[Contraception and abortion: an update in 2015]. / Contraception et interruption de grossesse.

Family doctors can play an important role in preventing unplanned pregnancies. This article addresses the different contraceptives methods available in Switzerland, which are classified in 2 groups and recommends using the GATHER approach (Greet, Ask, Tell, Help, Explain, Return) to promote compliance. LARC (long acting reversible contraceptives) can be recommended to any woman who needs a reliable birth control method. These contraceptives require minimum effort for high efficiency. Further explanation regarding the use of an emergency contraception must be provided when short action contraceptives are chosen. Switzerland's abortion rate is one of the lowest in the world. Medical abortion tends to be more and more prominent. Under certain circumstances, it can be self-administered at home.

Rationally designed strings of promiscuous CD4(+) T cell epitopes provide help to Haemophilus influenzae type b oligosaccharide: a model for new conjugate vaccines.

The age-related and T cell-independent immunological properties of most capsular polysaccharides limit their use as vaccines, especially in children under 2 years of age. To overcome these limitations, polysaccharide antigens have been successfully conjugated to a variety of carrier proteins, such as diphtheria toxoid or tetanus toxoid (TT) and the diphtheria mutant (CRM197) to produce very successful glycoconjugate vaccines. The increasing demand for new conjugate vaccines requires the availability of additional carriers providing high and long-lasting T helper cell immunity. Here we describe the design and construction of three recombinant carrier proteins (N6, N10, N19) constituted by strings of 6, 10 or 19 human CD4(+) T cell epitopes from various pathogen-derived antigens, including TT and proteins from Plasmodium falciparum, influenza virus and hepatitis B virus. Each of these epitopes is defined as universal in that it binds to many human MHC class II molecules. When conjugated to Haemophilus influenzae type b (Hib) oligosaccharide, these carriers elicit a potent anti-Hib antibody response in mice. In the case of the N19-Hib conjugate, this response is at least as good as that observed with CRM197-Hib, a conjugate vaccine currently used for mass immunization. We also show that some of the universal epitopes constituting the recombinant carriers are specifically recognized by two human in vitro systems, suggesting that T cell memory is provided by the selected epitopes. The data indicate that rationally designed recombinant polyepitope proteins represent excellent candidates for the development and clinical testing of new conjugate vaccines.

Identification of vaccine candidates against serogroup B meningococcus by whole-genome sequencing.

Neisseria meningitidis is a major cause of bacterial septicemia and meningitis. Sequence variation of surface-exposed proteins and cross-reactivity of the serogroup B capsular polysaccharide with human tissues have hampered efforts to develop a successful vaccine. To overcome these obstacles, the entire genome sequence of a virulent serogroup B strain (MC58) was used to identify vaccine candidates. A total of 350 candidate antigens were expressed in Escherichia coli, purified, and used to immunize mice. The sera allowed the identification of proteins that are surface exposed, that are conserved in sequence across a range of strains, and that induce a bactericidal antibody response, a property known to correlate with vaccine efficacy in humans.

Quasi-total-body exposure to an oxygen-ozone mixture in a sauna cabin.

We have investigated the effects of quasi-total-body exposure of healthy volunteers to either an oxygen-ozone mixture (O(2)-O(3)) or to oxygen (O(2)) alone during a short period in a sauna cabin. The subjects underwent both an experimental and a control examination, separated by a 3.5-month interval. Body mass, blood pressure, body temperature changes, electrocardiograms, venous blood gas and haemocytometric analyses, total antioxidant status and plasma levels of protein thiol groups, thiobarbituric acid reactive substances (TBARS), plasma cytokine, hepatic enzymes and creatine were determined before, immediately after the 20-min period in the cabin and then 0.5, 1.0 and 24 h afterwards. We observed statistically significant variations of body temperature, venous partial pressure of O(2) values, TBARS and plasma levels of interleukin 8, particularly after O(2)-O(3) exposure. The increase in TBARS plasma levels concomitant with protein oxidation has been tentatively interpreted as being attributable to the transcutaneous passage of some reactive O(2) species, which should be considered if this approach is to be used as a biological response modifier. However, in the present study no adverse effects were noted after one session.

A competitive enzyme-linked immunosorbent assay for measuring the levels of serum antibody to Haemophilus influenzae type b.

