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Background: Research shows that only around half of all survivors of child sexual abuse (CSA) disclose the abuse during childhood and adolescence. This is worrying, as CSA is related to substantial suffering later in life. The proportion of children and adolescents who have been exposed to CSA is significantly higher in Child and Adolescent Psychiatry (CAP) than in the general population. Healthcare professionals report that uncovering CSA is a complex and challenging task. However, we know little about how they proceed when uncovering CSA. More knowledge of healthcare personnel's experience is therefore necessary to facilitate and increase CSA disclosure. The study aims to explore how CAP healthcare professionals in Norway proceed when assessing and detecting CSA, how they experience this work, and what hinders or facilitates their efforts., Methods: The study employed a mixed method approach. Data was collected through an anonymous online survey, generating both quantitative and qualitative data. The sample consisted of 111 healthcare professionals in CAP, of whom 84% were women, with a mean age of 40.7 years (range 24-72; sd = 10.8). Mean years of CAP clinical experience were 8.3 years (range 0-41; sd = 7.5). The quantitative data was analysed using descriptive statistics, correlations, and independent sample t-tests, while the qualitative data was analysed using a team-based qualitative content analysis., Results: The results showed that detection of CSA was viewed as an important, but complex task in CAP, and the existing procedures were deemed to be insufficient. The therapists mostly felt confident about how to proceed when they suspected or detected CSA, yet they seldom detected CSA. In their initial assessment they applied standardised procedures, but if their suspicion of possible CSA persisted, they seemed to rely more on clinical judgement. Specific challenges and facilitators for CSA detection were identified, both in the individual and in the organisation., Conclusions: The study highlights the challenges and complexities healthcare professionals and the CAP system face when assessing CSA, which may account for the low detection rate. The results show that healthcare professionals believe room for clinical autonomy and targeted competence development may improve CSA detection. Additionally, the findings suggest a need for CAP to define roles and responsibilities within and between agencies., (© 2024. The Author(s).)

The polychaete Capitella is a typical member of the 'thiobiome', and is commonly used as an eutrophication indicator species in environmental assessment studies. To deal with a sulfide-rich and poisonous surrounding, cells in close contact with the environment, and thus able to play a major role in detoxication and survival, are circulating cells. This work aimed to morpho-functionally describe the circulating coelomic cells of Capitella from the English Channel inhabiting the sulfide-rich mud in Roscoff Harbor. In general, worms have three types of circulating cells, granulocytes involved in bacterial clearance and defense against microorganisms, eleocytes with an essentially trophic role and elimination of cellular waste, and erythrocytes which play a role in detoxification and respiration via their intracellular hemoglobin. By combining diverse microscopic and cellular approaches, we provide evidence that Capitella does not possess granulocytes and eleocytes, but rather a single abundant rounded cell type with the morphological characteristics of erythrocytes i.e. small size and production of intracellular hemoglobin. Surprisingly, our data show that in addition to their respiratory function, these red cells could exert phagocytic activities, and produce an antimicrobial peptide. This latter immune role is usually supported by granulocytes. Our data highlight that the erythrocytes of Capitella from the English Channel differ in morphology and bear more functions than the erythrocytes of other annelids. The simplicity of this multi-task (or polyvalent) single-cell type makes Capitella an interesting model for studies of the impact of the environment on the immunity of this bioindicator species., Competing Interests: Declarations of competing interest The authors declare no potential conflicts of interests with respect to the research, authorship or publication of this article., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Adolescents with ADHD often struggle on many areas of their lives and have a high risk of adverse outcomes and negative life trajectories. Multimodal treatment including psychosocial interventions is recommended but evidence regarding effect of such interventions is still limited., Materials and Methods: This study was a follow-up study of adolescents participating in a randomised controlled trial (RCT) of a group intervention based on cognitive behavioural therapy (CBT). Participants were adolescents diagnosed with ADHD and still impaired by their symptoms after standard treatment including psychoeducation and medication. All participants were interviewed by telephone one year after inclusion, and outcome measures included both quantitative and qualitative measures., Results: There were 100 adolescents included in the study. We found no significant differences between treatment and control group on measures of ADHD-symptoms, self-efficacy, overall problems, global psychosocial functioning, or symptom severity at one-year follow-up. Still, participants in the intervention group reported on positive gains and that they learned a lot about ADHD and themselves., Conclusions: The intervention delivered in this trial failed to show a treatment effect on symptom level when added to standard care. Participants did however report on positive gains and felt they learned a lot. More research is needed to explore how the programme and delivery of treatment might be improved, and which patients might benefit the most from this type of interventions.

