Biomedical subjects

T Hoffmann

Publications and source records attributed to T Hoffmann.

At least 91 records · Page 5Linked to original sources

PubMed1996

Clinical and pathomorphological investigation of spontaneously occurring periodontal disease in dogs.

Diagnostic evaluation of, and treatment planning for, different periodontal diseases of dogs are a common problem in veterinary dentistry. Therefore the aim of the present study was to propose a classification for inflammatory periodontal disease. One hundred and twenty-three male and female poodles (from two to over 12 years) were used for the clinical investigations; 120 periodontitis teeth were analysed for pathomorphological signs of inflammation. The recorded data made it possible to differentiate five forms of gingivitis (gingivitis simplex, hyperplastica, ulcerosa, granulomatosa and desquamativa) and three forms of periodontitis (juvenile, rapidly progressive adult and chronic adult periodontitis). The pathomorphological investigations allowed a differentiation between acute progressive inflammation, chronic nonprogressive inflammation, chronic proliferative inflammation and chronic resorptive inflammation. It is concluded that standardised diagnoses as well as the pathomorphological background may be helpful tools for providing effective treatment-strategies.

Animals↗

PubMed1996

Epidemiology of periodontal disease in poodles.

The periodontal condition of 123 poodles was assessed using the parameters of depth of gum pocket, attachment level, bleeding on probing, and plaque and calculus accumulation. No dogs were free of periodontal disease. The number of deep pockets and dramatic loss of attachment ranged from 0.1 teeth in young dogs to 4.2 teeth in old animals. A sixfold increase in deep pocketing took place in dogs between the ages of six and seven, followed by a twofold increase in the number of teeth missing in dogs within the 10 to 11 year age group. First premolars, incisors and second molars were at highest risk. No correlation between dietary conditions and disease incidence could be detected. It is concluded that, by concentrating their diagnostic efforts on those age groups and types of teeth at highest risk, clinicians could improve the effectiveness of their treatment interventions.

Age Distribution↗

PubMed1996

[Bradykinin antagonism in ischemia and reperfusion of the pancreas].

INTRODUCTION: Acute pancreatitis is characterized by two different courses of the disease, edematous and hemorrhagic-necrotizing pancreatitis. The pathogenesis and causes for the progression of pancreatitis are unknown. Ischemia/reperfusion with formation of oxygen free radicals, activation of leukocytes and consecutive failure of the microcirculation has gained attention as a causative factor. Furthermore, the degree of microcirculatory injury correlates with the severity of pancreatitis. The aim of the study was to investigate the influence of long term reperfusion after ischemia of the pancreas for 2 hours on morphological changes and enzyme release of the pancreas in rats. Since the characteristic features of postischemic pancreatic reperfusion injury are kinin-mediated we employed the bradykinin B2-receptor antagonist HOE 140 to inhibit the progression of postischemic changes of the pancreas. METHODS: Under ether anesthesia Sprague-Dawley rats (n = 28) were laparotomized, the 4 supplying arteries of the pancreas were isolated (gastroduodenal artery, left gastric artery, splenic artery and caudal pancreaticoduodenal artery) and occluded with microvascular clips for 2 hours. At the end of ischemia the abdomen was closed and the animals were allowed to awake. 15 minutes before end of ischemia an osmotic minipump filled with NaCl (ischemia group NaCl), phosphate buffer (ischemia group phosphate buffer) or HOE 140 dissolved in phosphate buffer (ischemia group HOE 140) was placed intraperitoneally. Control animals underwent sham operation without vessel occlusion; the osmotic minipump was filled with 0.9 % NaCl. Five days after ischemia the animals were sacrificed for histology. Amylase concentration and peripheral leukocyte count were determined at baseline and daily after operation. RESULTS: After ischemia of 2 hours during reperfusion of 5 days all 14 animals developed histopathological changes as seen in hemorrhagic-necrotizing pancreatitis with a mortality rate of 50 %. These morphological changes were associated with a significant increase (p < 0.05) of pancreas amylase concentration from 1850 +/- 149 U/L before ischemia to a maximum of 3934 +/- 435 U/L at 1st postoperative day and decreased to 1518 +/- 399 U/L at 4th postoperative day. Leukocyte count increased significantly (p < 0.05) from 10 x 10(12)/L to 31 x 10(12)/L. In control animals as well as in animals receiving HOE 140, morphological and enzyme changes typical for acute pancreatitis were absent, leukocyte count increased only slightly. CONCLUSION: Ischemia of the pancreas of 2 hours with ensuing reperfusion of 5 days induces morphological and biochemical changes as observed in hemorrhagic-necrotizing pancreatitis. Organ dysfunction after ischemia/ reperfusion can be effectively inhibited by administration of the bradykinin-antagonist HOE 140.

