Plasma Of Healthy Controls Research Articles

Human Herpesvirus 7 in Patients with Pityriasis rosea

Clinical evidence suggests a viral etiology for pityriasis rosea (PR). To evaluate human herpesvirus (HHV)-6 and HHV-7 as candidates for the etiology of PR. Blood and skin tissue from 12 patients with acute PR, and 12 patients with other dermatoses were studied, as well as blood samples from 25 healthy persons. Serum interferon (IFN)-alpha and IFN-gamma were analyzed by ELISA. Analysis of morphological changes in cocultured peripheral blood mononuclear cells (PBMC) and electron microscopy (EM) to identify viral particles were performed. Polymerase chain reaction (PCR) with specific primers for HHV-6 and HHV-7 DNA sequences was performed on the plasma and PBMC of patients and healthy controls and on the skin of patients with PR and other skin diseases. PR plasma contained detectable IFN-alpha and IFN-gamma, whereas plasma from controls did not. PBMC from PR patients showed ballooning cells and syncytia after 7 days in culture whereas PBMC from controls and recovered PR patients did not. This cytopathic effect was also documented in a PR patient who relapsed and in Sup-T1 cell cultures inoculated with the cell-free supernatant from centrifuged cultured PBMC; in this supernatant, herpesvirus, virions were detected by EM, PCR identified HHV-7 DNA in PBMC, plasma and skin from all patients with active PR and in the PBMC only of 5 patients tested 10-14 months later. Weaker signals of HHV-7 DNA were detected in PBMC of 11 controls, but not in their plasma. Skin was negative for HHV-7 in all control specimens. Although the detection of HHV-7 DNA in PBMC and tissues does not prove directly a causal role, HHV-7 DNA in cell-free plasma corresponds to active replication which supports a causal relationship. We propose that PR is a clinical presentation of HHV-7 reactivation.

Anomalous phospholipid n-6 polyunsaturated fatty acid composition in idiopathic calcium nephrolithiasis.

Anomalies in the erythrocyte transport of anions and cations have been described in idiopathic calcium oxalate nephrolithiasis and seem to play a pathogenetic role in this disease. In consideration of the hypothesis that the complex array of ion flux cell abnormalities is an epiphenomenon of an anomaly in the composition of cell membranes, this study investigated cell-membrane lipid composition. In idiopathic calcium oxalate renal stone formers, in which ion transport abnormalities were present, and in healthy control subjects, plasma and erythrocyte membrane lipid composition, the erythrocyte oxalate exchange, and Na/K/2Cl cotransport activity were evaluated. Furthermore, in stone formers, the effect of a 30-day fish-oil diet supplementation on plasma lipids, erythrocyte oxalate exchange, oxaluria, and calciuria was investigated. The effect of archidonic acid released by phospholipase A2 on anion-carrier phosphorylation and activity in erythrocytes was evaluated as well. Patients had a lower content of linoleic and higher concentration of archidonic acids in both plasma and erythrocyte membrane phospholipids, and an increased archidonic/linoleic acid ratio. The archidonic acid level correlated with the erythrocyte oxalate exchange and sodium cotransport activity. Fish-oil supplementation lowered calcium and oxalate urine excretion, and normalized the erythrocyte oxalate exchange. Phospholipase A2 increased the erythrocyte anion-carrier protein phosphorylation and the oxalate exchange. This study shows that idiopathic calcium nephrolithiasis in the patient group reported here is characterized by a systemic defect in phospholipid archidonic acid levels that might provide an answer to the link between genetic background, dietary habits, and renal lithiasis.

Disturbed motilin and cholecystokinin release in the irritable bowel syndrome.

The irritable bowel syndrome (IBS) is associated with motor abnormalities in the small intestine and colon. Neuropeptides may have an important role in initiating and regulating the intestinal motility. Motilin has been proposed to initiate the peristaltic reflex in the small intestine and cholecystokinin the gastrocolic reflex. In 18 patients with IBS and 11 healthy control subjects plasma motilin and cholecystokinin (CCK) concentrations were measured after intraluminal stimulation of water and a fat-rich meal. The IBS patients had reduced motilin secretion after both water intake and the fat meal. In contrast, the fat meal elicited an exaggerated and prolonged CCK release in the IBS patients. Disturbed motilin and CCK release may partly be responsible for the intestinal dysmotility in the IBS patients.

