Prothrombotic polymorphisms in patients with Raynaud's phenomenon and migraine.

We have investigated the prevalence and possible association of inherited prothrombotic risk factors in patients with primary Raynaud's phenomenon (PRP) and migraine. We performed genotypic analysis of FVLeiden, prothrombin G20210A, methyltetrahydrofolate reductase C677T and FXIII-A V34L mutations in these patients. Two hundred patients with primary Raynaud's phenomenon of Hungarian origin with migraine (57 female, one male, mean age of 43.8 ± 11.5 years) or without migraine (101 female, 41 male, mean age of 41.8 ± 14.5 years) were included in this study. Duration of PRP among migrainous patients was significantly longer than patients without migraine. The prevalence of methyltetrahydrofolate reductase T677 allele among patients with migraine was significantly higher than in patients without migraine (odds ratio 2.1, 95% CI: 1.4-3.3, p = 0.001). The prevalence of other thrombosis-associated alleles did not differ between patients with or without migraine. FVLeiden mutation, prothrombin G20210A mutation, and FXIII-A V34L polymorphism have no apparent effect on the occurrence of migraine in PRP.

Monoclonal antibodies in the treatment of lung cancer.

BACKGROUND: Lung cancer is an aggressive disease and its conventional therapy is far from success. There is a strong need for new, better approaches to improve survival, symptom control and quality of life. METHODS: The authors searched the literature for indexed articles published over the past 30 years from Pubmed concentrating on all possibilities of monoclonal antibodies in the therapy of tumours and especially of lung cancer. RESULTS: The search resulted in more then 200 published articles. Important major reports of the pre-clinical/clinical investigations of monoclonal antibodies in the therapy of tumours, with an emphasis on lung cancer were reviewed, screened and tracked for other relevant publications and the yielded data were summarized and systematized. CONCLUSION: It is concluded, that immunotherapy and the reviewed use of monoclonal antibodies in the therapy of tumours (including lung cancer) certainly carries a hope. However, studies of this topic are in a wide range of phases, from experiments to clinical trials, thereby their results are not comparable with each other. Based on the data available though the authors feel that active immunization with monoclonal antibodies as anti-idiotype vaccines, and antibody targeting with immunoconjugates (immunotoxins, radioimmunoconjugates and chemoimmunoconjugates) are the most promising methods. Radioimmunoguided surgery and immunoguided focal ablation are also valuable. Anti-growth factor monoclonal antibodies are the most evaluated agents so far. They certainly have an objective effect, though they are still not the 'magic bullets', waited for by many clinicians. The use of monoclonal antibodies against the escape mechanisms of tumours can be a good auxiliary method. There are too little data on the value of antibodies directly targeting tumour cells and on combined passive immunotherapy. Due to constant research, other modalities, such as prodrug activation, T cell activation, the use of intrabodies, T bodies, and conjugated antibody fragments might also prove to be valuable.

Plasma endotoxin level of healthy donors.

The plasma level of endotoxin was determined in 116 healthy blood donors. After a routine physical and laboratory investigations the endotoxin level was determined with Limulus amebocyte lysate assay (LAL-test) by the chromogenic kinetic method of Bio-Whittaker Co. (USA). Its sensitivity was 0.005-50 EU/ml. The plasma level of endotoxin in most of the healthy donors was less than 1 EU/ml (in the range of 0.01-1.0 EU/ml), but always measurable. The average +/- S.D. was 0.128 +/- 0.215 EU/ml. Because of the high standard deviation and high range of values, the data were distributed into two groups with the means of 0.05 +/- 0.022 EU/ml and 0.294 +/- 0.186 EU/ml. The difference between the groups was significant (p < 0.001). In conclusion, endotoxin can be measured in plasma of healthy individuals.

[Schoenlein-Henoch purpura in adulthood (gastrointestinal manifestation and endoscopy)]. / Die Schoenlein-Henoch-Purpura bei erwachsenen. (Gastrointestinale Manifestation und Endoskopie).

