Quantitative and qualitative assessment and clinical meaning of molecules removed with BK membranes.
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Biomedical subjects
Publications and source records attributed to F Galli.
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The effects of nitric oxide (NO) or related molecules on the binding of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to the red blood cell (RBC) membrane were investigated. It was demonstrated that submillimolar concentrations of the NO donor sodium nitroprusside (SNP) not only strongly inactivated GAPDH by S-nitrosylation of the enzyme thiols but also decreased the binding affinity of GAPDH for the RBC membrane. In fact, the incubation with SNP for 60 min at 30 degrees C and at a concentration > 50 microM induced the dissociation of the native GAPDH from the white unsealed membranes (standard ghosts) in a concentration-dependent manner with a partial recovery of the enzyme activity and thiols when SNP concentrations higher of 1 mM were used. Binding experiments under saturating conditions indicate a Ka value for the nitrosylated GAPDH of 3.5 +/- 0.8 x 10(6) M-1, which was more than 50% less than the Ka value of 7.6 +/- 0.6 x 10(6) M-1 observed for the native enzyme. These data were also confirmed in reassociation experiments under nonsaturating conditions. Dithiothreitol (DTT), which at concentrations of less than 1 mM catalyzed the S-nitrosylation of GAPDH and the consequent modification of the binding properties described above, the concentrations higher than 5 mM restored both the enzyme activity and the binding properties. Furthermore, the enzyme-membrane association induced before the incubation step afforded at least partial protection from the loss of titrable thiols and from the inactivation induced either spontaneously or by SNP. Taken together, these data not only confirm the key role of the active site cysteine residues in the catalytic function of GAPDH but also suggest that they may be involved in the NO-dependent regulation of GAPDH binding to the RBC membrane.
OBJECTIVE: Headache is a notable problem in clinical practice and a frequent symptom in childhood and adolescence. The main aim of the present study is to analyze the evolution of migraine and tension headache (TH) using an 8-year follow-up. METHOD: 100 subjects (F60, M40; mean age 17.9; SD 2.6; range 12-26), randomly selected among all patients first seen in 1988 at the Headache Center, were directly contacted. We employed IHS criteria both in 1988 (the data were taken by the clinical charts) and 1996. We took into account changes in headache types and improvement, unchanging, worsening or remission of headache. This analysis was made with regard to gender differences and age at onset of headache, too. The chi-squared test is employed. FINDINGS: High tendency to remit (34%) or improve (45%) was recorded. A worsening situation was seen in 6% and an unchanging situation in 15%. In 1988, we had 57% migraine without aura (MwoA), 7% migraine with aura, 28% episodic tension-type headache (ETTH), and 8% chronic TH (CTTH). In 1996, we saw 30% MwoA, 2% MwA, 31% ETTH, and 3% CTH. Migraine shows a lower tendency to remit than TH (28.1% vs 44.4%). MwoA persists in the same form in 43.8% and becomes ETH in 26.3%. ETTH persists in the same form in 26.3% and changes in MwoA in 10.7%. Of headache-free subjects, we recorded a high tendency to remit (34%) and improve (95%); 13 were females (21.7%) and 21 were males (52.5%). The course of headache is not related to age at onset. CONCLUSION: Headache with juvenile onset changes its characteristics over time, with a high tendency to remit (mostly in males) or improve. The implications for pathophysiology and the role of hormonal factors are called into question.
UNLABELLED: Migraine with juvenile onset changes over time. The existence of prognostic factors is a point of focus. A strict relationship between migraine or tension-type headache (TTH) and psychiatric factors has been suggested, but the exact role and the influence on evolution of headache is unknown. OBJECTIVE: To analyze the evolution of migraine and TTH and psychiatric comorbidity (P-Co) from 1988 to 1996. MATERIAL AND METHOD: 100 subjects (40M, 60F; mean age 17.9 years; SD 2.7 years; range 12-26 years) were examined at our Center. The International Headache Society (IHS) criteria were employed. Psychometric tests and clinical interviews aided psychiatric diagnosis (DSM-III-R). SCID (Structured Clinical Interview for DSM-III-R) was employed in 1996. Chi square and logistic regression are used for statistical analysis. FINDINGS: Migraine and TTH change their clinical characteristics, with a high tendency to remission (mostly in males). The presence of P-Co in 1988 is related to a worsening or unchanging situation in 1996. Headache-free subjects did not present any psychiatric disorders in 1996. Anxiety disorders in 1988 are related to enduring of headache. Migraine shows comorbidity with anxiety disorders and depression. CONCLUSION: P-Co is a notable problem in clinical practice. Diagnostic, prognostic, and treatment implications require a systematic assessment of P-Co.