A competitive ELISA method is described for the measurement of total antibodies to the capsular polysaccharide of Haemophilus influenzae type b (HibCPS) in human sera. The competitive method showed an excellent correlation to the radioantigen binding assay (RABA, or Farr assay) and improved correlation of sera with low titers with respect to the more conventional noncompetitive method. Overestimation of samples in the low concentration range was no longer observed with the competitive ELISA method. The free HibCPS competition allowed us to eliminate the day-to-day background variation typical of some sera; thus, only values representing the true anti-HibCPS response were determined. The use of precoated microplates, which could be stored up to 8 months, greatly improved the speed of the procedure. An overall correlation coefficient of 0. 9660 was found when 407 serum samples with a wide variety of anti-HibCPS antibody levels were tested with the competitive ELISA and RABA. The regression line was very close to the ideal line, with a slope of 1.0045 and an intercept of -0.1996. A subset of 96 serum samples representative of all pre- and postimmunization samples was used to compare the competitive ELISA with a previously described ELISA method. The competitive method performed in two laboratories in different countries showed a better correlation with the RABA. The correlation factors were 0.9770 and 0.9816, respectively, while a factor of 0.9547 was found with the previously described noncompetitive procedure, which was better for this method than previously reported (r = 0.917). Therefore, the competitive ELISA is proposed for the assay of anti-HibCPS titers in sera from vaccinated subjects.

Pulmonary catabolism of interleukin 6 evaluated by lung perfusion of normal and smoker rats.

Cytokines such as interleukin 6 are involved in the pulmonary inflammation arising as a result of smoking. By use of isolated and perfused lung preparations we have evaluated the role of the lungs in the catabolism of human recombinant interleukin 6 both in normal rats and in rats subjected to an acute cigarette smoking episode. When interleukin 6 was incorporated into the lung perfusion medium, neither control nor smoke-exposed rat lungs cleared the cytokine and only 0.1 +/- 0.2% of the total dose was recovered in the bronchoalveolar lavage fluid. When, on the other hand, the same amount of interleukin 6 was instilled into the bronchoalveolar tree, concentrations of the cytokine in the perfusate increased progressively so that after 3 h up to 70.1 +/- 9.8% and 40.9 +/- 22.5% of the administered dose, as measured by immunoenzymatic test, had been transferred from the bronchial lumen to the perfusion medium of either control or smoker rat lungs, respectively, indicating significantly (P < or = 0.05) different behaviour of the cytokine in the two experimental groups. Total recoveries of the administered interleukin 6 evaluated in smoke-exposed rat lungs were 55.3 +/- 23.2%, significantly lower than those for control rat lungs (83.9 +/- 11%). Determination of biological activity gave values always lower than those measured by immunoenzymatic test, indicating loss of biological activity during the transalveolar transit. It appears that the transfer of interleukin 6, especially in smokers, is almost exclusively unidirectional, from the alveolar space to the plasmatic pool with degradation during the transalveolar passage.

Plasma levels of immunosuppressive mediators during cardiopulmonary bypass.

The aim of this study was to evaluate plasma levels of two mediators with immunosuppressive properties, complement fraction C3a (C3a) and transforming growth factor-beta(1) (TGF-beta(1)), during extracorporeal circulation. The proliferation index after phytohaemagglutinin (PHA) stimulation of isolated peripheral blood mononuclear cells was also investigated. Sixteen patients undergoing hypothermic (n = 8, group 1) and normothermic (n = 8, group 2) cardiopulmormry bypass (CPB) were enrolled in this study. As a control, we evaluated four patients undergoing thoracovascular operations without CPB. Blood samples were collected before CPB but after anaesthesia, every 30 min during CPB, at the end of CPB and 10 min after protamine administration. Both C3a and TGF-beta(1) increased significantly during CPB and after protamine administration in the hypothermic as well as the normothermic group. In the latter case the increase of C3a and TGF-beta(1), although more prominent, was not significantl higher than in the former group. Conversely, the proliferation, index of peripheral mononuclear cells had already decreased 30 min after CPB was started and remained depressed throughout the CPB time. These results suggest a possible role of C3a and TGF-beta(1) in the immunological changes occurring during extracorporeal circulation.

Pharmacokinetics and catabolism of tumor necrosis factor-alpha in rat lungs.

