Latencia al parto tras cerclaje de emergencia en gestación múltiple / Latency to delivery after emergency physical-examination indicated cerclage in twin pregnancy

Introducción: Las guías clínicas actuales recomiendan el uso del cerclaje de emergencia (CE) como tratamiento de la insuficiencia cervical con exposición de membranas en gestaciones únicas. Sin embargo, el CE en gestación múltiple es tema de controversia dado que no existen ensayos clínicos randomizados que demuestren su eficacia. Algunos estudios retrospectivos sugieren que el CE también podría prolongar la gestación en embarazos múltiples. El objetivo de nuestro estudio es evaluar los resultados de las gestaciones múltiples que se sometieron a un CE en nuestro centro. Materiales y métodos: Se diseñó un estudio retrospectivo que incluyó los CE realizados en gestaciones gemelares en nuestro centro entre 2007-2016. No fueron tributarias de CE gestaciones con malformaciones fetales, monocoriales-monoamnióticas, triple o superior y finalizaciones activas de la gestación. Variables primarias: latencia al parto espontáneo y edad gestacional al parto. Variables secundarias: mortalidad neonatal, ingreso en UCI neonatal, rotura prematura de membranas pretérmino, corioamnionitis y fallo del cerclaje. Resultados. El estudio incluyó 17 pacientes. La edad gestacional mediana (rango intercuartil) al parto fue de 27,1 (24,5-32,3) semanas y la latencia mediana (rango intercuartil) al parto fue de 43 (21-64) días. Hubo 4/17 (23,5%) casos de parto antes de las 24 semanas de gestación y 2/26 (7,7%) de muerte neonatal. Discusión: Estos resultados muestran que la latencia al parto después del CE en gestación múltiple es remarcable, por lo que podría ser considerado como una opción terapéutica. Sin embargo, se requiere evidencia basada en estudios randomizados para hacer una recomendación firme

Introduction: Current guidelines support the use of physical-examination indicated cerclage (PEIC) as a treatment for cervical insufficiency and membrane exposure in single pregnancies. However, PEIC in twin pregnancies is a controversial issue as no data from random clinical trial are available to demonstrate its efficacy. Few studies suggest that PEIC may prolong pregnancy also in twin pregnancies. The aim of this study was to evaluate the results of twin pregnancies that underwent a PEIC in our health centre. Material and methods: A retrospective review was performed on women that underwent a PEIC from 2007-2016 in our centre. Women were not eligible if they were carrying foetuses with major foetal anomalies, more than two foetuses or monochorionic-monoamniotic pregnancies, or three or more foetuses or requesting an elective termination of pregnancy. Primary outcomes: latency to spontaneous delivery and gestational age (GA) at delivery. Secondary outcomes: neonatal mortality and Neonatal Intensive Care Unit admission, preterm premature rupture of membranes (PPROM), chorioamnionitis and cerclage displacement. Results: The study included a total of 17 women. The median (inter-quartile range) gestational age at delivery was 27.1 (24.5-32.3) weeks, and median (inter-quartile range) latency, from cervical cerclage to delivery, was 43 (21-64) days. There were 4/17 (23.5%) cases of delivery before 24 weeks of pregnancy, and 2/26 (7.7%) cases of neonatal death. Discussion: These results suggest that latency to delivery after PEIC in twins is remarkable. Therefore, it could be considered as an optional management. Nevertheless, evidence based on random clinical trial is required to make firm recommendations on its formal use

Cervical consistency index and risk of Cesarean delivery after induction of labor at term.

