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Biomedical subjects

H Lindner

Publications and source records attributed to H Lindner.

At least 19 recordsLinked to original sources

Alteration in proportions of histone H1 variants during the differentiation of murine erythroleukaemic cells.

We have investigated the changes in the relative amounts of histone H1 zero and all five H1 variants during the differentiation in vitro of Friend erythroleukaemic cells. Three different agents were used as inducers of differentiation: dimethyl sulphoxide, hexamethylenebisacetamide and sodium butyrate. By applying a combination of reverse-phase h.p.l.c. and one-dimensional gel electrophoresis we observed that, during differentiation in vitro, (1) the relative amount of each subtype changes upon induction and that (2) dimethyl sulphoxide and hexamethylenebisacetamide produce a similar histone H1 pattern with a strong increase in histones H1 zero and H1c, a modest increase in histone H1e and a decrease in the relative amounts of histone H1a, H1b and H1d, whereas butyrate induces a different pattern, particularly with respect to both histones H1c and H1e: H1c increased slightly, and H1e strongly, during differentiation. These results are compared with changes in the histone H1 pattern during differentiation in vivo in the mouse [Lennox & Cohen (1983) J. Biol. Chem. 258, 262-268] and in the rat [Pina, Martinez & Suau (1987) Eur. J. Biochem. 164, 71-76], and similarities and deviations are discussed.

Acetamides

Specificity of Zea mays histone deacetylase is regulated by phosphorylation.

Mono Q ion exchange high performance liquid chromatography (HPLC) reveals that the main histone deacetylase activity (HD1) of germinating Zea mays embryos consists of multiple enzyme forms. Chromatography of HD1 after treatment with alkaline phosphatase yields two distinct histone deacetylase forms (HD1-A, HD1-B). The same is true for chromatography after phosphatase treatment of a total cell extract. One of these enzyme forms (HD1-A) is subject to phosphorylation, which causes a change in the substrate specificity of the enzyme, as shown with HPLC-purified individual core histone species; the substrate specificity for H2A increases more than 2-fold after phosphorylation, whereas the specificity for H3 decreases to about 60%. The total histone deacetylase activity is quantitatively released from isolated nuclei after extraction with moderate ionic strength buffers; no significant residual enzyme activity could be detected in the nuclear matrix.

Animals

Separation of phosphorylated histone H1 variants by high-performance capillary electrophoresis.

High-performance capillary electrophoresis (HPCE) was used to separate successfully distinct phosphorylated derivatives of individual histone H1 variants. With an untreated capillary (50 cm x 75 microns I.D.) the electrophoresis was performed in about 15 min. Inconvenient interactions of these highly basic proteins with the capillary wall were eliminated by using 0.1 M sodium phosphate buffer (pH 2.0) containing 0.03% hydroxypropylmethylcellulose. Under these experimental conditions the histone H1 variants H1b and H1c obtained from mitotic enriched NIH 3T3 fibroblasts and isolated by reversed-phase high-performance liquid chromatography were clearly separated in their non-phosphorylated and different phosphorylated forms. This result was confirmed by acid-urea gel electrophoresis, comparison with non-phosphorylated histones H1b and H1c, isolated from quiescent NIH 3T3 cells, and incubation of multi-phosphorylated histone H1b with alkaline phosphatase and subsequent acid-urea and capillary electrophoresis. The results illustrate that the application of HPCE to the analysis of histone modifications provides a new alternative to traditional gel electrophoresis.

Alkaline Phosphatase

High-performance capillary electrophoresis of core histones and their acetylated modified derivatives.

By using high-performance capillary electrophoresis, we have successfully separated rat liver core histones into several subfractions. Inconvenient interactions of the highly basic proteins with the capillary wall were eliminated by a phosphate buffer system containing 0.03% hydroxyprophylmethylcellulose. Sample amounts of a few nanolitres were analysed within about 20 min. Multiacetylated histones H4 and H3 from induced Friend erythroleukaemic cells prepurified by h.p.l.c. were clearly separated into their non-acetylated and distinct acetylated forms. Our results illustrate that the application of capillary zone electrophoresis on its own or in combination with h.p.l.c. to the analysis of histones provides an important new alternative to traditional gel electrophoreses.