BACKGROUND: The present study was designed to test the biocompatibility of a new vitamin E-modified multi-layer membrane (CL-E filter), as well as its ability to protect against oxygen free radicals during hemodialysis (HD). METHODS: We investigated, both in vitro and in vivo, the bioreactivity of the filter with respect to the blood antioxidants and its ability to prevent lipoperoxidation. The effects on the leukocyte respiratory burst were also studied. Cuprammonium rayon was used as a comparison material (CL-S filter). RESULTS: The in vitro results demonstrated that, under controlled conditions, CL-E is able to preserve blood antioxidants, and particularly vitamin E, from the spontaneous consumption observed in the incubation with CL-S filters and in control incubations. In accordance with this observation, the rate of the oxidative demolition of lipids either in plasma and red blood cells (RBC) or from rat brain homogenate decreased after the exposure to CL-E filters in comparison with the CL-S filter. Moreover, in the absence of any significant cytotoxic effects due to both the types of material studied, the production of oxygen free radicals and nitric oxide (NO) by leukocytes was higher after their in vitro exposure to CL-S, but was quite similar to that of the control leukocytes after exposure to CL-E. In vivo, a one-month treatment with the CL-E filter increased plasma vitamin E by 84.3% with respect to treatment with CL-S; this gain slightly decreased to 68.9% when CL-E treatment was prolonged to three months. In the RBC, vitamin E was found to have increased by 76.7% and 113.4% at one and three months, respectively. Plasma glutathione (GSH) levels determined at three months were significantly increased from 0.10 +/- 0.02 to 0.33 +/- 0.12 mumol/ml, while the erythrocyte GSH was only slightly increased. The leukocyte function estimated as responsiveness to soluble chemical stimuli in CL-S-treated patients was significantly improved both qualitatively and quantitatively after CL-E treatment. The presence of an increased number of mononuclear cells undergoing programmed cell death (apoptosis) in CL-S-treated patients (18.8 +/- 1.7% vs. a control value of 6.5 +/- 2.3%) as well as the apoptogenic effect of their plasma in vitro on U937 cells was significantly corrected after CL-E treatment (mean decrease in apoptotic mononuclear cells at 24 hours of culture, 25.5% and 27.1% at 1 and 3 months, respectively). The anti-apoptogenic effect of CL-E treatment showed a close dependence on the increase in vitamin E in the blood cell compartment. CONCLUSIONS: This study suggests that this vitamin E-modified membrane can be considered a highly biocompatible material, the antioxidant properties of which can exert a site-specific and timely scavenging function against oxygen free radicals in synergy with a hypostimulatory action on the PMN respiratory burst.
Allergic-type reactions experienced by patients on chronic hemodialysis are frequently reported in the literature, often related to compounds released during the hemodialysis sessions, in particular ethylene oxide (ETO). In these studies, dialysis patients seem to have higher values of IgE than those observed in a reference population. The aim of our work was to investigate IgE-related parameters of 126 dialysis patients in comparison with two control groups composed of healthy subjects and predialysis patients. IgE values were not significantly different in nonallergic dialysis patients, with a geometric mean (X) of 27.5 kU/l, when compared to healthy controls (X = 38 kU/l) and predialysis subjects (X = 40.7 kU/l). Higher values of IgE (X = 74.1 kU/l) were detected in allergic dialysis patients. However, only 3 patients, all without detectable antibodies against ETO, complained of anaphylactic symptoms during dialysis sessions in a 4-year surveillance period. At variance, 6 symptom-free patients carried IgE antibodies against ETO (range 0.7-15 kU/l), usually with high total IgE. Our results suggest a reduced frequency of hypersensitivity reactions during dialysis sessions. Furthermore, uremia does not alter IgE reactivity in the predialysis period or after long-term maintenance dialysis treatment.
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A comparison of the oxyradical exposure during hemodialysis (HD) carried out with vitamin-E-modified cellulose (CL-E) or conventional membranes, studying red blood cell (RBC) and plasma lipoperoxidation and RBC glutathione metabolism, was done. In this preliminary characterization of a new and original approach to the prevention of free radical damage in HD, the results obtained indicate that lipoperoxidation in plasma and RBC is decreased and therefore oxidative damage can be significantly decreased using CL-E dialysis membranes instead of conventional membranes.