OBJECTIVE: To evaluate the association between the cervical consistency index (CCI) and the risk of Cesarean delivery after planned induction of labor (IOL) at term. METHODS: This was a prospective observational study of women with a term singleton pregnancy admitted for IOL due to maternal or fetal indication. Ultrasonographic images were obtained before IOL and CCI was calculated offline once recruitment was completed. The main outcome was defined as Cesarean delivery due to failed IOL or arrest of labor. Cesarean deliveries indicated due to maternal or fetal compromise (Van Dillen's grade 1 or 2) were excluded from analysis. Univariate statistical analysis was performed using Fisher's exact test and Student's t-test for categorical and continuous variables, respectively. Multivariate analysis was performed using logistic regression, including CCI and other variables related to the main outcome. Intraclass correlation coefficients were used to estimate intra- and interobserver agreement. RESULTS: Of 510 women admitted for IOL during the study period and for whom image quality was adequate, 46 were excluded due to emergency Cesarean delivery leaving 464 pregnancies for analysis. Cesarean section due to failed IOL or arrest of labor was performed in 100/464 (21.6%) pregnancies. The mean CCI of women who underwent Cesarean delivery was not significantly different from that in those who had vaginal delivery after IOL (70.1 ± 12.3% vs 70.0 ± 13.1%; P = 0.94). Multivariate analysis also showed absence of statistical association between CCI and Cesarean delivery for failed IOL or arrest of labor. Intraclass correlation coefficients for intra- and interobserver agreement were 0.81 (95% CI, 0.66-0.89) and 0.86 (95% CI, 0.75-0.92), respectively. CONCLUSION: CCI does not seem to be associated with the risk of Cesarean delivery after IOL. Copyright © 2018 ISUOG. Published by John Wiley & Sons Ltd.

Quantitative analysis of cervical texture by ultrasound in mid-pregnancy and association with spontaneous preterm birth.

OBJECTIVE: New tools are required to improve the identification of women who are at increased risk for spontaneous preterm birth (sPTB). Quantitative analysis of tissue texture on ultrasound has been used to extract robust features from the ultrasound image to detect subtle changes in its microstructure. This may be applied to the cervix. The aim of this study was to determine if there is an association between quantitative analysis of cervical texture (CTx) on mid-trimester ultrasound and sPTB < 37 + 0 weeks' gestation. METHODS: This was a single-center nested case-control study of a prospective cohort of 677 consecutive women with singleton pregnancy assessed between 19 + 0 and 24 + 6 weeks' gestation. Women at increased risk for sPTB were included unless they received treatment to prevent sPTB. Women who delivered < 37 + 0 weeks (sPTB) were considered as cases and were matched in a 1: 10 ratio with randomly selected contemporary controls who delivered at term. For each woman, one ultrasound image of the cervix was obtained for which quality was assessed, cervical length (CL) measured offline and a region of interest in the midportion of the anterior cervical lip delineated for use in local binary patterns analysis of CTx. A learning algorithm was developed to obtain the combination of CTx features best associated with sPTB based on feature transformation and discriminant analysis regression. The ability of the learning algorithm to predict sPTB was evaluated using a leave-one-out cross-validation technique, which produced a CTx-based score for each participant. Receiver-operating characteristics (ROC) curves were produced and sensitivity, specificity, and positive and negative likelihood ratios were calculated for the optimal cut-off based on the ROC curve. The results were compared with those obtained for CL. Investigators studying the images were blinded to pregnancy outcome at all times. RESULTS: Images from 310 women (27 cases and 283 controls) were of sufficient quality and included in the study. Median CTx-based score was significantly lower in cases compared with controls (-1.01 vs -0.07, P ≤ 0.0001). CTx-based score maintained its significant association with sPTB after adjusting for possible confounders (history of sPTB, conization or Müllerian malformation, and CL < 25 mm). CTx-based score was a better predictor of sPTB (AUC, 0.77; 95% CI, 0.66-0.87) than was CL (AUC, 0.60; 95% CI, 0.47-0.72) (P = 0.03). Median CL was similar for cases and controls (37.7 vs 38.6 mm, P = 0.26), although cases were more likely to have CL < 25 mm (18.5% vs 0.4%, P < 0.001). CONCLUSION: Quantitative analysis of CTx enables the extraction of information relevant to sPTB from ultrasound images to generate a CTx-based score that is associated independently with sPTB. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

Mid-trimester sonographic cervical consistency index to predict spontaneous preterm birth in a low-risk population.

