired properties to be done. Ehrlich ascites tumor cells are fused by means of an electric field. Under too harsh external field strength or pulse length conditions, however, membrane breakdown leads to a loss of cellular cytoplasm. Addition of the high polymer Ficoll to the fusion medium increases its density and osmotic pressure, thus preventing cytoplasm from running out and, therefore, maintaining cell viability. Fusion between cells of different sizes is made possible or facilitated by means of Ficoll, as big cells do not lose their cytoplasm under conditions required for enhancing membrane permeability of the small cells. In presence of the proteolytic enzyme pronase, addition of Ficoll to the fusion medium further raises the fusion percentage compared to the exclusive addition of pronase. The fusion of cells of different densities is also greatly facilitated and its percentage increased by addition of Ficoll, thus obviating the necessity to perform electrofusion under conditions of microgravity.

Animals

DNA regions associated with the nuclear matrix of Ehrlich ascites cells expose single-stranded sites after deproteinization.

Ehrlich ascites cells were pulse-labeled with [3H]thymidine and subjected to prolonged labeling with [14C]thymidine. The isolated nuclei were digested with the restriction endonuclease BspRI and then processed to yield a 'matrix fraction' and a 'non-matrix fraction'. The DNA fragments purified from these fractions and from whole digested nuclei were examined for nitrocellulose-binding sites before and after digestion with single-strand-specific (S1) nuclease. Both, pulse-labeled and long-time-labeled fragments, isolated from the matrix fraction, exhibited a significantly increased content of nitrocellulose-binding sites. The major portion of these sites were rendered non-binding by digestion with single-strand-specific nuclease and consisted most probably of structures exposing relatively small stretches of non-base-paired DNA. The nature of the minor portion of binding sites which was insensitive to single-strand-specific nuclease is not clear. Both types of binding sites are possible candidates for mediating the attachment of DNA to the nuclear matrix.

Animals

[Results of a double-blind study on the effectiveness of ERU (extractum radicis Urticae) capsules in conservative treatment of benign prostatic hyperplasia].

In 50 patients with prostatic hyperplasia the effect on symptomatology and objective findings of ERU-capsules versus placebo was investigated in a double-blind study over a 9 week treatment period. Admitted to the study were patients in phases I and II who had been referred to the clinic in order to evaluate up the indication for operation. Concerning subjective complaints there was an improvement in dysuric symptoms in both patient groups. The evaluation of the objective parameters showed significant differencies. There was a statistically highly significant (p = 0.0005) decrease of the sex hormone binding globulin in the group of patients treated with ERU as well as a significant improvement of the micturition volume and maximum urinary flow. The improvement of the average flow in the ERU group was not significant. The increase of residual urine volume in both groups seems not to be significant according to the Covariance-analysis and is difficult to interpret. It is assumed, that, up to a certain grade, for a selected group of patients, mainly in the phase of beginning decompensation, length and dosage of therapy were possibly not sufficiently adjusted.

Aged

[Use of the gnotobiotic rat for determination of the in vivo activity of Bifidobacterium beta-galactosidase].

Under the conditions of in vitro cultivation, the height of the beta-galactosidase activity of Bifidobacterium spec. is essentially influenced by the composition of the culture medium. The use of gnotobiotic (germ-free and monoassociated with Bifidobacterium longum) rats permitted to differentiate in the chyme between beta-galactosidase activities of mucosal and microbial origin. In germ-free animals, the chyme in the small intestine and the colon contains nearly 10-20% of the activity measured in the mucosa (in each case expressed as g on a wet-weight basis). Monoassociation with B. longum does not affect the lactose-splitting activity of the chyme in the small intestine, but increases the activity of the chyme in the colon to twice the value of the mucosal activity. In the monoassociated animals, feeding of lactose leads to a further multiple increase of the chymal beta-galactosidase activity in the caecum, colon and faeces.

Animals