Acetylation

Enzymes involved in the dynamic equilibrium of core histone acetylation of Physarum polycephalum.

DEAE-Sepharose chromatography of extracts from plasmodia of the myxomycete Physarum polycephalum revealed the presence of multiple histone acetyltransferases and histone deacetylases. A cytoplasmic histone acetyltransferase B, specific for histone H4, and two nuclear acetyltransferases A1 and A2 were identified; A1 acetylates all core histones with a preference for H3 and H2A, whereas A2 is specific for H3 and also slightly for H2B. Two histone deacetylases, HD1 and HD2, could be discriminated. They differ with respect to substrate specificity and pH dependence. For the first time the substrate specificity of histone deacetylases was determined using HPLC-purified individual core histone species. The order of acetylated substrate preference is H2A much greater than H3 greater than or equal to H4 greater than H2B for HD1 and H3 greater than H2A greater than H4 for HD2, respectively; HD2 is inactive with H2B as substrate. Moreover histone deacetylases are very sensitive to butyrate, since 2 mM butyrate leads to more than 50% inhibition of enzyme activity.

Acetylation

Premenstrual symptoms: the role of irrational thinking.

104 women, between the ages of 18 and 45 years, were surveyed to investigate the relationship between premenstrual symptomatology, as measured by the Modified Menstrual Distress Questionnaire, and irrational thinking, as measured by the General Attitude and Belief Scale. The women who reported greater premenstrual symptomatology also reported significantly higher scores in the "need for comfort" irrationality subscale. This indicated that these women had particular difficulty dealing with hassles and the resulting feelings of tension and irritability in the premenstruum. It was suggested that the absence of significant effects for other rationality-irrationality subscales could be associated with testing at different times during the menstrual cycle. Irrationality, like other conditions (such as anxiety and depression) prevalent in the premenstruum, could change in intensity across phases of the menstrual cycle.

Adult

[Morphology and therapy of tracheal cysts].

Following a short representation of etiology, morphology and classification of cysts of trachea the symptoms, diagnosis and therapy were described. 7 cases of the own patients material in 27 years are presented extensively.

Bronchogenic Cyst

[Radiotherapy of small cell bronchial cancer].

In the combined modality treatment of the small carcinoma of the lung chemotherapy is obligatory. Radiotherapy plays a decisive role, especially in limited disease tumors. Numerous randomized studies show that the rate of locoregional recurrence is reduced, whereas the survival rate is raised by the irradiation of the thorax. Dose size, target volume, fractionation as well as adaptation to the chemotherapy regimes are still a matter for discussion. In extensive disease stage, radiotherapy generally plays a role only as a palliative treatment modality. Undoubtedly, the prophylactic cranial irradiation diminishes the frequency of brain metastases. Regarding the potential neurological late toxicity we must carefully pay attention to the dosage, the fractionation, and the indication to this treatment. Half and total body irradiation only should be considered in prospective studies.

Carcinoma, Small Cell

[A basic psychodiagnostic program for internists].

The increasing frequency of disturbances of the health, in which psychic and/or social influential factors are significant, reflects also in the number of patients of the specialist of internal medicine who is working in the outpatient department, the hospital and in a subspeciality. This requires from the specialists of internal medicine, if he will satisfy the claims of his patients for a comprehensive diagnostics and effective therapy, from the very beginning and in all phases of the care the involvement of a basic psychodiagnostics. Hereby the problem in question is an enhanced internal establishment of the anamnesis under psychodiagnostic aspect, which serves for the clarification of psychic and/or social influences on signs and symptoms, the experience of the and the course of the disease. An orientating systematization and a way to the positive diagnostics of neuroses, psychosomatic diseases or disturbances and further functional syndromes in the field of internal medicine are presented which demand the graduated psychotherapeutic measure.

Humans

Separation of rat tissue histone H1 subtypes by reverse-phase h.p.l.c. Identification and assignment to a standard H1 nomenclature.

