[The psychological experience of the mothers whose babies were born deformed in the neonatological ward of the academic hospital center of Treichville (Côte-d'Ivoire)]. / Vécu psychologique des mères de nouveau-nés malformés dans le centre hospitalier et universitaire de Treichville (Côte-d'Ivoire).

OBJECTIVE: When a baby is born deformed, his birth breaks up plans his family has made for his life as well as for the family itself. So, our objective was to describe the experience undergone by the mothers who gave birth to babies with serious deformities. METHOD: A prospective and descriptive study was carried out during 12 months about 35 mothers whose babies were born seriously deformed. The data were collected from newborn's medical file and from a well-conducted interview with volunteered mothers. RESULTS: The incidence of the major deformities was 1.4% and the antenatal diagnosis was carried out on only 6 cases. The announcement of deformities in the 30 cases discovered at birth by the clinical staff brought about a shock among mothers. In fact, 28 mothers were disappointed, while 23 experienced fear and 10 others felt guilty. Six mothers thought in terms of aborting, followed by 25 mothers who without any psychological aid felt lonely. Furthermore, 30 mothers suffered from a lack of communication with nurses and with medical staff concerning deformities. The anxiety of mothers was linked, respectively to fear of their husband (33 cases), to the social environment (35 cases) and to the health care (10 cases). CONCLUSION: Mothers facing congenital deformity experienced painful and violent emotional distress. Therefore, the emphasis must be put on the improvement of the neonatal diagnosis and of relationship between physicians and parents in order to achieve a close support toward mothers.

A new short-acting non-depolarizing muscle relaxant (SZ1677) without cardiovascular side-effects.

BACKGROUND: In order to facilitate rapid tracheal intubation, the development of a rapid onset, short duration, non-depolarizing muscle relaxant without cardiovascular side-effects would be a significant accomplishment in the field of anesthesiology. The aim of the present study was to test the action of a new non-depolarizing muscle relaxant (SZ1677) on neuromuscular transmission, muscarinic (M2, M3) receptors and cardiovascular reactions and to compare it with clinically used muscle relaxants. METHODS: Neuromuscular transmission was studied by recording muscle contractions elicited by indirect electrical stimulation, using (i). in vitro isolated phrenic nerve-hemidiaphragm preparation of mice, rats and guinea pigs and (ii). in vivo sciatic nerve-anterior tibial muscle preparation of anesthetized rats, guinea pigs and cats. Cardiovascular effects of muscle relaxants were evaluated on the grounds of their effects on changes of blood pressure and heart rate induced by electrical stimulation of the right vagal nerve in anesthetized cats. To study postsynaptic antimuscarinic affinity of muscle relaxants on M3 receptors, oxotremorine-induced contractions of longitudinal muscle strip of guinea pig ileum were registered in their presence and absence. RESULTS: One of more than 120 newly synthesized non-depolarizing muscle relaxants compounds, 1-3[alpha-hydroxy-17beta-acetyloxy-2beta-(1,4-dioxa-8-azaspiro[4,5]dec-8-yl)-5alpha-androstane-16beta-il] -1-(2-propenyl)pyrrolidinium bromide (SZ1677), excelled with its advantageous pharmacological properties: relatively short duration of action, no accumulation and lack of unwanted side-effects. Pharmacodynamic studies show that SZ1677 is a non-depolarizing neuromuscular blocking agent with a relatively short duration and rapid onset of action in a variety of laboratory animal species. It is without cumulative effect, does not reduce blood pressure, and fails to produce tachycardia. Significant cardiac vagal blocking effects were not observed even at concentrations or dosages of 8 times the ED90. This compound, unlike many other muscle relaxants, does not have atropine-like effects on human atrial tissue; it does not increase the release of NA from sympathetic innervation in the heart. In all practical ways, at least from the vantage point of the preclinical study, SZ1677 compares favorably with all presently available short-acting muscle relaxants, including rapacuronium. CONCLUSION: In experiments, SZ1677 proved to be a short-acting neuromuscular blocking compound having a large safety margin between the doses required to produce neuromuscular block and those likely to lead to cardiovascular side-effects.

Determination of circular dichroism and ultraviolet spectral parameters of norgestimate- and other Delta(4)-3-ketosteroid oxime isomers via normal phase HPLC method.