Amylases↗

PubMed1995

Inhibition of dipeptidyl peptidase IV (DP IV) by anti-DP IV antibodies and non-substrate X-X-Pro- oligopeptides ascertained by capillary electrophoresis.

Dipeptidyl peptidase IV (DP IV)-catalyzed hydrolysis of the NH2-X-Pro-containing N-terminal dodecapeptide of IL-2 was studied using free zone capillary electrophoresis as an alternative peptidase assay. In contrast to the conventional DP IV substrate glycyl-prolyl-p-nitroanilide (Gly-Pro-pNA), the hydrolysis of this peptide by DP IV was found to be significantly inhibited by anti-DP IV antibodies. Inhibition of DP IV was also observed with a number of non-substrate oligopeptides containing an N-terminal X-X-Pro- structure, including the HIV Tat protein. For Met-IL-2(1-6), we determined a competitive inhibition with an inhibition constant of ca. 100 microM.

Amino Acid Sequence↗

PubMed1995

The anaerobic life of Bacillus subtilis: cloning of the genes encoding the respiratory nitrate reductase system.

The Gram-positive soil bacterium Bacillus subtilis, generally regarded as an aerobe, grows under strict anaerobic conditions using nitrate as an electron acceptor and should be designated as a facultative anaerobe. Growth experiments demonstrated a lag phase of 24 to 36 hours after the shift from aerobic, to the onset of anaerobic respiratory growth. Anaerobically adapted cells grew without further lag phase after their transfer to fresh anaerobic growth medium. The cells change their morphology from rods to longer filament-like structures when moved from aerobic to anaerobic respiratory growth conditions. Surprisingly, anaerobically grown B. subtilis lost the capacity for sporulation. An investigation of the molecular basis of the switch between aerobic and anaerobic growth was initiated by the cloning of the genes encoding the respiratory nitrate reductase from B. subtilis. Oligonucleotides deduced from conserved amino acid sequence regions of eubacterial respiratory nitrate reductases and related enzymes were used for the isolation of the genes. Four open reading frames with significant homology to the E. coli respiratory nitrate reductase operons (narGHIJ, narZYWV) were isolated and termed narGHJI. A chromosomal knock-out mutation of the B. subtilis nar operon totally abolished nitrate respiration.

Amino Acid Sequence↗

PubMed1995

Irradiation of bone metastases in breast cancer patients: a randomized study with 1 year follow-up.

The results from a prospective randomized trial comparing two different radiation schedules for treatment of painful bone metastases in women with recurrent breast cancer are presented. A total of 217 patients with painful bone metastases were randomized to either 30 Grey (Gy) in ten fractions, five fractions a week (5F/W) or 15 Gy in three fractions 2F/W. The effect of treatment was evaluated by pain assessment, the radiological response and the degree of side-effects. The patients were rated at start of treatment and after 1, 3, 6 and 12 months. No difference between the two radiation regimes was found, neither in achieved pain relief, improvement in level of activity and medication, nor was there any difference in radiological response and side-effects from treatment. Both regimes resulted in a significant improvement in both pain score and level of activity 1 month after treatment, an improvement which persisted during the follow-up period. We conclude that 15 Gy given in three fractions 2F/W is as effective as 30 Gy in ten fractions 5F/W, but more convenient to the patient and of less cost to society.