Antibodies against CD4 + lymphocytes in plasma of HIV-infected patients are related to CD4 cell depletion in vivo

The role of autoimmune phenomena in the pathogenesis of AIDS is not well understood. Antibodies against CD4 + lymphocytes are frequently detectable in HIV-infected patients. However, the relevance of these antibodies is unknown. In this study anti-CD4 cell antibodies in plasma of HIV-infected patients were compared to patients' CD4 cell count. Lymphocytes of a healthy donor were incubated with plasma of patients and controls. Antibodies against CD4 + lymphocytes were detected using anti-human Ig antibodies and monoclonal anti-CD4 antibodies simultaneously. The degree of staining was visualized by flow cytometry. The experiments revealed that 60% of HIV-infected patients harbored anti-CD4 cell antibodies in their plasma. Anti-CD4 cell antibodies were not detectable in plasma of healthy controls. Patients with anti-CD4 cell antibodies in their plasma presented with significantly lower numbers of circulating CD4 + lymphocytes ( P = 0.025). The degree of antibody reactivity was negatively correlated to patients' CD4 cell counts ( P = 0.02). In conclusion there is evidence for an association between plasma anti-CD4 cell antibodies and CD4 cell depletion. Whether this association represents an epiphenomenon or a pathogenetic relevant pathway needs to be investigated in further studies.

Tryptophan and its metabolites in patients on continuous ambulatory peritoneal dialysis and following renal transplantation

Plasma tryptophan (Trp) concentration in its total (TTrp) and non-protein-bound free form (FTrp) and its metabolite 5-hydroxyindole-3-acetic acid (5-HIAA) as well as muscle Trp contents (MTrp) were studied in 12 uraemic patients undergoing continuous ambulatory peritoneal dialysis (CAPD), 10 renal transplant patients, and 10 healthy control subjects. The CAPD patients exhibited signs of muscle weakness, as assessed by dynamometry, and signs of protein malnutrition with a decreased ratio of alkali-soluble protein relative to DNA in muscle as well as low serum albumin concentration. In the uraemic patients TTrp was markedly reduced, whereas in the transplant patients it did not differ from the controls. The FTrp, which was separated by the process of equilibrium dialysis, was less in the uraemic (P < 0.01) as well as transplant patients (P < 0.01) than in the controls. The plasma FTrp/TTrp ratio was increased in the uraemic patients (40 +/- 8%) but less in the transplant patients (16 +/- 4%), as compared to the controls (25 +/- 5%). The uraemic patients had increased plasma concentrations of 5-HIAA, whereas this metabolite could not be found in the plasma of renal transplant patients and healthy controls. MTrp was increased by an average of 33% in the uraemic patients whereas it did not differ between the transplant patients and the controls. The results indicate that the abnormal Trp metabolism in uraemic patients is to a large extent corrected by a successful renal transplantation.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of the antidepressant drug tianeptine on plasma and platelet serotonin of depressive patients and healthy controls

We have examined the effects of a single 12.5 mg dose and of 12 weeks treatment up to 37.5 mg daily with tianeptine, a new antidepressant drug that potentiates in vivo the uptake of serotonin (5-HT). On day 0, tianeptine reduced plasma 5-HT concentration. This acute effect occured also on subsequent examination days. However, long-term treatment tended ( P < 0.06) to increase basal plasma 5-HT concentrations, in covariation with decreases of MADRS (Montgomery-Asberg Depression Rating Scale) and HARS (Hamilton Anxiety Rating Scale). Platelet 5-HT increased only in elderly patients, probably due to the higher plasma concentration of the drug in this group than in younger patients. These results show that the acute effects of therapeutic doses of tianeptine are consistent with an enhancement of the 5-HT uptake. However, long-term treatment does not result in a decreased plasma 5-HT, as might be expected from the acute effects of the drug.