Executive functional deficits (EFDs) play an important role in functional impairment in adolescents with attention deficit/hyperactivity disorder (ADHD). More knowledge of executive function (EF) profiles and informant discrepancies will guide clinicians and provide tailored treatment advice. The objectives of this study were to use teacher, parent, and self-reported EF ratings to describe (a) problem profiles and (b) the correlation and agreement between informants. This study included 100 adolescents aged 14-18 years with ADHD still experiencing clinically impairing symptoms despite standard treatment including medication. EFs were measured using the Behavior Rating Inventory of Executive Function (BRIEF). Agreement between informants was quantified using Pearson correlation and informant discrepancies were analyzed using paired samples t-test. Overall, the results indicated considerable EFDs in the study population. Correlation and agreement varied between the informants. Agreement was highest between adolescents and their parents, especially for female participants, and lowest between male participants and their teachers. Teachers reported the highest level of EFDs, whereas adolescents generally self-reported EFDs at a lower level than both parents and teachers. Identifying and tailoring treatment for EFDs might improve future prognosis for adolescents with ADHD, however, self-awareness of these difficulties is a challenge that needs to be considered when planning interventions.

Background: High latitude seeps are dominated by Oligobrachia siboglinid worms. Since these worms are often the sole chemosymbiotrophic taxon present (they host chemosynthetic bacteria within the trophosome organ in their trunk region), a key question in the study of high latitude seep ecology has been whether they harbor methanotrophic symbionts. This debate has manifested due to the mismatch between stable carbon isotope signatures of the worms (lower than -50‰ and usually indicative of methanotrophic symbioses) and the lack of molecular or microscopic evidence for methanotrophic symbionts. Two hypotheses have circulated to explain this paradox: (1) the uptake of sediment carbon compounds with depleted δC 13 values from the seep environment, and (2) a small, but significant and difficult to detect population of methanotrophic symbionts. We conducted 16S rRNA amplicon sequencing of the V3-V4 regions on two species of northern seep Oligobrachia (Oligobrachia webbi and Oligobrachia sp. CPL-clade), from four different high latitude sites, to investigate the latter hypothesis. We also visually checked the worms' symbiotic bacteria within the symbiont-hosting organ, the trophosome, through transmission electron microscopy., Results: The vast majority of the obtained reads corresponded to sulfide-oxidizers and only a very small proportion of the reads pertained to methane-oxidizers, which suggests a lack of methanotrophic symbionts. A number of sulfur oxidizing bacterial strains were recovered from the different worms, however, host individuals tended to possess a single strain, or sometimes two closely-related strains. However, strains did not correspond specifically with either of the two Oligobrachia species we investigated. Water depth could play a role in determining local sediment bacterial communities that were opportunistically taken up by the worms. Bacteria were abundant in non-trophosome (and thereby symbiont-free) tissue and are likely epibiotic or tube bacterial communities., Conclusions: The absence of methanotrophic bacterial sequences in the trophosome of Arctic and north Atlantic seep Oligobrachia likely indicates a lack of methanotrophic symbionts in these worms, which suggests that nutrition is sulfur-based. This is turn implies that sediment carbon uptake is responsible for the low δ 13 C values of these animals. Furthermore, endosymbiotic partners could be locally determined, and possibly only represent a fraction of all bacterial sequences obtained from tissues of these (and other) species of frenulates., (© 2023. The Author(s).)

Background: A goal has been set to establish paediatric high dependency units (PHDUs) in Norwegian hospitals. We sought to describe the patient population in one such unit, and to investigate whether the need for admission to an intensive care unit (ICU) changed after the unit was established., Material and Method: Information was obtained from electronic patient records, patient administration systems and the quality register for the PHDU at the Department of Paediatrics and Adolescent Medicine at Haukeland University Hospital. All patients admitted to the PHDU within five years of its opening in 2017 were included. A comparison was made between ICU patients in the five years before and the five years after the establishment of the PHDU., Results: The PHDU had a total of 851 patient admissions in the period 2017-2021, increasing from 125 in 2017 to 247 in 2021. This accounted for 3.5 % and 7.6 % of the total number of patients admitted to the Department of Paediatrics and Adolescent Medicine in these years, respectively. The ICU had 185 paediatric patient admissions in the period 2012-2016 and 187 in the period 2017-2021, which constituted 0.9 % and 1.1 % of all patients admitted to the Department of Paediatrics and Adolescent Medicine during the periods, respectively. After the start-up of the PHDU, a lower proportion of patients were admitted to the ICU in the diagnostic groups diabetic ketoacidosis (15 % in 2017-2021 versus 20 % in 2012-2016) and sepsis (12 % in 2017-2021 versus 19 % in 2012-2016)., Interpretation: The introduction of a PHDU was not associated with a simultaneous reduction in the number of ICU admissions overall. For diabetic ketoacidosis and sepsis, however, the proportion of transfers to the ICU was reduced.

