Preliminary insights into the incorporation of rosemary extract (Rosmarinus officinalis L.) in fish feed: influence on performance and physiology of gilthead seabream (Sparus aurata).

Gilthead seabream (Sparus aurata) were fed a basal (control) diet and four experimental diets (R600, R1200, R1800 and R2400), containing 600, 1200, 1800 and 2400 mg kg(-1), respectively, of rosemary extract (Rosmarinus officinalis L.). At 4 and 12 weeks from the beginning of the ongrowing period, the fish were sacrificed, blood was drawn to obtain plasma and the liver and intestines were dissected. Growth and feed intake were unaffected by rosemary extract addition. A histological examination of the intestine revealed no differences among the dosages, while the liver showed a sharp decrease in hepatic steatosis in diets supplemented with rosemary extract. Furthermore, plasma alanine aminotransferase was lower with these diets at the end of the ongrowing period. Rosemary extract reduced the plasma levels of glucose and triglycerides on week 4 and glucose and HDL/LDL cholesterol ratio on week 12, suggesting better transport and energy metabolism of the lipids. Overall, the most evident effect of rosemary extract was observed with the 600 mg kg(-1) dose.

Description of series of 10 patients with hypnic headache: Discussion of the diagnostic criteria.

INTRODUCTION: Hypnic headache is a rare primary headache. The diagnostic criteria of the International Headache Classification (IHS) for this condition are discussed, as they have been modified in the new edition of the 2013. PATIENTS AND METHODS: The clinical characteristics, and fulfilment of the criteria of the IHS classification in a series of 10patients diagnosed in our Headaches Clinic, are analyzed. RESULTS: The mean age of onset of symptoms was 52.1years (SD: 13.4; range: 28-69). The pain was reported as oppressive in 60% of the patients, and as sharp in 30%. The headache was described as holocranial in 60% and hemicranial in 40%. They occurred exclusively during night-time sleep in 80% of the patients. The mean duration of headache was 136.5minutes (range: 10-480). The mean number of days per month was 16.4 (range: 3-30), and 50% had less than 15 headache days per month. No patient had autonomic manifestations, 70% had phonophobia, 50% had photophobia, and 50% had both. All of them (100%) had a VAS score equal to or higher than 8. CONCLUSIONS: The criteria for hypnic headache of the new classification are best suited to the characteristics of these patients. Our results show the major changes in the criteria: pain is not always dull, headache frequency may be less than 15days a month, and it can occur in people under age 50. There may be phonophobia or photophobia only, or both. Although it does not form part of the diagnostic criteria, the pain intensity in our series was higher than described.

Post-stroke depression: an update.

INTRODUCTION: Post-stroke depression (PSD) is the most common mood disorder following a stroke, and also the main factor limiting recovery and rehabilitation in stroke patients. In addition, it may increase mortality by up to ten times. DEVELOPMENT: PSD occurs in 1 in 3 stroke patients and more than half of all cases are neither diagnosed nor treated. Several mechanisms, including biological, behavioral, and social factors, are involved in its pathogenesis. Symptoms usually occur within the first three months after stroke (early onset PSD), and less frequently at a later time (late onset PSD). Symptoms resemble those of other types of depression, although there are some differences: PSD patients experience more sleep disturbances, vegetative symptoms, and social withdrawal. For PSD diagnosis, we recommended vigilance and use of specific diagnostic tools such as the Patient Health Questionnaire-2 (PHQ-2). The treatments of choice are selective serotonin reuptake inhibitors (SSRI). However, there are still many unanswered questions in the treatment of PSD, such as the best time to start treatment or the effects of antidepressants on cognition and motor function, among others. CONCLUSIONS: Neurologists play a pivotal role in the care and management of patients recovering from stroke. They must be familiar with methods for early detection and treatment of PSD, as this can facilitate a patient's functional recovery and social reintegration, and improve quality of life for patients and their families.

Trends in the use of statins after ischaemic stroke: Have clinical practices changed?