H1 histones from rat liver and rat testis were separated by reverse-phase h.p.l.c. Within 40 min six subfractions (H1(0), H1b, H1a, H1d, H1e + H1c and H1c) and seven subfractions (H1(0), H1b, H1a, H1d, H1e + H1c, H1c and H1t) respectively were isolated by using a linear acetonitrile gradient. Each individual H1 subtype was identified either by comparing the H1 variants (contained in both tissues but in different quantities) or by SDS/PAGE and acetic acid/urea/PAGE. Moreover, all H1 variants were characterized by amino acid analyses. The amino acid compositions of rat histone subfractions H1(0), H1b and H1e were determined for the first time. It was possible to classify unambiguously the H1 subfractions obtained by h.p.l.c. by following the standardized H1 nomenclature for electrophoretic systems recommended by Lennox, Oshima & Cohen [(1982) J. Biol. Chem. 257, 5183-5189]. Incorrect assignments that have been made in various publications are discussed.

Acetates

Autoantibodies against different histone H1 subtypes in systemic lupus erythematosus sera.

The H1 histones represent the most heterogenous class of histone proteins. In this study, we analyzed the specificity of human antibodies against 6 H1 subtypes. H1 histones from rat organs were separated by reverse-phase high performance liquid chromatography and used as antigens in immunoblotting experiments. Sera containing anti-histone H1 antibodies were obtained from patients with systemic lupus erythematosus. Of the 9 sera tested, 2 reacted with only 1 H1 subtype. The other sera recognized different combinations of H1 subtypes. Only 1 serum reacted with all 6 H1 subtypes. Histones H1.5 and H1.1 were the subtypes most frequently recognized by the human autoantibodies. Our data indicate that human anti-H1 antibodies represent a heterogenous population, directed mainly against epitopes localized in the variable region of the H1 molecule.

Antibody Specificity

Primary adjuvant whole abdominal irradiation in ovarian carcinoma.

Eighty-four patients with an ovarian carcinoma Stage I-III received an adjuvant whole abdominal irradiation (WAI) with pelvic boost postoperatively. Surgery included a bilateral salpingo-oophorectomy, hysterectomy, and omentectomy in 59% of the patients; in 41% surgery was less radical. For the whole abdominal irradiation we used the moving-strip method on 43 patients. The open-field technique was used on the other patients. Median dose of WAI was 22.5 Gy and median pelvic dose was 45.5 Gy. After a median follow-up of 68.5 months, a 5-year survival rate of 64 +/- 5.7% was determined, as well as a 5-year NED rate of 61.2 +/- 5.7%. Five-year survival rate was 80.1 +/- 7.4% in Stage I, 64.1 +/- 9.7% in Stage II, and 35.4 +/- 11.6% in Stage III. Five-year survival depended on tumor rest significantly. There was a trend to better prognosis when surgery was complete and grade was G1 or G2. The risk factor according to Dembo proved to be the most reliable prognostic factor: the 5-year survival rates were 75.0 +/- 6.3% for patients with intermediate risk and 20.1 +/- 10.4% for those with high risk (p = 0.001). Side effects were generally well tolerated. We only saw one serious complication, a radiation-induced small bowel obstruction.

Abdomen

[Postoperative adjuvant whole abdomen irradiation in ovarian cancer].

Seventy-nine patients with ovarian carcinoma stage I-III postoperatively received a whole abdominal irradiation (median dose: 22.5 Gy) followed by pelvic boost irradiation (median pelvic dose: 45 Gy) as the sole adjuvant therapy. Surgery preceding the irradiation was radical only in 59 percent of the patients. Whole abdominal irradiation was performed either in the moving-strip-technique or the open-field-technique. The median follow-up time was 57 months. The overall and the progression-free five-year survival rate of the whole group is 66 +/- 6% and 63 +/- 6%, respectively. We have not seen a clear-cut dependence of the survival rates on the completeness of the surgery, on tumor histology or grading. In a statistically significant way, the survival rates depend on the stage, the presence of tumor rests, and, above all, on the risk group. The overall five-year survival rate for the intermediate-risk patients is 78 +/- 7% and 22 +/- 11% for high risk patients. There aren't any randomized studies concerning the optimal adjuvant therapy for intermediate-risk patients. Because of the reproduceable results and the infrequent toxicity, whole abdominal irradiation seems to be a treatment equivalent to other adjuvant therapies for these patients.

Adult

Identification of ADP-ribosylated histones by the combined use of high-performance liquid chromatography and electrophoresis.

