Identification of the sex-determining region in flathead grey mullet (Mugil cephalus).

Elucidation of the sex-determination mechanism in flathead grey mullet (Mugil cephalus) is required to exploit its economic potential by production of genetically determined monosex populations and application of hormonal treatment to parents rather than to the marketed progeny. Our objective was to construct a first-generation linkage map of the M. cephalus in order to identify the sex-determining region and sex-determination system. Deep-sequencing data of a single male was assembled and aligned to the genome of Nile tilapia (Oreochromis niloticus). A total 245 M. cephalus microsatellite markers were designed, spanning the syntenic tilapia genome assembly at intervals of 10 Mb. In the mapping family of full-sib progeny, 156 segregating markers were used to construct a first-generation linkage map of 24 linkage groups (LGs), corresponding to the number of chromosomes. The linkage map spanned approximately 1200 cM with an average inter-marker distance of 10.6 cM. Markers segregating on LG9 in two independent mapping families showed nearly complete concordance with gender (R2  = 0.95). The sex determining locus was fine mapped within an interval of 8.6 cM on LG9. The sex of offspring was determined only by the alleles transmitted from the father, thus indicating an XY sex-determination system.

In vitro effect of cortisol and urotensin I on arginine vasotocin and isotocin secretion from pituitary cells of gilthead sea bream Sparus aurata.

This study aimed at determining whether in vitro secretion of two neuropeptides, arginine vasotocin (AVT) and isotocin (IT), from pituitary cells of gilthead sea bream Sparus aurata was affected by cortisol and urotensin (UI). Pituitary cells were exposed to 1·4 × 10(-8) , 1·4 × 10(-7) and 0·4 × 10(-6) M cortisol and 10(-12) , 10(-10) and 10(-8) M UI for 6, 24 and 48 h, respectively. AVT and IT contents were determined in the culture media by high-performance liquid chromatography (HPLC). An increase in AVT secretion and a decrease in IT secretion were observed at all cortisol doses. UI increased AVT secretion after 6 h of incubation at all doses. After 24 h, however, only the highest dose of UI still displayed an effect. IT secretion was not influenced by UI. It was thus demonstrated that cortisol does influence AVT and IT secretion from S. aurata pituitary cells, while UI regulates AVT secretion, as a component of hypothalamic-pituitary-interrenal (HPI) axis in this species.

Enzyme replacement therapy for mucopolysaccharidosis VI: long-term cardiac effects of galsulfase (Naglazyme®) therapy.

Characteristic cardiac valve abnormalities and left ventricular hypertrophy are present in untreated patients with mucopolysaccharidosis type VI (MPS VI). Cardiac ultrasound was performed to investigate these findings in subjects during long-term enzyme replacement therapy (ERT) with recombinant human arylsulfatase B (rhASB, rhN-acetylgalactosamine 4-sulfatase, galsulfase, Naglazyme®). Studies were conducted in 54 subjects before ERT was begun and at specific intervals for up to 96 weeks of weekly infusions of rhASB at 1 mg/kg during phase 1/2, phase 2, and phase 3 trials of rhASB. At baseline, mitral and aortic valve obstruction was present and was significantly greater in those ≥12 years of age. Mild mitral and trace aortic regurgitation were present, the former being significantly greater in those <12 years. Left ventricular hypertrophy, with averaged z-scores ranging from 1.6-1.9 SD greater than normal, was present for ages both <12 and ≥12 years. After 96 weeks of ERT, ventricular septal hypertrophy regressed in those <12 years. For those ≥12 years, septal hypertrophy was unchanged, and aortic regurgitation increased statistically but not physiologically. Obstructive gradients across mitral and aortic valves remained unchanged. The results suggest that long-term ERT is effective in reducing intraventricular septal hypertrophy and preventing progression of cardiac valve abnormalities when administered to those <12 years of age.

High-efficiency solution-processed Cu2ZnSn(S,Se)4 thin-film solar cells prepared from binary and ternary nanoparticles.