During a 15-year-period 62 adult patients were admitted with diagnosis of Schoenlein-Henoch purpura in our hospital. 25 female and 37 male patients ranged from 30-87 years (mean: 59.5 years) presenting with cutaneous, joint, renal and particularly abdominal involvement were investigated retrospectively. During the course of the disease, all patients developed purpuric rash (100 %), 14 (22,5 %) patients had joint symptoms and renal involvement occurred in 12 (19,3 %) patients. In this study, we discuss 15 (24 %) patients with gastrointestinal symptoms appearing in Henoch's purpura. Analysis of the gastrointestinal clinical features revealed: Abdominal pain 13 (86 %), massive colorectal bleeding 3 (20 %), occult blood loss 10 (66 %) vomiting 6 (40 %) and diarrhea in 3 (20 %) patients. Surgical consultation was obtained for 4 of the 15 patients and laparotomy was performed in 2 patients. All the patients underwent lower and upper endoscopic examination, in 3 cases the authors saw purpuric mucosal lesions in duodenum and in 8 patients were also found coin-like elevated lesions, additionally, biopsy from colonic lesions showed leukocytoclastic vasculitis. It is concluded that endoscopy may play a very important role in the diagnosis and treatment of Schoenlein-Henoch purpura.

[Current pharmacotherapy of Parkinson's disease]. / A Parkinson-kór korszerú gyógyszeres kezelése.

Parkinson's disease (PD) is one of the most common neurodegenerative disorders. The morbidity is about 1-2 of 1000 under 65 years, but it increases to 1-2 of 100 in the population over 65 years. PD became the model of degenerative disorders either concerning the etiopathogenesis or the therapeutic possibilities. Recently the therapy is focused mostly on pharmaceutics, although the treatment of PD is complex. The new drugs improve the quality of life, and there is some evidence that some of them may slow down the progression of the disease. These drugs may provide a neuroprotective therapy, and so a better life expectancy.

[Plasma endotoxin level in healthy blood donors]. / Egészséges véradók plazma endotoxin szintje.

The plasma level of endotoxin was determined in 116 healthy blood donors in this laboratory. After a routine physical and laboratory investigations the endotoxin level was determined with Limulus amebocyte lysate assay (LAL test) by the chromogenic kinetic method of Bio-Whittaker Co. (USA). Its sensitivity was 0.005-50 EU/ml. The plasma level of endotoxin in most of healthy donors was less than 1 EU/ml (in the range of 0.01-1.0 EU/ml), but always measurable. The average +/- S. D. was 0.128 +/- 0.215 EU/ml. Because of the high standard deviation and high range of values, the data were distributed to two groups with the means of 0.05 +/- 0.022 EU/ml and 0.294 +/- 0.186 EU/ml. The difference between the groups was significant found (p < 0.001). In conclusion, endotoxin can be measured in plasma of healthy individuals.

X-irradiation-induced changes of the prelysosomal and lysosomal compartments and proteolysis in HT-29 cells.

As a consequence of external and internal ionizing radiation, lysosome-like bodies have been observed to increase both in size and number in some cell types. We investigated this process by morphological methods (electron microscopy, cationized ferritin uptake, acid phosphatase histochemistry, morphometry) in cultured HT-29 cells. In parallel with these studies, we measured the rate of protein degradation on the basis of 14C-valine release from prelabeled cellular proteins. We found that at 2 and 4 Gy doses of X-irradiation the volume of the vacuolar (probably lysosomal) compartment increased without detectable changes of acid phosphatase activity. A 2 Gy irradiation dose did not change protein degradation rate. However, 4 Gy caused a significant inhibition of 14C-valine release from prelabeled proteins. Our results indicate, that the radiation induced expansion of the lysosomal compartment is not necessarily accompanied by increased lytic activity of HT-29 cells.

Radiodetoxified endotoxin-induced tolerance. Effect on endotoxin lethality and macrophage arachidonic acid metabolism.

Endotoxin (LPS) tolerance produces changes in macrophage mediator production which is thought to be responsible for the acquired LPS resistance. Detoxification of LPS by gamma irradiation has been reported to diminish certain noxious properties while retaining its tolerance inducing actions. We compared the efficacy of LPS and radiodetoxified (RD)-LPS from Escherichia coli O101 on stimulating rat peritoneal macrophage arachidonic acid (AA) metabolism, measured by thromboxane (TXB2). Changes in macrophage production of these mediators were also assessed after tolerance induction. LPS tolerance was induced by i.p. injection of LPS, RD-LPS or vehicle on day 1 (100 micrograms/kg, i.p.) and day 2 (500 micrograms/kg, i.p.). On day 5 or 4 weeks after pretreatment, peritoneal macrophages were harvested for in vitro studies, or rats were tested for lethality resistance. Macrophages were incubated +/- LPS (0.1 ng to 50 micrograms/ml), lipid A (1 or 10 micrograms/ml) or Ca+2 ionophore A23187 (10 microM) for determination of TXB2 production. Minimum effective concentrations of LPS and RD-LPS for stimulation (P < 0.05) of TXB2 were 100 ng/ml and 1 microgram/ml, respectively. Maximal stimulation of TXB2 occurred at 10 micrograms/ml of LPS or RD-LPS. Macrophages from LPS or RD-LPS tolerized rats were refractory to stimulated TXB2 with LPS or RD-LPS (0.1 ng to 50 micrograms/ml). The suppressed in vitro macrophage TXB2 production was apparent 4 weeks after rats were tolerized with LPS or RD-LPS. In subsequent mortality studies, LPS challenge of control or tolerance rats at day 5 in vivo with Salmonella enteritidis LPS (15 mg/kg, i.v.) produced a 90% mortality in control rats (N = 22), versus 13% mortality in the LPS pretreated group (N = 23) and a 20% in the RD-LPS pretreated group (N = 10) (P < 0.05 vs control). However, this lethality resistance was not apparent at 4 weeks after LPS or RD-LPS pretreatment. Both LPS and RD-LPS appear to be equipotent in inducing macrophage alterations, and in lethality resistance during LPS tolerance induction. However, these observations suggest that during LPS tolerance suppression of LPS-stimulated AA in peritoneal macrophage metabolism persists longer than acquired lethality resistance.