Adult↗

PubMed1995

Biodegradation of cyclic and substituted linear oligomers of poly(3-hydroxybutyrate).

Cyclic oligo(3-hydroxybutyrate), oligo(3-HB), was synthesized and purified, resulting in oligolides that contained three to seven (R)-3-hydroxybutyrate units (triolides up to heptolides). In addition, linear 3-HB octamers obtained as either tert-butyl or methyl esters were substituted with different end groups at the hydroxy end. The hydroxy terminus was replaced by either a benzyloxy, trifluoroacetoxy, crotonyloxy (S)-3-hydroxybutyryloxy, or fluorenylmethylcarbonyloxy (FMOC) group. P(3-HB) hairpin loops occurred on the surface of certain regions of the polymer, especially of lamellar crystallites. Cyclic 3-HB oligomers provide a model system for these loops. It is assumed that they provide attachment points for the depolymerizing enzymes. All of the (R)-oligolides tested were degraded except the (R)-triolide. Triolides were not degraded, suggesting that enzymatic attack was prevented presumably by steric hindrance on the rigid ring system. Unsubstituted linear octamers were degraded. Biodegradation was prevented when the hydroxy terminus was protected by the FMOC group, but was not dependent on a free hydroxy terminal group; all other protecting groups did not prevent degradation. Substitution of the carboxy end of a methyl or tert-butyl ester group did not influence biodegradation.

Biodegradation, Environmental↗

PubMed1994

Further studies on the presence of functional tissue factor activity on the subendothelium of normal human and rabbit arteries.

Although tissue factor (TF) activity has been observed on the subendothelial surface of rabbit aorta and human umbilical cord, immunofluorescent and in situ hybridization methods have failed repeatedly to demonstrate TF in the intima of human blood vessels. In the present study, TF activity on everted, de-endothelialized arteries was studied by two methods. One utilized a flow system and measured fibrin deposition and fibrinopeptide A formation. The other utilized a newly developed rotating probe system and measured the conversion of factor X to factor Xa in the presence of factor VIIa and Ca+2. The study attempted to control, or assess, the possibility that functional TF could have been exposed on the vessel surface by the procedures used to prepare the arterial segments. By both methods, TF activity was detected on the subendothelium of rabbit aortae and human umbilical arteries, and was unaffected by the length of storage or by inclusion of actinomycin D in the storage buffer. TF activity was also observed in the subendothelium of adult human ileo-colic, internal mammary, and renal arteries, studied by the rotating probe method. The latter may underestimate TF activity, as some of the factor Xa formed appears to bind to the subendothelial surface. TF activity (Xa formation) was detected on the luminal surface (subendothelium) of non-everted arteries, but increased activity was observed after eversion of the vessel. The source of the subendothelial TF, and its presence in normal subendothelium in vivo, requires further study. In addition, if any of the TF activity observed in this study was derived from injured endothelial or myointimal cells during preparation of the everted vessel segments, the techniques described could serve as a useful model for studying TF-induced thrombosis and factor Xa formation on injured blood vessels, and for evaluating the anti-thrombotic properties of TF-inhibitors.

Animals↗

PubMed1994

Foreign DNA sequences are received by a wild-type strain of Aspergillus niger after co-culture with transgenic higher plants.

Different transgenic plants of Brassica napus, Brassica nigra, Datura innoxia and Vicia narbonensis expressing the hph gene under the control of the 35s promoter were co-cultivated with mycelial material of Aspergillus niger in microcosms under sterile conditions. A significantly higher number of hygromycin B-resistant colonies of re-isolated fungi was obtained if compared with co-cultures with non-transgenic plants. The hph gene and other foreign sequences could be detected in some of the resistant strains only for a short time after selection, indicating a rapid loss of foreign DNA. A more stable transgenic strain was obtained after co-culture with transgenic plants of D. innoxia including a high number of hph copies in their genome. DNA with detected pUC sequences was prepared to transform E. coli DH5 alpha. One of the recovered plasmids is shown to include pieces of the plant-transforming vector and a foreign sequence. The 35s-regulated expression of genes is studied in A. niger.