Background: Efforts are directed both towards prevention and early detection of Child sexual abuse (CSA). Yet, only about 50% of CSA survivors disclose before adulthood, and health professionals rarely are the first disclosure recipients. Increasing the detection rate of CSA within the context of Child and Adolescent Psychiatry (CAP) therefore represents a significant secondary prevention strategy. However, facilitating CSA disclosure when the survivor is reluctant to tell is a highly complex and emotionally demanding clinical task. We therefore argue that efforts to increase detection rates of CSA within CAP need to rest on knowledge of how both survivors and health professionals experience addressing CSA., Method: Using meta-ethnography as method, we present separate sub-syntheses as well an overarching joint synthesis of how survivors and health professionals experience addressing CSA., Results: Results show how both survivors and health professionals facing CSA disclosure feel deeply isolated, they experience the consequences of addressing CSA as highly unpredictable, and they need support from others to counteract the negative impact of CSA., Conclusion: The results indicate that adapting the organization of CAP to knowledge of how the survivors and health professionals experience addressing CSA is critical to facilitate earlier disclosure of CSA within CAP.

The European flat oyster ( Ostrea edulis L.) is a native bivalve of the European coasts. Harvest of this species has declined during the last decades because of the appearance of two parasites that have led to the collapse of the stocks and the loss of the natural oyster beds. O. edulis has been the subject of numerous studies in population genetics and on the detection of the parasites Bonamia ostreae and Marteilia refringens. These studies investigated immune responses to these parasites at the molecular and cellular levels. Several genetic improvement programs have been initiated especially for parasite resistance. Within the framework of a European project (PERLE 2) that aims to produce genetic lines of O. edulis with hardiness traits (growth, survival, resistance) for the purpose of repopulating natural oyster beds in Brittany and reviving the culture of this species in the foreshore, obtaining a reference genome becomes essential as done recently in many bivalve species of aquaculture interest. Here, we present a chromosome-level genome assembly and annotation for the European flat oyster, generated by combining PacBio, Illumina, 10X linked, and Hi-C sequencing. The finished assembly is 887.2 Mb with a scaffold-N50 of 97.1 Mb scaffolded on the expected 10 pseudochromosomes. Annotation of the genome revealed the presence of 35,962 protein-coding genes. We analyzed in detail the transposable element (TE) diversity in the flat oyster genome, highlighted some specificities in tRNA and miRNA composition, and provided the first insight into the molecular response of O. edulis to M. refringens . This genome provides a reference for genomic studies on O. edulis to better understand its basic physiology and as a useful resource for genetic breeding in support of aquaculture and natural reef restoration., Competing Interests: The authors declare no conflicts of interest., (© 2022 The Authors. Evolutionary Applications published by John Wiley & Sons Ltd.)

Background: Adolescents with ADHD are at increased risk of adverse outcomes and a negative life trajectory into adulthood. Evidence regarding treatment specifically tailored for the needs of this age group are still limited. High dropout rates, discontinuation of medication and treatment resistance are common issues in this population, and the patient perspective on new treatment options is therefore important. In this study, we aimed to investigate treatment satisfaction and feasibility of a group CBT program for adolescents with ADHD. We further aimed to identify any baseline characteristics predicting satisfaction., Materials and Methods: This study was part of a larger RCT of group CBT as add-on treatment for adolescents aged 14-18 years (Mean age 15.9 years, SD 1.3) with ADHD in Norway. Satisfaction and feasibility in the treatment group ( n  = 48) were measured by completion of an evaluation questionnaire, attendance of group sessions and a group-leaders checklist. Predictors of satisfaction were analysed using linear regression., Results: Overall satisfaction was very high with a significant age effect, the eldest participants being most satisfied. Attendance rate was high with few dropouts and medical adherence during the treatment period was good. Group-leaders generally self-evaluated adherence to treatment manual positively but addressing resistance towards homework as challenging., Conclusions: The participants were very satisfied with the group CBT treatment. Treatment options that are accepted and well-liked by the targeted population have the potential of reducing resistance towards treatment, improving future health and adherence to medication. The program is considered suitable for a clinical setting and may represent a feasible treatment supplement for adolescent ADHD.

Half of all of the elements in the Universe that are heavier than iron were created by rapid neutron capture. The theory underlying this astrophysical r-process was worked out six decades ago, and requires an enormous neutron flux to make the bulk of the elements 1 . Where this happens is still debated 2 . A key piece of evidence would be the discovery of freshly synthesized r-process elements in an astrophysical site. Existing models 3-5 and circumstantial evidence 6 point to neutron-star mergers as a probable r-process site; the optical/infrared transient known as a 'kilonova' that emerges in the days after a merger is a likely place to detect the spectral signatures of newly created neutron-capture elements 7-9 . The kilonova AT2017gfo-which was found following the discovery of the neutron-star merger GW170817 by gravitational-wave detectors 10 -was the first kilonova for which detailed spectra were recorded. When these spectra were first reported 11,12 , it was argued that they were broadly consistent with an outflow of radioactive heavy elements; however, there was no robust identification of any one element. Here we report the identification of the neutron-capture element strontium in a reanalysis of these spectra. The detection of a neutron-capture element associated with the collision of two extreme-density stars establishes the origin of r-process elements in neutron-star mergers, and shows that neutron stars are made of neutron-rich matter 13 .