INTRODUCTION: The role of statins after ischaemic stroke changed with the publication of the SPARCL study in 2006. We analyse how this has influenced the prescription of statins in this patient population. METHODS: We conducted a retrospective study of patients discharged with ischaemic stroke at the Virgen Macarena, Virgen del Rocío, and Valme hospitals in Seville (Spain) over two periods: 1999-2001 and 2014-2016. RESULTS: The study included 1575 patients: 661 (42%) were women and mean age (standard deviation) was 69 (10) years. Patients from the later period are older (68 [10] vs 71 [11]; P = .0001); include a higher proportion of women; and present higher rates of dyslipidaemia, hypertension, and diabetes. At discharge, statins were used in 18.7% of patients (vs 86.9% in the first period; P = .0001), with high-intensity statins prescribed in 11.1% of cases (vs 54.4%; P = .0001). In both periods, atorvastatin was the most commonly prescribed statin (80 mg: 6% vs 42.7%; 40 mg: 5.1% vs 11.1%). In the first period, the use of statins and high-intensity statins was correlated with hypercholesterolaemia, and inversely correlated with age. In the second period, statin use was correlated with hypertension and hypercholesterolaemia, and high-intensity statin use was correlated with ischaemic heart disease and inversely correlated with age. CONCLUSION: There has been a clear change in the prescription of statins to patients with ischaemic stroke at discharge. However, many patients remain undertreated and the use of these drugs needs to be optimised.

Analysis of headache management in Andalusia.

INTRODUCTION AND OBJECTIVE: Headache is a frequent reason for consultation between primary care physicians, emergency services physicians, and neurology specialists; however, it is not always well managed. The Andalusian Society of Neurology's Headache Study Group (SANCE) aimed to analyse headache management at different levels of care. MATERIAL AND METHODS: We conducted a descriptive cross-sectional study with data gathered through a retrospective survey in July 2019. Participants completed a series of structured questionnaires on different social and work-related variables from 4 different groups of healthcare professionals (primary care [PC], emergency departments, neurology departments, headache units). RESULTS: A total of 204 healthcare professionals completed the survey: 35 emergency department physicians, 113 PC physicians, 37 general neurologists, and 19 neurologists specialising in headache. Eighty-five percent of PC physicians reported prescribing preventive drugs, which were maintained for at least 6 months (59%), with flunarizine and amitriptyline being the most commonly used. Most patients attended at neurology consultations (65%) are referred by PC physicians, with changes in the headache pattern being the main reason for referral (74%). Healthcare professionals across all levels of care showed great interest in headache and in receiving training in headache management (97% of PC physicians, 100% of emergency services physicians, 100% of general neurologists). CONCLUSIONS: Migraine sparks great interest among healthcare professionals from different levels of care. Our results also reveal a lack of resources for headache management, which is reflected in the long waiting times. Other means of bilateral communication between different levels of care should be explored (eg, e-mail).

Pharmacological therapy in the secondary prevention of ischemic stroke in the oldest-old patients: has it improved in recent decades?

INTRODUCTION: Population aging has caused an increase in strokes in very elderly patients (VEP). We assess how secondary prevention of ischemic stroke has changed in VEP in recent decades. METHOD: Retrospective study of discharges due to ischemic stroke in the Virgen Macarena, Virgen del Rocio and Valme hospitals in Seville (Spain), during the periods 1999-2001, 2014-16 and 2019-2020. VEP were considered those with ≥80 years. RESULT: We studied 1806 patients, 349 (19.3%) were VEP. Over the years, VEPs have doubled (13.5% vs. 25.9% and 28% p = 0.0001) and age has increased (83.3 ±â€¯3 vs. 84.1 ±â€¯3 vs. 85.2 ±â€¯4 p = 0.001). Comparing the periods, the VEPs have more hypertension (69.9% vs. 84.8% vs. 84.6%; p = 0.0001) and dyslipidemia (12% vs. 41.7% vs. 52.3%; p = 0.0001) and have prescribed more antihypertensives (69.1% vs. 86.7% vs. 92.3%; p = 0.0001), statins (5.3% vs. 78% vs. 81.5%; p = 0.0001) and anticoagulants (16.5% vs. 19.4% vs. 53.1%; p = 0.001), increasing the number of antihypertensives (1 ±â€¯0.9 vs. 1.6 ±â€¯0, 9 vs. 1.9 ±â€¯0.8 drugs p = 0.0001), and high-intensity statins (2.3% vs. 42.7 vs. 69.2% p = 0.0001). Comparing the VEPs with the younger ones, there were no differences in antihypertensive treatment in any period, there were differences in antithrombotic treatment in the first period, and with statins the differences were maintained until the end. CONCLUSIONS: In the last 20 years the number of VEPs has doubled, exceeding a quarter of the discharges. Although there is improvement in secondary stroke prevention in VEPs, there is room for improvement.