A new solution-based method to fabricate Cu(2)ZnSn(S,Se)(4) (CZTSSe) thin films is presented. Binary and ternary chalcogenide nanoparticles were synthesized and used as precursors to form CZTSSe thin films. The composition of the CZTSSe films can be easily controlled by adjusting the ratio of the nanoparticles used. The effect of compositional adjustment on device performance is illustrated. Laboratory-scale photovoltaic cells with 8.5% total-area efficiency (or 9.6% active-area efficiency) were demonstrated without anti-reflective coatings. Material characterization data revealed the formation of a bilayer microstructure during thermal processing and suggested a path forward on device improvement.

GnRHa-mediated stimulation of the reproductive endocrine axis in captive Atlantic bluefin tuna, Thunnus thynnus.

A controlled-release implant loaded with GnRH agonist (GnRHa) was used to induce spawning in Atlantic bluefin tuna (Thunnus thynnus) during two consecutive reproductive seasons. The fish were implanted underwater and sampled between days 2 and 8 after treatment. At the time of GnRHa treatment, females were in full vitellogenesis and males in spermiation. There was a rapid burst of pituitary luteinizing hormone (LH) release at day 2 after treatment in GnRHa-treated fish, and circulating LH remained elevated up to day 8 after treatment. In contrast, control fish had significantly lower levels in the plasma, but higher LH content in the pituitary, as observed in many other cultured fishes that fail to undergo oocyte maturation, ovulation and spawning unless induced by an exogenous GnRHa. Plasma testosterone (T) and 17ß-estradiol (E(2)) were elevated in response to the GnRHa treatment in females, while 11-ketotestosterone (11-KT) but not T was elevated in males. Even though oocyte maturation and ovulation did occur in GnRHa-induced fish, no significant elevations in 17,20ß-dihydroxy-4-pregnen-3-one (17,20ß-P) or 17,20ß,21-trihydroxy-4-pregnen-3-one (20ß-S), in either the free, conjugated or 5ß-reduced,3α-hydroxylated forms was observed in fish sampled within 6 days after treatment. Interestingly, a significant peak in plasma free 17,20ß-P levels occurred in both males and females at day 8 after treatment. Histological sections of the ovaries in these females contained oocytes at the migrating germinal vesicle stage, suggesting the role of this hormone as a maturation-inducing steroid in Atlantic bluefin tuna. In conclusion, the GnRHa implants activated effectively the reproductive endocrine axis in captive Atlantic bluefin tuna broodstocks, through stimulation of sustained elevations in plasma LH, which in turn evoked the synthesis and secretion of the relevant sex steroids leading to gamete maturation and release.

Glycolipids from a colloid chemical point of view.

Glycolipids are a group of compounds with a broad range of applications. Two types of glycolipids (alkylpolyglycosides and gangliosides) were examined with regard to their physicochemical properties. Despite their structural differences, they have in common that they are amphiphilic molecules and able to aggregate to form monolayers, bilayers, micelles, lyothropic mesophases or vesicles. The structures of glycolipid micelles were investigated by different experimental techniques in addition to molecular dynamic simulations. The knowledge of the physicochemical properties of gangliosides enables a better understanding of their biological functions. Structural features were obtained for the monosialogangliosides GM1, GM2 and GT1b from bovine brain by means of mass spectrometry. Further the aggregation behaviour was determined by small-angle neutron and dynamic light scattering experiments. Interaction studies of these compounds were carried out by means of surface plasmon resonance using gangliosides incorporated liposomes. They were used as model membranes that interact with the lectins WGA, RCA and HPA. The interaction of lectins immobilized to a modified silicon surface was investigated by in-situ ellipsometry.

Enhancement of antitumor activity of glutamine antagonists 6-diazo-5-oxo-L-norleucine and acivicin in cell culture by glutaminase-asparaginase.

Mouse P388 and L1210 leukemia cells grown in vitro were found to be 4 to 10 times more sensitive to 6-diazo-5-oxo-L-norleucine and 3 to 5 times more sensitive to Acivicin than were 3T3 and C57BL x DBA/2 F1 embryonic fibroblasts. The combined actions of succinylated Acinetobacter glutaminase-asparaginase and 6-diazo-5-oxo-L-norleucine or Acivicin produced synergistic inhibition of nucleic acid synthesis in P388 tumor cells. An uptake system for Acivicin is described. Its properties in P388 and 3T3 cells are similar in their strong temperature dependence, utilization of the "L" transport system, presumably competitive inhibition by glutamine, similar Km's (about 200 microM), and potent inhibition by p-chloromercuribenzene sulfonate, NA+. However, Acivicin uptake was inhibited in 3T3 (but not in P388) cells by KCN or 2,4-dinitrophenol. At equilibrium in P388 cells, the intracellular level of Acivicin was approximately 57-fold greater than was the extracellular concentration. The accumulated Acivicin was not metabolized by P388 cells, nor does exchange of 3H label into water occur. Rapid efflux of Acivicin occurred with both cell lines at 37 degrees, but efflux from 3T3 cells was greatly diminished at 0 degrees. The rate of efflux was accelerated by including glutamine or unlabeled Acivicin in the extracellular medium.

