Speciation of antimony(III) and antimony(V) in cell extracts by anion chromatography/inductively coupled plasma mass spectrometry.

An analytical method for the separation and quantification of Sb(III) and Sb(V) using anion chromatography with ICP-MS is presented. The optimum conditions for the separation of the antimony species were established with 15 mmol/L nitric acid at pH 6 as eluent system on a PRP-X100 column. The retention times for antimony(V) and antimony(III) were 85 s and 300 s with detection limits of 0.06 microg/L and 0.29 microg/L, respectively. The proposed method was applied to cell extracts of Leishmania donovani, which were incubated with antimony(III) and antimony(V). Some metabolism seemed to occur within the cells.

Stage-specific activity of pentavalent antimony against Leishmania donovani axenic amastigotes.

The standard treatment of human visceral leishmaniasis involves the use of pentavalent antimony (SbV) compounds. In recent years increasing numbers of clinical failures of treatment with SbV have been reported, probably due to the development of parasite resistance to this compound. The mode of action and mechanisms of resistance to SbV have not been fully elucidated. In the present study an axenic amastigote culture was used to study the in vitro responses of Leishmania donovani to SbV. Susceptibility to both sodium stibogluconate and meglumine antimoniate was found to be stage specific. Amastigotes were 73 to 271 times more susceptible to SbV than were promastigotes. As opposed to SbV, trivalent antimony (SbIII) was similarly toxic to both developmental stages. When promastigotes were transformed to amastigotes, susceptibility to meglumine antimoniate developed after 4 to 5 days, upon the completion of differentiation. In contrast, with transformation from amastigotes to promastigotes, resistance to meglumine antimoniate was acquired rapidly, within 24 h, before the completion of differentiation. The culture of promastigotes at an acidic pH (5.5) or at an elevated temperature (37 degrees C) alone did not lead to the appearance of SbV susceptibility, emphasizing the requirement of both these environmental factors for the development of SbV susceptibility. A previously isolated sodium stibogluconate (Pentostam)-resistant L. donovani mutant (Ld1S.20) is also resistant to meglumine antimoniate, indicating cross-resistance to SbV-containing compounds. In contrast, no cross-resistance was found with SbIII, suggesting a mechanism of SbV resistance different from that described in Leishmania tarentolae. These data show that L. donovani susceptibility to SbV is parasite intrinsic, stage specific, and macrophage independent.

Pseudohyperphosphataemia incidence in an automatic analyzer.

A routine blood analysis, using the Hitachi 717 analyser, of an asymptomatic patient with multiple myeloma revealed a phosphate concentration of 6.2 mmol/l (reference range 0.8-1.4 mmol/l). There was no clinical or biochemical evidence for secondary hyperphosphataemia. Two additional myeloma patients with a normal renal function were found to have serum phosphate concentrations of 5 and 4.7 mmol/l. Globulin-depleted sera of these patients were found to have phosphate levels within the normal range as assayed by a Hitachi 717. All these patients were found to have normal inorganic phosphate levels when a SMAC autoanalyser was used, thus indicating spurious readings by the Hitachi 717. The incidence of pseudohyperphosphataemia in 298 patients with normal renal function and hyperglobulinaemia was 8%. To test the direct effect of globulin on phosphate analysis by the Hitachi 717, globulin was precipitated from serum of myeloma and non-myeloma patients by ammonium sulphate. The analysed data showed a positive correlation between globulin concentration and the spurious phosphate levels. Furthermore, even when inorganic phosphate was completely removed from the tested samples, spurious phosphate readings were detected in the presence of globulins from either myeloma or non-myeloma patients by Hitachi 717. It can be concluded that the ammonium molybdate method for determining inorganic phosphate in the Hitachi 717 gives spuriously high phosphate levels in the presence of a high serum globulin concentration.

Ultrastructure of rabbit endosalpinx and glycoprotein pattern of the oviduct fluid in the presence of copper intrauterine devices.