Reversed-phase high-performance liquid chromatography (HPLC) was employed for analysing mono- and oligo(ADP-ribosyl)ated histones. Under the chromatographic conditions described, the ADP-ribosylated histones showed similar retention times to the unmodified histones, although the molecular weight and the charge of the proteins are significantly altered by their modification. The simultaneous elution of unmodified and labelled modified histones was detected by two types of gel electrophoresis and by autoradiography. In addition, the HPLC fractions did not display overlapping ladders of the multiply modified histones, as is commonly seen in one-dimensional electrophoretic analyses of unfractionated material. Hence individual bands could be unambiguously assigned. After in vitro labelling of isolated rat liver nuclei, the following ADP-ribosylated and unmodified histones were identified by HPLC and gel electrophoresis: histone H1(0), four histone H1 subfractions, histone H2A.1, histone H2A.2, oxidized histone H2A.2, histone H2A.X, histone H2A.Z, histone H2B, three histone H3 variants and histone H4.

Adenosine Diphosphate Ribose

[Treatment of otherwise incurable tumor diseases in childhood using whole-body hyperthermia and chemotherapy].

Conventional methods of treatment having failed in 17 children (aged 9/12 to 16 5/12 years) with incurable solid malignant tumours underwent whole-body hyperthermia (41.8-42.0 degrees C, for 2-3 h), hyperglycaemia (20-25 mmol/l) and polychemotherapy. Five children had neuroblastoma (stage 4), three Wilm's tumour (stage 4 or 5, unfavourable histology), five skeletal sarcoma with metastases, three inoperable malignant liver tumour, and one brainstem tumour of unknown histology. Whole-body hyperthermia was induced by extracorporeal blood warming in an haemodialysis apparatus under neuroleptic analgesia, thermistors measuring the temperature in the oesophagus, rectum, trachea and skin. There were on average four treatment sessions (between one and ten, total 58), a week apart. The result could be assessed in 12 children: one persisting complete remission (19 months-metastasising renal rhabdoid tumour), eight partial or incomplete remissions, and three nonresponders (osteogenic sarcoma; Ewing sarcoma; brainstem tumour). If the risk can be satisfactorily judged the method is useful and of bearable toxicity. The results point to a high antitumour effectiveness of combined hyperthermia and chemotherapy.

Adolescent

Separation of Friend erythroleukaemic cell histones and high-mobility-group proteins by reversed-phase high-performance liquid chromatography.

A procedure for the rapid separation of histones and high-mobility-group (HMG) proteins from Friend erythroleukaemic cells (line F4N) by reversed-phase high-performance liquid chromatography is reported. By using a Nucleosil 300-5 C4 column and a multistep water-acetonitrile gradient containing 0.1% trifluoroacetic acid, the HMG-1 and HMG-2 proteins, several H1 subfractions including H1(0), H4, H2B, two H2A variants and two H3 subfractions were separated. Under changed conditions, by applying a varied acetonitrile gradient system, even two H2B variants were fractionated. The methods described seem to be a real alternative to the time-consuming polyacrylamide gel electrophoresis.

Cell Line

A new h.p.l.c. isolation procedure for chicken and goose erythrocyte histones.

Total chicken erythrocyte histones were separated by reversed-phase h.p.l.c. using a multi-step acetonitrile gradient in a very short time (35 min). The proteins were eluted in the following order: H1, H5, H2B, H2A.2, H4, H2A.1 and H3.2. Applying a special gradient system adapted for the separation of very-lysine-rich histones, chicken erythrocyte H5 was resolved into two subfractions. Their electrophoretic mobilities were identical in both SDS and acetic acid/urea/Triton polyacrylamide-gel electrophoresis, but different in free-flow electrophoresis. Amino-acid-sequence analyses revealed that the two components only differ with respect to position 15, one having glutamine in that position and the other arginine. A separation of histones prepared from goose erythrocytes disclosed no H5 subfractionation. Furthermore, histones obtained from anaemic-chicken blood were analysed by the above-mentioned h.p.l.c. conditions. An alteration in the relation of H1 to H5 was detected, but no further differences in the number and quantity of the histones and histone variants were observed as compared with the corresponding proteins processed from normal-chicken blood.

Anemia