OBJECTIVES: To investigate the effectiveness of mid-trimester sonographic cervical consistency index (CCI) for the prediction of spontaneous preterm birth (sPTB) in low-risk pregnancies and to compare its performance with that of mid-trimester sonographic cervical-length (CL) measurement. METHODS: This was a prospective cohort study of women with a singleton pregnancy examined by ultrasound at 19 + 0 to 24 + 6 weeks' gestation. All women underwent transvaginal ultrasound examination of the cervix, but CCI and CL were measured, offline, only in women without a risk factor for sPTB. Staff and participants were blinded to CL and CCI results. CCI was obtained by calculating the ratio between the anteroposterior diameter of the uterine cervix at maximum compression and at rest. The primary outcome was prediction of sPTB before 37 + 0 weeks. Receiver-operating characteristics (ROC) curves were produced and sensitivity and specificity were calculated for the optimal cut-off based on the ROC curve and for the 1st , 5th and 10th centiles of CCI and CL. Intraclass correlation coefficients (ICC) and Bland-Altman plots were used to estimate intra- and interobserver agreement and reliability for measurement of CCI and CL. RESULTS: Of the 749 women who underwent ultrasound examination of the cervix, 532 were included for analysis. The rates of sPTB before 37 + 0 and before 34 + 0 weeks were 4.1% (22/532) and 1.3% (7/532), respectively. The rates of short cervix < 25 mm and ≤ 20 mm were 0.9% (5/532) and 0.4% (2/532), respectively. The area under the ROC curve (AUC) with regard to predicting sPTB before 37 + 0 weeks was 0.84 (95% CI, 0.75-0.93) for CCI compared with 0.68 (95% CI, 0.56-0.81) for CL (P = 0.03). The optimal cut-off based on the ROC curve was 64.6% for CCI (sensitivity, 77.3%; specificity, 82.7%) and that for CL was 37.9 mm (sensitivity, 72.7%; specificity, 61.2%). The AUC with regard to predicting sPTB before 34 + 0 weeks was 0.87 (95% CI, 0.71-1.0) for CCI compared with 0.71 (95% CI, 0.47-0.94) for CL (P = 0.25). The optimal cut-off based on the ROC curve was 63.6% for CCI (sensitivity, 85.7%; specificity, 84.0%) and that for CL was 37.9 mm (sensitivity, 85.7%; specificity, 61.3%). Intraobserver ICC was > 0.90 both for CCI and CL, while interobserver ICC was 0.89 for CCI and 0.90 for CL. CONCLUSIONS: Second-trimester CCI is a better predictor of sPTB < 37 weeks in low-risk pregnancies than is CL. External validation is needed as well as studies assessing the value of CCI as a screening tool in unselected and high-risk populations. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

Antigen characterization from second instars of oestrid bot flies for the detection of anti-Cephenemyia stimulator antibodies by ELISA in roe deer (Capreolus capreolus).

A study to determine the most appropriate antigen for use in the serodiagnosis of Cephenemyia (Diptera: Oestridae) infestation in roe deer (Capreolus capreolus) was carried out using immunoenzymatic tests. Serum samples from 43 roe deer from northern Spain were obtained post-mortem and corresponding numbers of bot fly larvae established. Three antigen complexes were tested, including Cephenemyia stimulator Clark excretory/secretory antigens (CsES), C. stimulator somatic antigens (CsSA) and Oestrus ovis L. (Diptera: Oestridae) excretory/secretory antigens (OoES). In addition, the composition of each antigen was analysed using an electrophoresis system. Cephenemyia stimulator larvae were found in 25% of roe deer; the mean intensity of infection was 24.3 larvae per infested animal. In the antigen analysis, CsSA showed four exclusive bands of molecular weight (17-19, 62, 65 and 67-70 kDa). A positive correlation between immunoglobulin G (IgG) values and total number of larvae was found with CsES and CsSA. The highest sensitivity value, negative predictive value and negative likelihood ratio were obtained using CsES. The highest specificity value, positive likelihood ratio and kappa value were achieved with CsSA. The predictive values of the enzyme-linked immunosorbent assay (ELISA) using CsES and CsSA reached statistical significance and seroprevalence values were 26-44%. The use of ELISA with CsES and CsSA seems promising in the non-invasive diagnosis of Cephenemyia infestation in roe deer.

