Prevention of preterm delivery with vaginal progesterone in women with preterm labour (4P): randomised double-blind placebo-controlled trial.

OBJECTIVE: To evaluate the effectiveness of 200 mg of daily vaginal natural progesterone to prevent preterm birth in women with preterm labour. DESIGN: Multicentre, randomised, double-blind, placebo-controlled trial. SETTING: Twenty-nine centres in Switzerland and Argentina. POPULATION: A total of 385 women with preterm labour (24(0/7) to 33(6/7) weeks of gestation) treated with acute tocolysis. METHODS: Participants were randomly allocated to either 200 mg daily of self-administered vaginal progesterone or placebo within 48 hours of starting acute tocolysis. MAIN OUTCOME MEASURES: Primary outcome was delivery before 37 weeks of gestation. Secondary outcomes were delivery before 32 and 34 weeks, adverse effects, duration of tocolysis, re-admissions for preterm labour, length of hospital stay, and neonatal morbidity and mortality. The study was ended prematurely based on results of the intermediate analysis. RESULTS: Preterm birth occurred in 42.5% of women in the progesterone group versus 35.5% in the placebo group (relative risk [RR] 1.2; 95% confidence interval [95% CI] 0.93-1.5). Delivery at <32 and <34 weeks did not differ between the two groups (12.9 versus 9.7%; [RR 1.3; 95% CI 0.7-2.5] and 19.7 versus 12.9% [RR 1.5; 95% CI 0.9-2.4], respectively). The duration of tocolysis, hospitalisation, and recurrence of preterm labour were comparable between groups. Neonatal morbidity occurred in 44 (22.8%) cases on progesterone versus 35 (18.8%) cases on placebo (RR: 1.2; 95% CI 0.82-1.8), whereas there were 4 (2%) neonatal deaths in each study group. CONCLUSION: There is no evidence that the daily administration of 200 mg vaginal progesterone decreases preterm birth or improves neonatal outcome in women with preterm labour.

cDNA cloning of blue gourami (Trichogaster trichopterus) prolactin and its expression during the gonadal cycles of males and females.

BACKGROUND: The blue gourami fish (Trichogaster trichopterus) provides a unique model for the study of reproduction endocrinology in teleost fish. Its oocyte development may be controlled easily, and the vitellogenic and final maturation phases may be separated artificially in the laboratory. Moreover, this gourami exhibits exclusive parental behavior. AIM: The aim of the present study was to clone and sequence the blue gourami PRL (bgPRL) cDNA in order to enable the determination of its mRNA levels in the male and female blue gourami during the gonadal cycles. MATERIALS AND METHODS: bgPRL was cloned by extracting total RNA from freshly excised pituitaries of gourami fish, followed by cDNA synthesis, rapid amplification of cDNA ends (RACE)-PCR and finally, sequencing. bgPRL mRNA expression was determined by realtime PCR, and results were normalized with 18S RNA. RESULTS: When bgPRL was compared to PRLs of other fish, it had the most homology with PRL of Perciformes and the least with those of Anguilliformes. bgPRL was expressed during the entire gonadal cycle in males and females. The average levels of PRL mRNA in juvenile and low vitellogenetic females were lower than in mature females (at high vitellogenesis and maturation), but the differences were not significant. On the other hand, the PRL mRNA levels in mature reproductive males (nestbuilders) and non-reproductive (non-nest-builders) were significantly higher in comparison to young males. CONCLUSIONS: The results of this study imply that PRL has a possible role in the endocrine control of gonadal development in fish, in addition to its role in reproductive behavior.

Brain gonadotropin releasing hormone3 expression variation during oogenesis and sexual behavior and its effect on pituitary hormonal expression in the blue gourami.