Aspergillus niger↗

PubMed1993

Dipeptidyl peptidase IV (CD 26) and aminopeptidase N (CD 13) catalyzed hydrolysis of cytokines and peptides with N-terminal cytokine sequences.

A number of natural cytokines are characterized as having dipeptidyl peptidase (DP) IV susceptible N-terminal peptide sequences. Here we demonstrate that oligopeptides with sequences analogous to the N-terminal part of human IL-1 beta, IL-2, TNF-beta and murine IL-6 were hydrolyzed by purified DP IV and aminopeptidase N (AP-N). The rate of DP IV-catalyzed hydrolysis of these peptides was negatively correlated with their chain length. In contrast to these results, no degradation was found under our conditions for the intact recombinant cytokines, IL-1 alpha, IL-1 beta, IL-2, G-CSF and for natural IL-2, independent of whether DP IV and AP-N were used separately or in combination.

Amino Acid Sequence↗

PubMed1993

Glycosylation of human prolactin regulates hormone bioactivity and metabolic clearance.

To analyze the role of individual glycosylation pattern on PRL biopotency, monomeric prolactin (PRL), secreted by human prolactinoma cells in culture, was isolated by gel filtration and separated by affinity chromatography on Concanavalin A-Sepharose or Lentil-Agarose. These lectins allowed the isolation of PRL glycoforms containing either biantennary, mannose-rich or fucosylated complex carbohydrate structures, respectively. Endoglycosidase treatment and carbohydrate content of PRL was found to be consistent with N-linked oligosaccharides of mannose-rich structure and complex units terminated in sialic acid. Mannose-rich PRL and PRL with biantennary oligosaccharides promoted cell growth of rat lymphoma cells to a diminished extent compared to non-glycosylated PRL (NG-PRL), indicating that the two major types of carbohydrate structure are able to decrease the intrinsic bioactivity of PRL. Metabolic clearance of the various forms of PRL in rats was also found to be highly dependent upon hormone glycosylation. The various glycosylated forms (G-PRLs) proved to be totally eliminated from the circulation within 60 min, faster than NG-PRL 10% of which was still present at that time. Mannose-rich or biantennary G-PRLs were differently cleared in a biphasial fashion with a similar rapid phase of about 2 min followed by distinct slow phases of 12 and 27 min, respectively. The presence of fucose did not alter this distribution. In contrast, NG-PRL was eliminated with a half-time of approximately 5 min, followed by a very slow disappearance over several h. It thus appeared that glycosylation increased the metabolic clearance rate of PRL from 0.13 +/- 0.07 ml/min for NG-PRL to 0.47 +/- 0.12 ml/min for PRL with biantennary carbohydrate chains and 0.8 +/- 0.2 ml/min for the hormone with mannose-rich oligosaccharides. The distribution of PRL to target and elimination organs was also found to be different according to the carbohydrate structure present in the hormone. NG-PRL and mannose-rich G-PRL showed higher incorporation in liver than biantennary G-PRL which was preferentially eliminated by the kidney. Altogether, the current data show that addition of oligosaccharides to PRL as well as carbohydrate structure contribute to modulate both the duration of the hormone in the blood and its distribution to different organs. It is proposed that glycosylation may selectively down-regulate PRL action at individual target tissues.

Animals↗

PubMed1993

Reaction of acetaldehyde with proteins: formation of stable fluorescent adducts.