[Cryptogenic new-onset super-refractory status epilepticus following SARS-CoV-2 vaccination. A case report]. / Estado epiléptico superrefractario de nueva aparición criptógeno tras vacunación contra el SARS-CoV-2. A propósito de un caso.

INTRODUCTION: New-onset super-refractory status epilepticus (NOSRSE) is a neurological emergency characterised by the development of status epilepticus in a patient without epilepsy or any known prior neurological disease and with no clear structural, toxic or metabolic cause, which recurs after 24 hours of induced coma. The most common identifiable cause is inflammatory-autoimmune. Consequently, we present a case of NOSRSE related to SARS-CoV-2 vaccination as an opportunity to investigate the dysimmune origin of this pathology. CASE REPORT: We report the case of a 40-year-old male who presented at the emergency department with fever and headache with no clear source of infection. His personal history included bacterial meningitis in childhood without any sequelae and protein S deficiency without treatment at the time, as well as vaccination with ChAdOx1 nCoV-19 21 days earlier. He was initially diagnosed with a urinary tract infection and treated with cefuroxime. Two days later, he was taken back to the emergency department with confusional symptoms and tonic-clonic seizures. He did not respond to midazolam and finally required sedation and orotracheal intubation for refractory status epilepticus. While in hospital, he required a number of lines of antiepileptic drugs, ketamine, a ketogenic diet, immunotherapy and plasmapheresis in order to successfully limit NOSRSE. The aetiological study offered normal results for serology, antineuronal antibodies in serum and cerebrospinal fluid, transthoracic echocardiography, testicular ultrasound and computed tomographic angiography. Only the control MRI scan showed a diffuse and bilateral alteration of the right hemispheric cortex and thalamic pulvinar as the only finding. CONCLUSION: It is crucial to report suspected adverse reactions associated with SARS-CoV-2 vaccination, thereby allowing continued monitoring of the risk/benefit ratio of vaccination.

TITLE: Estado epiléptico superrefractario de nueva aparición criptógeno tras vacunación contra el SARS-CoV-2. A propósito de un caso.Introducción. El estado epiléptico superrefractario de nueva aparición (NOSRSE) es una emergencia neurológica caracterizada por el desarrollo de estado epiléptico en un paciente sin epilepsia ni enfermedad neurológica previa conocida y sin clara causa estructural, tóxica o metabólica, que recurre tras 24 horas del coma inducido. La causa identificable más frecuente es la inflamatoria-autoinmune. En consecuencia, planteamos un caso de NOSRSE relacionado con la vacunación para el SARS-CoV-2 como una oportunidad de indagar el origen disinmune de esta patología. Caso clínico. Varón de 40 años que acude al servicio de urgencias refiriendo fiebre y cefalea sin claro foco infeccioso. Entre sus antecedentes personales destacamos una meningitis bacteriana en la infancia sin secuelas y un déficit de proteína S sin tratamiento en ese momento, así como vacunación con ChAdOx1 nCoV-19 21 días antes. Fue inicialmente diagnosticado de infección del tracto urinario y tratado con cefuroxima. Dos días después, se le llevó de nuevo a urgencias con cuadro confusional y crisis tonicoclónicas, sin respuesta al midazolam, y requirió finalmente sedación e intubación orotraqueal por estado epiléptico refractario. Durante su ingreso requirió múltiples líneas de antiepilépticos, quetamina, dieta cetógena, inmunoterapia y plasmaféresis para conseguir limitar el NOSRSE. El estudio etiológico ofrecía normalidad de los resultados de serología, anticuerpos antineuronales en el suero y líquido cefalorraquídeo, ecocardiografía transtorácica, ecografía testicular y angiotomografía computarizada. Únicamente la resonancia magnética de control mostró una alteración difusa y bilateral de la corteza hemisférica y pulvinar talámica derecha como único hallazgo. Conclusión. Es crucial notificar las sospechas de reacciones adversas asociadas a la vacunación frente al SARS-CoV-2, permitiendo así una supervisión continuada de la relación riesgo/beneficio de ésta.