To gain a better insight regarding the roles of gonadotropin releasing hormone3 (GnRH3) in the regulation of reproduction in the suborder Labyrinthici, GnRH3 expression was investigated during the reproduction cycle of the male and female blue gourami (Trichogaster trichopterus). The full-length blue gourami brain GnRH3 gene was sequenced (EMBL acc. no. EU107388) and was found to be expressed in both the brain and pituitary of the blue gourami. High mRNA levels were detected only in the brain of females with oocytes in the maturation stage. Correspondingly, significantly greater mRNA levels of GnRH3 were detected in mature males than in immature ones. In primary cultures of dispersed pituitary cells, GnRH3 significantly increased betaFSH and betaLH subunit mRNA levels in cells from both females and males, whereas GH gene transcription was affected differently by GnRH3 in females, as compared to males. Thus, we propose that GnRH3 can act differentially in the blue gourami females and males. In females, GnRH3 may be involved in the final maturation stage of the oocyte and induces betaFSH, betaLH and GH gene expression; in males, it is engaged in sexual behavior and spermatogenesis regulation via betaFSH and betaLH stimulation and dowregulation of GH transcription.

Cloning of Russian sturgeon (Acipenser gueldenstaedtii) growth hormone and insulin-like growth factor I and their expression in male and female fish during the first period of growth.

In this study, the GH and IGF-I of the Russian sturgeon (rs), Acipenser gueldenstaedtii, were cloned and sequenced, and their mRNA gene expression determined. In addition, to improve our understanding of the GH function, the expression of this hormone was assessed in young males and females. Moreover, IGF-I expression was quantified in young males and compared to that in older ones. The nucleotide sequence of the rsGH cDNA was 980 bp long and had an open reading frame of 642 bp, beginning with the first ATG codon at position 39 and ending with the stop codon at position 683. A putative polyadenylation signal, AATAAA, was recognized 42 bp upstream of the poly (A) tail. The position of the signal- peptide cleavage site was predicted to be at position 111, yielding a signal peptide of 24 amino-acids (aa) and a mature peptide of 190 aa. When the rsGH aa sequence was compared with other species, the highest degree of identity was found to be with mammalians (66-70% identity), followed by anguilliformes and amphibia (61%) and other fish (39-47%). The level of rsGH mRNA was discovered to be similar in pituitaries of females and males of 5 age groups (1, 2, 3, 4, and 5- yr-old). In females and males, the levels did not change dramatically during the first 5 yr of growth. The partial nucleotide sequence of the rsIGF-I was 445 bp long and had an open reading frame of 396 bp, beginning with the ATG codon at position 50. The position of the signal-peptide cleavage site was predicted to be at position 187, yielding a signal peptide of 44 aa. The highest level of IGF-I mRNA expression was recorded in the kidney of adult sturgeons. The IGF-I mRNA expression levels in the intestine, pituitary gland, and liver were not significantly different. Low levels of expression were found in the brain, heart, and muscle. In most tissues, there was no significant difference between mRNA levels of one and 5-yr-old fish. In conclusion, based on the GH-sequence analysis, A. gueldenstaedtii is genetically distant from other teleosts. The expression of the GH mRNA was similar in males and females, and its level remained constant during the first 5 yr of growth. While the IGF-I mRNA expression differed amongst various tissues, the level in each tissue was similar in 1 and 5-yr-old fish.

Blue gourami (Trichogaster trichopterus) gonadotropic beta subunits (I and II) cDNA sequences and expression during oogenesis.

We have cloned two cDNAs from the pituitary gland of blue gourami (Trichogaster trichopterus), coding for the beta subunits of the gonadotropin hormones GtH-I and GtH-II. The two cDNAs were sequenced and subjected to sequence analysis. We have found that the deduced amino acid sequences of both cDNAs were most similar to their striped bass counterparts. The beta GtH-I subunits of blue gourami and striped bass shared 73% of their residues, and the beta GtH-II subunits 84%. The cloning of the cDNAs of beta GtH-I and beta GtH-II has enabled us to measure the expression of their respective mRNAs in the pituitaries of female blue gourami at different stages of the reproductive cycle. The highest levels of beta GtH-I and beta GtH-II mRNA were found in specimens classified as high vitellogenic and in females that were at the final stages of oocyte maturation.