The oxime formation reaction of therapeutical progestogen (levonorgestrel, levonorgestrel acetate, norethisterone), androgen (methyltestosterone, testosterone phenylpropionate) and anabolic (nortestosterone phenylpropionate) Delta(4)-3-ketosteroids has been investigated. The ketosteroid-hydroxylamine reaction was monitored by reversed phase HPLC system. It was established, that under the experimental conditions applied the oxime formation was complete within 2 h. The reaction leads to the formation of Z and E oxime isomers. The isomers of norgestimate (levonorgestrel 17-acetate oxime) and other Delta(4)-3-ketosteroid oximes have been separated by a new normal phase HPLC method. The identification (elution order assignation) and determination of the formation ratio of the isomers have been performed by 1H NMR spectroscopy on the basis of the chemical shift differences of 4-H signals. The on-line CD and UV spectra of the pure oxime isomers were recorded and then molar ellipticities and absorbances of the isomers were calculated in the wavelength range of 200-300 nm via parameter estimation method.

[Tandem MS/MS combined with different ionization techniques in the study of steroid structures]. / Szteroidok tömegspektrometriás szerkezetvizsgálata különbözó ionizációs módszerek és MS/MS technikák kombinált alkalmazásával.

The various mass spectrometric ionisation techniques involve different ionisation energy. Their parallel application may provide complementary information for the structure. By the use of soft ionisation methods, especially when combined with tandem mass spectrometric techniques, stereospecific dissociation can also be detected, as the fragmentation processes of low energy differences result in significant abundance differences. The EI spectra of the stereoisomer compounds 1 and 2 are compared to their FAB-daughter ion spectra. This latter method leads to higher differences in the water elimination, indicating that the beta-hydroxi-isomer is more stable. ESI studies, using the "in-source CID" technique and also combined with MS/MS investigation have provided direct information about the position of substituents for the bis-quaternary salts of amino androstane derivatives, pipecuromium bromide [ARDUAN (3)] and its related compound 4. Due to the complementary information of the different ionisation methods used in mass spectrometry, the exact and detailed structural study requires the combined application of the various ionisation techniques together with MS/MS methods.

Estimation of impurity profiles of drugs and related materials Part 18. Impurities and degradation products of mazipredone.

Reversed-phase HPLC methods using C-18 and C-8 columns as well as various isocratic and gradient systems with aqueous ammonium acetate, methanol and acetonitrile are described for the separation of the impurities of mazipredone (11beta,17-dihydroxy-21-(4-methyl-1-piperazinyl)-pregna-1,4-diene- 3,20-dione hydrochloride). These methods were used also for the estimation of the hydrolytic and oxidative degradation pathways of mazipredone in 0.1 M hydrochloric acid and sodium hydroxide at 80 degrees C. With the aid of HPLC-(APCI)-MS and HPLC-diode-array UV techniques 15 impurities and degradation products have been identified.

[Mass spectrometry of pipecuronium bromide and some related compounds]. / Tömegspektrometriás vizsgálatok a pipecuronium bromid és rokon vegyületei körében.

The mass spectral behaviour of the bis-quaternary salt pipecuronium bromide and some related Cn+ A-n type ammonium salts (n = 1, 2, 3) has been studied and compared. Under EI-MS conditions the evaporation of the samples preceded by in situ dealkylation led to formation of alkyl bromide and amine bases. It has been demonstrated that this technique, completed with quantitation of the relative amount of the decomposition products is applicable to get structure information about the basis part as well as the number and positions of the quaternary centres and the substituents at these centres. The FAB mass spectra of these mono-, bis- and this quaternary ammonium salts were found to be very interesting and provide direct structural information about the salt. They exhibit primary Cn+ intact cations and (Cn+ A-n + 1)+ single charged cluster ions and also fragments characteristic of partial structures at the quaternary centres.

[Synthesis of a new neuromuscular blocking agent, pipecuronium bromide (Arduan)]. / Egy új neuromuszkuláris blokkoló, a pipecuronium bromid (ARDUAN) szintézise.

A series of bisquaternary ammonio steroids having androstane skeleton have been prepared some of which possessed high neuromuscular blocking activity. One of the series 3 alpha, 17 beta-diacetoxy-2 beta, 16 beta-bis (4,4-dimethyl-1-piperazinyl)-5 alpha-androstane dibromide (19, pipecuronium bromide, ARDUAN) has proved to be a clinically useful agent of long duration of action without side effects. The preparation of pipecuronium bromide and its metabolites and the impurities are also described. The structure of 19 and related compounds was elucidated by spectrometric methods IR, NMR and MS.

Glycosidase activities in alveolar macrophages from guinea pigs stimulated with a glyco-proteic complex extracted from Klebsiella pneumoniae.

