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Biomedical subjects

K Hoffmann

Publications and source records attributed to K Hoffmann.

At least 19 recordsLinked to original sources

Two new cyclophilins from Fusarium sambucinum and Aspergillus niger: resistance of cyclophilin/cyclosporin A complexes against proteolysis.

Two new peptidyl-prolyl-cis/trans-isomerases were purified to homogeneity from Fusarium sambucinum and Aspergillus niger. They belong to the class of cyclosporin A binding proteins (cyclophilins) and have molecular masses of about 18 kDa. As has been shown for other cyclophilins, the isomerase activity of the enzymes is inhibited by cyclosporin A in the nanomolar range. Furthermore binding of cyclosporin A prevents proteolytic digestion of the cyclophilin/cyclosporin complexes by the endoproteases GluC, LysC and alpha-chymotrypsin, in contrast to the free cyclophilins, which are readily cleaved by these proteases. We could also observe this protection for cyclophilins from sheep thymus and from the cyclosporin producing fungus Tolypocladium inflatum.

Amino Acid Isomerases

Nuclear factor I (NF I) binds to an NF I-type site but not to the CCAAT site in the human alpha-globin gene promoter.

Methylation interference and missing contact analyses demonstrate that nuclear factor I (NF I) recognizes an NF I-like site (5'-GGG(N)6GCCAG-3') within the alpha-globin promoter rather than the adjacent CCAAT box. Consistent with this, mutations within the CCAAT box do not alter significantly the affinity and specificity of the interaction whereas elimination of the 5'-GGG-3' half-site of the recognition sequence reduces the DNA binding strength of NF I by 2 orders of magnitude down to the range of unspecific interaction. On the other hand, the mutated alpha-globin promoter sequence that is no longer bound by NF I, although it retains an intact CCAAT box, interacts specifically with a protein component from nuclear extracts of HeLa cells. From these results we conclude that NF I is not the factor that interacts with the CCAAT box and that the second half of the canonical 5'-TGG(N)6GCCAA-3' NF I binding site cannot be regarded as identical with the CCAAT promoter element, as suggested previously.

Autoradiography

Human-immunodeficiency-virus-type-1-encoded Vpu protein is phosphorylated by casein kinase II.

Vpu as a human-immunodeficiency-virus-type-1-encoded 81-amino-acid integral-membrane protein was expressed in Escherichia coli using the inducible ptrc promoter of an ATG fusion vector. Recombinant Vpu is associated with membranes of E. coli and could be partially solubilized by detergents. Recombinant Vpu was phosphorylated in vitro with purified porcine casein kinase II (CKII) as well as with a CKII-related protein kinase found in cytoplasmic extracts of human and hamster cells. Recombinant Vpu associated with E. coli membranes has turned out to be the best substrate for in vitro phosphorylation with CKII. This reaction can be inhibited by heparin and the ATP analogue 5,6-dichloro-1-(beta-D-ribofuranosyl)benzimidazole (DRB), both known to be potent inhibitors of CKII. Radiolabelled gamma ATP and gamma GTP were used as phosphate donors in vitro phosphorylation of recombinant Vpu. In vivo phosphorylation of Vpu in HIV-1-infected H9 cells was also inhibited by DRB. We concluded therefrom that the Vpu protein is phosphorylated by the ubiquitous CKII in HIV-1-infected human host cells. Two seryl residues in the sequence of Vpu (position 52 and 56) correspond to the consensus S/TXXD/E for CKII. These potential phosphorylation sites are located within a well-conserved dodecapeptide of Vpu (residues 47-58), which is found in different HIV-1 strains as well as in a Vpu-like protein of SIVCPZ. Monoclonal and polyclonal antibodies directed against two different epitopes of Vpu were used for immunoprecipitation of Vpu from HIV-1-infected cells and for detection of Vpu in Western blot analyses. Vpu from HIV-1-infected cells as well as recombinant Vpu expressed in E. coli were determined by SDS/PAGE using 6 M urea to be 9 kDa, which corresponds to the calculated molecular mass of Vpu.

Adenosine Triphosphate

FK-506 binding protein from Tolypocladium inflatum: resistance of FKBP/FK-506 complex against proteolysis.