Immunoglobulin DNA Analysis as a Marker of Clonality in the Follow-up of Patients with Hairy Cell Leukemia Treated with Alpha-Interferon.

In fourteen patients with hairy cell leukemia (HCL) the configuration of the immunoglobulin (Ig) heavy chain genes was used as a marker of clonality, to monitor the response of the neoplastic population to treatment with alpha-interferon (a-IFN). In agreement with the morphological, hematological and immunological data, twelve of them showed, after a variable length of therapy, a complete disappearance of rearranged bands in peripheral blood cells. In one patient, who was treated less intensively, the molecularly-defined neoplastic population was still present on two consecutive determinations, whilst in the last patient persistence of disease was repeatedly documented despite prolonged A-IFN treatment. Three further cases were analyzed sequentially: in two, no rearranged bands could be found at repeated determinations; the third, who was in complete remission whilst on 3 × 10(6) U of α-IFN every other day, showed recurrence of disease nine months later when on a maintenance protocol with 3 × 10(6) U/weekly. Nine bone marrow specimens were also analyzed following treatment with α-IFN. In four a monoclonally rearranged band could still be detected, while in another four, reversal of fibrosis and hemopoietic recovery wits coupled with the absence of a molecularly recognizable neoplastic clone. In the last (case, persistence of disease paralleled the findings in the peripheral blood cells. These data indicate that α-IFPJ is capable of producing a specific cytolytic effect on the leukemic population in HCL, which in some cases may lead to complete clonal remissions. Analysis at the DNA level may represent a valuable tool towards monitoring the clinical course of HCL patients and for optimal individual therapeutic scheduling.

Cloning of European eel (Anguilla anguilla) FSH-beta subunit, and expression of FSH-beta and LH-beta in males and females after sex determination.

The European eel (Anguilla anguilla) is a catadromic teleost species with a complex life cycle, both in sea and freshwater environments. The sex determination phase of gonadal development occurs in a freshwater environment. Polymorphism occurs in increasing rates with respect to gender. While males stop growing at approximately 150 g, females continue to grow to being much larger. In this study, we cloned the cDNA FSH-beta subunit of the European eel (A. anguilla), and measured the mRNA levels of FSH-beta and LH-beta in males and females after sex determination. The FSH-beta subunit cDNA consisted of 1068 bp, encoding a 127 amino acid peptide. A comparison between European and Japanese eels of the FSH-beta amino acid sequence showed 98% similarity.

Growth hormone, gonad development, and steroid levels in female carp.

The pituitary and plasma growth hormone (GH) levels of female carp (Cyprinus carpio L.) were measured in fish of various sizes and degrees of maturity, and were matched against the levels of 17 beta-estradiol (E2), testosterone (T), 17 alpha-hydroxyprogesterone (17-P), and progesterone (P) in the ovary and plasma. The short-term action of the above hormones and 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one (17,20-P) on the release of GH was examined in vitro in primary culture pituitary cells. The gonadosomatic index (%GSI) increased rapidly in specimens when they had attained 900 +/- 50 g body weight (BW). The pituitary and plasma GH levels increased between 150 and 600 g BW (when oocytes reached the stage at which lipoprotein appeared in the cytoplasm), but at 900 g BW (with oocytes in vitellogenesis) the plasma GH dropped, while pituitary GH remained high. E2 increased with BW, reaching its maximum at 600 and 900 g BW in the ovary and plasma, respectively. Similar patterns were found in the levels of T and P, both hormones reaching their maximum levels at 900 g BW. The level of 17-P was very low and did not increase in proportion to BW. The application of various concentrations of different steroids on a primary culture of pituitary cells led to release of GH. The highest degrees of release were obtained from 10(-6) and 10(-7) M E2, 10(-6) M T, 10(-7) M 17-P and 10(-8) M 17,20-P. In all these cases, hormone treatment effected higher release of GH than was found in the control. A model of the relationship between GH and the steroids associated with maturation is proposed.