The role of the lungs in the catabolism of tumor necrosis factor alpha (TNF-alpha) either in normal rats, or in rats subjected to an acute cigarette smoking episode has been evaluated by using isolated and perfused lung preparations. After administration of TNF-alpha into the lung perfusion medium, there was no clearance of the cytokine in both control and smoker rat lungs and only 0.2 +/- 0.1% of the administered dose was recovered in the bronchoalveolar lavage fluid. When TNF-alpha was instilled into the bronchoalveolar tree, concentrations of the cytokine in the perfusate increased progressively so that after 3 h up to 68.8 +/- 8% and 52.7 +/- 11.4% of the administered dose had been transferred from the bronchial lumen to the perfusion medium of either control or smoker rat lungs, respectively, the latter values being significantly lower (p < or = 0.05) than those obtained in control lungs. Moreover, total recoveries of TNF-alpha evaluated in smoker rat lungs (65.5 +/- 10.2%) were also significantly lower than those observed in control rat lungs (82.8 +/- 7.1%). In conclusion, it appears that transfer of TNF-alpha is almost exclusively unidirectional, from the alveolar space to the plasma pool with partial degradation during the transalveolar passage. These results may be useful when attempting to deliver TNF-alpha by aerosol.

Pulmonary catabolism of interferon-gamma evaluated by lung perfusion of both normal and smoke-exposed rats.

The role of the lungs in the catabolism of rat recombinant interferon-gamma, either in normal rats or in rats subjected to an acute cigarette smoking episode, was evaluated using an isolated and perfused lung preparation. After administration of interferon-gamma into the lung perfusion medium, there was no clearance of the cytokine in either control or smoke-exposed rat lungs, and only 0.1 +/- 0.2% of the total dose was recovered in the bronchoalveolar lavage fluid. When the same amount of interferon-gamma was instilled into the bronchial alveolar tree, concentrations of the cytokine in the perfusate increased progressively so that after 3 h up to 71.2 +/- 4.3 and 62 +/- 5.7% of the administered dose, as measured by ELISA test, had been transferred from the bronchial lumen to the perfusion medium of either control or smoke-exposed rat lungs, respectively, the latter values being significantly lower (p < or = 0.05) than those obtained in control lungs. Moreover, total recoveries of interferon-gamma evaluated in smoke-exposed rat lungs (78.4 +/- 8.6%) were also significantly lower than those observed in control rat lungs (91.4 +/- 11.8%). Biologic activity evaluations on the same samples gave values significantly lower than those obtained using ELISA, indicating a partial loss of biologic activity during transalveolar transit. In conclusion, it appears that the transfer of interferon-gamma is almost exclusively unidirectional from the alveolar space to the plasmatic pool, with partial degradation during transalveolar passage.

Maternal and neonatal plasma cytokine levels in relation to mode of delivery.

After birth, host defences must be recruited to manage the transition from an almost sterile to a normal environment. The present study was undertaken to evaluate the relationship between cytokine plasma levels and phagocyte burst in mothers and neonates during the peripartal period. Plasma levels of interleukin (IL)-1, IL-6, tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, and granulocyte-macrophage colony-stimulating factor (GM-CSF) and whole blood superoxide anion (.O2-) generation were evaluated in 27 healthy mothers, 16 undergoing vaginal delivery (VD) and 11 elective caesarean section (ECS) and in their babies. Blood specimens were taken from the mothers at the beginning of labour, during labour, immediately after delivery and 4 days later in the VD group, and before anaesthesia, immediately after delivery and 4 days later in the ECS group; neonatal samples were taken at birth (cord blood) and 4 days later. After delivery by VD, these mothers had higher plasma levels of IL-1 beta, IL-6, IFN-gamma and higher .O2- generation than those delivered by ECS. IL-6 plasma levels and .O2- generation were higher in babies born by VD than in those born by ECS. A statistically significant correlation between IL-6 plasma levels and .O2- release was observed in cord blood of babies born by VD (r = 0.69; p < 0.006). The study demonstrates that labour plays an important role in modulating host defences in the newborn.

Studies on the biological effects of ozone: 6. Production of transforming growth factor 1 by human blood after ozone treatment.

After exposing human whole blood from normal volunteers to ozone concentrations ranging from 22 to 156 micrograms/ml, we have shown that, upon incubation of up to 8 hours, there is a significant release of transforming growth factor beta (TGF-beta 1). In comparison to TGF-beta 1, TGF-beta 2 production is not influenced by ozone concentrations. In line with our previous findings it appears that blood, in the presence of heparin and 5mM Ca,2+ allows a consistent production of tumor necrosis factor a (TNF alpha) and the release of low and non-hazardous levels of free hemoglobin. These data support the contention that autohemotherapy performed after treating blood with ozone followed by reinfusion into the donor, may represent a valuable therapeutic approach for achieving immunoregulatory effects.