Gastrointestinal nematode infections in roe deer (Capreolus capreolus) from the NW of the Iberian Peninsula: assessment of some risk factors.

Intestinal contents of 218 roe deer hunted in the northwest (NW) of the Iberian Peninsula during the 2008-2009 hunting seasons were examined in order to provide information on the gastrointestinal (GI) nematode prevalence and intensity of infection and the possible influence of some environmental and intrinsic factors such as climatic conditions, age and sex. All the animals studied harboured GI nematodes, and a total of 20 different species belonging to ten genera were identified. Spiculopteragia spiculoptera/Spiculopteragia mathevossiani, Ostertagia leptospicularis/Ostertagia kolchida and Nematodirus filicollis were the most common. This is the first citation for Chabertia ovina, Cooperia pectinata, Cooperia punctata, Cooperia oncophora, Haemonchus contortus, Nematodirus spathiger, Oesophagostomum venulosum, Teladorsagia trifurcata, Trichostrongylus capricola, Trichostrongylus colubriformis, Trichostrongylus vitrinus and Trichuris capreoli in roe deer from the Iberian Peninsula. Prevalence and intensity were significantly higher in the abomasum, where infections with more than one GI nematode species were the most common; in the other intestinal segments infections with only one GI nematode species were the most prevalent. When considering the influence of the different risk factors on the prevalence of GI nematodes, the highest prevalence for most of the genera were observed in roe deer from coastal areas, where climatic conditions are more favourable for the development and survival of third stage larvae in the environment. Regarding the sex of the animals, the prevalence was, in general, higher in males than in females, probably due to behavioural and physiological sex-related differences. On the contrary, no differences were found in relation to the age of the animals. This study reveals that roe deer from the NW of the Iberian Peninsula are widely and intensely infected with gastrointestinal nematodes, which probably affect the health status of these ungulates.

Toxocara canis larvae viability after disinfectant-exposition.

The effect of three routinely used disinfectants on the embryonary development of Toxocara canis eggs was evaluated both in vivo and in vitro. In the in vitro experiment, T. canis eggs were treated with the ethanol, sodium hypochlorite, and one commercial mix of benzalconium chloride and formaldehyde, and the embryonary development was assessed. After a period of 24 days incubation, ethanol was the best disinfectant because it prevented the development of the T. canis larvae 2 in the eggs, and sodium hypochlorite caused degeneration in 50% eggs. By using the commercial mix, 25% T. canis eggs developed to 2nd stage larvae. In the in vivo experiment, the embryonated eggs treated with the disinfectants were inoculated to mice, and their brain tissues were examined for larval presence on the 24th day postinfection. In addition, a control group was set up for comparison with the infected groups. No injury or T. canis larvae were observed in mice infected with sodium hypochlorite-treated eggs, opposite to that recorded in the animals infected with the commercial disinfectant-treated eggs. These results showed that both ethanol and sodium hypochlorite are very appropriate because of their full efficacy against infective T. canis eggs. Disinfection of kennels, animal shelters, cages, and veterinary clinics with one of these products to eliminate T. canis eggs and to avoid contamination is strongly recommended.

Larval development of Neostrongylus linearis (Nematoda, Protostrongylidae) in the mollusc Cochlicella barbara infected and maintained in a subhumid area (north-west Spain) and its possible influence on the infection of small ruminants.