Symbiosis between Bathymodiolus and Gammaproteobacteria allows these deep-sea mussels to live in toxic environments such as hydrothermal vents and cold seeps. The quantity of endosymbionts within the gill-bacteriocytes appears to vary according to the hosts environment; however, the mechanisms of endosymbiont population size regulation remain obscure. We investigated the possibility of a control of endosymbiont density by apoptosis, a programmed cell death, in three mussel species. Fluorometric TUNEL and active Caspase-3-targeting antibodies were used to visualize and quantify apoptotic cells in mussel gills. To control for potential artefacts due to depressurization upon specimen recovery from the deep-sea, the apoptotic rates between mussels recovered unpressurised, versus mussels recovered in a pressure-maintaining device, were compared in two species from hydrothermal vents on the Mid-Atlantic Ridge: Bathymodiolus azoricus and B. puteoserpentis. Results show that pressurized recovery had no significant effect on the apoptotic rate in the gill filaments. Apoptotic levels were highest in the ciliated zone and in the circulating hemocytes, compared to the bacteriocyte zone. Apoptotic gill-cells in B. aff. boomerang from cold seeps off the Gulf of Guinea show similar distribution patterns. Deep-sea symbiotic mussels have much higher rates of apoptosis in their gills than the coastal mussel Mytilus edulis, which lacks chemolithoautotrophic symbionts. We discuss how apoptosis might be one of the mechanisms that contribute to the adaptation of deep-sea mussels to toxic environments and/or to symbiosis., Competing Interests: The authors have declared that no competing interests exist.

We provide the first detailed identification of Barents Sea cold seep frenulate hosts and their symbionts. Mitochondrial COI sequence analysis, in combination with detailed morphological investigations through both light and electron microscopy was used for identifying frenulate hosts, and comparing them to Oligobrachia haakonmosbiensis and Oligobrachia webbi, two morphologically similar species known from the Norwegian Sea. Specimens from sites previously assumed to host O. haakonmosbiensis were included in our molecular analysis, which allowed us to provide new insight on the debate regarding species identity of these Oligobrachia worms. Our results indicate that high Arctic seeps are inhabited by a species that though closely related to Oligobrachia haakonmosbiensis, is nonetheless distinct. We refer to this group as the Oligobrachia sp. CPL-clade, based on the colloquial names of the sites they are currently known to inhabit. Since members of the Oligobrachia sp. CPL-clade cannot be distinguished from O. haakonmosbiensis or O. webbi based on morphology, we suggest that a complex of cryptic Oligobrachia species inhabit seeps in the Norwegian Sea and the Arctic. The symbionts of the Oligobrachia sp. CPL-clade were also found to be closely related to O. haakonmosbiensis symbionts, but genetically distinct. Fluorescent in situ hybridization and transmission electron micrographs revealed extremely dense populations of bacteria within the trophosome of members of the Oligobrachia sp. CPL-clade, which is unusual for frenulates. Bacterial genes for sulfur oxidation were detected and small rod shaped bacteria (round in cross section), typical of siboglinid-associated sulfur-oxidizing bacteria, were seen on electron micrographs of trophosome bacteriocytes, suggesting that sulfide constitutes the main energy source. We hypothesize that specific, local geochemical conditions, in particular, high sulfide fluxes and concentrations could account for the unusually high symbiont densities in members of the Oligrobrachia sp. CPL-clade., Competing Interests: The authors have declared that no competing interests exist.

Vesicomyid bivalves form dense clam beds in both deep-sea cold seeps and hydrothermal vents. The species diversity within this family raises questions about niche separation and specific adaptations. To compare their abilities to withstand hypoxia, we have studied the structure and function of erythrocyte hemoglobin (Hb) and foot myoglobin (Mb) from two vesicomyid species, Christineconcha regab and Laubiericoncha chuni, collected from the Regab pockmark in the Gulf of Guinea at a depth of 3,000 m. Laubiericoncha chuni possesses three monomeric globins, G1 (15,361 Da), G2 (15,668 Da), and G3 (15,682 Da) in circulating erythrocytes (Hb), and also three globins, G1, G3, and G4 (14,786 Da) in foot muscle (Mb). Therefore, globins G2 and G4 appear to be specific for erythrocytes and muscle, respectively, but globins G1 and G3 are common. In contrast, C. regab lacks erythrocyte Hb completely and possesses only globin monomers G1' (14,941 Da), G2' (15,169 Da), and G3' (15,683 Da) in foot muscle. Thus, these two vesicomyid species, C. regab and L. chuni, show a remarkable diversity in globin expression when examined by electrospray ionization mass spectrometry. Oxygen-binding affinities reveal extremely high oxygen affinities (P50 < 1 Torr, from 5° to 15°C at pH 7.5), in particular L. chuni globins, which might be an advantage allowing L. chuni to dig deeply for sulfides and remain buried for long periods in reduced sediments.