Video-assisted thoracoscopic vascular ring division in infants and children.

OBJECTIVES: This study evaluated our early experience with video-assisted thoracoscopic vascular ring division and compared this approach with division by means of a conventional open thoracotomy. BACKGROUND: Video-assisted thoracoscopic techniques reduce surgical trauma and have been applied to several adult thoracic procedures; however, pediatric applications have been limited. We developed instruments and techniques for video-assisted thoracoscopic vascular ring division in the pediatric population. METHODS: We compared patient characteristics, operative results and postoperative hospital courses of all patients undergoing vascular ring division by a video-assisted approach with a historical control group of all patients undergoing division by an open thoracotomy between January 1991 and December 1992. RESULTS: Eight patients (median age 5 months, range 40 days to 5.5 years; median weight 6.2 kg, range 1.8 to 17.1) underwent video-assisted thoracoscopic vascular ring division. Four had a double aortic arch with an atretic left arch and a left ligamentum, and four had a right aortic arch with aberrant left subclavian artery and a left ligamentum. All eight had successful ring division with symptomatic relief and no mortality. A limited thoracotomy was performed in three patients to divide patent vascular structures, and the hospital period was prolonged in one because of chylothorax. These eight patients were compared with a historical cohort of eight pediatric patients having vascular ring division performed by a conventional thoracotomy. The two groups did not differ in age, weight, intensive care unit or postoperative hospital stay, duration of intubation or thoracostomy tube or hospital charges. Total operating room time was longer for the group undergoing video-assisted operation. CONCLUSIONS: Early results for video-assisted thoracoscopic vascular ring division are comparable to those of the conventional surgical approach. With further refinement in technique and instrumentation, video-assisted surgical intervention may become a viable alternative to open thoracotomy for management of the symptomatic vascular ring.

In-situ X-ray absorption spectroscopy study of Pt and Ru chemistry during methanol electrooxidation.

Methanol electrooxidation in a 0.5 M sulfuric acid electrolyte containing 1.0 M CH3OH was studied on 30% Pt/carbon and 30% PtRu/carbon (Pt/Ru = 1:1) catalysts using X-ray absorption spectroscopy (XAS). Absorption by Pt and Ru was measured at constant photon energy in the near edge region during linear potential sweeps of 10-50 mV/s between 0.01 and 1.36 V vs rhe. The absorption results were used to follow Pt and Ru oxidation and reduction under transient conditions as well as to monitor Ru dissolution. Both catalysts exhibited higher activity for methanol oxidation at high potential following multiple potential cycles. Correlation of XAS data with the potential sweeps indicates that Pt catalysts lose activity at high potentials due to Pt oxidation. The addition of Ru to Pt accelerates the rate of methanol oxidation at all potentials. Ru is more readily oxidized than Pt, but unlike Pt, its oxidation does not result in a decrease in catalytic activity. PtRu/carbon catalysts underwent significant changes during potential cycling due to Ru loss. Similar current density vs potential results were obtained using the same PtRu/carbon catalyst at the same loading in a membrane electrode assembly half cell with only a Nafion (DuPont) solid electrolyte. The results are interpreted in terms of a bifunctional catalyst mechanism in which Pt surface sites serve to chemisorb and dissociate methanol to protons and carbon monoxide, while Ru surface sites activate water and accelerate the oxidation of the chemisorbed CO intermediate. PtRu/carbon catalysts maintain their activity at very high potentials, which is attributed to the ability of the added Ru to keep Pt present in a reduced state, a necessary requirement for methanol chemisorption and dissociation.

Gaseous ammonia pretreatment lowers the required energy input for fine milling-enhanced enzymatic saccharification of switchgrass.