"Full moon" endoscopic sign in intraventricular neurocysticercosis.

INTRODUCTION: Despite improvements in sanitation, diagnosis and treatment, neurocysticercosis is still a public health problem in many countries. In symptomatic patients, there is a broad spectrum of clinical manifestations. When cysticerci are lodged in the ventricles or the subarachnoid space, the flow of cerebrospinal fluid can be obstructed and lead to hydrocephalus and intracranial hypertension. The endoscopic view may be useful as a diagnostic tool. PATIENTS: This report clearly shows a common endoscopic pattern in 4 selected patients with ventricular cysticercosis (2 third ventricle/2 lateral ventricle). The endoscopic view of the cysts in the ventricles resembles a "full moon". This analogy helped to identify the features of cysticerci with intact walls and the vesicular stage, malleable due to its cystic content and having an irregular surface, as evidence of the microscopic structure of the cyst wall in a cysticercus. CONCLUSIONS: This finding is not seen in other intraventricular cysts or tumors that can actually be considered as an additional diagnostic criterion among the definitive findings to establish the diagnosis of cysticercosis, since it involves direct endoscopic visualization of a cysticercus under histopathological demonstration. Additionally, the endoscopic approach can be used as primary treatment for these cases, following the minimally invasive approach principle.

Neurophysiology training in the Neurology Specialist Education Program in Spain.

INTRODUCTION: The training period in neurophysiology is a substantial part of the Neurology Specialist Program in Spain. The National Neurology Committee (La Comisión Nacional de Neurología (CNN), which is the body reporting to the Ministries of Health and Education, must ensure compliance to the Program. MATERIAL AND METHODS: During the first trimester of 2008, the CNN sent a questionnaire, in which there was a question asking about this training period, to each of the managers of the 69 teaching units accredited for neurology training in Spain, for them to answer. RESULTS: Of the 69 questionnaires issued, 49 were received completed, which was a response rate of 71%. The neurophysiology training period of the neurology specialist program in Spain was carried out in the same hospital in 44 teaching unit (90%): the remaining 5 sent their neurology trainees to 4 different hospitals. The Unit that carried out the neurophysiology training period was incorporated into the Neurology Department in 27 (55%) cases, and the formula was mixed in 3 (6%). A total of 69% of tutors were satisfied with the training, but was 90% in the hospitals where the unit was integrated into Neurology, and was 65% where this relationship did not exist. The neurologists in training were informed about EEG in 49% of education units, performed EMG/ENG 57%, and informed about evoked potentials in 35% after their training period. CONCLUSIONS: Although the level of satisfaction is high, the level of responsibility assumed by the neurologists in training during their rotation into neurophysiology does not appear to comply to the demands laid out in the training program, particularly in these units not integrated into Neurology Departments.

Diagnosis and management of immunodeficiencies in adults by allergologists.

Primary immunodeficiencies (PIDs) are genetic diseases that cause alterations in the immune response and occur with an increased rate of infection, allergy, autoimmune disorders, and cancer. They affect adults and children, and the diagnostic delay, morbidity, effect on quality of life, and socioeconomic impact are important. Therapy (gamma-globulin substitution in most cases) is highly effective. We examine adult PIDs and their clinical presentation and provide a sequential and directed framework for their diagnosis. Finally, we present a brief review of the most important adult PIDs, common variable immunodeficiency, including diagnosis, pathogenesis, clinical signs, and disease management.

Trends in the use of statins after ischaemic stroke: Have clinical practices changed? / Tendencias en el uso de estatinas tras un ictus isquémico. ¿Hemos cambiado nuestra práctica clínica?