Siboglinids are symbiotic polychete annelids having hemoglobins as essential oxygen- and sulfide-carriers for their endosymbiotic bacteria. We analyzed the structure of the hemoglobins from two species of siboglinids: the monilifera Sclerolinum contortum and the frenulata Oligobrachia webbi (i.e. haakonmosbiensis) from Norwegian cold seeps. Measured by Multi-Angle Laser Light Scattering (MALLS), Sclerolinum shows a 3190+/-50 kDa hexagonal bilayer hemoglobin (HBL-Hb) and a 461+/-46 kDa ring-Hb, just as vestimentifera, whereas Oligobrachia has a 409+/-3.7 kDa ring-Hb only. Electrospray Ionization-Mass Spectrometry (ESI-MS) showed Sclerolinum HBL-Hb composed of seven monomeric globins (15-16 kDa), three disulfide-bonded globin heterodimers and three linkers. The heterodimers always contain globin-b (15814.4+/-1.5 Da). Sclerolinum ring-Hb is composed of globins and dimers with identical masses as its HBL-Hb, but lacks linkers. Oligobrachia ring-Hb has three globin monomers (14-15 kDa) only, with no disulfide-bonded dimers. Comparison of Sclerolinum hemoglobins between Storegga and Haakon Mosby Mud Volcano, using the normalized height of deconvoluted ESI-MS peaks, shows differences in globin monomers abundances that could reflect genetic differences or differential gene expression between distinct seep populations. The discovery of HBL-Hb in Sclerolinum is a new element supporting the hypothesis of monilifera being phylogenetically more closely related to vestimentifera, than to frenulata.

Peroxiredoxins (PRDXs) are a superfamily of thiol-dependent peroxidases found in all phyla. PRDXs are mechanistically divided into three subfamilies, namely typical 2-Cys, atypical 2-Cys, and 1-Cys PRDXs. To reduce peroxides, the N-terminal peroxidatic Cys of PRDXs is first oxidized into sulfenic acid. This intermediate is reduced by forming a disulfide bond either with a resolving Cys of another monomeric entity (typical 2-Cys) or of the same molecule (atypical 2-Cys). In 1-Cys PRDXs, the resolving Cys is missing and the sulfenic acid of the peroxidatic Cys is reduced by a heterologous thiol-containing reductant. In search of a homolog of human 1-Cys PRDX6 in Arenicola marina, an annelid worm living in intertidal sediments, we have cloned and characterized a PRDX exhibiting high sequence homology with its mammalian counterpart. However, A. marina PRDX6 possesses five Cys among which two Cys function as peroxidatic and resolving Cys of typical 2-Cys PRDXs. Thus, A. marina PRDX6 belongs to a transient group exhibiting sequence homologies with mammalian 1-Cys PRDX6 but must be mechanistically classified into typical 2-Cys PRDXs. Moreover, PRDX6 is highly expressed in tissues directly exposed to the external environment, suggesting that this PRDX may be of particular importance for protection against exogenous oxidative attacks.

The vestimentiferan annelid Riftia pachyptila forms dense populations at hydrothermal vents along the East Pacific Rise at a depth of 2600 m. It harbors CO(2)-assimilating sulfide-oxidizing bacteria that provide all of its nutrition. To find specific host transcripts that could be important for the functioning of this symbiosis, we used a subtractive suppression hybridization approach to identify plume- or trophosome-specific proteins. We demonstrated the existence of carbonic anhydrase transcripts, a protein endowed with an essential role in generating the influx of CO(2) required by the symbionts. One of the transcripts was previously known and sequenced. Our quantification analyses showed a higher expression of this transcript in the trophosome compared to the branchial plume or the body wall. A second transcript, with 69.7% nucleotide identity compared to the previous one, was almost only expressed in the branchial plume. Fluorescent in situ hybridization confirmed the coexpression of the two transcripts in the branchial plume in contrast with the trophosome where only one transcript could be detected. An alignment of these translated carbonic anhydrase cDNAs with vertebrate and nonvertebrate carbonic anhydrase protein sequences revealed the conservation of most amino acids involved in the catalytic site. According to the phylogenetic analyses, the two R. pachyptila transcripts clustered together but not all nonvertebrate sequences grouped together. Complete sequencing of the new carbonic anhydrase transcript revealed the existence of two slightly divergent isoforms probably coded by two different genes.

A fully self-contained model of homochirality is presented that contains the effects of both polymerization and dissociation. The dissociation fragments are assumed to replenish the substrate from which new monomers can grow and undergo new polymerization. The mean length of isotactic polymers is found to grow slowly with the normalized total number of corresponding building blocks. Alternatively, if one assumes that the dissociation fragments themselves can polymerize further, then this corresponds to a strong source of short polymers, and an unrealistically short average length of only 3. By contrast, without dissociation, isotactic polymers becomes infinitely long.

