Cholesterol-independent targeting of Golgi membrane proteins in insect cells

Distinct lipid compositions of intracellular organelles could provide a physical basis for targeting of membrane proteins, particularly where transmembrane domains have been shown to play a role. We tested the possibility that cholesterol is required for targeting of membrane proteins to the Golgi complex. We used insect cells for our studies because they are cholesterol auxotrophs and can be depleted of cholesterol by growth in delipidated serum. We found that two well-characterized mammalian Golgi proteins were targeted to the Golgi region of Aedes albopictus cells, both in the presence and absence of cellular cholesterol. Our results imply that a cholesterol gradient through the secretory pathway is not required for membrane protein targeting to the Golgi complex, at least in insect cells.     

Staphylococcal enterotoxin A-induced fever is associated with increased circulating levels of cytokines in rabbits

Rabbits were injected intravenously with 10 to 100 ng of staphylococcal enterotoxin A (SEA) per kg, and colonic temperatures were monitored. The febrile responses were compared with circulating levels of interferon (IFN), tumor necrosis factor (TNF), interleukin-1 (IL-1), IL-2, and IL-6 just before the injection of SEA. Both colonic temperatures and circulating levels of IFN, TNF, and IL-2 started to rise at 1 to 2 h and reached their peak levels at 3 to 5 h after SEA injection. Both the fever and the increased circulating levels of IFN, TNF, and IL-2 produced by SEA were decreased by pretreatment with indomethacin (a cyclo-oxygenase inhibitor) (15 mg/kg, intraperitoneally), anisomycin (a protein synthesis inhibitor) (15 mg/kg, subcutaneously), or dexamethasone (an effective anti-inflammatory and immunosuppressive agent) (4 mg/kg, intravenously) in rabbits. Rabbits were injected intravenously with 30 ng of SEA per kg on four consecutive days, and colonic temperatures were monitored. Compared to rabbits that received the single injection of SEA, rabbits that received four consecutive injections of SEA showed a lesser increase in circulating levels of IFN, TNF, and IL-2 as well as colonic temperatures in response to an intravenous dose of SEA (30 ng/kg). The data suggest that the prevention of the febrile response elicited by SEA by indomethacin, anisomycin, or dexamethasone is due to prevention by these compounds of the increase in the circulating levels of IFN, TNF, and IL-2. The pyrogenic hyporesponsiveness to repeated injection of SEA is associated with decreased production of these circulating cytokines.     

Functional switch from facilitation to inhibition in the control of glutamate release by metabotropic glutamate receptors

We have investigated the role of metabotropic glutamate receptors linked to phosphoinositide hydrolysis in the control of glutamate release in cerebrocortical nerve terminals. The activation of these receptors with the agonist 3,5-dihydroxyphenylglycine enhanced intra-synaptosomal diacylglycerol and facilitated both the depolarization-induced increase in the cytosolic free Ca2+ concentration and the release of glutamate. However, 5 min after receptor activation, a second stimulation of the pathway with the agonist failed to produce diacylglycerol and to facilitate glutamate release. Interestingly, during the period in which the diacylglycerol response was desensitized, a strong agonist-induced inhibition of Ca2+ entry and glutamate release was observed. This change in the presynaptic effects of 3,5-dihydroxyphenylglycine is reversible since 30 min after the first stimulation, the agonist-induced inhibition of release disappeared, whereas both the production of diacylglycerol and the facilitation of glutamate release were recovered. The tonic elevation of the extracellular glutamate concentration from basal levels (0.8 microM) up to 5 microM also produced the switch from facilitation to inhibition in the receptor response. The existence of this activity-dependent switch in the presynaptic control of glutamate release suggests that release facilitation is limited to conditions under which an appropriate clearance of synaptic glutamate exists, probably to prevent the neurotoxic accumulation of glutamate in the synapse.     

Impact of HIV counselling and testing on HIV seroconversion and reported STD incidence among male factory workers in Harare, Zimbabwe

OBJECTIVES: To assess the impact of HIV counselling and testing on HIV seroconversion and incidence of reported sexually transmitted diseases (STDs) among male factory workers in Harare, Zimbabwe. DESIGN: Prospective, observational study among men recruited to participate in a future workplace based AIDS prevention intervention. METHODS: Participants provided STD histories and blood for HIV antibody testing at enrolment and six month intervals during visits to factories. Participants received HIV test results, post test counselling, and free STD services at the project clinic. RESULTS: Between March 1993 and June 1995, 2,414 men were enrolled with 85% follow up. Overall HIV sero-incidence was 2.60 per 100 person-years; the incidence of reported STDs was 10.19 per 100 person-years. Men who obtained their HIV test results had significantly higher HIV sero-incidence and incidence of reported STDs compared to men who did not obtain their results (IRRs: 1.87, 3.47, respectively). Among men who obtained their HIV test results, a non-significant 40% decrease in HIV sero-incidence was observed after obtaining test results compared to before obtaining results (p = 0.18). The incidence of reported STDs, however, increased by 30% after obtaining HIV test results (p = 0.10). CONCLUSIONS: Decreased HIV sero-incidence in the face of increased reported STD incidence suggests that timely treatment of STDs may decrease the risk of acquiring HIV even in the absence of behaviour change. In populations with high rates of HIV and STDs, the greatest benefit of HIV counselling and testing may be achieved by simultaneously offering STD screening and treatment services.     