Growth hormone, sexual maturity and steroids in male carp (Cyprinus carpio).

Samples of pituitary, blood plasma and gonad were taken from male carp. The growth hormones (GH) in the pituitary and plasma were measured in fish of various body weights (BW) and degrees of gonad development, and compared with the levels of 17 beta-estradiol (E2), testosterone (T), 17 alpha-hydroprogesterone (17-P), 11-ketotesterone (11-KT) and progesterone (P) in the testes and plasma. The gonadosomatic index increased rapidly with BW from 100 to 600 g, and then decreased at 900 g. The pituitary GH did not change with BW, but plasma GH was higher in fish weighing 300 +/- 50 and 600 +/- 50 g, than in fish weighing 900 +/- 50 g. In fish weighing 150 +/- 50 to 300 +/- 50 g, the level of T rose significantly in the testes (2.27 ng g-1) and plasma (1.3 ng g-1); E2 was very low in both testes (0-30 pg g-1) and plasma (11-28 pg ml-1), increasing as BW rose from 150 to 600 g. The level of P rose mainly at BW of 300 +/- 50 and 600 +/- 50 g: from 0 to 25 ng g-1 in the testes and from 0 to 17 ng ml-1 in the plasma. The level 17-P rose from 2.5 to 20 ng g-1 in the testes at 600 +/- 50 g BW, but no significant change was recorded in the plasma. The level of 11-KT rose significantly in the tests of fish at 300, 600 and 900 g (0.5-6 ng g-1). The application of different steroids (E2, T and 17-P) on a primary culture of pituitary cells led to the release of GH. Release was significantly higher (P < 0.05) after 4 h at steroid concentrations of 10(-6) M.

[Acute leukemia. Case material and therapeutic results]. / Leucemia acuta. Casistica e risultati terapeutici

In the five years period 1969-1973, 22 cases of acute lymphoblastic (ALL) and 75 of acute myeloid leukaemia (AML) were observed in adults. In ALL, complete remission was obtained in 61% of cases. The median duration of remission worked out to be 3 months in the group of patients who died and more than 6 months in the patients still alive. The mean survival time was 7,8 and more than 10,6 months respectively. The various regimens of treatment used in the phase of induction appeared to be equally effective, whilst cyclic chemotherapy was found to be preferable in maintaining remission. As far as AML is concerned, complete remission was achieved in 25% of the patients with stem cells leukaemia, 50% in myeloblastic leukaemia and 33% in mielyomoocytic leulaemia. The median duration of remission was 3,5 and 9 months respectively. No case of promyelocytic leukaemia and erythroleukaemia achieved complete remission. The mean survival time was 3,4 months in stem cells leukaemia, 6 months in myeloblastic leukaemia, 6,6 months in myelomonocytic leukaemia, 0,4 months in promyelocytic leukaemia and 2,5 months in erythroleukaemia. In stem cells leukaemia, the highest frequency of remission was found in patients aged 13 to 20 years, whilst in myeloblastic and myelomonocytic leukaemia the most favourable results were obtained in patients aged 50 to 70 and 20 to 50 respectively. The longest mediam duration of remission and the best survival time was obtained in myelomonocytic leukaemia. The best rate of remission was achieved in the patients whose initial leukocyte count ranged from 10.000 to 50.000/mm-3. No patient with an initial WBC count above 50.000/mm-3 entered complete remission. The frequency of complete remission worked out to be much higher in the patients treated with cyclic chemotherapy according to the Hammersmith protocol, except in the case of stem cells leukaemia.