A 12-kDa peptidyl-prolyl-cis/trans-isomerase was purified 225-fold to homogeneity from the cyclosporin producing fungus Tolypocladium inflatum. The enzyme is highly sensitive to the immunosuppressant FK-506 but not to cyclosporin and thus belongs to the class of FK-506 binding proteins (FKBP). Interestingly the FKBP/FK-506 complex is resistant against proteolytic digestion by the endoproteases GluC and LysC, in contrast to the free FKBP, which is readily cleaved by these proteases. This protection may play a role in the effects of FK-506 in the living cell.

Amino Acid Isomerases

Physiopathology of the knee joint after distal iliotibial band transfer.

In an experimental study of 14 cadaver knee joints, the pressure load on the joint surface after distal iliotibial band transfer was measured using Fuji Prescale foils. With an intact anterior cruciate ligament, increases of up to 153% for the average pressure load and of 225% for the total pressure in the lateral compartment were found in relation to the fixation point chosen. At point P3--slightly dorsal to the insertion of the lateral collateral ligament--the area loaded with maximum pressure increased to six-fold. Fixation at the transition of the lateral femoral condyle to the femoral shaft at the start of the linea aspera was associated with the least pressure increases in both the lateral and the medial compartments. Under all experimental conditions, lateral extra-articular stabilization with fixation at the insertion of the fibular collateral ligament was shown to be associated with significantly higher load increases. While a shift of pressure load to the dorsal third was seen in the lateral compartment, the mid-third remained the focus of the pressure load in the medial compartment. After transection of the anterior cruciate ligament and iliotibial band transfer at the "over-the-top" point, a significant shift of pressure towards the medial compartment was seen, while the lateral pressure load decreased. Medially, the area loaded with peak pressure remained constant, while the corresponding area in the lateral joint space showed a highly significant decrease to nearly one-third of normal. After additional bilateral meniscectomy this tendency was even more pronounced.(ABSTRACT TRUNCATED AT 250 WORDS)

Anterior Cruciate Ligament

Pattern of pineal melatonin concentrations during prolonged exposure to short photoperiods in photosensitive and photoinsensitive Djungarian hamsters.

In the Djungarian hamster (Phodopus sungorus) there is strong evidence that the nightly decompression or lengthening, respectively, of the pineal melatonin pattern is involved in conveying the inhibitory influence of short photoperiods. Some hamsters, however, are insensitive to photoinhibitory influences; other late-responding hamsters show a delayed reaction of several months. The daily pattern of pineal melatonin proves that the mechanisms determining long- or short-day patterns become effective in the morning and evening hours when melatonin concentrations of photoinhibited hamsters are clearly elevated, while those of long-day hamsters are reduced to negligible low values. At two decisive time points in the morning and evening, male and female early-responding photosensitive hamsters were killed after 5, 10, 15, and 20 weeks in short photoperiods, nonresponding hamsters after 10 and 20 weeks, and late-responding after 20 weeks. In all early-responding hamsters changes in gonadal stage, body weight, and pelage color corresponded to the decompression of the nightly melatonin pattern. During prolonged exposure to short photoperiods a decrease of mean melatonin concentrations in the morning and an increase of evening concentrations was observed in both sexes. The lengthening of nightly elevated melatonin concentrations was missing in photoinsensitive hamsters. In late-responding hamsters this decompression seems to be delayed, as melatonin concentrations of female late responders indicate. In conclusion, the lengthening of the melatonin pattern seems to predict a photoperiodic reaction of the effector system. The lengthened pattern of photosensitive hamsters may not be unchangeable, as it may be subjected to modifications during prolonged exposure to short photoperiods.

Animals

Malignant melanoma in 20-MHz B scan sonography.

In order to assess the value of 20-MHz sonography in the pre-operative diagnosis of malignant melanomas, 54 melanomas were examined. The pre-operative ultrasound scans were compared with corresponding histological sections from the excised tumours. A computer-aided measurement of the tumour thickness (sonometry) and the internal echo density (densitometry) was performed in the ultrasound scans. The melanomas appeared as largely echolucent zones. There was a significant correlation between the tumour thickness measured in the ultrasound scan and those measured in the histological sections (r = 0.938, p less than 0.001). As a rule the tumour thickness determined by sonometry were greater than the histometrically determined values. Subtumoral inflammatory infiltrate and other hypo-echoic structures in the region of the tumour are possible causes of the discrepancies. Some of those hypo-echoic structures can be identified and disregarded in B scan measurement. Even if a definite differential diagnosis is not possible on the basis of the ultrasound scan alone, 20-MHz sonography provides additional information which can be of use in surgical planning.