Immunological properties of apoB-containing lipoprotein particles in human atherosclerotic arteries

In this study, we have documented immunochemical properties of apolipoprotein (apo) B-containing particles (LpB) extracted from human atherosclerotic lesions obtained during vascular reconstructive surgery of patients. These properties were compared to those of particles purified from corresponding atherosclerotic plasma and healthy control plasma. LpB immunoreactivities were tested in solid phase competitive binding radioimmunoassays using five anti-apoB monoclonal antibodies (MAb) for which epitopes have been previously located on the protein. The regions encompassed amino acids 405 to 539 (MAb B1), 1854 to 1879 (MAb B4), 3506 (MAb BA11), and 4355 (MAb BL3). The fifth antibody (MAb BL5) recognizes a conformationally expressed epitope. LpB from lesions presented a significantly decreased immunoreactivity as compared to LpB from respective plasma except for the epitope recognized by MAb BA11 located precisely in the low density lipoprotein (LDL) receptor binding site. The accessibility of the four sequential epitopes was similar on LpB from atherosclerotic and healthy plasma while it was decreased for the conformational one in LpB from atherosclerotic samples. These altered immunoreactivities were not related to changes in chemical composition of LpB as this was quite comparable in all preparations. With regard to electronegativity, apoB fragmentation, immunological accessibility, and size distribution of the particles, changes seem to increase in the following order from healthy plasma, atherosclerotic plasma, and the corresponding lesions. The results confirm some structural characteristics of oxidatively modified particles from human atherosclerotic lesions and to a lesser degree from respective plasma, but more specifically demonstrate a global conformational change in LpB from lesions, this change being perhaps initiated in the plasma.

Basal and stimulated plasma levels of pancreatic amylin indicate its co-secretion with insulin in humans

Amylin is a 37-amino acid pancreatic polypeptide, probably involved in the pathophysiology of Type 2 (non-insulin-dependent) diabetes mellitus. We have determined amylin in human plasma by extraction-based radioimmunoassay (Sep-Pak C18). Of 23 healthy control subjects plasma amylin was determined as 11.9 +/- 3.5 ng/l. Of 27 patients with Type 2 diabetes receiving insulin the amylin levels were lower, and in 16 patients with Type 2 diabetes on oral medication they were higher than in the control subjects; 8.2 +/- 4.4 ng/l (p less than 0.01) vs 18.8 +/- 9.9 ng/l (p less than 0.05). In 14 Type 1 (insulin-dependent) diabetic patients we found extremely low mean amounts of amylin: 2.9 +/- 1.9 ng/l (p less than 0.002). Thus, basal amylin appears to be associated with the capacity to release insulin. An oral glucose load stimulated the release of amylin, this was more pronounced in patients with Type 2 diabetes than in healthy subjects. An excellent correlation of mean amylin with mean insulin concentrations was obtained (r = 0.949). In patients with Type 2 diabetes amylin was reduced congruent to decreases in C-peptide during a hyperinsulinaemic, euglycaemic glucose clamp experiment (r = 0.971 for linear correlation between C-peptide levels and amylin). We conclude, that amylin and insulin are co-secreted in humans, and that the amylin release is under feedback-control by insulin.

Action of Oren-gedoku-to on Platelet Aggregation in Vitro

Oren-gedoku-to is a Chinese medicine with hypotensive activity, hemostatic actions and the ability to augment cerebral blood flow. It is therefore expected to be useful in patients with hypertension and cerebrovascular accident (CVA). These effects of Oren-gedoku-to on platelet aggregation and coagulation-fibrinolysis were evaluated. A significant decline in platelet aggregation rate in both healthy control and patient plasma in response to Oren-gedoku-to was found. Maximal platelet aggregation time shortened, however, in healthy controls, but was prolonged in 8 of 20 patients with CVA, in particular those with cerebral infarction.

Midazolam and nitrazepam in the maternity ward: milk concentrations and clinical effects.

