Blurring borders: innate immunity with adaptive features.

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.

Still waiting for the toll?

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.

Anticipating innate immunity without a Toll.

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.

Compound 48/80 is a selective and powerful inhibitor of calmodulin-regulated functions.

Compound 48/80, a condensation product of N-methyl-p-methoxyphenethylamine with formaldehyde, is composed of a family of cationic amphiphiles differing in the degree of polymerization. Compound 48/80 was found to be a potent inhibitor of the calmodulin-activated fraction of brain phosphodiesterase and red blood cell Ca2+-transport ATPase, with IC50 values of 0.3 and 0.85 micrograms/ml, respectively. However, the basal activity of both enzymes is not at all suppressed by the drug at concentrations up to 300 micrograms/ml. Inhibition of Ca2+ transport into inside-out red blood cell vesicles by compound 48/80 follows a similar pattern in that basal, calmodulin-independent, transport is also not affected by the drug. Kinetic analysis revealed that the stimulation of Ca2+-transport ATPase induced by calmodulin is inhibited by compound 48/80 according to a competitive mechanism. It was demonstrated that the inhibitory constituents of compound 48/80 bind to calmodulin in a Ca2+-dependent fashion. Comparison of the specificity of several anti-calmodulin drugs showed that compound 48/80 is the most specific inhibitor of the calmodulin-dependent fraction of red blood cell Ca2+-transport ATPase that has been described hitherto. In addition, compound 48/80 was found to be a rather specific inhibitor of the calmodulin-induced activation of Ca2+-transport ATPase when compared with the stimulation induced by an anionic amphiphile or by limited proteolysis. Half-maximal inhibition of the activity stimulated by oleic acid or mild tryptic digestion required 8- and 32-times higher concentrations of compound 48/80, respectively, compared with the calmodulin-dependent fraction of the ATPase activity. Moreover, calmodulin-independent systems as rabbit skeletal muscle sarcoplasmic reticulum Ca2+-transport ATPase or calf cardiac sarcolemma (Na+ + K+)-transport ATPase are far less influenced by compound 48/80 as compared with trifluoperazine and calmidazolium. Because of its high specificity compound 48/80 is proposed to be a promising tool for studying calmodulin-dependent processes.

Induction of histamine release and calmodulin antagonism are two distinct properties of compound 48/80.

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.

Earthworm leukocytes kill HeLa, HEp-2, PC-12 and PA317 cells in vitro.

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.

Ambroxol improves the broncho-spasmolytic activity of clenbuterol in the guinea-pig.

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.

The variation of the mean spinability of human cervical mucus throughout the various stages of reproductive life. Automatic measurement and statistical study.

The evolution of the mean spinability value of human cervical mucus was studied by means of an automatic device throughout the menstrual cycle and pregnancy and during and after the onset of menopause. The automatic measurement brings to light the major drawbacks presented by manual methods: values previously obtained should be considered as underestimated. The mean spinability value recorded from cyclic mucus exhibits a very significant increase at mid-cycle. Graphical analysis demonstrates that only small differences in the mean spinability value occur as pregnancy proceeds. A progressive and regular decrease in the mean spinability value can be recorded from the very beginning of the menopause. However, the spinability values observed in premenopausal mucus are noticeably higher than those recorded after the onset of the menopause. The numeric data furnished by the automatic measurement are discussed in terms of statistics.

[Study of the relation of gynecologic and obstetric findings and psychological disorders of pregnancy and the puerperium]. / Etude des relations entre données gynéco-obstétricales et troubles psychiques de la grossesse et du post-partum.

The relationship between gynaecological and obstetrical parameters and psychological troubles in pregnancy and after delivery are a matter of much debate. This arises partly because of different methods of studying the conditions and because of epidemiological studies that have been carried out on this subject. This longitudinal prospective study has been carried out on 211 women who were delivered in the maternity department of Louis Mourier Hospital. The figure for anxiety (in the de Bonis scale of greater than or equal to 20) was 13 to 15% at different times in the pregnancy, of depression (in the CESd scale greater than or equal to 16) was between 11 and 14.5%. 34.5% of those women who had delivered suffered from "baby blues", while 20% had symptoms of depression (CESd greater than or equal to 16 in the first week post-partum) and 6% had a quite obvious syndrome of depression (MADRS greater than or equal to 20). Finally, 18 to 24.5% of women had a major episode of depression (according to the DSM III test) in the year following delivery. As far as risk factors were concerned there was a statistically significant correlation between anxiety and depression in pregnancy and induction of labour, as well as difficulties with the third stage of labour and with an Apgar score of less than 9 in the infant. There was poor correlation between "baby blues" and gynaecological and pathological changes except where vaginal delivery was not spontaneous and where large episiotomies were carried out. On the other hand, women who had pathological labours or cesareans or a general anaesthetic for the third stage had more troubles than others in the year following delivery. Finally, women who showed signs of anxiety or depression in pregnancy or had "baby blues" after delivery were less likely to be able to breast feed their children.

[Late cerclage]. / Le cerclage tardif.

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.