Dimethyl sulfoxide-induced apoptosis in human leukemic U937 cells

The effects of two oral contraceptives, containing gestodene and either 20 micrograms or 30 micrograms ethinylestradiol, on hemostatic parameters was investigated in a six-month randomized study involving a total of 40 healthy women between the ages of 18 and 30 years. A large number of hemostatic parameters were measured, which were categorized as either pro-coagulatory, anti-coagulatory, profibrinolytic, anti-fibrinolytic or indicative of fibrin turnover. Additionally, tissue plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI-1) were measured before and after venous occlusion and delta and ratio values calculated. Pro-coagulatory factors as well as reaction products reflecting in vivo coagulatory activity (thrombin-antithrombin III complex, prothrombin fragment 1 + 2) were found to increase. Among the anti-coagulatory parameters, only protein S concentration and protein S activity decreased, most notably in the 30 micrograms EE group. There was a corresponding increase in fibrinolytic activity reflected by reaction products of in vivo fibrinolysis (plasmin-antiplasmin 2-complex, fibrin-degradation products). Measurement of t-PA and PAI-1, before and after venous occlusion, revealed that the fibrinolytic response was more pronounced in the 20 micrograms EE group. There was also an increase in the threshold of fibrinolytic inhibition (ratio PAI-1) in both groups, which was less pronounced in the 20 micrograms EE group. Apart from isolated measurements, all parameters remained within their normal ranges and values returned to baseline in the follow-up cycle. It is concluded that both preparations had a balanced effect on the hemostatic system stimulating both pro-coagulant and fibrinolytic activity. No statistically significant differences were observed between the two groups; however, there was a trend towards greater fibrinolytic capacity in the 20 micrograms EE group.     

Levocabastine versus cetirizine for perennial allergic rhinitis in children

OBJECTIVE: To compare the efficacy and safety of levocabastine nasal spray asid cetirizine oral for the treatment of perennial allergic rhinitis in children. MATERIAL AND METHODS: In this randomized, prospective experimental, open clinical trial. We studied 30 children with ages between 6 and 16 years with perennial allergic rhinitis. Group 1, 17 subjects (7 female, 10 male) received cetirizine once daily, 5 mg children weientig less dian 30 k asid 10 mg in children weighing more trw' 30 k during 15 days. Group 2, 13 subjects (7 male, 6 female) received levocabastine 2 puffs BID on each nostril during tbe same time. A nasal symptoms score, nasal peal: flow vid eosinophils in a nasal smear were performed before and after treatment. RESULTS: There were no statistical differences in age, weight, height and arid duration of symptoms. Both groups showed improvement of symptoms via nasal peak flow with no differences between them (intergroup); nasal eosinophils remained unchanged. We for third statistical differences pre vid postreatment in each group (intragroup): Group 1, nasal congestion p = 0.002, ocular itch p = 0.01, sneezing p = 0.007, nasal secretion p = 0.01, nasal itch O = 0.009, total points O = 0.0005. Group 2, nasal congestion O = 0.02, ocular itch p = 0.05, sneezing p = 0.01, nasal secretion p = 0.01, nasal itch p = 0.04, total points p = 0.005. Significant differences were found in nasal peal' flow in Group 1 (p = 0.01) but no differences in eosinophils between file two groups. Side effects: 3 subjects in Group 1 (drowsiness, 1 appetite increase said 1 rhinorrea with epistaxis) vide 1 in Group 2 sensation of facial edema. CONCLUSION: Bofil drugs are effective the clinical relief of symptoms of perennial allergic rhinitis in children vied levocabastine has less side effects.     

A membrane-permeant, bioactivatable derivative of Ins(1,3,4)P3 and its effect on Cl(-)-secretion from T84 cells

The synthesis of rac-2,5,6-tri-O-butyryl-myo-inositol 1,3,4-trisphosphate hexakis(acetoxymethyl) ester [Bt3-Ins(1,3,4)P3/AM, 1], a membrane-permeant derivative of myo-inositol 1,3,4-trisphosphate [Ins(1,3,4)P3] is reported. 1 inhibited calcium-mediated chloride secretion of T84 cells, suggesting a regulatory link of Ins(1,3,4)P3 and the biosynthesis of the known inhibitor myo-inositol 3,4,5,6-tetrakisphosphate.