Effect of irradiation on the GTP binding kinetics of chicken embryo brain plasma membranes.

The membrane bound form of the catalytic subunit of adenylate cyclase of chicken embryo brain has been found earlier to be rather radioresistant [28]. The radiation induced changes of G proteins of membrane preparations of 19 day old chicken embryo brains were investigated in this work. The activation of catalytic subunit of adenylate cyclase by G protein dependent activators (Gpp/NH/p and NaF) was found elevated at lower radiation doses (0-400 Gy), while both basal enzyme activity (measured without any activator) and the activity measured in the presence of activators decreased at higher doses (above 800 Gy). Heterogeneity of GTP binding sites measured with 3-H-Gpp/NH/p a GTP-ase resistant GTP analogue was observed in the case of control (with Kd1 = 0.0663 +/- 0.034 mumol/l, Bmax = 0.0079 +/- 0.0022 nmol/ml and Kd2 = 2.038 +/- 0.4779 mumol/l, Bmax2 = 0.0291 +/- 0.0017 nmol/ml). The lower affinity high capacity binding sites seemed to be more radiosensitive, than the higher affinity sites. A marked decrease was observed in the number of low affinity binding sites above 200 Gy and these low affinity binding sites practically disappeared after irradiation with 400 Gy. At high doses (above 1600 Gy) the catalytic subunit was damaged, too. On the basis of the decrease of low affinity binding sites together with an increase in activation of the catalytic subunit via G proteins one can conclude that it is caused by radiation induced damage of Gi protein that can be more radiosensitive, than Gs protein.

Effect of radioprotectors on the aggregability, cAMP and thiol content of blood platelets.

Aminothiol radioprotectors (MEA, AET, WR-1065) were found to be inhibitors of platelet aggregation. The aim of this study has been to clarify the potential involvement of the adenylate cyclase-cAMP system in this effect. In vitro aminothiol compounds (> 1 mmol/l) inhibited platelet function as well as AC-activity, i.e. significantly reduced their cAMP content. In less than 1 mmol/l concentration WR-1065 elevated the cAMP content of PGI2-stimulated platelets, while basal cAMP level mainly remained unchanged. On the effect of WR-1065 the level of non-protein thiols (NP-SH) markedly elevated in the platelets, while their protein-bound thiol content (PB-SH) either decreased or remained at control level. The phosphorothioate WR-2721 was effective only after in vitro or in vivo dephosphorylation. Although the amount of NP-SH slightly increased, the PB-SH content decreased in the platelets isolated after intravenous administration of WR-2721. Moreover, platelet-poor plasma samples of WR-2721 treated animals also inhibited both the AC-activity and function of control platelets, too. Based on these results it is suggested that aminothiol radioprotectors non-specifically inhibit platelet functions forming mixed disulfides with endogenous, mostly protein-bound thiols. The slight elevation of AC-activity caused by low doses of WR-1065 also suggest that the AC-complex consists of unevenly sensitive subunits (G-proteins, catalytic subunit).

Adenylate cyclase-the more membrane associated, the less radiosensitive.

A dose dependent but not parallel decreases were observed both in SH content and catalytic activity of "free" catalytic subunit after irradiation (0-3200 Gy), while SH groups of membrane-associated adenylate cyclase were insensitive (under 3200 Gy). An initial "radioactivation" of membrane-associated enzyme was found under 800 Gy, then an inhibition above 1600 Gy. The SH alkylating agent, N-ethylmaleimide resulted in a complete inactivation, both of membrane associated form of adenylate cyclase and "free" catalytic subunit with similar inactivation profiles. These data indicate that in the radiosensitivity or "radioprotection" of adenylate cyclase, its membrane association/integration might play a more important role than the SH groups themselves.