[Association of leukemias and tumors. Studies of 502 cases of leukemia]. / L'associazione leucemie-tumori. Indagine su 502 casi di leucemia

Nineteen association of leukaemia and tumour were noted in a series of 502 cases of leukaemia: 12/180 (6.6%, compared with 4.7% of 5136 cases in the liteature) for Chr. L.L. (hypogammaglobulinaemia, reduction in single Ig. serious herpes zoster and the T-lymphocyte nature of leukaemia were not more frequent in these associations); 2/102 (1.9%, compared with 2.6% of 1267 cases in the literure) for Chr. M.L.; 5/220 (2.2%, compared with 2.19% of 1138 cases in the literature) for A.L. The mean age of the overall leukaemia series was virtually the same for A.L. (47 yr in a group composed of subjects aged over 12 yr) and Chr. M.L. (48 yr), with the same incidence of association (2.2 and 1.9%), whereas it was 64 yr and 6.6% incidence in Chr. L.L. The bilogarithmic increase in the incidence of tumours with age may itself explain the higher incidence of Chr. L.L. associations. The duration of leukaemia and the age of incidence must be taken into account in any discussion of the significance of such associations.

[Adenopathies in chronic myeloid leukemia (CML). Study of 161 cases]. / Adenopatie nella leucemia mieloide cronica (LMC). Studio su 161 casi.

161 cases of CML have been studied. Clinically significant adenopathies were present in 3,2% of the patients at the moment of diagnosis, and in the subsequent course they appeared in 7% of them. The behaviour of adenopathies showed to be unrelated to: --splenomegaly; --blastic metamorphosis in the peripheral blood or in the marrow (which they often preceded from 3 to 26 months); --hematological sensibility to cytostatic therapy; and furthermore they often acted as the most important clinical and therapeutic problem. From the cyto-histological point of view three features have been observed: 1) blastic metamorphosis in a lymphnode showing features of myeloid metaplasia; 2) blastic invasion in a lymphnode without any sign of myeloid metaplasia; 3) malignant lymphoma. Cytological examination of imprints and ultrastructural studies, besides the usual histological investigations, proved to be useful for the definition of the above mentioned features. In lymphomatous forms, together with the study of the cariotype and the research of the Ph' chromosome, the performance of immunocytological investigations is also necessary.

Evidence of a reproduction-related function for pituitary adenylate cyclase-activating polypeptide-related peptide in an Anabantidae fish.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is synthesized from a precursor, which includes the PACAP-related peptide (PRP; formerly known as GHRH-like peptide). PRP can act as a hypophysiotropic factor in teleosts by stimulating GH secretion. However, no information points to this peptide as a regulator of reproduction. Recently, the blue gourami PRP-PACAP cDNA was cloned and found to be expressed in the brain. Thus, the aims of the present study were to investigate the PRP-PACAP gene expression pattern during sexual behavior and oogenesis, and to learn its effect on pituitary hormonal transcription in pituitary cells. Examination of the PRP-PACAP expression profile during sexual behavior and oogenesis revealed that PRP-PACAP mRNA levels were higher in mature non-reproductively active males than in nest builders and juveniles; and higher in females with oocytes in the final maturation stage than in vitellogenic individuals. Stimulation of pituitary cells with blue gourami PRP (bgPRP) caused an increase in ßLH subunit transcription levels only in females, whereas in males, it only brought about a rise in GH mRNA levels. These data were further supported by the presence of PRP receptor in the pituitary cells. Therefore, we propose that as a hypophysiotropic factor in the blue gourami, bgPRP may act differently on the gonadotropic axes in females and males, up-regulating gonadotropin mRNA in females and GH mRNA in males. This research provides a basis for the further understanding of the integrative network that regulates growth and reproduction, which may contribute to hormonal treatments and manipulations in aquaculture.

Osmoregulation in red blood cells of Bufo viridis.

The effect of hyperosmotic solution of NaCl, urea and mannitol on Bufo viridis red blood cells were studied. The percentage of water content in B. viridis red blood cells decreased significantly in NaCl and mannitol hypertonic solutions compared to urea hypertonic solution. The urea concentration found in red blood cells in a urea hypertonic solution was significantly higher than in red blood cells acclimated to NaCl and mannitol hypertonic solutions. The Na+ concentration was significantly lower in red blood cells immersed in urea hypertonic solution than in red blood cells immersed in hypertonic NaCl and mannitol solutions. However, the K+ concentration increased at a similar rate in three different hypertonic solutions.

