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Adaptive immunity has often been considered the penultimate of immune capacities. That system is now being deconstructed to encompass less stringent rules that govern its initiation, actual effector activity, and ambivalent results. Expanding the repertoire of innate immunity found in all invertebrates has greatly facilitated the relaxation of convictions concerning what actually constitutes innate and adaptive immunity. Two animal models, incidentally not on the line of chordate evolution (C. elegans and Drosophila), have contributed enormously to defining homology. The characteristics of specificity and memory and whether the antigen is pathogenic or nonpathogenic reveal considerable information on homology, thus deconstructing the more fundamentalist view. Senescence, cancer, and immunosuppression often associated with mammals that possess both innate and adaptive immunity also exist in invertebrates that only possess innate immunity. Strict definitions become blurred casting skepticism on the utility of creating rigid definitions of what innate and adaptive immunity are without considering overlaps.

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Multicellular organisms including invertebrates and vertebrates live in various habitats that may be aquatic or terrestrial where they are constantly exposed to deleterious pathogens. These include viruses, bacteria, fungi, and parasites. They have evolved various immunodefense mechanisms that may protect them from infection by these microorganisms. These include cellular and humoral responses and the level of differentiation of the response parallels the evolutionary development of the species. The first line of innate immunity in earthworms is the body wall that prevents the entrance of microbes into the coelomic cavity that contains fluid in which there are numerous leukocyte effectors of immune responses. When this first barrier is broken, a series of host responses is set into motion activating the leukocytes and the coelomic fluid. The responses are classified as innate, natural, non-specific, non-anticipatory, non-clonal (germ line) in contrast to the vertebrate capacity that is considered adaptive, induced, specific, anticipatory and clonal (somatic). Specific memory is associated with the vertebrate response and there is information that the innate response of invertebrates may under certain conditions possess specific memory. The invertebrate system when challenged affects phagocytosis, encapsulation, agglutination, opsonization, clotting and lysis. At least two major leukocytes, small and large mediate lytic reactions against several tumor cell targets. Destruction of tumor cells in vitro shows that phagocytosis and natural killer cell responses are distinct properties of these leukocytes. This has prompted newer searches for immune function and regulation in other systems. The innate immune system of the earthworm has been analyzed for more than 40 years with every aspect examined. However, there are no known entire sequences of the earthworm as exists in these other invertebrates. Because the earthworm lives in soil and has been utilized as a successful monitor for pollution, there are studies that reveal up and down regulation of responses in the immune system after exposure to a variety of environmental pollutants. Moreover, there are partial sequences that appear in earthworms after exposure to environmental pollutants such as cadmium and copper. There are now attempts to define the AHR receptor crucial for intracellular signaling after exposure to pollutants, but without linking the signals to changes in the immune system. There are several pathways for signal transduction, including JAK/STAT, TOLL, TRAF PIP3, known in invertebrates and vertebrates. For resistance to pathogens, conserved signal transduction components are required and these include a Toll/IL-1 receptor domain adaptor protein that functions upstream of a conserved p38 MAP kinase pathway. This pathway may be an ancestral innate immune signaling pathway found in a putative common ancestor of nematodes, arthropods and even vertebrates. It could also help us to link pollution, innate immunity and transduction in earthworms.

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Earthworm innate immunity depends upon small and large leukocytes (coelomocytes) that synthesize and secrete humoral antimicrobial molecules (e.g. lysenin, fetidin, eiseniapore, coelomic cytolytic factor [CCF]; Lumbricin I). Small coelomocytes (cytotoxic) are positive (CD11a, CD45RA, CD45RO, CDw49b, CD54, beta(2)-m and Thy-1 [CD90]; CD24; TNF-alpha) but negative using other mammalian markers. Large coelomocytes (phagocytic) are uniformly negative. Specific earthworm anti-EFCC 1, 2, 3, 4 mAbs are negative for Drosophila melanogaster hemocytes and mammalian cells but positive those of earthworms. Coelomocytes contain several lysosomal enzymes involved in phagocytosis and a pattern recognition molecule (CCF) that may trigger the prophenoloxidase cascade a crucial innate immune response. Earthworms and other invertebrates possess natural, non-specific, non-clonal, and non-anticipatory immune response governed by germ line genes. Toll and Toll-like receptor signaling is essential for phagocytosis and antimicrobial peptide synthesis and secretion in insects and vertebrates but has not yet been shown to be essential in earthworm innate responses.