BACKGROUND: Fine milling of dry lignocellulosic biomass, without prior chemical pretreatment, can produce a high percent theoretical yield of sugars during subsequent enzymatic saccharification. However, the high sugar yields, necessary for a commercial biofuels process, are costly, with the milling energy input, necessary to achieve such yields even exceeding the energy content of the biomass. In this study, we show that low moisture gaseous ammonia pretreatment of switchgrass, in advance of the milling step, significantly reduces the milling energy required to give high sugar titers. RESULTS: We have found that the increase in monomeric sugar yields upon enzymatic saccharification of ball-milled, but not chemically treated switchgrass, is more closely tied to the formation of crystallites of cellulose with a negative linear dependence on the coherent domain size than to a decrease in particle size or to an increase in surface area of the biomass. The milling energy required to reach ~80 % of theoretical yield of glucose under these conditions is intolerably high, however, approximating two times the energy content of the biomass. Two different low moisture content ammonia pretreatments, prior to milling, significantly reduce the required milling energy (four to eightfold, depending on the pretreatment). These involve either heating the biomass at 150-160 °C for 1 h at 10 wt% gaseous ammonia or incubating at room temperature for 9 days at 20 wt% gaseous ammonia, the latter mimicking potential treatment during biomass storage. We have tested this combination of pretreatment and milling on switchgrass using a variety of milling methods, but mostly using ball and attritor milling. In the case of the high-temperature gaseous ammonia treatment followed by attritor milling, the increase in the monomeric sugar yield upon saccharification shows a negative linear dependence on the second or third power of the cellulose crystalline coherent domain size, implying that the surfaces as well as the ends of the cellulose fibrils are accessible to cellulolytic enzymes. CONCLUSIONS: The combination of knife milling, low moisture gaseous ammonia pretreatment followed by attritor milling that costs only ~5 % of the energy content of the biomass for a total energy input of ~11 % of the biomass energy content, is capable of delivering high sugar titers upon enzymatic saccharification. These results show, therefore, how to better integrate a mechanochemical step into the pretreatment of switchgrass in a commercial biomass to biofuels conversion process.

Possible interactions between gonadotrophs and somatotrophs in the pituitary of tilapia: apparent roles for insulin-like growth factor I and estradiol.

The unique organization of the teleost pituitary, in which cells are grouped according to their characteristic hormone, makes this a suitable model for studying pituitary paracrine interactions. In a number of fish, including tilapia, there are variations in the circulating levels of the gonadotropins and GH, which are elevated during the reproductive season, suggesting interactions between the reproductive and growth axes. The aim of this study was to investigate paracrine interactions between the gonadotrophs and somatotrophs in the tilapia pituitary. Initially, dispersed pituitary cells were separated on a density gradient in which the gonadotrophs were found in the least dense fractions, and the somatotrophs were concentrated in the densest fraction. After 4 days in culture, cells in the least dense fractions showed characteristic cytoplasmic extensions not seen in the somatotrophs, which appeared small and failed to form aggregates; somatotrophs were found, however, attached to other non-GH cells. Staining of the nuclei with 4,6-diaminidino-2-phenyl-dihydrochloride revealed that the isolated somatotrophs had undergone nuclear condensation and fragmentation typical of apoptosis. Addition of either estradiol or human recombinant insulin-like growth factor I (IGF-I; 10 nM) to the somatotroph cultures increased the number of cell aggregates and reduced the number of condensed or fragmented nuclei. Immunocytochemical studies on pituitary sections revealed IGF-I immunoreactivity in regions of the proximal pars distalis that stain with gonadotropin IIbeta antisera and also in regions of the rostral pars distalis characteristic of corticotrophs; immunoreactive IGF-I was never seen in the region of the somatotrophs. Incubation of cells from the different fractions with testosterone (10 nM; 24 h) revealed that cells of the least dense fractions, which were rich in gonadotrophs, possessed aromatizing ability, which was absent in the somatotroph-enriched fraction. These results suggest that estradiol and IGF-I, both generated from cells other than the somatotrophs, may exert antiapoptotic effects and thus possibly control the size of this population of cells.

Brotizolam, a triazolothienodiazepine, in insomnia.