The stability and conservation properties of a recently proposed polymerization model are studied. The achiral (racemic) solution is linearly unstable once the relevant control parameter (here the fidelity of the catalyst) exceeds a critical value. The growth rate is calculated for different fidelity parameters and cross-inhibition rates. A chirality parameter is defined and shown to be conserved by the nonlinear terms of the model. Finally, a truncated version of the model is used to derive a set of two ordinary differential equations and it is argued that these equations are more realistic than those used in earlier models of that form.

The symbiotic tubeworm Riftia pachyptila needs to fuel its chemoautotrophic symbiotic bacteria with inorganic carbon. CO(2) is transported from the surrounding water to the bacteriocytes located in the trophosome, through the branchial plume and the body fluids. Previous studies have demonstrated the implication of carbonic anhydrase (CA) and proton pumps (ATPases) at various steps of CO(2) transport. The present study describes the expression pattern of cytosolic CA using an RNA probe and its histochemical and immunocytochemical localization in the trophosome and branchial plume of RIFTIA: Immunolocalization of V-H(+)ATPase and Na(+)K(+)-ATPase were also performed and related to CA localization. In the branchial plume, CA is expressed and localized in the most apical region of the branchial epithelium, close to the surrounding water. V-H(+)ATPase is mostly colocalized with CA and both enzymes probably allow CO(2) entry against the concentration gradient while regulating intracellular pH. Na(+)K(+)-ATPase is mostly restricted to the basal part of epithelial cells and probably participates in CO(2) transport to the body fluids. In the trophosome lobules, cytosolic CA is expressed and found in bacteriocytes and peritoneal cells. Hypotheses on the role of CA in bicarbonate and CO(2) interconversion to fuel the symbiotic bacteria are discussed.

We have used an antibody specific for phosphotyrosine to investigate protein phosphorylation on tyrosine during hormone-induced maturation of starfish oocytes. Analysis of immunoprecipitates from cortices of in vivo labeled Marthasterias glacialis oocytes revealed the presence of labeled phosphotyrosine-containing proteins only after hormone addition. Six major phosphoproteins of 195, 155, 100, 85, 45, and 35 kDa were detected. Total activity in immunoprecipitates increased until first polar body emission and was greatly reduced upon completion of meiosis but some proteins exhibited different kinetics. The labeling of the 155-kDa protein reached a maximum at germinal vesicle breakdown, while the 35-kDa appeared later and disappeared after polar body emission. Similar results were obtained with Asterias rubens oocytes. In vitro phosphorylation of cortices showed that tyrosine kinase activity is a major protein kinase activity in this fraction, the main endogenous substrate being a 68-kDa protein. The proteins phosphorylated on tyrosine in vitro were almost similar in extracts from oocytes treated or not with the hormone.

The distribution of melanin-concentrating hormone (MCH) in the central nervous system of the frog Rana ridibunda was determined by the indirect immunofluorescence technique using antibodies against synthetic salmon MCH, generated in rabbits. The most prominent group of MCH-like containing perikarya was detected in the preoptic nucleus. Comparatively, a moderate number of cell bodies was observed in the dorsal infundibular nucleus and in the ventral thalamic area. Brightly immunofluorescent nerve bundles were found in the preoptic nucleus and in the ventral infundibular nucleus, coursing towards the internal zone of the median eminence and the pituitary stalk. An intense network of immunofluorescent fibers was localized in the neural lobe of the pituitary. The subcellular localization of MCH-like material was studied in the neurohypophysis using the immunogold technique. It was demonstrated that MCH-like material was contained in dense core vesicles (80-90 mm in diameter) within specific nerve terminals. The present findings indicate that, in amphibians, MCH-like peptide is located in specific hypothalamic neurons. Our data suggest that MCH may be released by neurohypophyseal nerve endings as a typical neurohormone.

By means of the peroxidase-anti-peroxidase technique, the distribution of neuropeptide tyrosine (NPY) and its C-terminal flanking peptide (C-PON) has been studied on serial sections of the brain and pituitary of the frog Rana ridibunda. Throughout the brain, NPY and C-PON-immunoreactive perikarya exhibited a remarkable co-distribution. These two peptides were found to be co-located within the same cell bodies in various brain regions including the dorsal and ventral pallium, the dorsal and ventral infundibular nuclei and the preoptic nucleus. The distribution of NPY- and C-PON-containing fibers in the brain and pituitary was similar. Sequential double immunohistochemical staining using the indirect immunofluorescence method, showed that NPY and C-PON were actually located within the same nerve processes throughout the frog brain and in the intermediate lobe of the pituitary. These studies indicate that the deduced C-PON sequence is present within the frog precursor to NPY and is formed in vivo in the frog brain. Like NPY, C-PON is transported distally in nerve terminals and is likely released with NPY in various regions of the brain and in the intermediate lobe of the pituitary.