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Earthworm coelomic fluid contains biologically active molecules and leukocytes that participate in phagocytosis, encapsulation. Presumably they synthesize and secrete several effector modulators of innate immune responses such as antibacterial molecules, cytotoxic proteins and cytokines. Several lytic molecules have been detected in coelomic fluid previously but it is not yet clear which are actually released from the coelomocytes. Our aim was to analyze the cytotoxic effects of coelomocytes on mammalian target cells and to provide evidence that the lytic factors originate from coelomocytes. Cell-free coelomic fluid, supernatants of short-term cultured coelomocytes, and lysates from coelomocytes--derived by mechanical and detergent extraction--were used in cytotoxicity assays performed on different mammalian standard tumor cell lines and mouse fibroblasts. We used native and denaturized (using proteinase K, and trypsin digestions, or heat-inactivation) coelomocyte lysates (CCL). The viability controls of targeted cells were made by measuring photometrically and analyzing by inverted microscopy. According to our results the coelomic fluid, the supernatant of cultured coelomocytes, and the CCL significantly decreased ratios of living cells compared to controls in a dose-dependent manner. Our experiments performed with CCLs suggest that coelomocytes are responsible for the productions of cytotoxic components presumably proteins.

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Earthworm leukocytes (coelomocytes) are responsible for innate cellular immune functions such as phagocytosis and encapsulation against parasites and pathogens. Microbial killing results from the combined action of the phagocytic process with humoral immune factors such as agglutinins (e.g., lectins), lysosomal enzymes (e.g., acid phosphatase, lysozyme), and various cytotoxic and antimicrobial molecules. There is also evidence of weak adaptive immune responses against foreign transplants. This study focused on aspects of the innate immune response. First, anti-human acid phosphatase (anti-AcP) polyclonal antibody characterized different acid hydrolase patterns in coelomocytes. Second, flow cytometry identified a strongly immunoreactive coelomocyte population. Third, ultrastructural and cytochemical analyses revealed acid phosphatase in discrete granules (lysosomes) of effector hyaline and granular coelomocytes but not in mature chloragocytes. Coelomocytes were exposed to bacteria to assess how phagocytosis influences: (a) the production of acid phosphatase using Western blot, and (b) release of acid phosphatase using ELISA from cell-free coelomic fluid. Fourth, after phagocytosis, acid phosphatase levels differed between controls and experimentals. Fifth, we found a 39-kDa molecule that reacted intensely with anti-AcP. Our results suggest that effector earthworm coelomocytes may not eliminate pathogens only by phagocytosis but also by extracellular lysis.

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OBJECTIVES: To compare maternity ward professionals' and patients' views regarding the length of stay in the maternity after a normal delivery and to explore working relationships with ambulatory health professionals. METHODS: Three surveys RESULTS: General professional agreement on a minimum of 4 days; few contacts with ambulatory health professionals. One third of the women found their length of stay excessive. Most did not appear to gain new skills after 3 days, nor to encounter major difficulties once they returned to their home. DISCUSSION: and conclusion. It will be essential to organize effective working relationships between the maternity ward and ambulatory health professionals to ensure appropriate follow-up after an earlier discharge from the hospital, in agreement with the capabilities and expectations of a majority of women.

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BACKGROUND: Actinomycosis is uncommonly observed in a pelvic localization. The presence of an intrauterine device is an important risk factor. CASE REPORT: A 50-year-old woman presented a tumoral mass in the pelvis with secondary liver involvement. Pathology examination corrected the initial diagnosis of advanced stage pelvic neoplasia to actinomycosis. Rapid improvement was achieved with penicillin. DISCUSSION: Differential diagnosis is difficult but essential in case of pelvic actinomycosis as misdiagnosis can have disastrous surgical consequences. Medical treatment is indicated. Actinomycosis should be entertained as a possible diagnosis in all cases associating a pelvic tumor and inflammation in women wearing an intrauterine device.