The properties of stable acetaldehyde-protein adducts, using bovine serum albumin as a model protein, were investigated. Upon prolonged incubation at 37 degrees C and pH 7.4, the reaction of acetaldehyde and albumin yielded stable adducts that exhibited fluorescent properties. Reaction mixtures of acetaldehyde with polylysine or ethylamine also formed fluorescent products with similar fluorescent spectral properties like acetaldehyde-albumin adducts, indicating that the amino groups of protein alone can generate fluorescent products on reaction with acetaldehyde. When reactions of acetaldehyde with albumin or polylysine were conducted at 22 degrees C, stable binding reached a maximum after 24 hr of incubation and essentially remained at this level during the remaining 216 hr of incubation, and minimal-to-no fluorescence was associated with this binding. At 37 degrees C, stable binding was greater and increased continuously over the entire 216 hr of incubation. After an initial lag period of 24 to 48 hr, increases in fluorescence intensity paralleled the increases in stable binding. The presence of sodium cyanoborohydride, which reduces Schiff bases, in the reaction mixtures prevented fluorescence, indicating that Schiff bases are intermediates in the formation of fluorescent products. Both stable binding and fluorescence intensities were minimally affected by exhaustive dialysis (up to 144 hr), indicating that the fluorescent products were quite stable. These results suggest that an initial reaction of a Schiff base with another acetaldehyde molecule via an aldol condensation reaction gives rise to the formation of a crotonaldehyde Schiff base derivative. This reactive intermediate could then undergo further condensation reactions and form advanced conjugated products, some of which could be fluorescent.

Acetaldehyde↗

PubMed1993

Graft rejection by a population of primed CDw52- host T cells after in vivo/ex vivo T-depleted bone marrow transplantation.

We investigated a case of graft rejection after in vivo/ex vivo T-depleted BMT in a patient who had received a HVG-matched, GVH one locus-mismatched, MLC-negative graft from his cousin. In vivo/ex vivo T cell depletion was performed with Campath 1G (CP1G) and Campath 1M (CP1M), respectively. We identified a failure of CP1G to eradicate a CDw52- (Campath-negative) host T cell population as the main cause of treatment failure. The analysis also suggests that significant host-versus-donor reactivity prior to transplant, as detected by limiting dilution analysis, also contributed to graft rejection. The rejecting T cells were bifunctional in that they showed cytotoxic activity and were capable of inhibiting haemopoietic progenitor growth by producing inhibitory lymphokines.

Adult↗

PubMed1993

Frequency of bone marrow T cells responding to HLA-identical non-leukemic and leukemic stimulator cells.

Grafted immunocompetent cells are considered responsible for GVHD as well as for the elimination of residual leukemic cells ('graft-versus-leukemia reactivity', GVLR) in leukemic patients after allogeneic BMT. Clinical and experimental investigations have given contradictory answers to the question whether GVHD and GVLR are two manifestations of the same process or separate immunologic processes. We have addressed this question by analysing the primary in vitro response of BM-derived proliferating and cytotoxic T lymphocyte precursors (PTLp and CTLp) in HLA identical relative pairs (n = 17). PTLp frequency estimation reveals strong responses (> 1 in 5000) on non-leukemic as well as leukemic stimulation in a majority of cases. CTLp amount variably to 10-100% of the proliferating precursor cells. Preliminary specificity analyses show that on non-leukemic stimulation about 90% of colonies exhibit exclusive lysis of the non-leukemic target. At the same time, on leukemic stimulation, about 75% of cytolytic colonies are exclusively reactive against leukemic targets without crossreactivity against nonleukemic targets from the same patient. Our data show that primary in vitro responses in HLA identical sibling pairs may be as strong as those against allo MHC antigens. In addition CTL specifically lysing leukemic or non-leukemic targets may represent an in vitro model of the immunologic non-identity of GVHD and GVLR.

Bone Marrow↗

PubMed1993

Evidence that the arg1744 gly1745 asp1746 sequence in the GPIIb-IIIa-binding domain of von Willebrand factor is involved in platelet adhesion and thrombus formation on subendothelium.