Twelve batches of 100 Cochlicella barbara (Mollusca, Helicidae) were deposited monthly, from January to December in plots (0.5 x 0.5 m) with faeces of sheep containing first-stage larvae of Neostrongylus linearis in order to be naturally infected, and another 12 batches acted as uninfected controls. Every 2 weeks dead molluscs were removed from the plots and three to five specimens were examined to study larval development in the snail. Infected and control snails did not differ in their mortality, which may be associated with the low rate of infection of snails. The highest number of molluscs that were infected and the maximum level of infection were observed in batches deposited in summer months. There was a positive correlation between the percentage of infected snails and temperature. Molluscs harboured higher number of infectious third-stage larvae (L3) from mid-spring to mid-autumn and especially in summer months; thus under these climatic conditions and periods small ruminants are more likely to get infected by N. linearis.

Parasitological analysis of Leonese royalty from Collegiate-Basilica of St. Isidoro, León (Spain): helminths, protozoa, and mites.

The royal burial chamber of what is today the Collegiate-Basilica of St. Isidoro in León, Spain, built and remodeled between the 10th and 13th centuries and in the 20th century renamed the Kings' Pantheon, has 13 royal tombs that were opened in the presence of the Abbot-Prior of the Collegiate to enable a group of researchers to obtain all possible information from the royal remains. Several samples were sent to the Parasitology Unit of the Animal Pathology (Animal Health) Department at the Veterinary Faculty of León (Spain). In all the tombs, eggs and remains of nonparasitic mites were observed. In a piece of linen cloth from the bottom of 1 tomb, an Anoplocephala perfoliata egg was found. Furthermore, 4 mummified bodies were found. In 2 of these, those belonging to Infantes María and Fernando, Ascaris lumbricoides eggs were found and in the latter Trichuris trichiura eggs. We have not found in the literature reviewed any records of studies of this kind carried out in Spain.

Efficacy of moxidectin 1% injectable and 0.2% oral drench against natural infection by Dictyocaulus filaria in sheep.

Two separate trials (I and II) with 34 and 32 Churra ewes, respectively, and distributed into two groups, have been carried out to evaluate the efficacy of two different formulations of moxidectin at a dose rate of 0.2mg/kg body weight (b.w.) against natural infection by Dictyocaulus filaria in sheep. Trial I was designed to evaluate a 1% moxidectin injectable formulation, whereas in trial II a 0.2% moxidectin oral drench formulation was used. The efficacy was measured on the basis of the reduction of the faecal larval counts and of adult worm recoveries at slaughter. In each trial, a group of animals was treated on day 0 with moxidectin 1% injectable or moxidectin 0.2% oral drench and the other group acted as untreated control. When the faecal larval counts was compared within the treated groups, the efficacy was over 95% until day +13, and 100% at the remainder of the sampling dates after the application of injectable moxidectin, whereas in trial II, the larvae per gram (lpg) of faeces increased until the first sampling time post treatment (p.t.), day +6, and zero counts were recorded for all animals by the following days. On the basis of adult worm recoveries at necropsy, the efficacy of the treatment was 100% in both trials, however, adult worms were detected at slaughter for all control sheep. These results indicate that moxidectin 1% injectable and moxidectin 0.2% oral drench, administered at 0.2mg/kg b.w., were 100% effective against D. filaria infection in sheep. No adverse reactions to the treatments were observed in the animals.

Efficacy of moxidectin 1% injectable against natural infection of Sarcoptes scabiei in sheep.

Thirty ewes naturally infected with Sarcoptes scabiei var. ovis, were allocated into three groups of 10 animals each. Animals in groups B and C were treated on day 0 and on days 0 and +10, respectively, with moxidectin 1% injectable at a dose of 0.2mg moxidectin/kg body weight (BW). Group A remained untreated. Seven days before treatment, the geometric mean of Sarcoptes scabiei var. ovis per square centimeter of skin in groups A, B and C were not significantly different. From the day of treatment to the end of the trial, the average number of mites/cm(2) increased in untreated animals and decreased in groups B and C, but these values were higher for group C. Active lesions produced by S. scabiei var. ovis consistently increased during the trial in the untreated animals; in group B the minimum count occurred on day +56 this reduction being more evident in group C (no lesions on days +49 and +56). Also in this group, the number of cured animals was 100%, therefore, the application of two treatments with moxidectin (group C) showed higher efficacy than a single treatment (group B). Body condition score decreased in the three experimental groups along the trial. All animals were individually weighed on days -1, +28 and at the end of the trial. No adverse reactions were observed in the animals treated with 0.2mg moxidectin/kg BW.