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UNLABELLED: AN UNUSUAL INFECTION: Actinomycosis, uncommon in a pelvic localization, is a severe condition not well known to gynecologists. It is caused by Actinomyces israeli and is closely associated with long-term use of an intrauterine device. In the pelvic localization, the disease generally presents as a pseudoneoplastic formation. DIFFICULT DIAGNOSIS: Diagnosis is generally not established clinically. Pathology provides positive diagnosis. The germ cannot be isolated easily as it does not survive standard bacteriology sampling. Monoclonal antibodies may be helpful. MEDICAL TREATMENT: Intravenous penicillin G followed by at least 6 months oral penicillin is generally successful in eradicating the infection. Indications for surgery should be limited to diagnostic procedures (pathology specimen), drainage in case of abscess formation, and removal of an obstacle compressing the digestive or urinary tract.

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Radiolabeling of [Tyr8]-substance P ([Tyr8]-SP) with the (125)I-isotope was performed by use of the chloramine-T technique. The primary formed radiolabeled product, having been quantitatively converted to the corresponding sulfoxide yielding [(125)I]-[Tyr8]-(Met11-->O)-SP completely lacked any binding to proteins rich in SP receptor populations. However, after reductive treatment with mercaptoethanol for about 2 h, a complete reconstitution of the Met11 thioether structure was observed. The reduced peptide, consisting of [(125)I]-[Tyr8]-(Met11)-SP was separated from its by-products by reversed-phase high-performance liquid chromatography on octadecylsilyl silica gel with 100 mM triethyl ammonium formate buffer containing 22% acetonitrile (pH 2.2). The labeled SP derivative prepared by this two-step synthesis was obtained in 73% overall yield related to the [Tyr8]-SP starting material and exhibited a specific activity of 1.9-10(6) Ci/M. In contrast to [(125)I]-[Tyr8]-->(Met11-->O)-SP, satisfactory receptor-binding was now observed with the [(125)I]-->[Tyr8]-(Met11)-SP derivative.

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The effects of ambroxol on the spasmolytic action of clenbuterol were investigated on acetylcholine-induced bronchospasm in guinea-pigs. Ambroxol (50 mg kg-1 day-1) or vehicle was administered orally for 14 days. Approximately 45 min after the final dose on day 14, the animals were anaesthetized and the spasmolytic effects of clenbuterol (3, 6 or 12 micrograms kg-1 injected intravenously) were determined by use of acetylcholine (40 micrograms kg-1, i.v.)-induced bronchoconstriction. For both vehicle- and ambroxol-treated animals, a positive linear relationship was observed between the log-dose of clenbuterol and the percent inhibition of bronchospasm. The calculated ED25 of clenbuterol (i.e., the dose producing 25% inhibition of the acetylcholine-induced bronchospasm) was 3.98 micrograms kg-1 (3.29 to 4.82 micrograms kg-1, 95% confidence interval) in the presence of ambroxol and 5.81 micrograms kg-1 (4.98 to 6.79 micrograms kg-1) in the absence of ambroxol. The linear regressions with or without ambroxol differed from each other (P < 0.001) but ran parallel (covariance analysis), enabling us to calculate a relative potency, the value of which was 1.46 (1.16 to 1.84). These results demonstrate that the spasmolytic activity of clenbuterol is significantly improved in animals pretreated with ambroxol.

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Conditions for the labeling of insulin with radioactive iodine isotopes were investigated by means of incorporation of non-radioactive 127I into the peptide. Either the chloramine-T (CT) or lactoperoxidase-hydrogen peroxide (LPO) technique was applied and reversed-phase high-performance liquid chromatography (RP-HPLC) was used for analysis of the reaction products. The LPO method provided the 127I-labeled peptide within 15-30 min, whereas the CT alternative yielded the labeled substrate even within 15 s. However, the latter reaction can only be controlled in a reproducible manner with difficulty and undesired side-reactions became increasingly prominent when the reaction time of 15 s was exceeded for only a few seconds. In another experiment, the LPO technique was applied for radiolabeling insulin with 125I. The product was first purified by size-exclusion chromatography (SEC) and then subjected to RP-HPLC. SEC yielded two peaks. The smaller one, which eluted at a slightly higher Kd value (accounting for about 14% of total radioactivity) predominantly consisted of material eluting at the column's void volume under the conditions of RP-HPLC, whereas the main SEC fraction (accounting for about 86% of total radioactivity) yielded a single peak, as shown by HPLC. The radioactive material attributable to the main SEC fraction revealed the expected receptor-binding properties, as evidenced by displacement experiments with non-radioactive insulin, as well as the action of tetradecanoyl phorbol acetate on the binding characteristics and thus indicating formation of a labeled hormone retaining biological activity.