INTRODUCTION: The role of statins after ischaemic stroke changed with the publication of the SPARCL study in 2006. We analyse how this has influenced the prescription of statins in this patient population. METHODS: We conducted a retrospective study of patients discharged with ischaemic stroke at the Virgen Macarena, Virgen del Rocío, and Valme hospitals in Seville (Spain) over two periods: 1999-2001 and 2014-2016. RESULTS: The study included 1575 patients: 661 (42%) were women and mean age (standard deviation) was 69 (10) years. Patients from the later period are older (68 [10] vs 71 [11]; P=.0001); include a higher proportion of women; and present higher rates of dyslipidaemia, hypertension, and diabetes. At discharge, statins were used in 18.7% of patients (vs 86.9% in the first period; P=.0001), with high-intensity statins prescribed in 11.1% of cases (vs 54.4%; P=.0001). In both periods, atorvastatin was the most commonly prescribed statin (80mg: 6% vs 42.7%; 40mg: 5.1% vs 11.1%). In the first period, the use of statins and high-intensity statins was correlated with hypercholesterolaemia, and inversely correlated with age. In the second period, statin use was correlated with hypertension and hypercholesterolaemia, and high-intensity statin use was correlated with ischaemic heart disease and inversely correlated with age. CONCLUSION: There has been a clear change in the prescription of statins to patients with ischaemic stroke at discharge. However, many patients remain undertreated and the use of these drugs needs to be optimised.

Optimization of Free Phytoprostane and Phytofuran Production by Enzymatic Hydrolysis of Pea Extracts Using Esterases.

Given the growing interest in phytoprostanes (PhytoPs) and phytofurans (PhytoFs) in the fields of plant physiology, biotechnology, and biological function, the present study aims to optimize a method of enzymatic hydrolysis that utilizes bacterial and yeast esterases that allow the appropriate quantification of PhytoPs and PhytoFs. To obtain the highest concentration of PhytoPs and PhytoFs, a response surface methodology/Box-Behnken design was used to optimize the hydrolysis conditions. Based on the information available in the literature on the most critical parameters that influence the activity of esterases, the three variables selected for the study were temperature (°C), time (min), and enzyme concentration (%). The optimal hydrolysis conditions retrieved differed between PhytoPs (21.5 °C, 5.7 min, and 0.61 µg of enzyme per reaction) and PhytoFs (20.0 °C, 5.0 min, and 2.17 µg of enzyme per reaction) and provided up to 25.1- and 1.7-fold higher contents relative to nonhydrolyzed extracts. The models were validated by comparing theoretical and experimental values for PhytoP and PhytoF yields (1.01 and 1.06 theoretical/experimental rates, respectively). The optimal conditions were evaluated for their relative influence on the yield of individual nonesterified PhytoPs and PhytoFs to define the limitations of the models for obtaining the highest concentration of most considered compounds. In conclusion, the models developed provided valuable alternatives to the currently applied methods using unspecific alkaline hydrolysis to obtain free nonesterified PhytoPs and PhytoFs, which give rise to more specific hydrolysis of PhytoP and PhytoF esters, reducing the degradation of free compounds by classical chemical procedures.

Impact of SARS-CoV-2 infection on neurodegenerative and neuropsychiatric diseases: a delayed pandemic? / Influencia de la infección SARS-CoV-2 sobre enfermedades neurodegenerativas y neuropsiquiátricas: ¿una pandemia demorada?

INTRODUCTION: SARS-CoV-2 was first detected in December 2019 in the Chinese city of Wuhan and has since spread across the world. At present, the virus has infected over 1.7 million people and caused over 100 000 deaths worldwide. Research is currently focused on understanding the acute infection and developing effective treatment strategies. In view of the magnitude of the epidemic, we conducted a speculative review of possible medium- and long-term neurological consequences of SARS-CoV-2 infection, with particular emphasis on neurodegenerative and neuropsychiatric diseases of neuroinflammatory origin, based on the available evidence on neurological symptoms of acute SARS-CoV-2 infection. DEVELOPMENT: We systematically reviewed the available evidence about the pathogenic mechanisms of SARS-CoV-2 infection, the immediate and lasting effects of the cytokine storm on the central nervous system, and the consequences of neuroinflammation for the central nervous system. CONCLUSIONS: SARS-CoV-2 is a neuroinvasive virus capable of triggering a cytokine storm, with persistent effects in specific populations. Although our hypothesis is highly speculative, the impact of SARS-CoV-2 infection on the onset and progression of neurodegenerative and neuropsychiatric diseases of neuroinflammatory origin should be regarded as the potential cause of a delayed pandemic that may have a major public health impact in the medium to long term. Cognitive and neuropsychological function should be closely monitored in COVID-19 survivors.