Studies on the biological effects of ozone: 4. Cytokine production and glutathione levels in human erythrocytes.

We have investigated the effect of various concentrations of ozone on human blood aiming to correlate the production of cytokines with depletion of reduced glutathione and hemolysis. As erythrocytes constitute the bulk of blood cells and represent the main target of ozone they have been taken as a useful marker of its oxidative activity. It appears that a transient exposure (30 sec) of blood of up to 78 micrograms ozone per ml of blood does not depress the production of cytokines even though there is a slight increase of hemolysis and a small decrease of intracellular reduced glutathione. In contrast either a constant (up to 30 sec) exposure to an ozone flux or a high ozone concentration (108 micrograms/ml) markedly decreases reduced glutathione levels and depresses cytokine production.

Studies on the biological effects of ozone: 3. An attempt to define conditions for optimal induction of cytokines.

Ozonization of blood, normally carried out with citrated blood, may be fine for the autohemotherapy of ischemic diseases but it may be at a loss when employed in viral diseases or in immunodeficiencies. We have shown that heparin, used as an anticoagulant, with the addition of 5 mM CaCl2 favors production of cytokines by leukocytes with only a modest increase in hemolysis. High plasmatic levels of glucose, glutathione, and ascorbic acid decrease cytokine's yield because these compounds act as antioxidants and quench the inducing activity of ozone. Autohemotherapy with heparinized and Ca(2+)-supplemented blood has not revealed any side effects in volunteers.

Presence of interferon-gamma and interleukin-6 in colostrum of normal women.

Colostrum and blood samples were obtained on postpartum day 2 and 3 from 17 lactating, healthy women. After delipidation and molecular sieving fractionation of colostrum, interferon-gamma (IFN-gamma) and interleukin-6 (IL-6) could be readily measured by using a sensitive immunoassay. Antiviral activity could be also measured in some colostrum samples suggesting that interferon was biologically active. On the contrary, corresponding plasma samples showed negligible activity. These results expand previous data showing the presence of IL-1, tumor necrosis factor (TNF-alpha), and IL-6 in normal colostrum and are in line with the concept of a basal cytokine production in physiological conditions. All of these cytokines probably act on the oropharyngeal and gut-associated lymphoid tissue of the newborn and favor the development and maturation of the immune system.

Effects of acute cigarette smoke exposure on macrophage kinetics and release of tumour necrosis factor alpha in rats.

Some biological parameters before and after an acute episode of cigarette smoking in rats have been evaluated. The carboxyhaemoglobin levels depended either on the number of cigarettes, or on the time of exposure to cigarette smoke and returned to pre-smoking values in about 2 h. The evaluation of the kinetics of alveolar and peritoneal macrophages in rats after a smoking session of three cigarettes within an hour, indicated that alveolar macrophages in the bronchoalveolar lavage fluid significantly increased 8 h after the smoking, whereas the number of peritoneal macrophages remained practically constant. The incubation of these cells for various times at 37( degrees )C in a humidified atmosphere, resulted in a spontaneous release, 24 h thereafter, of variable amounts of tumour necrosis factor alpha (TNFalpha), which remained practically constant during the following days. Neither alveolar macrophages of control rats, nor peritoneal macrophages of both control and smoking rats were able to release TNFalpha. Moreover, after lipopolysaccharide induction of alveolar macrophages of both control and smoking rats, an increased release of TNFalpha was observed, indicating that these cells were in an active state.

Chronic cigarette smoking enhances spontaneous release of tumour necrosis factor-alpha from alveolar macrophages of rats.

Some biological effects of chronic cigarette smoking (two cigarettes for 2 h, daily for 4 months) in rats were evaluated. During the smoking period, body weight of smoker rats was always significantly lower than that of control rats. Immediately after the last smoking session the carboxyhaemoglobin concentration in the blood was about 8.5% and the polymorphonuclear cells in the bronchoalveolar fluid increased significantly. At the same time, enzymatic analyses on the supernatants of bronchoalveolar fluid revealed a significant increase of beta-glucuronidase in the smoker group. Alveolar macrophages, collected 0, 8 and 24 h after the last smoking session, significantly increased the generation of superoxide anion and, after incubation for 24 h at 37( degrees ) C in a humidified atmosphere, released significantly high amounts of TNF-alpha. When challenged with lipopolysaccharide, alveolar macrophages of smoker rats released much more TNF-alpha but, in such a case, TNF-alpha release was about one half of that observed in the control group. Peritoneal macrophages of both control and smoker rats were unable either to generate high levels of superoxide anion or to release significant amounts of TNF-alpha. The results clearly demonstrated the activated state of alveolar macrophages and the resting state of peritoneal macrophages.