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OBJECTIVES: Evaluation (usefulness and safety) of programmed autologous transfusion in obstetrics. SITE. Blood Transfusion Centre, Hôpital Louis-Mourier, F 92700 Colombes. PATIENTS: Prospective study of 150 patients for whom blood withdrawal was planned during the last month of pregnancy. Entry criteria were either a risk of haemorrhage or persistent patient request. PROTOCOL: Two withdrawals were planned during the last month of pregnancy at the out-patient clinic at a one-week interval. The autologous units were transfused per-partum in case of haemorrhage and/or post-partum in case of anaemia. RESULTS: One hundred pregnant women entered the protocol (43 had a risk of haemorrhage). Both preplanned withdrawals were made in 60 of these patients. Per-partum transfusions were necessary in only 7 patients including 4/43 with a risk factor (9%) and 3/57 with no risk factor (5%). Post-partum transfusions were made in 22 other patients. Consequently, 117 of the 160 units collected were not used (73%). CONCLUSION: Despite good tolerance (5% incidence) due to the known problems in evaluating the risk of haemorrhage and the small percentage of patients without risk factors who were transfused per-partum, we have decided to reserve this protocol for patients with an authentically identified risk of haemorrhage (placenta praevia, cesarean section, uterine scar tissue).

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OBJECTIVE: To determine the relationship between bacterial colonization of the amniotic fluid sampled by amniocentesis and premature rupture of the membranes. METHODS: A prospective multicentric study conducted over a one year period in 6 maternity wards in the suburban area of Paris. Thirty-six women with premature rupture of the membranes were studied. RESULTS: There were 11 patients (30%) with contaminated fluid at the first amniocentesis. Twenty-five patients had sterile fluid. In 7 patients with prolonged rupture for more than one week, repeated amniocentesis. Twenty-five patients had sterile fluid. In 7 patients with prolonged rupture for more than one week, repeated amniocenteses were used to follow bacterial colonization. In one patient, Proteus mirabilis in the amniotic fluid was eradicated by adapted antibiotic therapy. In patients with sterile amniotic fluid, there was no secondary colonization. In the 11 cases with colonized liquid, the vaginal swab could only be considered as positive in 4 cases. Amniocentesis was able to discover 7 bacterial colonizations of the amniotic fluid in patients with an indeterminant vaginal swab. Likewise, C-reactive protein levels were raised only in 26% of the cases with a colonized amniotic fluid.

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Several authors have found that caesarean section might be a risk factor for post-partum depression, while others consider that mood disturbances are not a specific response to delivery and can be observed in women undergoing hysterectomy. This longitudinal prospective study was carried out in 156 in-patient women at the Maternity department of Louis Mourier hospital, near Paris. These women were divided into three groups: 52 with normal delivery, 63 with caesarean section and 41 with gynecological surgery. Seventeen per cent of the women who had delivered suffered from baby-blues. The figure for anxiety (De Bonis scale > or = 20) was 23% in the 1st group, 41% in the 2nd group and 54% in the 3rd group. The figure for mood disturbances (according to DSM III criteria) was 4% in the 1st group, 16% in the 2nd group and 21% in the 3rd group. There was no significant difference between the 3 groups in figures for anxiety and depression during the first 3 months after delivery or surgery, but anxiety and depression ratings seem to fall faster and more steadily in women who had surgery.