Adolescent

A seventeen kilodaltons peptidyl-prolyl cis-trans isomerase of the cyclosporin-producer Tolypocladium inflatum is sensitive to cyclosporin A.

A peptidyl-prolyl cis-trans isomerase (PPIase) was purified to homogeneity about 24-fold from the cyclosporin-producing fungus Tolypocladium inflatum. The molecular mass of the enzyme was in the range of 17 kdaltons. Remarkably, the enzyme could be inhibited by cyclosporin A in the nanomolar range as has been shown for numerous other cyclophilins from eukaryotic organisms. This indicates, that Tolypocladium inflatum must possess a self protection system in order to survive in the presence of cyclosporin.

Amino Acid Isomerases

[Ultrasound diagnosis of skin tumors].

Twenty-one patients with skin tumors were examined before and after operation using special high-frequency ultrasound B-scanners. The different histological types were related to sonographic phenotypes, and the extent of tumors in the ultrasound image was comparable with that seen on the histometric image. The extent and invasion depth of skin tumors were assessed correctly by ultrasound.

Basal Cell Carcinoma

Number of helper T cells and phytohemagglutinin stimulation correlate in cancer patients.

Mononuclear cells from 12 normal controls (co), 10 advanced untreated (c1), and 6 advanced treated cancer patients (c2) have been isolated. The numbers of mononuclear cells bearing Leu1, Leu2, Leu3, Leu2/HLA-DR and LeuM3 were measured with a fluorescence-activated cell sorter. Only the quantity of helper T cells (Leu3) was decreased in cancer patients (co: 0.89, cl: 0.32, c2: 0.44 x 10(9)/l). Expression of all other markers, including activated suppressor T cells (Leu2/HLA-DR), did not differ significantly from the control. The proliferation of the lymphocytes was determined in a phytohemagglutinin-culture assay. The cancer groups showed a significantly decreased response (co: 95.8 x 10(9), cl: 28.7 x 10(9), c2: 25.7 x 10(9) cpm). These values correlated with the number of helper T cells but not with the suppressor T cells. Monocytes of cancer patients absorbed significantly more immunoglobulins than the monocytes of controls. The addition of indomethacin or isoprinosine to phytohemagglutinin-culture assay increased the proliferation of lymphocytes from both the cancer patients and normal controls.

Adult

20-MHz B-mode ultrasound in monitoring the course of localized scleroderma (morphea).

Ultrasonographic methods have recently provided us with the means for objective and non-invasive monitoring of the dynamics of chronic skin diseases. We examined 34 patients with localized scleroderma (morphea) using a 20-MHz B-mode ultrasound scanner (DUB 20, Taberna pro Medicum, Lüneburg). In patients with plaque-type and linear band-type localized scleroderma intraindividual comparison of sclerotic skin with corresponding areas of healthy skin showed thickening of the corium. The increase in corium thickness was between 2% and 251%. The extent of the difference in corium thickness between sclerotic and healthy skin depended on the location-originally thin skin showed a greater degree of sclerosis. We also frequently found enhanced reflexes in the lower corium and hyperechoic, widened bands of connective tissue traversing the subcutaneous fatty tissue from the corium-subcutis border in the direction of the muscle fascia. 20 patients were examined several times in the course of one year. In nine patients we found ultrasonographic evidence of regression (decrease in thickness 26%) and in nine the ultrasound examination showed progression (increase in thickness 28%). 20-MHz B-mode ultrasound imaging is a suitable non-invasive method for monitoring the course and treatment of localized scleroderma. Its routine use is strongly recommended.

Adolescent

Statistical evaluation of the evidential value of human hairs possibly coming from multiple sources.

A concept of the mathematical evaluation of human hair evidence is derived. This concept can be realized in a special computer program, the output of which is an incrimination probability. The problems of not knowing the true number of sources and the correct partition of hairs corresponding to their sources are solved from the point of view of avoiding an unjustified incrimination.

Forensic Medicine

Long-term effects of acetazolamide and sodium chloride loading on cisplatin nephrotoxicity in the rat.