1. In a randomized study of 22 patients in a maternity ward, the residual concentrations of two hypnotics, midazolam 15 mg p.o. and nitrazepam 5 mg p.o., in early breast milk and plasma were measured 7 h after intake on day 2 to day 6 postpartum. Milk pH, milk fat and binding to plasma proteins were also investigated. Sleep variables were scored on questionnaires. 2. No measurable (less than 10 nmol l-1) concentrations of drug in milk were found in the group receiving 15 mg midazolam at night, either after the first night or after the fifth night. Additional investigations in two mothers demonstrated that midazolam and its hydroxymetabolite disappeared rapidly from milk with undetectable levels after 4 h. The mean (s.d.) milk to plasma ratio for midazolam was 0.15 (0.06) in six paired samples. It may be assumed that practically no midazolam is transferred via early milk to the baby if the baby is nursed more than 4 h after tablet intake. 3. Milk nitrazepam concentrations increased significantly from the first (30 nmol l-1) to the fifth morning (48 nmol l-1) in the group receiving 5 mg nitrazepam at night. The mean (s.d.) milk to plasma ratio of nitrazepam after 7 h was 0.27 (0.06) in 32 paired samples, and did not vary from day 1 to day 5. Plasma protein binding of nitrazepam in puerperal women was found to be lower than that in plasma of healthy controls. The average amount of nitrazepam received by the breast-fed baby in the morning was calculated to increase from 1 to 1.5 micrograms 100 ml-1 breast milk, from days 1 to 5. In the mothers nitrazepam was associated with better hypnotic effect, but a higher incidence of complaints than midazolam. 4. Milk pH, assuming anaerobic conditions, was found in 10 women to average 6.91 +/- 0.09 (s.d.) on days 2-6 postpartum, which is less than previously reported. 5. It is concluded that both hypnotics may be used safely for a few days in the maternity ward. However, possible long-term effects in the suckling infant of small doses of benzodiazepines ingested with breast milk remain to be investigated.

Distribution of selenium and glutathione peroxidase in plasma compared in healthy subjects and rheumatoid arthritis patients.

The distribution of selenium and glutathione peroxidase activity in plasma was compared in samples from healthy adult controls and patients with rheumatoid arthritis. Plasma was separated by gel filtration, and selenium was measured in the eluted fractions by means of graphite furnace atomic absorption spectrometry with Zeeman background correction. Most of the selenium in plasma of healthy controls was seen in proteins that migrated close to immunoglobulin G, and that had an apparent molecular weight at the peak of 174 kDa. One major peak of glutathione peroxidase activity with an apparent molecular weight of 99 kDa was unmatched by any significant peak in selenium content. Thus, there was only limited correspondence between the elution patterns of selenium and glutathione peroxidase activity. Also, in patients with active rheumatoid arthritis, selenium was distributed amongst plasma proteins with a wide range of molecular weights. The major selenium peak was less pronounced in patients, but the amount of selenium in other parts of the chromatogram was not different from that in controls. Further work is necessary to define the occurrence of different selenoproteins in plasma in patients and in healthy subjects.

Determination of prostaglandin E 2 in plasma of schizophrenics and healthy controls by gas chromatography-negative-ion chemical ionization mass spectrometry and radioimmunoassay

Determination of prostaglandin E 2 in plasma of schizophrenics and healthy controls by gas chromatography-negative-ion chemical ionization mass spectrometry and radioimmunoassay

Serum levels of pyridostigmine in myasthenia gravis: methods and clinical significance

Correlation studies on patients with myasthenia gravis are reported in which clinical assessment of fatigue and neurophysiological findings are compared to blood levels of pyridostigmine. Measurements using a high-pressure liquid chromatography method (HPLC), give reproducible results. The levels of pyridostigmine in the serum or plasma of healthy controls and of patients show no essential differences. Components of coffee, tea, chocolate and cigarettes can markedly disturb the chromatography by adding additional peaks, so that interpretation becomes difficult or impossible. Blood levels can be measured approximately one hour after oral intake of 60 mg pyridostigmine. Concentrations rise for two to four hours and then decline exponentially. The half-life of pyridostigmine was between 156 and 210 minutes. Despite identical oral dosages, the concentration differed intraindividually and interindividually among patients. While the blood level does not reach its maximum value for 1-1 1/2 to 3 hours, the maximum clinical and neurophysiological effect of pyridostigmine appears 30-60 minutes after ingestion. Variable distribution of cholinesterase inhibitors over the different compartments (blood, synaptic region) is assumed to cause this temporal lag. If the total amount of pyridostigmine is divided into 4-5 doses, the concentration profiles over the course of a day are relatively stable. There is no significant correlation between the variations in blood level throughout one day, and changes in myasthenic symptomatology. Effects of pyridostigmine can be measured at levels as low as 5 ng/ml; at levels above 40 ng/ml further improvement can be detected only rarely. Blood levels were lower if corticosteroids were administered simultaneously; azathioprine had no influence on blood levels. Blood levels assays allow better differentiation of cholinergic and myasthenic crises and the identification of disturbed absorption and interactions with other medications.