Urea tolerance and osmoregulation in Bufo viridis and Rana ridibunda.

1. The tolerance and adaptation to urea solutions by terrestrial green toads (Bufo viridis) and semi-aquatic frogs (Rana Ridibunda) were studied. 2. the green toad showed tolerance to urea solution of 800 mM and the frogs showed tolerance only to about 400 mM urea solution. 3. The plasma concentrations of both species was hyperosmotic to the external medium in all the different urea solutions. 4. Blood osmolality, urea, Na+ and Cl- concentrations of B. viridis were always higher than in R. ridibunda. 5. The urea concentration in muscle of R. ridibunda was higher than the urea concentration in muscle of B. viridis. 6. The muscle tissue weight loss of B. viridis was significantly lower than R. ridibunda.

Effect of dietary carbohydrate source on soluble protein glucose concentration and enzyme activity (aldolase) of European eels (Anguilla anguilla L.).

This study was undertaken to determine the influence of dietary carbohydrate sources: wheat meal, bread meal, soluble corn starch, native potato starch and sorghum meal, on soluble protein, enzyme activity (aldolase) and glucose concentration in muscle and liver of European eels (Anguilla anguilla). There was less soluble protein in both muscle and liver of eels fed 30% wheat meal or bread meal than the other experimental groups. However, eels fed 30% bread meal or soluble corn starch had a higher glucose concentration in muscle and liver than the other experimental groups. High enzyme activity (aldolase) was found in the liver of eels fed 30% wheat meal, bread meal or soluble starch.

Oxygen consumption of a terrestrial toad (Bufo viridis) and semi-aquatic frog (Rana ridibunda).

1. Oxygen consumption was measured before and after dehydration at different ambient temperatures (Ta) in the terrestrial toad Bufo viridis and the semi-aquatic frog Rana ridibunda. 2. The metabolic rates at Tas between 14 and 27 degrees C were almost the same for R. ridibunda and B. viridis. 3. The metabolic rate at higher Tas (between 27 and 36 degrees C) was higher in R. ridibunda than in B. viridis. This situation was found before and after dehydration. 4. A similar situation was found with CO2 production, which was higher at high Tas in R. ridibunda compared with B. viridis.

Sensitivity to maturation-inducing steroids and gonadotropin in the oocytes of blue gourami Trichogaster trichopterus (Anabantidae, Pallas, 1770).

This study examined in vitro the effect of 17 alpha-hydroxyprogesterone (17-P), 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17,20-P), and human chorionic gonadotropin (hCG) on vitellogenesis and the induction of germinal vesicle breakdown (GVBD) in oocytes of the female Trichogaster trichopterus (Pallas, 1770) after various in vivo treatments: (a) in nonreproductive condition; (b) in reproductive condition together with males; (c) injected with hCG; (d) injected with extract of carp gonadotropin (cGtH). Neither 17-P nor 17,20-P affected vitellogenesis or GVBD in oocytes from females in nonreproductive condition. However, the development of maturational competence was observed in oocytes obtained from fish in the other three treatments, 17,20-P having a significantly higher effect than 17-P. In vivo injection with hCG raised the percentage of vitellogenesis and caused some GVBD, while the addition of 17-P or 17,20-P in vitro significantly increased GVBD. Similarly, females maintained with males for 48 hr showed GVBD in all the in vitro cultures treated with 17,20-P. Injection in vivo with cGtH caused sensitivity to both 17,20-P and hCG treatment in vitro, leading to ovulation. It may be concluded that, since injection with hCG and cGtH, like the presence of the male, leads to maturational competence in the female, male presence causes the secretion of these maturation-inducing hormones.