HIV-1: a case of RT67 deletion in a multi-treated non responder patient.

The early detection of mutations in the HIV-1 polymerase is a key point in the management of anti-retroviral therapy. While nucleotide substitutions and insertions have been well and frequently desribed in literature as linked to drug resistance, deletions have been rarely observed and desribed (ART67, Imamichi et al.). The aim of this study is to describe a case of deletion of three nucleotides in the RT gene (ART67) of a multi-treated HIV-1 infected patient. As this deletion has not been detected by the oligoprobe assay, the phenotyping test was used to support therapy but without an appreciable success in terms of viral load. Then a sequencing based genotyping system was used to analyse the viral polymerase and a novel deletion was found at codon 67 of RT gene.

Congenital deficiency of alpha-2-adrenoceptors on human platelets: description of two cases.

The biochemistry and functionality of platelets from two related subjects (mother and son) with alpha-2-adrenoceptor-deficient platelets has been evaluated. Radioligand binding experiments with the specific alpha-2-adrenergic-receptor antagonist, 3H-yohimbine, showed a drastic reduction of alpha-2-adrenoceptors in platelets from both subjects in comparison with the control values. Electron microscopy studies revealed a normal morphology and a normal number of alpha granules and dense bodies. Levels of adenine nucleotides; 5-hydroxytryptamine; B-thromboglobulin; platelet-factor-4 and thromboxane A2 production were within normal limits. Platelet aggregation and 5-hydroxytryptamine production in response to adrenalin (at concentrations up to 50 microM) were absent, whereas ADP, AA, PAF, collagen and thrombin-induced aggregation, secretion, Ca++ flux and thromboxane A2 production were normal. The inhibitory effect caused by different concentrations of prostacyclin on Ca++ flux, aggregation, secretion and thromboxane A2 production of platelet functionally lacking of alpha-2-adrenoceptor was not distinguishable from control platelets and platelets preincubated with yohimbine.

Absence of (-) [3H]desmethoxyverapamil binding sites on human platelets and lack of evidence for voltage-dependent calcium channels.

The two major pathways for Ca2+ entry into cells are potential-sensitive channels and receptor-operated channels. The main object of this investigation was to identify which mechanism regulates Ca2+ entry into human platelets. Platelet stimulation with thrombin, adenosine diphosphate, platelet activating factor and arachidonic acid resulted in a concentration-dependent 2.5-3-fold increase in cytoplasmic free calcium concentration over the basal levels (140 +/- 32 nM or 104 +/- 21 respectively) as measured with the fluorescent dyes Quin-2 and Fura-2. Adrenaline and collagen had no effect in promoting intracellular Ca2+ increase as measured with Quin-2 and little effect when measured with Fura-2. Incubation of Quin-2-loaded platelets with the calcium antagonists verapamil and diltiazem, which are known to inhibit Ca2+ entry from voltage-gated channels in many types of cells, over the concentration range 10(-8) - 10(-4) M did not alter significantly either the resting or the cytoplasmic free Ca2+ after stimulation of platelets by several agonists. Moreover, the calcium antagonists exhibited little or no effect on aggregation and 5-hydroxytryptamine secretion induced by platelet activating factor, adenosine diphosphate, collagen or arachidonic acid in whole blood, platelet-rich plasma or washed platelets when employed at concentration ranges as above. Similar results were obtained in washed thrombin-stimulated platelets. High doses of verapamil (but not diltiazem) inhibited platelet aggregation and secretion in response to adrenaline. Direct radioligand binding studies with (-)[3H]desmethoxyverapamil showed that platelet membranes have no receptors for this drug, suggesting that Ca2+ entry occurs in human platelets via a pathway different from potential-sensitive Ca2+ channels.(ABSTRACT TRUNCATED AT 250 WORDS)

In vivo platelet activation in lower limbs thrombophlebitis. A longitudinal study.

Platelet activation in vivo was studied in patients with thrombophlebitis of the lower limbs. The parameters considered were the platelet aggregate ratio (PAR) and the beta-thromboglobulin (beta-tg) level, which were repeatedly evaluated from the disease onset up to 3 months later, during anticoagulating and antiaggregating therapy. A significant decrease of PAR was found, along with a significant rise of the beta-tg level at the onset of the disease, and these values slowly returned to normal on therapy course. The same parameters exceeded the normal range again when the patients arbitrarily suspended any drug assumption. The possible significance and implications of these findings are discussed.