The antiheparin effect of α1-acid glycoprotein: Influence on heparin tolerance during the acute phase reaction

In 10 patients with acute phase reaction, the plasma concentrations of fibrinogen, α 1-antitrypsin and α 1-acid glycoprotein ( α 1-acid GP) were increased, but only the concentration of the acid glycoprotein was negatively correlated with the heparin thrombin clotting time. The antiheparin effect of highly purified, non-desialyzed α 1-acid GP from pooled plasma of patients with acute phase reaction and from healthy controls, was identical when compared at the same protein concentration. The present findings indicate that α 1-acid GP contributes to the increased heparin tolerance regularly observed during the acute phase reaction of disease. Unexpectedly, removal of α 1-acid GP by precipitation with rabbit antihuman α 1-acid GP antibodies failed to reduce the heparin tolerance in patient and healthy control plasma, probably because rabbit immunoglobulins per se possess heparin neutralizing activity.

Fatty acid profiles in mental disease: Part 1. Linolenate variations in schizophrenia

Lipids were isolated from 6 patients suffering from schizophrenia and also from 6 age-matched healthy controls. The extracted lipids were fractionated into erythrocyte and plasma lipids. Column and thin-layer chromatography on silica gel were employed further to isolate the extracts into sub-classes within the major neutral and polar lipid classifications. The fatty acid composition of each sub-class was then monitored by gas chromatography after transmethylation. A significantly higher proportion of linolenic acid (18:3w3) was found in schizophrenics when compared with the controls. The change was reflected in both the neutral lipids and phospholipids from plasma and erythrocytes of the patients. On average, 8.7 ± 2.6 and 3.9 ± 1.5 mole% linolenate/100 mg lipid extract were obtained from the plasma of patients and healthy controls, respectively, while 8.6 ± 2.3 and 3.0 ± 1.2 mole/% linolenate/100 mg lipid were recorded from the red blood cells. The other fatty acids investigated did not show such significant differences between patient and healthy subject. A net increase in the amount of total fatty acid was recorded in the patients and it is thought that either linolenate alone or linolenate together with other polyunsaturated fatty acids not considered here are responsible for these observations. Correlations of these findings with the pathology of schizophrenia are discussed.

CARNITINE DEFICIENCY INDUCED DURING INTERMITTENT HÆMODIALYSIS FOR RENAL FAILURE

Carnitine concentration was measured in plasma, muscle, and dialysate before and after hæmodialysis in patients with renal failure and in plasma and muscle of healthy controls. In eight of the nine patients carnitine concentration in muscle after hæmodialysis was only 10% of the concentration in controls. Plasma-carnitine varied in patients before dialysis and in all of them was reduced by dialysis. The loss of carnitine into the dialysate (190-2100 μmol/treatment) greatly exceeded the normal loss in urine for most of patients, and was only partly compensated for. In some patients normal or high plasma-carnitine and low concentrations in muscle indicated that the carnitine-concentrating mechanisms in the muscle cell had failed. The reduction in carnitine will interfere seriously with normal cellular functions and this may help to explain the clinical syndrome of cardiomyopathy and cardiac failure which has been observed in some patients treated for a long time with intermittent hæmodialysis.

A simple and sensitive radiochemical assay for plasma guanase.

A simple and sensitive radiochemical micro-assay has been developed for the determination of plasma guanase activity. The method is based upon the measurement of the conversion of 14C-labeled guanine to xanthine, catalysed by the enzyme guanase (guanine aminohydrolase;EC 3.5.4.3). Using this method, the catalytic activity in the plasma of adult healthy controls was 0.040 +/- 0.09 nmol/h.mg protein (x +/- s). In children under 5 years of age higher levels of enzyme activity were demonstrated. In adult patients with liver disease plasma guanase activities were found to be 3 to 7-fold increased as compared to the normal adult mean value.