Red blood cells and plasma reduced and oxidized glutathione levels, glutathione peroxidase (GSH-Px) activity, thiobarbituric acid reactants (TBAR) of both chronic ambulatory peritoneal dialysis (CAPD) patients and a matched control group were investigated in this study. Oxidized and reduced pyridinic nucleotides in red blood cells (RBC), in which NADPH is a direct expression of hexose monophosphate shunt function, were also studied. The results obtained indicate that RBC and plasma are exposed to oxidative stress in CAPD. This condition is characterized by a decreased GSH/GSSG ratio, particularly evident in RBC as a consequence of the GSSG accumulation. Lipid peroxidation is increased, as indicated by raised TBAR levels, and reduced pyridinic nucleotides are decreased. Increased GSH-Px levels and unmodified or slightly increased GSH content were observed in the RBC but not in plasma, which showed decreased GSH and unmodified peroxidase activity. Peroxidase correlated positively with TBAR levels in the RBC lysates. In a subgroup of patients treated with erythropoietin (vs. untreated patients and controls) no differences were observed in the glutathione-related parameters studied. These data suggest that a mechanism for adaptation to oxidative conditions may be present in CAPD and its effects on RBC integrity are discussed in comparison with the hemodialysis conditions previously studied.
Pruritus is a common symptom in uraemic patients and its aetiology is poorly understood, although several factors have been implicated, including high histamine plasma levels. The aim of the study was to evaluate plasma levels of histamine in patients on maintenance haemodialysis in relation to the presence of itching (35 pruritic patients = group A; 50 non-pruritic patients = group B). Furthermore, we compared the values obtained with those of a healthy control group (64 subjects). We observed increased plasma histamine levels in uraemic patients compared with healthy controls. However, no relationship was found between plasma histamine values and itching because groups A and B showed overlapping values and there was no correlation between the intensity of pruritus and plasma histamine concentrations in group A patients. In conclusion, we suggest that there is no evidence for plasma histamine playing a significant part in uraemic pruritus.
The authors have investigated the efficacy of i.m. therapy with interferon (IFN) in the treatment of female genital condylomatosis: 94 consecutive patients underwent a randomized therapy with two different IFNs: beta-IFN and alpha-2A-IFN (3,000.000 UI i.m. on alternate days for 4 weeks). A total and/or partial response was observed in 76.5% of cases (72/94). Observed response was unrelated with used IFN, independently of type and seat of treated lesion. There was only an important difference between the two treatments in the incidence of side-effects and drop-out (31% and 6.3% with alpha-2A-IFN and 14.8% and 2.1% with beta-IFN). Systemic therapy with IFN is a good alternative to destructive techniques in the treatment of HPV-associated pathology of female lower genital tract.
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Between 1987 and 1991 219 patients (1.3% of all hospitalized patients) with acute infectious diarrhea were investigated retrospectively. 52% of the patients were hospitalized and 48% were outpatients. In 55% the inducing diarrhea microorganism could be identified. The most frequently detected pathogens were endemic Salmonella sp. and Campylobacter jejuni (65%). Imported diarrheas (Shigella sp. and parasites) were rare, as only 15% of the patients had a history of travel. All Clostridium difficile infections were associated with antibiotic treatment. The stool examinations for bacteria were positive in 92/160 patients (57%). Stool examinations for occult blood or fecal leukocytes were highly useful in detecting the infectious agent. In 70% of the patients with positive occult blood test or 81% of the patients with fecal leukocytes, an infectious agent in the stool was found. All patients recovered with few complications. 31% of the patients were treated by antibiotics because of septic disease or pathogenic parasites.
The activity and some kinetic properties of RBC Na+,K(+)-ATPase (EC 3.6.1.37) were investigated in essential hypertensives (EH; 40 subjects) and normotensives (NT; 20 subjects). A decrease in ouabain-sensitive 86Rb uptake as well as ouabain-sensitive ATPase activity was found in EH. [Na+]i and [K+]i of EH did not show any statistical difference from NT. Na+,K(+)-ATPase showed a reduced Mg2+ activation and the apparent Km value for Mg2+ was 2-fold increased in the EH group. The influence of temperature on the Na+,K(+)-ATPase showed a reduced modulation and a minor activity peak at 37 degrees C in the patients, consequently the calculated activation energy of the enzyme was increased at temperatures lower than 40 degrees C. Increased RBC adenylate energy charge (EC) was observed in EH when compared with NT. A negative correlation between EC and total Na+,K(+)-ATPase activity was found when all subjects were compared and also in both groups, showing a possible pump involvement in the regulation of the RBC metabolic flux in EH. These data provide evidence about some modifications in active Na+,K+ transport and in EC in RBC which allows a further characterization of membrane cation fluxes in EH.