Will neurological care change over the next 5 years due to the COVID-19 pandemic? Key informant consensus survey. / ¿Va a cambiar la neurología tras la pandemia de COVID-19 en los próximos 5 años? Estudio de enfoque mediante informadores clave.

INTRODUCTION: The COVID-19 pandemic will give rise to long-term changes in neurological care, which are not easily predictable. MATERIAL AND METHODS: A key informant survey was used to enquire about the changes expected in the specialty over the next 5 years. The survey was completed by heads of neurology departments with broad knowledge of the situation, having been active during the pandemic. RESULTS: Despite a low level of consensus between participants, there was strong (85%) and moderate consensus (70%) about certain subjects, mainly the increase in precautions to be taken, the use of telemedicine and teleconsultations, the reduction of care provided in in-person consultations to avoid the presence of large numbers of people in waiting rooms, the development of remote training solutions, and the changes in monitoring visits during clinical trials. There was consensus that there would be no changes to the indication of complementary testing or neurological examination. CONCLUSION: The key informant survey identified the foreseeable changes in neurological care after the pandemic.

[Development of a program of nutritional education and valuation of the change of healthful nourishing habits in a population of students of Obligatory Secondary Education]. / Desarrollo de un programa de educación nutricional y valoración del cambio de hábitos alimentarios saludables en una población de estudiantes de Enseñanza Secundaria Obligatoria.

INTRODUCTION: Adolescence is a critical period for the acquisition and configuration of healthy dietary habits and lifestyle for the young future, which will likely persist throughout the adulthood. Paediatric and juvenile obesity is a public health problem which control necessarily implies prevention and nutritional education. OBJECTIVES: To evaluate the nutritional status of the adolescents and determine the proportion with overweight or obesity, and to establish a Nutritional Intervention Programme and analyse the improvement in the pattern of dietary habits among the adolescents. METHODS: The study has been carried out in a population of 372 Obligatory Secondary Education (OSE) students from the Institute of Secondary Education of Gandía (Valencia). RESULTS: 37.8% of the adolescents have improved the level of their diet quality. Those consuming a high quality diet have increased from 30.0% to 58.6%. Also significant is the number of students that have taken up having breakfast and those having discontinued taking industrial bakery with this meal. The decrease in the number of adolescents going to fast food places and of those that have discontinued eating candies regularly is statistically significant. The data from the KIDMED index show that 47.4% (p < 0.001) of de the students have improved the quality of their diet and in none of them it has worsened. DISCUSSION: Before starting the programme, 30% of the students followed a high quality diet comparable to the traditional Mediterranean Diet, and after the education programme, this percentage increased to 58.6%. Forty-seven point four percent of overweighed or obese students receiving the Nutritional Education and Intervention have improved their diet quality and the percentage of those following a high quality diet varied from 28.9% to 71.0%.

Knowing Patients Better After a Stroke and Secondary Prevention.

The habits and lifestyles are the fundamental factors in the control of cardiovascular risk. Patients who have had a cerebrovascular accident (CVA) have a high risk of having a new event with similar characteristics. The exponentially growing success, penetration and adherence of the new communication technologies, based on applications (APPs), allows to use them to obtain information and influence the risk factors. We propose that empowering patients in their disease can make a more efficient management of it. For this reason, we designed and developed a system which integrates a mobile application and a web application. This system also makes use of peripheral devices to monitor patients and allow the automatic acquisition of information to enable the characterization of this kind of patients in relation to habits and lifestyle. At the same time, the system can also empower these patients with their disease to do secondary prevention.

Energy intake and macronutrient selection in sharpsnout seabream (Diplodus puntazzo) challenged with fat dilution and fat deprivation using encapsulated diets.