Previous studies have strongly suggested that the initial attachment (adhesion) of platelets to subendothelium or collagen at high shear rates is mediated by the binding of von Willebrand factor (vWf) to platelet glycoprotein Ib (GPIb). The finding in the present study that incubating human umbilical artery subendothelium with a monoclonal antibody to the GPIb binding site of vWf markedly inhibited platelet adhesion is fully consistent with this hypothesis. Platelet adhesion (and thrombus formation) also requires GPIIb-IIIa, and previous studies have suggested that the binding of vWf to GPIIb-IIIa may be involved in this process. To explore this further, we utilized two monoclonal antibodies (152B-20 and 152B-6) that recognize the arg1744 gly1745 asp1746 (RGD) sequence of vWf. These antibodies selectively inhibit vWf binding to GPIIb-IIIa and have no cross-reactivity with other RGD-containing proteins that can bind to GPIIb-IIIa. When added to citrated human blood, these antibodies (152B-20 in particular) inhibited platelet adhesion and thrombus formation on rabbit subendothelium at a shear rate of 2600 sec-1 but not at 400 sec-1. The results of the study thus provide direct evidence that the binding of vWf to GPIIb-IIIa is important for platelet adhesion and thrombus formation on subendothelium at high shear rates and that the arg1744 gly1745 asp1746 sequence in the mature vWf is involved in this process.

Amino Acid Sequence↗

PubMed1992

Prolactin isoforms secreted by human prolactinomas.

Prolactin (hPRL) secreted by human prolactinoma cells in culture was purified by gel filtration, lectin affinity chromatography and gel electrophoresis in order to identify the different isoforms of the hormone and to test their respective immunoreactivities and bioactivities. The nonglycosylated hPRL (NG-hPRL), unbound to lectins, was the major form and was a species (NG1-hPRL), of 23,000 (M(r)) apparent molecular weight. The lectin-bound glycosylated hPRL (G-hPRL) consisted of three forms, G1-, G2- and G3-hPRL, of identical molecular weights (25,000 M(r)). Endoglycosidase treatment indicated that these three forms differed by the heterogeneity of their carbohydrate chains. These G-PRLs proved to be 68% less immunoreactive and 50% less bioactive than NG-hPRL. It is concluded from these data that, in prolactinomas, the main variant of the hormone is the nonglycosylated form of PRL.

Antibodies, Monoclonal↗

PubMed1992

Immunoradiometric analysis of circulating human glycosylated and nonglycosylated prolactin forms: spontaneous and stimulated secretions.

The monoclonal antibodies (MAbs) obtained in mice immunized with human PRL coupled to an anti-PRL MAb were screened for their ability to distinguish the glycosylated (G-) and nonglycosylated (NG-) forms of PRL. The 431-29 MAb exhibited high affinity binding for NG-PRL but little or no cross-reactivity to G-PRL. Using this antibody in conjunction with other MAbs which equally recognized both forms, we developed 2 immunoradiometric assays which were used to determine the amount of G- and NG-PRL in plasma. In 85 normal subjects, NG-PRL baseline levels averaged 6.6 +/- 3 micrograms/L, and represented 76 +/- 8% of the total PRL immunoreactivity. In 74 pregnant women, this proportion was significantly higher during the last 2 trimesters (84 +/- 4% and 85 +/- 6%), as compared to the first trimester (76 +/- 7%). In 6 healthy volunteers studied over 24 h, 79% of the NG-PRL peaks detected using the cluster algorithm occurred concomitantly to a G-PRL peak. The mean NG-PRL/PRL ratio was significantly higher during NG-PRL pulses (81 +/- 9%) than during valleys (71 +/- 12%). Similarly, this ratio was significantly increased during TRH or metoclopramide stimulated PRL secretion (to 88 +/- 7% and 86 +/- 6%, respectively). We conclude that 1) NG-PRL is the predominant immunoassayable form of PRL in plasma; 2) both G- and NG-PRL are cosecreted but NG-PRL is the main PRL form released during spontaneous or pharmacologically induced PRL secretion.

Adult↗

PubMed1992

Defective function of blood granulocytes in patients with diabetes mellitus-associated marginal periodontitis.

In ten periodontitis patients suffering from type I diabetes mellitus, phagocytic activity, aggregation and chemiluminescence generation of blood granulocytes were determined. Compared to controls with clinically healthy periodontal conditions, the phagocytosis of zymosan particles and the aggregation response induced by the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine were significantly decreased, whereas aggregation induced by the platelet-activating factor, a potent mediator of inflammation, was significantly enhanced.

Adult↗