An increased resistance of laboratory animals to pulmonary infections following per os administration of a glyco-proteic complex extracted from Klebsiella pneumoniae has been reported. This was associated with an increased phagocytic capacity of alveolar macrophages (AM). In this report, the effect of treating guinea pigs with this extract on the alveolar macrophage (AM) glycosidase machinery has been studied. AM were collected by bronchoalveolar lavage, the cells were pelleted by centrifugation and AM were purified by adherence on plastic dishes. Sialidase, beta-galactosidase, beta-glucuronidase and N-acetyl-beta-D glucosaminidase activities were measured in the AM homogenate. In order to evaluate an extracellular release of these enzymes, they were also assayed in the cell free lavage fluid. Lactic dehydrogenase (LDH) activity was assayed as a control for cell lysis. In treated animals, the total number of cells as well as the number of AM increased by 25% (ns). The protein concentration was slightly reduced in the cell homogenate and unchanged in the lavage fluid. The only significant change was a decreased sialidase activity, in AM homogenate (p less than or equal to 0.01) and in lavage fluids (ns). The LDH activity was not increased in the lavage fluids.

Sialidase activity and antibodies to sialidase-treated autologous erythrocytes in bronchoalveolar lavages from patients with idiopathic pulmonary fibrosis or sarcoidosis.

Sialidases catalyse the hydrolysis of terminal sialic acid of the carbohydrate moiety of glycoconjugates. Sialic acids play a key role in the expression or masking of antigenic sites and in cell-cell interactions. As an example, removal of sialic acid from the human erythrocyte membrane unmasks underlying molecules such as the specific carbohydrates (Gal-GalNac) of the so-called T or Thomsen-Friedenreich cryptic antigen. A consequence of this, is the recognition of that antigen by natural serum antibodies. Since the T antigen has been shown to be present in the lung, we have investigated the possible presence of sialidase and of specific antibodies to sialidase-treated cells in bronchoalveolar lavage fluids (BALF) from patients with pulmonary sarcoidosis or idiopathic pulmonary fibrosis (IPF). By using a fluorogenic substrate (4-methyl umbelliferyl-alpha-D-N-acetyl sodium neuraminate), we were able to detect a sialidase activity in BALF from eight out of nine patients with IPF and from ten out of thirty-five patients with sarcoidosis. BALF from normal volunteers and serum from both patients and normal volunteers were devoid of activity. BALF sialidase has an optimum pH activity of 5.4, it is not inhibited by EDTA and has a molecular weight close to 21 kD. BALF anti-T antibodies (galactose specific) were detectable in minute amounts in only one out of the nine normal volunteers. By contrast, they were frequently present in BALF from sarcoidosis (77%) or IPF (66%) patients and sarcoidosis patients had a higher mean activity. No correlation was observed between the enzymatic and antibody activities.

Determination of the complement component C2 by ELISA in human serum and bronchoalveolar lavage fluids.

In order to measure the concentration of the human complement component C2 in various biological fluids, an enzyme linked immunosorbent assay (ELISA) was developed. This assay was highly sensitive and allowed to detect as few as 400 pg of C2 in a sample volume of 150 microliters (i.e. 2.6 ng/ml). This is a 10- to 15-fold increase in sensitivity with regard to the conventional hemolytic test. As assessed by an immunoblot analysis, our anti-C2 antiserum was able to detect native C2 as well as the cleavage fragments C2a and C2b generated upon complement activation through the classical pathway. Thus, complement activation involving the classical pathway can easily be evidenced by comparing functional (hemolytic) and immunochemical (ELISA) C2 assays which respectively do not and do reveal activated C2. When C2 was assayed in either normal human serum or bronchoalveolar fluids, in both ELISA and hemolytic tests, a highly significant correlation was observed between the two assays (P less than or equal to 0.01). The specific C2 activity (i.e. functional hemolytic activity/ng C2 assayed in ELISA) was higher in serum than in bronchoalveolar lavage fluids from both normal volunteers and patients with pulmonary diseases.

Bronchoalveolar lavage fluid and serum complement activity in pulmonary sarcoidosis.

In this work, using bronchoalveolar lavage fluids (BALF), we demonstrated the presence of complement within airways by assaying hemolytic activity of the whole classical pathway (CH50) and by measuring the complement component C2 (C2H50). Patients with sarcoidosis, patients with idiopathic pulmonary fibrosis (IPF), and healthy control subjects were compared. No CH50 activity was found in BALF from healthy control subjects (n = 9), but some activity (mean, 20 CH50) was associated with IPF (n = 7). Complement activities ranged from 40 to 554 CH50 in patients with sarcoidosis (n = 27). During the treatment, complement activity decreased in BALF from the few patients in our series who received corticotherapy. C2 hemolytic activity was detected in BALF from the normal control group (in the absence of CH50 activity). In the sarcoidosis and IPF groups when CH50 was present, the variations in the C2/CH50 ratio were studied. The high ratio observed in BALF from patients with sarcoidosis and a chronic derangement of alveolar structure suggests either an increased C2 production or an alternative complement pathway (C2-independent) activation within their lungs.