The objective of this study was to examine peripheral vascular function before and after cardiac transplantation and to assess the effect of immunosuppressive therapy on peripheral vascular reactivity. Peripheral vascular function abnormalities present in congestive heart failure may be reversed with cardiac transplantation, but immunosuppressive therapy may alter these changes in the peripheral vasculature. Venous occlusion plethysmography was used to study peripheral vascular function in nine patients with severe congestive heart failure who underwent cardiac transplantation. Forearm blood flow and forearm vascular resistance were measured in patients with congestive heart failure in response to cold stimulation, maximal hyperemia, and hand grip exercise (1) before transplantation; (2) 24 to 36 hours posttransplantation before the commencement of cyclosporine; (3) 6 to 8 days posttransplantation in the presence of therapeutic cyclosporine levels; and (4) 6 weeks posttransplantation. Venous capacitance was also measured. After cardiac transplantation, mean arterial pressure increased and remained elevated. Forearm blood flow initially increased after transplantation but subsequently decreased with cyclosporine. Cold-induced reflex sympathetic activation decreased immediately after transplantation but was significantly enhanced with cyclosporine. The maximal vasodilatory response following ischemic cuff occlusion and with 5 minutes of isometric hand grip exercise increased significantly after transplantation and remained improved at 6 weeks. Thus after cardiac transplantation, peripheral vasodilator function improves and is not altered by cyclosporine. However, with cyclosporine therapy resting forearm vascular resistance increases and reflex sympathetic vasoconstriction is enhanced, suggesting that cyclosporine may potentiate adrenergic-mediated peripheral vasoconstriction and thus may contribute to posttransplant hypertension.
OBJECTIVES: The purpose of this study was to test the hypothesis that active compression-decompression would improve resuscitation success in human subjects after cardiac arrest. BACKGROUND: Active compression-decompression cardiopulmonary resuscitation is a new method that improves cardiopulmonary hemodynamic function in animal models and humans after cardiac arrest. METHODS: We conducted a prospective randomized clinical trial in patients with in-hospital cardiac arrest. Patients were assigned to receive standard manual or active compression-decompression cardiopulmonary resuscitation. The primary study end points were spontaneous return of circulation, 24-h survival and survival to hospital discharge. RESULTS: Fifty-three consecutive patients after cardiac arrest undergoing 64 resuscitation attempts were studied (30 women, 23 men; mean [+/- SD] age 71 +/- 13 years, range 38 to 96). Spontaneous return of circulation was observed in 24 (47%) of 53 patients and was increased in patients receiving active compression-decompression compared with those receiving standard manual cardiopulmonary resuscitation (15 [60%] of 25 vs. 9 [32%] of 28, respectively, p = 0.042); 24-h survival was increased (12 [48%] of 25 vs. 6 [21%] of 28, respectively, p = 0.041); and there was a trend toward improved survival to hospital discharge (6 [24%] of 25 vs. 3 [11%] of 28, respectively, p = 0.198) when active compression-decompression was compared with standard manual cardiopulmonary resuscitation. CONCLUSIONS: Active compression-decompression cardiopulmonary resuscitation improves return of spontaneous circulation and 24-h survival after in-hospital cardiac arrest. Active compression-decompression cardiopulmonary resuscitation appears to be a beneficial adjunct to standard manual cardiopulmonary resuscitation.
Reduced and oxidized glutathione and pyridine coenzymes, glutathione-related enzymes and Cu,Zn-superoxide dismutase (Cu,Zn-SOD) were investigated in the RBC of patients with chronic renal failure (CRF) and in age- and sex-matched controls. The effects of hemodialysis (HD) were also studied. A defective RBC redox state was shown in the CRF group based on a decreased GSH/GSSG ratio and NADPH levels. Increased activities of glutathione transferase (GSH-S-T) and Cu,Zn-SOD were observed before HD. Dialysis apparently restores the levels of antioxidant enzymes and at the same time strongly affects the redox state. Thus we can speculate that HD can generate severe redox impairment inducing damage in RBC and plasma antioxidant enzymes. Increased erythrocyte GSSG and GSM-S-T levels coupled with a reduced hexose monophosphate shunt (HMPS) function may be useful indexes of oxidative stress in uremic anemia.