Bronchopulmonary helminths of roe deer (Capreolus capreolus) in the northwest of Spain.

From 1993 to 1995, 148 hunter killed roe deer (Capreolus capreolus L.) from game reserves in Lugo (northwestern Spain) were examined for lungworm infection. The overall prevalence of infection was high (66.2%). Infections were comprised to two species, Varestrongylus capreoli and Dictyocaulus noerneri with a prevalence of 62.0 and 18.2%, respectively. Significant differences in the prevalence and larval burdens in lungs and faecal samples were observed between different locations. The highest overall prevalence of lungworm infection was observed in the large reserve at Ancares (78%) in comparison to that in smaller reserves (61.7%). Considering that the species recorded in this study are highly host-specific, it is unlikely that roe deer represent a significant reservoir of lungworm transmissible to domestic ruminants in northwestern Spain.

Dynamics of the antibody response in cattle infested by Hypoderma (Diptera: Oestridae) after a treatment with injectable and pour-on ivermectin.

The dynamics of the anti-Hypoderma antibody response after a treatment with injectable and pour-on ivermectin against first instars of Hypoderma sp. were determined in naturally infested cows. Cattle were randomly assigned to three groups: group 1, which served as an untreated control; group 2, treated with pour-on ivermectin at a dose of 500 microg/kg of body weight, and group 3, which received 200 microg/kg of body weight of ivermectin by subcutaneous injection. Both ivermectins showed total efficacy against L-1 of Hypoderma sp. In untreated control animals, grubs became detectable in the 2 mo after treatment. Indirect enzyme-linked immunosorbent assay (ELISA) showed a significant increase in group 3 antibody levels 1 mo after treatment, whereas in group 2 the increase was not significant. Following the peak of anitibody activity in group 3, antibody levels declined at a faster rate than in those treated topically. In both treated groups, cattle remained serologically positive for 4 mo after ivermectin treatment. Thus, serological surveillance programs for cattle grubs conducted during this posttreatment period will not be influenced by treatment with ivermectin. The persistence of antibody levels after larval destruction makes the indirect ELISA unsuitable for the prediction of the efficacy of treatment.

Insulin and IGF-I receptors in trout adipose tissue are physiologically regulated by circulating hormone levels.

In fish, insulin is believed to act on adipose tissue to promote lipid accumulation, but a direct role for insulin in fish adipose tissue lipogenesis has yet to be demonstrated. To investigate the role of insulin and insulin-like growth factor I (IGF-I) in fish adipose tissue function, we have investigated the presence and the regulation of insulin and IGF-I receptors in adipose tissue of brown trout (Salmo trutta). Receptors for insulin and IGF-I were detected in trout adipose tissue, with IGF-I receptors being more abundant (two- to tenfold) and having a higher affinity (twofold) than insulin receptors. In contrast to the situation in mammals, arginine treatment, which elevates the levels of insulin and IGF-I in plasma, increased the number of insulin receptors 1.7-fold and the number of IGF-I receptors 2.3-fold. When plasma levels of insulin and IGF-I were decreased by fasting, insulin receptor numbers fell 3.6-fold and IGF-I receptor numbers fell 2.2-fold. These results demonstrate for the first time the presence of specific insulin and IGF-I receptors in adipose tissue of ectothermic vertebrates and suggest that adipose tissue may be a target for the actions of insulin and IGF-I in fish.

Regulation of plasma insulin-like growth factor-I levels in brown trout (Salmo trutta).