[Anxiety and depressive disorders in the postpartum period in pregnant females]. / Etude des troubles anxieux et dépressifs du post-partum chez les femmes enceintes.

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.

[Pregnancy type nephropathies experimentally induced in rats]. / Néphropathies de type gravidique obtenues expérimentalement chez le rat

The theory behind our work is the following: toxaemia may be due to the accidental passage of fetal proteins into the maternal organism. Particularly suspect are alpha-fetoproteins from the fetal blood or the amniotic fluid. The experiments were carried out on rats by injecting amniotic fluid or fetal serum into the rats by different routes and rhythms and in different doses. Arterial blood pressure readings showed that the levels were consistently higher than those of controls. The study of the kidneys anatomo-pathologically shows the following triad: intense congestion, tubular lesions, connective tissues affected. These lesions are reversible. The discussion concentrates on the similarities and differences between these findings and pathological changes found in kidneys in pregnancy.

[Pregnancy in women wearing a heart valve prosthesis: 5 case histories (author's transl)]. / Grossesse chez les femmes porteuses de prothèse valvulaire cardiaque. A propos de 5 grossesses.

The study of a personal series of five pregnancies and the review of 130 cases published enabled us to recall the physiological, clinical and therapeutic sides of the "pregnancy-heart valve prothesis" combination. Maternal mortality remains at 3%; morbidity, essentially in the form of haemorragic or thrombo-embolic accidents, comes to 45%. We insist on the importance of a pregnancy planned and accepted both by the cardiologist and the obstetrician as contraception is a difficult problem with these patients, classical oestroprogestative methods and intra-uterine devices being too much of a risk. An intake of a progestagen in small doses combines efficiency with innocuity.

[Placental localisation : screening and prognosis (author's transl)]. / La localisation placentaire : surveillance et pronostic.

Ultrasounds are the best means at present available for observing the placenta. It follows that it is necessary to appreciate the different phases that occur in placental development. Ultrasound will allow us to work out this evolution and to look for abnormalities that may develop. The placenta becomes an entity at about the 12th week of amenorrhoea. Precise localisation of the placenta is possible in the second trimester of pregnancy, but the final site of the placenta is not fixed in the uterus because different segments develop unevenly. All the same, it is necessary to make a prognosis of the risks of an abnormal insertion of the placenta at the end of pregnancy. It is therefore necessary to be precise in localising the placenta. There are several sources of error which have to be avoided: -- difficulty of exact localisation of the internal os of the cervix. -- the existence of thickening of the myometrium, which forms a reserve for uterine stretch.. These thickenings sometimes come at the same places as the placenta and it may be difficult to differentiate one from the other. -- the presence of placental folds. There are several different elements involving prognosis : -- the site of the lower edge of the placenta in relationship to the internal os. -- the site of the upper edge of the placenta in relationship to the fundus of the uterus. This, statistically, is the best index of prognosis, and so we suggest a classification. -- finally the volume of the myometrium between the lower edge of the placenta and the internal os gives an idea and allows us to work out how the lower segment can stretch. Finally, it is by combining all these elements in a study that a better prognosis can be worked out for the final site of the placenta in the uterus. It is only in the third trimester of pregnancy after the lower segment has formed that the final site of the placenta can be confirmed and that a decision can be taken on the way patient should be treated.

[Length of stay in maternity wards after normal delivery: diverging point of views]. / Durée du séjour en maternité après un accouchement normal: des points de vue divergents.

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.

[Megabladder. Microcolon. Prenatal ultrasonic diagnosis and a review of the literature. Apropos of a case]. / Mégavessie. Microcôlon. Diagnostic échographique anténatal et revue de la littérature. A propos d'un cas.

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.

[Premature detachment of ova diagnosed by echography. Apropos of 42 cases]. / Les décollements d'oeufs jeunes à l'échographie. A propos de 42 cas.

The authors noted, while carrying out ultrasound examinations, that there was many cases of abruption of oocytes at the beginning of pregnancy. This study was of 42 cases in 3 maternity units of Paris and its suburbs. It shows up how often, in pregnancies complicated bleeding in the first trimester, ultrasound can demonstrate such abruption. Ultrasound pictures can demonstrate the principal characteristics and the outcome of these abruptions, as well as their effect on the continuation of the pregnancy. In this way it establishes a correlation between the anatomy and the pathology and brings to the fore various hypothesis as to the aetiology of these pathological conditions.

[Programmed autologous transfusion in obstetrics. Blood samples in 100 patients in the last month of pregnancy]. / Transfusion autologue programmée en Obstétrique. 100 patientes prélevées dans le dernier mois de la grossesse.

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).

[Study of amniotic fluid bacterial colonization sampled by amniocentesis in cases with premature rupture of membranes. Prospective multicenter study]. / Etude de la colonisation bactérienne du liquide amniotique prélevé par amniocentèse en cas de rupture prématurée des membranes. Etude prospective multicentrique.

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.

[Pelvic actinomycosis: a case with secondary localization in the liver]. / Actinomycose pelvienne: un cas avec localisation secondaire hépatique.

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.