Production of tumor necrosis factor alpha by rat alveolar macrophages collected after acute cigarette smoking.

In this study we evaluated the effect of cigarette smoke on the activation of alveolar macrophages of the rat lungs exposed to an episode of acute passive cigarette smoking. Our experiments were carried out in rats that, after undergoing smoking (3 cigarettes within 1 h) showed a COHb increase of about 16%. The evaluation of the kinetics of alveolar and peritoneal macrophages, indicated that the number of alveolar macrophages in the bronchoalveolar lavage fluids significantly increased 8 h after the smoking session, whereas the number of peritoneal macrophages remained practically constant. Alveolar macrophages collected 0.8 and 24 h after smoking and incubated for 24 h at 37 degrees C in an atmosphere of 5% CO2 in air spontaneously released 5 +/- 1, 48 +/- 14 and 15 +/- 9 units of TNF-alpha per 10(6) cells, respectively. Moreover, neither alveolar macrophages collected from smokers, nor those collected from controls, released IFN, and both cytokines were also absent either in bronchoalveolar lavage and peritoneal lavage fluids or in plasma. Alveolar macrophages collected from controls rats, when challenged with lipopolysaccharide (LPS), released more TNF than those collected from smoke exposed rats. Thus, it seemed that macrophages of experimental animals were activated but at the same time were somewhat depressed and responded less well to LPS.

Studies on the biological effects of ozone: 5. Evaluation of immunological parameters and tolerability in normal volunteers receiving ambulatory autohaemotherapy.

Autohaemotherapy, after a bland treatment ex vivo of blood with ozone, is a fairly unknown medical procedure claimed to have therapeutic value in viral diseases and neoplasms. Having already shown that ozone acts as a mild inducer of cytokines, we have undertaken an investigation in normal rabbits and in normal volunteers aiming to evaluate eventual changes of some cytokine levels in plasma as well as of immunological parameters such as the Mx protein, neopterin, beta 2-microglobulin and of some acute-phase proteins after single or repeated autohaemotherapy. We have also evaluated the potential development of of side-effects. This study is the first one to show that autohaemotherapy can activate an immunological marker in normal subjects without procuring any toxic effects.

Effect of tuftsin and its retro-inverso analogue on the release of interferon (IFN-gamma) and tumor necrosis factor (TNF-alpha) by human leucocytes.

The aim of this work was to demonstrate whether natural tuftsin or a retro-inverso (r.i.) analogue may induce interferon (IFN) and tumor necrosis factor (TNF) in peripheral-blood-mononuclear-cells (PBMC). For this purpose tuftsin or its analogue were added at different molar concentrations to PBMC and the supernatants were tested for IFN and TNF activity. Both cytokines were released after 12 hours incubation with r.i. tuftsin at an optimum concentration of 10(-10) M. Under the same conditions no activity was observed in the presence of natural tuftsin. In comparison to natural tuftsin the stimulatory activity of this tuftsin analogue is likely to be due to its high stability.

Studies on the biological effects of ozone: 2. Induction of tumor necrosis factor (TNF-alpha) on human leucocytes.

The effect of ozone as a probable inducer of tumor necrosis factor (TNF-alpha) has been investigated on human blood and on Ficoll-purified blood mononuclear cells (PBMC). Samples were exposed at different ozone concentrations ranging from 2.2 to 108 micrograms/ml and incubated at 37 degrees C in an 95% air-5% CO2 atmosphere. At predetermined times, all cell supernatants were tested for TNF activity and some PBMC cultures were examined for DNA synthesis. We have shown that ozone concentration is critical in terms of TNF production and of cell mitogenesis and that, owing to the presence of erythrocytes, higher ozone concentrations are required to be effective in blood than in PBMC. Because ozonization of blood is a procedure followed in several European countries for the treatment of viral diseases and tumors, the release of factors with antiviral and immunomodulatory activities by leukocytes may explain the mechanism of action of ozone and of autohemotherapy.