Five antisera against insulin (Ins), glucagon (Glu), somatostatin (SRIF), met-enkephalin (met-enk), and serotonin (5-HT) were used for immunofluorescence detection of endocrine cells in pancreas and gastrointestinal tract (GIT) of the European eel (Anguilla anguilla L.) at three stages of development (leptocephalic larva, glass-eel, and adult eel). Comparable distribution of endocrine cells was observed for adults and glass-eels. In their pancreatic islets, positive immunoreactions were obtained only for Ins, SRIF, and Glu; this later was also present in the pancreatic ducts. 5-HT cells were present throughout the GIT. SRIF cells were situated mostly in the stomach and less in the intestine. Met-enk cells were abundant in the pyloric cecum, but less frequent in the intestinal mucosa. Glu cells were present only in the intestine. No insulin-immunoreactive cells could be detected in the GIT. The pancreatic islets of leptocephalic larvae exhibited a strong reaction for SRIF, a weak reaction for Glu, and none at all for Ins, met-Enk, or 5-HT. The GIT of these larvae contained numerous met-enk cells, mainly in the foregut. In the fore- and midgut, cells exhibited a weak fluorescence after treatment with Glu antiserum. No positive immunoreactive cells were observed with 5-HT, SRIF, or Ins antisera.

Gonadotropin-releasing-hormone (GnRH)-associated peptide (GnRH-AP) is a 56 amino acid neuropeptide derived from the GnRH prohormone. GnRH-AP corresponds to the C-terminal fragment flanking the GnRH peptide. Using an antiserum raised against human GnRH-AP [1-56], or against human GnRH, we have investigated the neuronal systems containing either peptide in the central nervous system of the frog Rana ridibunda by immunohistological techniques. A main group of GnRH-AP-containing perikarya was found in a dorsoventral orientation of the supra anterior preoptic area (SAPA) just in front of the preoptic recess. Fibers originating from these perikarya projected rostrally toward the medial septal nucleus and the diagonal band of Broca. A network of GnRH-AP-immunoreactive (ir) fibers runs caudally from the SAPA toward the ventral hypothalamus. A high density of GnRH-AP-ir terminals was found in the median eminence. A few positive fibers were detected in the neural lobe of the pituitary, particularly in the region bordering the pars intermedia. Labelling of consecutive sections by either GnRH-AP or GnRH antibodies showed that GnRH and GnRH-AP-like irs were contained in the same cells of the SAPA. The double-staining technique with electrophoretic elution confirmed the colocalization of GnRH and GnRH-AP within the same neurons. Such a coexistence indicates that frog GnRH originates from a high molecular weight precursor which is closely related to rat pro-GnRH. The relative preservation of the C-terminal sequence of the pro-GnRH during evolution suggests that GnRH-AP may possess intrinsic biological activity. The high density of GnRH-AP-containing neurons projecting through the external zone of the median eminence would support the concept that GnRH-AP is involved in the modulation of pituitary hormone secretion.

Coexistence of MCH- and alpha-MSH-like peptides in specific neurons of the frog hypothalamus has been investigated on serial frozen sections using the indirect immunofluorescence method. In the anterior region of the preoptic nucleus, perikarya containing MCH- and alpha-MSH-immunoreactive materials were co-distributed and the two peptides were generally co-sequestered within the same neurons. In contrast, alpha-MSH immunoreactive neurons of the ventral infundibular nucleus did not contain any MCH-like peptide. These data suggest that MCH and alpha-MSH are transported by the same nerve fibers originating from preoptic perikarya and are likely released together by axon terminals. Since MCH and alpha-MSH exert antagonistic hormonal activities on dermal melanophores, our results suggest that the two regulatory peptides may also interact in the central nervous system.

Distribution and quantification of PRL in the pituitary gland of the frog Rana ridibunda were investigated using a high-affinity antiserum raised against bullfrog prolactin (PRL). The immunoreactive PRL-producing cells were distributed throughout the pars distalis, the highest density of cells being observed in the rostral region of the adenohypophysis facing the neurointermediate lobe. The dorsal region of the pars distalis contained only a few scattered PRL-immunoreactive cells. At the electron microscopic level, PRL-containing cells were visualized using the immunogold procedure. PRL-immunoreactive material was exclusively stored in secretory granules (size ranging from 200 to 700 nm in diameter). Neither the rough endoplasmic reticulum nor the Golgi apparatus were immunolabeled. Using a radioimmunoassay method we have compared the displacement curves obtained with bullfrog PRL and acetic extracts from Rana ridibunda pituitary. The two binding curves were not completely parallel, suggesting the existence of slight variations of the amino acid sequences of PRL in the two species. The concentration of PRL in the green frog adenohypophysis appeared somewhat higher (35.3 +/- 8.8 micrograms/mg protein) than that in the bullfrog pituitary. These results validate the use of an antiserum to bullfrog PRL to investigate the regulation of PRL secretion in Rana ridibunda.