In this study we report that the use of a heterologous radioimmunoassay (RIA) is valid for the detection of insulin-like growth factor-I (IGF-I) levels in plasma of a variety of fish species. Parallelism between standard curves and plasma dilutions were observed and the standard curve obtained with mammalian IGF-I presented the same characteristics as that obtained with coho salmon recombinant IGF-I. The RIA was biologically validated since total plasma IGF-I values were significantly modified by different experimental conditions. Hyperinsulinemia induced either by arginine or insulin injection was accompanied by increases in IGF-I plasma levels in brown trout (Salmo trutta). In contrast, parallel decreases in insulin and IGF-I circulating levels were observed after 45 days of fasting and 20 days after a single streptozotocin injection. Administration of arginine in fasted fish led to a relative increase in insulin and IGF-I plasma concentrations, while arginine injection in fish previously treated with streptozotocin increased IGF-I levels only. The above data suggest that insulin, together with other factors, may act to increase the levels of IGF-I in plasma.

Insulin, insulin-like growth factor-I (IGF-I) and glucagon: the evolution of their receptors.

Insulin and glucagon, two of the most studied pancreatic hormones bind to specific membrane receptors to exert their biological actions. Insulin-like growth factors IGF-I and IGF-II are structurally related to insulin, although they are expressed ubiquitously. The biological functions of the IGFs are mediated by different transmembrane receptors, which includes the insulin, IGF-I and IGF-II receptors. The interaction of insulin, insulin related peptides and glucagon with the corresponding receptors has been studied extensively in mammals and continues to be so. At the same time, research on ectothermic animals has made enormous progress in the recent years. This paper summarizes current knowledge on insulin, IGF-I and glucagon receptors, from a comparative point of view with special attention to non-mammalian vertebrates. The review covers adult and mostly typical target tissues, and with very few exceptions, developmental aspects are not considered. Binding characteristics, tissue distribution and structure of insulin and IGF-I receptors will be considered first, because both ligands and receptors are structurally related and have overlapping functions. These sections will be followed by similar distribution of information on glucagon receptors. Readers interested in either structure or functions of insulin, IGFs and glucagon in nonmammalian vertebrates are referred to other reviews (Mommsen TP, Plisetskaya EM. Insulin in fishes and agnathans: history, structure and metabolic regulation. Rev Aquat Sci 1991;4:225-259; Mommsen TP, Plisetskaya EM. Metabolic and endocrine functions of glucagon-like peptides: evolutionary and biochemical perspectives. Fish Physiol Biochem 1993;11:429-438; Duguay SJ, Mommsen TP. Molecular aspects of pancreatic peptides. In: Sherwood NM, Hew CL, editors, Fish Physiology. vol 13. 1994:225-271; Plisetskaya EM, Mommsen TP. Glucagon and glucagon-like peptides in fishes. Int Rev Citol 1996;168:187-257.).

Influence of high-carbohydrate enriched diets on plasma insulin levels and insulin and IGF-I receptors in trout.

Insulin and insulin growth factor-I (IGF-I) binding to skeletal muscle semipurified receptors were assessed in rainbow trout (Oncorhynchus mykiss) fed with different enriched carbohydrate diets. The animals were fed for 2 months, either in spring, summer or autumn with a control diet (C, commercial diet containing 21% raw carbohydrates) or with two diets supplied with highly digestible carbohydrates (E1, 22% expanded wheat; and E2, 37% expanded wheat). Insulin and IGF-I receptors were semipurified by affinity chromatography (WGA-agarose). Fish fed with a carbohydrate enriched diet did not show lower growth rates than those fed with the control diet. Independently of the season, rainbow trout fed E1 and E2 presented higher insulin and glucose plasma levels as well as higher tissue glycogen reserves than fish fed C. An increase in the number of insulin receptors during the diet adaptation was observed especially in fish fed with E2. No differences in the affinity of receptors were observed. IGF-I specific binding in skeletal muscle was higher than that of insulin in all groups and in all seasons. Furthermore, IGF-I receptors showed the same tendency as insulin receptors, with increases in their number in experimentally fed fish, especially those fed with E2. Insulin and IGF-I receptors TKA increased only slightly, as a consequence of E1 and E2 diet adaptation. In conclusion, rainbow trout can be fed high-carbohydrate levels and show good rates of growth. This adaptation determines increases in circulating glucose and insulin, and muscle insulin receptors, which indicate an adaptation of the fish to higher levels of glucose supply. The response of IGF-I receptors also suggests a possible role in the regulation of metabolism.