Potential role of SH groups in the radiosensitivity of adenylate cyclase.

The role of oxidation of SH groups in the activity of adenylate cyclase and in radiosensitivity of the enzyme was investigated. Adenylate cyclase activity was measured in purified membrane preparation of 19 day old chicken embryo brains. N-ethyl-maleimide (NEM) and lead-acetate were used as SH inhibitors. Gamma irradiation was carried out with 60-Co source. NEM inhibition of adenylate cyclase was dose dependent and 50 per cent inhibition was observed at 40-50 microM NEM. Activity of adenylate cyclase was elevated at lower concentrations of lead-acetate (10 nM-100 microM) and was inhibited at higher concentrations (above 100 microM). The presence of 40 microM NEM did not alter the shape of lead acetate saturation curve of adenylate cyclase. Gamma irradiation in the dose range of 100-800 Gy elevated the adenylate cyclase activity measured in the presence of 5 mM NaF but did not alter the basal activity. Gamma irradiation did not have significant effect on NEM saturation of adenylate cyclase, while it altered slightly the lead acetate saturation curve.

Cyclic AMP level of lymphocytes in patients with systemic lupus erythematosus and its relation to disease activity.

The basal and stimulated intracellular cyclic AMP (cAMP) levels of peripheral blood mononuclear cells (PBMC) of 16 control subjects and 14 patients with systemic lupus erythematosus (SLE), all fulfilling the ARA criteria, were studied. No significant difference in basal cAMP level was observed between SLE patients and controls. SLE lymphocytes (both active and inactive) elicited a diminished response to aminophylline and prostaglandin E2 (PGE2). No correlation was seen between disease activity and either baseline cAMP levels or response to these stimulators. We suggest an intrinsic (not disease activity-related) impairment of the adenylate cyclase-dependent regulatory mechanism in the PBMC of SLE patients, which may result in a defective IL-2 production and IL-2 dependent biological functions.

A possible mechanism of the inhibitory effect of endotoxin on adenylate cyclase.

The possible mechanism of the inhibitory effect of endotoxin on adenylate cyclase was investigated. Plasma membranes were isolated from porcine thyroid gland and rat liver. The effects of endotoxin on adenylate cyclase activity was determined. The adenylate cyclase activity followed in the presence of various concentrations of guanosine-imidodiphosphate (5 x 10(-8)-10(-6) M) and NaF (0.1-10 mM) was markedly inhibited by endotoxin. Moreover, the basal activity of adenylate cyclase was inhibited similarly. The inhibition was concentration dependent. At 400 micrograms/ml of endotoxin a 60% inhibition of adenylate cyclase both without and in the presence of activators was detected. The inhibitory effects of endotoxin and its radiodetoxified derivative were similar. Neither of the two investigated endotoxin preparations tested had any effect on the 3H-guanosine-imidodiphosphate binding activity of adenylate cyclase (neither on the binding capacity, nor on the Kd value). Forskolin activated adenylate cyclase was also inhibited markedly by both endotoxin and radiodetoxified endotoxin. These results suggest that beyond the other membrane effects there is a probability of direct inhibitory effect of endotoxin on the catalytic subunit of adenylate cyclase.

Effect of X-irradiation on adenylate cyclase activity and cyclic AMP content of primary human fibroblasts.

Ionizing radiation provokes an increase of the cAMP level in several organs and body fluids. After reviewing the relevant literature we present the results of our own experiments on primary human fibroblasts. X-irradiation at doses of 0.5 and 2.5 Gy in vitro evoked a rapid and reversible increase of adenylate cyclase enzyme activity. A significant increase in cAMP level of these cells was also observed. Adenylate cyclase was usually localized basolaterally on the surface of unirradiated cells, while irradiation resulted in a modification of distribution, i.e., the enzyme activity also appeared in apical localization.

(-)Deprenyl in the treatment of decompensated Parkinson's disease.

Two main forms of declining motor performance are evident in Parkinson's disease: response fluctuations and "loss of benefit", i.e., the progression of the disease without "on-off" symptoms. (-)Deprenyl has a favourable beneficial effect in reducing the mild forms of response fluctuations. The addition of (-)deprenyl in such patients to the continuing substitution therapy prevents the development of more severe "on-off" manifestations. In severely disabled patients with irregular response swings or permanent akinesia the use of (-)deprenyl as an adjuvant drug cannot modify anymore the course of the disease.