Differences in carbohydrate specificities and complement-activating capacity of guinea pig and human antibodies to neuraminidase-treated autologous erythrocytes.

Guinea pig erythrocytes desialated by treatment with neuraminidase from Vibrio cholerae were lyzed in autologous serum through a natural-antibody-dependent activation of the classical complement pathway. Lysis was inhibited when a mannose, glucose, galactose or N-acetyl-glucosamine was added to the incubation mixture. Methyl-alpha- or -beta-D-galactopyranosides were poorly effective and N-acetyl-D-galactosamine was not effective at all. Inhibition of lysis by the carbohydrates was due neither to an anti-complementary effect nor to a modification of the osmotic pressure since: (a) they did not alter the total complement haemolytic activity of guinea pig serum, and (b) they did not inhibit lysis of desialated guinea pig erythrocytes in human serum through activation of the alternative complement pathway. The presence of mannose, glucose, galactose or N-acetyl-glucosamine in the incubation mixture resulted in an impaired fixation of natural auto-antibodies on antigenic sites, namely the T-antigen (Thomsen-Friedenreich), which were unmasked following membrane sialic acid removal. When tested under the same conditions, only small percentage of the normal human population showed the phenomenon of lysis of desialated erythrocytes in autologous serum. Lysis was not due to a particular susceptibility of erythrocytes from these individuals to complement-mediated lysis but to the presence in their serum of complement-activating anti-T antibodies. As expected, the activity of human anti-T antibodies was inhibited by galactose and N-acetyl-galactosamine, which are the immunodominant sugars of the human T-antigen. Mannose and glucose had no effect, and methyl- alpha- or - beta-D-galactopyranosides were almost as effective as galactose. The heterogeneity of the human population with regard to the complement-activating capacity of anti-T antibodies could be of significance for the individual response of the host to an infection by a neuraminidase-producing microorganism. That the immunodominant sugars of the T-antigen were different between humans and guinea pigs was further assessed by absorption experiments. We have demonstrated that guinea pig anti-T antibodies were not removed during contact with desialated human red cells which do not have the mannose specificity, whereas human antibodies were almost entirely retained on desialated guinea pig red cells which, beside mannose, express galactose. These results also suggest that guinea pig antibodies are mostly directed towards mannose and glucose.

Early cellular and biochemical alveolar responses following intra-tracheal inoculation with low dose of asbestos and quartz.

The study of broncho-alveolar lavage harvested from rats intratracheally dusted with chrysotile (0.5 mg), leached chrysotile (0.5 mg), crocidolite (0.5 mg) and quartz (0.5 and 5 mg) indicated: 1. A cytotoxic lysis of alveolar macrophages in relation to phagocytosis of dust particles in the following decreasing order: quartz, crocidolite, chrysotile. 2. Regarding cell intensity and duration while asbestos, even leached chrysotile, gave merely an early and transient response. This cell recruitment concerned mostly PMN leukocytes and at a less extent alveolar macrophage. 3. Cell recruitment was associated with an increased protein and phospholipids alveolar content. The increase of proteins came probably mostly from an inflammatory serum exudation. However, an increase synthesis of complement C3 and phospholipids is not excluded.

Auto-antibody dependent activation of the autologous classical complement pathway by guinea-pig red cells treated with influenza virus or neuraminidase: in vitro and in vivo study.

Guinea-pig erythrocytes that had been exposed to influenza A virus or Vibrio cholerae neuraminidase activated the classical complement pathway in autologous serum. Because all viral particles were eluted from the treated cells, activation was not dependent on anti-viral antibodies or on the particles themselves. After a threshold of 45-55% desialation, had been reached, the relative capacity of treated cells to activate complement increased very rapidly with desialation. Desialation unmasked sites on which natural auto-antibodies of the IgM class were fixed. Antibody fixation on the membrane led to C3b deposition on the cell membrane and activation of the classical complement sequence then cell lysis. The relevance of in vitro lysis of desialated cells to in vivo clearance of these cells is not certain because C4-deficient guinea-pigs were able to eliminate desialated cells from the blood stream as efficiently as did normal guinea-pigs. Nevertheless, membrane desialation occurring during myxovirus infection could lead to autoimmunity and tissue changes, as well as to recovery by eliminating virus-modified cells.