Sharpsnout seabream fed pure macronutrient capsules were challenged to fat dilution and fat deprivation in order to investigate the effects of fat level on energy intake regulation and macronutrient selection by fish, as they lack oropharyngeal chemosensory information from the diet. During the control phase, the fish were fed three individually encapsulated macronutrients, from which they composed a diet containing 67.36% protein (P), 19.08% carbohydrates (CH) and 13.57% fat (F), in terms of macronutrient weight intake percentage. During the second phase of the experiment, a lipid content reduction in F capsules from 55.0% to 13.4% did not significantly modify this selection pattern, energy ingestion or the number of capsules ingested of each macronutrient. During the third phase, in which they were subjected to fat deprivation, starting on almost the first day, the fish increased their total energy intake and total ingested number of capsules. These results reveal that fish are capable of distinguishing and selecting each of the three macronutrients contained in gelatine capsules, and that fish selection of a balanced diet from pure macronutrients is remarkably stable. Fish are capable of sustaining their macronutrient selection pattern and energy intake with very low amounts of fat in their diets (Phase 2). A certain instability in the initial P, CH and energy intake was only observed when fat was totally deprived (Phase 3), which resulted in higher values than those observed in Phase 1. In order to examine any possible effects of diet encapsulation, digestibility assays were performed in a second experiment. The fish were divided into two experimental groups and fed the same complete commercial diet, the only difference being the way it was presented to each group (pelleted or encapsulated). No statistical differences between the experimental groups were found with regards to both apparent digestibility coefficients and fish growth.

Myocyte nuclear factor, a novel winged-helix transcription factor under both developmental and neural regulation in striated myocytes.

A sequence motif (CCAC box) within an upstream enhancer region of the human myoglobin gene is essential for transcriptional activity in both cardiac and skeletal muscle. A cDNA clone, myocyte nuclear factor (MNF), was isolated from a murine expression library on the basis of sequence-specific binding to the myoglobin CCAC box motif and was found to encode a novel member of the winged-helix or HNF-3/fork head family of transcription factors. Probes based on this sequence identify two mRNA species that are upregulated during myocyte differentiation, and antibodies raised against recombinant MNF identify proteins of approximately 90, 68, and 65 kDa whose expression is regulated following differentiation of myogenic cells in culture. In addition, the 90-kDa form of MNF is phosphorylated and is upregulated in intact muscles subjected to chronic motor nerve stimulation, a potent stimulus to myoglobin gene regulation. Amino acid residues 280 to 389 of MNF demonstrate 35 to 89% sequence identity to the winged-helix domain from other known members of this family, but MNF is otherwise divergent. A proline-rich amino-terminal region (residues 1 to 206) of MNF functions as a transcriptional activation domain. These studies provide the first evidence that members of the winged-helix family of transcription factors have a role in myogenic differentiation and in remodeling processes of adult muscles that occur in response to physiological stimuli.

A 40-kilodalton protein binds specifically to an upstream sequence element essential for muscle-specific transcription of the human myoglobin promoter.

To define transcriptional control elements responsible for muscle-specific expression of the human myoglobin gene, we performed mutational analysis of upstream sequences (nucleotide positions -373 to +7 relative to the transcriptional start site) linked to a firefly luciferase gene. Transient expression assays in avian and mammalian cells indicated that a CCCACCCCC (CCAC box) sequence (-223 to -204) is necessary for muscle-specific transcription directed either by the native myoglobin promoter or by a heterologous minimal promoter linked to the myoglobin upstream enhancer region. A putative MEF2-like site (-160 to -169) was likewise necessary for full transcriptional activity in myotubes. Mutations within either of two CANNTG (E-box) motifs (-176 to -148) had only minimal effects on promoter function. We identified and partially purified from nuclear extracts a 40-kDa protein (CBF40) that binds specifically to oligonucleotides containing the CCAC box sequence. A mutation of the CCAC box that disrupted promoter function in vivo also impaired binding of CBF40 in vitro. These data suggest that cooperative interactions between CBF40 and other factors including MEF-2 are required for expression of the human myoglobin gene in skeletal muscle.