The protective effect of acetazolamide or sodium chloride loading on cisplatin nephrotoxicity was investigated in rats. After a single dose of cisplatin (5 mg kg-1 i.p.) kidney function was studied after 5, 28 and 84 days. Acetazolamide (75 mg kg-1 i.p.) was administered as a single dose prior (30 min) to the cisplatin injection. By the time of cisplatin administration, the rats were sodium depleted except the sodium-loaded group. Five days after the cisplatin administration all rats received a regular rat chow for the rest of the experiment. Cisplatin alone caused renal failure that could be observed for up to 12 weeks (ClCr 0.32 +/- 0.13 vs. 0.62 +/- 0.06 ml min-1 x 100 g BW-1) with polyuria (UVol 41.2 +/- 4.5 vs. 18.4 +/- 4.6 ml 24 h-1). Pretreatment with acetazolamide was the most protective manoeuvre tested. Five days after cisplatin, kidney function was significantly better than in rats treated with cisplatin alone (ClCr 0.21 +/- 0.06 vs. 0.03 +/- 0.01 ml min-1 x 100 g BW-1), after 28 days the only sign of nephrotoxicity was polyuria (UVol 28.9 +/- 3.7 vs. 19.0 +/- 2.6 ml 24 h-1) after 84 days no differences could be observed at all. Sodium chloride loading was less protective on cisplatin nephrotoxicity. Impaired renal function could still be observed after 12 weeks (ClCr 0.41 +/- 0.05 vs. 0.62 +/- 0.06 ml min-1 x 100 g BW-1) with no difference in comparison with the rats treated with cisplatin alone. However, since 12 rats died in the group having received cisplatin alone and only one rat in the high-salt group, sodium chloride loading is regarded as being advantageous over sodium depletion on cisplatin nephrotoxicity.

Acetazolamide

Distribution pattern of testicular sulphydryloxidase immuno-activity in the djungarian hamster (Phodopus sungorus) during photoperiodically induced involution and recrudescence.

The distribution pattern of testicular sulphydryloxidase (SOx) immuno-activity was investigated in the djungarian hamster during photoperiodically induced testicular involution and recrudescence. SOx immuno-activity, indicating functional integrity of labelled cells, did not change in pachytene spermatocytes and spermatids as long as these cells were present in the seminiferous epithelium. Its disappearance coincided with the degeneration of spermatocytes in phases IV and V of involution and reappeared during recrudescence, when the first spermatogenic wave had reached the pachytene stage. In tubules at phase VI of involution (showing maximal regression), the apical cytoplasm of Sertoli cells showed immuno-activity. This immuno-activity disappeared during recrudescence prior to the differentiation of pachytene spermatocytes. Changes in SOx immuno-activity resembled those of lactate dehydrogenase-X (LDH-X) in photo-inhibited testes or during puberty, indicating a close functional relationship which still remains to be elucidated. The data suggest that the hamster exposed to different photoperiods can be used as a suitable model to study the relationship between testicular morphology and function in different states of gonadal activity.

Animals

[Use of DNA polymorphism in forensic paternity evaluation].

Problematical paternity cases were additionally subjected to DNA-polymorphism analysis. 5 cases are reported, focusing on problems due to, 'silent' allele transmittance, relatively low probability for paternity, obvious occurrence of the extremely rare Rh gene complex CWc, involvement of brothers as putative fathers, non-paternity of a man although his W-value was 99.975%. The aim of this paper is to demonstrate the need for DNA-polymorphism analysis, if conventional blood group tests do not lead to a clear-cut decision. Extended typing of conventional blood group polymorphisms (except HLA) cannot compete with highly polymorphic DNA loci.

Adult

[1989 Bochum health education program for malignant melanoma].

The incidence rate of all skin cancer forms has been increasing during the last few decades. The incidence of malignant melanoma is important due to its very high malignancy and its growing spread especially among young patients. Malignant melanoma is mainly observed on the external skin and is therefore well suited for screening methods. During a campaign in the city of Bochum, 1226 patients were examined. A total of 9 malignant melanomas and 10 basal cell carcinomas were discovered. The statistical data show that most of the patients could be motivated to participate by regional media and not by supraregional media. The wide acceptance of our prevention week shows that prevention campaigns are feasible, important and necessary.

Adolescent