Appearance of insulin and insulin-like growth factor-I (IGF-I) receptors throughout the ontogeny of brown trout (Salmo trutta fario).

Insulin and IGF-I receptors were characterized in glycoprotein fractions prepared by affinity chromatography from different developmental stages of brown trout. The specificity of insulin and IGF-I binding was demonstrated by crossed-competition assays: unlabelled insulin displaced bound radiolabelled insulin at concentrations 45-fold lower than unlabelled IGF-I, whilst unlabelled IGF-I displaced bound radiolabelled IGF-I at concentrations 2,000-fold lower than unlabelled insulin. The affinity of these receptors did not change significantly during trout development. Insulin-specific binding was detectable 3 weeks after spawning, after which it increased to a maximum in fry weighing 0.4 g, and decreased progressively to adult levels. IGF-I specific binding was detectable in newly laid eggs and increased to a maximum during organogenesis in eyed eggs. It then decreased progressively during subsequent stages of development to adult levels. The apparent molecular weight (Mr) of the alpha-subunit of brown trout insulin and IGF-I receptors was smaller than that of the alpha-subunit of the rat insulin receptor. Receptor tyrosine kinase activity was stimulated in a dose-dependent manner by insulin and IGF-I. Insulin and IGF-I stimulated tyrosine kinase activity and reached a maximum of 201 +/- 17.6 and 240 +/- 29.6% of basal phosphorylation, respectively.

Insulin and insulin-like growth factor-I (IGF-I) binding in fish red muscle: regulation by high insulin levels.

Insulin and IGF-I binding to semi-purified red muscle receptors was characterized in brown trout, Salmo trutta and the common carp. Cyprinus carpio. The yield of glycoprotein obtained after semipurification of receptors with WGA-agarose affinity chromatography in microgram g-1 initial tissue was 210.6 +/- 21 micrograms g-1 in trout and 108.5 +/- 2.5 micrograms g-1 in carp. IGF-I specific binding (4.72 +/- 0.64%/10 micrograms glycoprotein) was 4-5-times higher than insulin binding (1.04 +/- 0.12%/10 micrograms glycoprotein) in trout red muscle. This difference in binding was due to a higher number and a greater affinity of the IGF-I (Kd, 0.21 +/- 0.03 nM) compared with the insulin (Kd, 0.67 +/- 0.06 nM) receptors in this tissue. Carp red muscle IGF-I binding (9.14 +/- 0.55%/10 micrograms glycoprotein) surpassed insulin binding (2.59 +/- 0.094%/10 micrograms glycoprotein) mainly because of a greater affinity of the IGF-I (Kd, 0.092 +/- 0.027 nM) compared with the insulin (Kd, 0.1515 +/- 0.0285 nM) receptor. IGF-I and insulin binding in carp red muscle were higher than in trout, as a consequence of a higher affinity of carp red muscle receptors. Arginine injection provoked acute hyperinsulinemia in both trout (23.3 +/- 1.01 ng ml-1) and carp (24.3 +/- 1.34 ng ml-1. Specific binding of insulin and IGF-I to the red muscle decreased 4 h after injection. In trout, a decrease of insulin and IGF-I binding of 47.0% and 63.3%, respectively was observed compared with controls, in carp, these values were 44.0% and 45.0%. The number of insulin and IGF-I receptors decreased (42-55%) but affinities did not change suggesting that receptor down-regulation is a consequence of high insulin levels.