Oviduct fluid was collected from 24 mature female rabbits in estrus fitted with a copper intrauterine device (IUD) in one of the uterine horns by laparotomy and examined for total sugar and protein content. The percentage of total sugar in the dry protein-containing material from the experimental side was 7.3 compared to 5.2 from the control side. The percentage of total protein was 67.0% of the total dry material from the experimental side, compared to 77.7% in the control side. Cellulose acetate electrophoresis confirmed these findings, demonstrating an increase in the strength of the periodic acid-Schiff stain bands of the glycoproteins from the copper IUD side and a weaker Ponceau-S stain of the protein bands. The ampullary-isthmic area and the uterotubal junction of both oviducts were examined with the scanning electron microscope. The secretory and ciliary cells on the copper IUD side were damaged in these two areas. These changes can be attributed to the presence and action of the copper ions in the oviduct milieu.

The prognostic value of serum acetylcholinesterase in myocardial infarction. Theoretical and clinical considerations.

The aim of the present work was to determine experimentally the prognostic value of reduced activity of serum acetylcholinesterase (ACE) in cases of acute myocardial infarction (AMI). In the first part of the research we studied the mechanism of this reduction by comparing the serum results with the levels of the enzyme in cardiac tissue in such cases. It was found that ACE activity is reduced in serum and also in cardiac tissue in AMI, in contrast to creatine kinase (CK) that is augmented in serum but reduced in cardiac tissue. This fact was interpreted as a "reduced cardiac flow of ACE," in contrast to the "augmented cardiac clearance of CK" in similar cases. In the second part of our research, 50 patients, admitted in our hospital for AMI, were examined and their blood analysed mainly for ACE and CK at brief intervals (every 2-3 days), during hospitalization, and thereafter every 1-2 weeks for 1-4 months. From the results of ACE activity it was possible to classify these variations into four groups, each of them having a defined prognostic value for the evolution of the AMI, a persistent reduced serum activity being interpreted as a bad, severe prognosis, with high morbidity or mortality (groups II and III). We suggest, therefore, that the determination of ACE in serum in cases of AMI, especially before discharge home of such patients, may be an additional useful laboratory test in such cases.

Clinical trial of bromhexine in Sjögren's syndrome.

Tear secretion and lysozyme tear content were measured in 30 patients with Sjögren's syndrome after treatment with oral bromhexine, 32 mg/day. In 21 patients (70%) there was a marked increase in tear secretion and in lysozyme content. In patients with keratoconjunctivitis sicca (KCS) good results in clarifying the mucoid eye discharge were obtained. A remarkable amelioration of xerostomia was also noted. Six other patients, serving as controls, were given a placebo and bromhexine. The placebo had no influence on the rate of tear secretion, while bromhexine caused it to increase in 70% of the controls. This side effects of bromhexine treatment encountered in the present study were negligible and transient. We consider bromhexine to be the drug of choice in the treatment of Sjögren's syndrome.

Abnormal chromosomal marker (D14 q+) in a patient with alpha heavy chain disease.

A patient with alpha heavy chain disease (alphaHCD), who showed an abnormal chromosomal marker (D14 q+) in 10% of the bone marrow cells, is described. The mesenteric lymph nodes, which showed reactive hyperplasia in the first biopsy, transformed later to a malignant lymphoma and finally to a plasma cell tumour. The small intestine revealed villous atrophy, diminished crypts, and intact surface epithelium. The ultrastructure of the goblet and epithelial cells appeared to be normal, and the microvilli were preserved except for circumscribed areas of destruction. The lamina propria was heavily infiltrated with mononuclear cells, mainly mature plasma cells. Alpha heavy chains (alphaHC) were found in the patient's saliva.

Lysozyme content of tears in patients with Sjögren's syndrome and rheumatoid arthritis.

In 37 patients and 143 control patients we estimated tear fluid lysozyme content by the Micrococcus lysodeikticus agar diffusion assay. We found no correlation between the titer of lysozyme in tear fluid and the rate of tear flow. Decrease in lysozyme production was found to be a sensitive indicator of the involvement of the lacrimal system in Sjögren's syndrome.

The danger of "yellow dyes" (tartrazine) to allergic subjects.

Oral administration of 50 mg tartrazine to 122 patients with a variety of allergic disorders caused the following reactions: general weakness, heatwaves, palpitations, blurred vision, rhinorrhoea, feeling of suffocation, pruritus and urticaria. There was activation of the fibrinolytic pathway as shown by reduction of plasminogen with high pre-kallikrein and low kallikrein values. Reduction in complement activity (CH50) was seen in three out of sixteen reactions.