The distribution of melanin-concentrating hormone (MCH) in the central nervous system of the dogfish Scyliorhinus canicula was determined by indirect immunofluorescence and peroxidase-anti-peroxidase techniques, using an antiserum raised against synthetic salmon MCH. Three groups of MCH-positive cell bodies were localized in the posterior hypothalamus. The most prominent cell group was detected in the nucleus sacci vasculosi. Scattered MCH-immunoreactive cells were observed in the nucleus tuberculi posterioris and in the nucleus lateralis tuberis. At the pituitary level, the caudal part of the median lobe of the pars distalis contained strongly MCH-positive perikarya. Some of these cells were liquor-contacting-type. Immunoreactive fibers originating from the hypothalamic perikarya projected throughout the dorsal wall of the posterior hypothalamus. Positive fibers were also detected within the thalamus and the central gray of the mesencephalon. The distribution of MCH-containing neurons was compared to that of alpha-MSH-immunoreactive elements using consecutive, 5-micron thick sections. Both MCH- and alpha-MSH-immunoreactive peptides were found in the same neurons of the nucleus sacci vasculosi. These data suggest that MCH and alpha-MSH, two neuropeptides which exert antagonistic activities on skin melanophores, may also act in a coordinate manner in the central nervous system of cartilaginous fish.

This study combines trace-metal analysis with an immunofluorescent detection of a methionine-enkephalin-like substance in the digestive gland of the shore crab, Carcinus maenas L. The crabs were taken from two sites: Saint Nazaire and Le Croisic, the first being polluted in comparison to the second. The experimental crabs were also taken in Le Croisic and contaminated with Cd, Pb, Cu, and Zn during 1 to 3 weeks in the laboratory. The immunohistological observations indicate a change in the localization of the immunofluorescent methionine-enkephalin-like substance in the cells of the tubules constituting the digestive gland. In crabs from the clean site, experimentally starved or not, the immunofluorescence appears mostly basal while it exhibits an apical localization in metal-contaminated crabs or crabs caught in the polluted area. The detected substance, the nature of which remains unknown, accompanies cytoplasmic secretory granules during their migration to the cellular apex preceding the apocrine secretion. This change of the immunoreactivity enables the detection of metal contamination but it is nonspecific and therefore, a general environment pollution could produce the same phenomenon. In the particular case of zinc, this alteration appears at a Zn concentration in seawater which does not disturb the natural level of this essential metal in the digestive gland of C. maenas.

Using the indirect immunofluorescence technique, methionine-enkephaline-like, alpha- and beta-endorphin-like peptides were detected on whole body sections of Mytilus edulis L. Met-enkephalin-like immunoreactivity was localized in the epithelium of the digestive tract, in the hepatopancreas, and in the nervous system. The immunoreactive cell bodies were very abundant in the anterior gastric epithelium, but sparse in the terminal portion of the digestive tract. By their basal processes the immunoreactive cells were in contact with a plexus of immunoreactive cells and fibers located in the connective tissue underlying the digestive epithelium. In the principal hepatopancreatic ducts, isolated cells showing met-enkephalin-like immunoreactivity were detected between the epithelial cells and the basal lamina. A few immunoreactive cells and fibers were observed between the hepatopancreatic tubules. The three pairs of nervous ganglia contained in their cortical layer numerous met-enkephalin-like immunoreactive perikarya. Their central area possessed fluorescent immunoreactive bundles of fibers extending to the commissures, the connectives, and the nerves. Met-enkephalin-like immunoreactive fibers were detected between the smooth muscle cells. At the surface of these smooth muscle cells, immunopositive met-enkephalin-like tapered nervous endings were observed. The alpha- and beta-endorphin antisera produced a positive immunoreaction in some gastric epithelial cells, in some perikarya of the pedal ganglia, and in some nervous fibers. The endorphin-like structures were far less abundant than the met-enkephalin-like structures, but very close to them.

The distribution of melanin concentrating hormone (MCH) in the central nervous system of the locust Locusta migratoria was studied by the indirect immunofluorescence technique, using antibodies against salmon MCH. Most MCH-immunoreactive perikarya were found in the optic lobes at both sides of the brain, dorsally with respect to the lamina ganglionaris. The same neurons also contain alpha-melanocyte-stimulating hormone (alpha-MSH)-like material. In addition, a moderate number of MCH-like neurons, which were devoid of alpha-MSH-immunoreactive substances, was observed in the pars intercerebralis. Bright immunofluorescent fibers were visualized in various regions of the central nervous system of the locust: the optic lobes, the ocelli, the proto-and deuterocerebrum, the subesophageal connectives and the corpora cardiaca. In the ventral nerve cord and the subesophageal ganglion, where alpha-MSH-like cell bodies were encountered, MCH immunoreactive perikarya were absent and immunoreactive fibers were scarce. The coexistence of MCH and alpha-MSH-immunoreactive material within the same specific neurons might indicate an evolutionary relationship of both peptides.