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Group B streptococcus (GBS) is an important cause of neonatal infection. Early-onset diseases are due to perinatal contamination. The epidemiology of late-onset infections is poorly known. Maternal colonization may be responsible for some of them. The relationships between neonatal colonization and late disease could be a colonization of the gut. The purpose of this 3 year-prospective study was to analyse the kinetics of gut colonization in neonates and the influence of antibiotherapy. One hundred and nineteen infants less than one month of age were included because of the presence of GBS in their gastric aspirates or GBS infection. Depending on the therapeutic strategy, the infants were separated into 3 groups: 1) amoxicillin plus aminoside greater than or equal to 10 days because of neonatal infection (28 infants), 2) same combination less than or equal to 5 days because a GBS infection was suspected but not confirmed (17 infants), 3) no antibiotics (77 infants). Fecal flora was regularly analysed by differential count. Antibiotics caused rapid disappearance of GBS from the gut. However, the same strain reappeared after stopping the antibiotics at a rate of 13.5%. Without antibiotics, GBS was implanted in 33% of cases. This difference of implantation rate is statistically significant (p less than 0.05). No GBS infection was observed in any infant after a follow-up examination of 6 months to 2 years. Among the clinical and bacteriological factors studied, adhesion only was correlated with the GBS implantation. These results allow to discuss therapeutic abstention in colonized infants without any signs of infection.

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The condition of "megacystis-microcolon" is a very rare condition which can now be diagnosed ante-natally due to ultrasound. The presence of a megacystis (large bladder) with increased levels of amniotic fluid makes it possible to eliminate other ante-natal causes of dilatation of the bladder and in particular when it is found in a female fetus (as it is in 80% of cases). At birth, one finds, apart from the massive dilatation of the bladder with very little effect on the upper urinary tract, a microcolon with complete absence of peristalsis. This last abnormality always leads to a catastrophic short term result because treatment is quite ineffective. Many histological studies have been carried out and these sometimes differ from one another but it is possible to think that the condition is due to pathology of neurogenic origin in the muscle of the bladder and the aetiology of this is unknown. Finally, because there have been a few familial cases, there may be a genetic element in this illness that has such a poor prognosis.

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Cervico-isthmial incompetence develops in the second trimester of pregnancy. There are problems as to the correct treatment. The authors report four cases where stitches were put in late using the technique of "parachute" cerclage. This made it possible for four live babies to be born. A search of the literature shows that after late cerclage in 60-90% of cases pregnancy continues normally. But it has to be realised that complications do occur and these are mainly chorio-amnionitis which carries a bad prognosis obstetrically and also premature rupture of the membranes. The authors recommend this manoeuvre in spite of the risks because of the benefits that can be obtained. The procedure should only be carried out if there is no cervico-vaginal infection as determined by negative swabs and tocolytic treatment should also be given.

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Histamine release induced by compound 48/80 from rat mast cells is not dependent on extracellular Ca2+. Preincubation of mast cells with trypsin has only little effects on histamine release induced by this polycation. This work also demonstrates that histamine release induced by compound 48/80 and its analogues in the absence of extracellular Ca2+ depends on membrane bound sialic acid of the mast cell. Neuraminidase treatment of the cells in the presence of extracellular Ca2+ leads to histamine liberation. These findings suggest that sialic acid residues of the mast cell membrane constitute the site at which polycations exert their stimulatory actions of histamine liberation.

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Compound 48/80, a mixture of oligomers, was fractionated by passing it in the presence of Ca2+ over a calmodulin-Sepharose column. The fraction not retained by the gel was shown by mass spectrometry to consist mainly of trimers, tetramers and pentamers. A second fraction consisting of hexamers and heptamers was eluted from the column at high ionic strength in the presence of Ca2+. Finally, in the presence of EGTA at high ionic strength, a third fraction eluted mainly consisting of higher oligomers (hexamers to dodecamers). The different fractions were characterized by testing their influence on calmodulin-sensitive Ca2+-transporting ATPase and their ability to elicit histamine release from mast cells. The third fraction showed the highest potency as calmodulin antagonist, however, the second fraction was the most potent in inducing histamine secretion. This would imply that the ability of compound 48/80 to evoke histamine release and to inhibit the function of calmodulin are distinct properties of the agent which are unrelated.

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