Sixty-three outpatients with chronic insomnia were treated for 3 weeks under double-blind conditions with either brotizolam (n = 29) at a dose of 0.25 mg or 0.5 mg or placebo (n = 34). A 3-day placebo period preceded and followed the double-blind treatment phase. Brotizolam consistently produced significantly more sleep improvement than placebo but also more adverse effects. In those patients switched abruptly from brotizolam to placebo, rebound insomnia was observed, being most marked at the first post-brotizolam placebo night.

Triazolam in insomniac family practice patients.

Triazolam, 0.5 mg, a benzodiazepine with hypnotic properties, was compared to secobarbital, 100 mg, and placebo in a 1-wk study conducted with 100 insomniac family practice patients. Considerable sensitivity to differential treatment effects was demonstrated for these family practice patients as well as for a research methodology that combines a crossover design, permitting preference ratings, with a between-patient design. In almost all sleep parameters, assessed with a variety of subjective techniques, triazolam and secobarbital were shown to be significantly more effective than placebo. Triazolam was consistently and often significantly indicated to be a more effective hypnotic, particularly for reducing nocturnal awakening, than secobarbital. Analysis of self-report emotional distress data revealed that present insomniac patients were slightly more emotionally symptomatic than other nonpsychiatric populations. Triazolam was followed by the greatest and secobarbital the least relief of emotional symptoms and triazolam emerged as an especially effective hypnotic for initially more depressed insomniac patients. Present findings suggest that type and degree of emotional symptomatology may affect the response of insomniac patients to hypnotics.

Balloon aortic valvuloplasty in the young adult with congenital aortic stenosis.

Although balloon aortic valvuloplasty (BAV) is effective in the acute management of valvar aortic stenosis (AS), sustained benefit of this technique has been shown to vary in differing age groups. The role of BAV in the young adult with congenital, nondegenerative AS is poorly defined. The catheterization results and follow-up echocardiographic data were reviewed for all patients (n = 18) between the ages of 17 and 40 years (mean 23 +/- 7) undergoing BAV at our institution between March 1986 and January 1992. Peak-to-peak systolic ejection gradient was reduced by 55%, from 85 +/- 29 to 38 +/- 17 mm Hg (p < 0.001). Aortic valve area increased from 0.9 +/- 0.2 to 1.1 +/- 0.3 cm2 (p = 0.003). Results of dilation were inadequate in 2 patients. There were no deaths, myocardial infarction, or embolic events. The 16 "effectively" dilated patients were followed with serial echocardiography for a period of 1 to 82 months, demonstrating persistent gradient relief in most patients (maximal instantaneous gradient at follow-up, 55 +/- 17 vs 79 +/- 22 mm Hg before dilation, p < 0.001). Aortic valve replacements were performed in 5 patients, 2 with unsuccessful initial dilations. At the time of the most recent echocardiogram, 8 of 16 patients remained "incident free," with no subsequent catheterization or surgical interventions, a maximal instantaneous gradient of < or = 55 mm Hg, no more than moderate aortic regurgitation, and preserved ventricular function.(ABSTRACT TRUNCATED AT 250 WORDS)

Tilapia glycoprotein hormone alpha subunit: cDNA cloning and hypothalamic regulation.

The cDNA encoding the glycoprotein alpha (GPalpha) subunit of tilapia (Oreochromis mossambicus) was partially cloned using RACE-polymerase chain reaction (PCR) technique. The amplified cDNA was found to be 583 bases long, and to consist of a portion of the signal peptide, the full sequence encoding the mature peptide (94 amino acids) and the 3' untranslated region. Northern blot analysis revealed a single band of approximately 600 bp. Alignment of the deduced amino acids of the mature protein showed that the tilapia GPalpha subunit shares more than 80% identity with that of other perciform fish (i.e. striped bass, sea bream and yellowfin porgy) and less than 70% with that of more taxonomically remote fish and other vertebrates. Exposure of dispersed tilapia pituitary cells to salmon gonadotropin-releasing hormone (sGnRH) elevated GPalpha mRNA levels via both PKC and cAMP-protein kinase A (PKA) pathways. The transcript levels were also regulated by pituitary adenylate cyclase activating polypeptide (PACAP) and neuropeptide Y (NPY), both acting through PKC and PKA pathways. Moreover, a combined treatment of PACAP or NPY with GnRH seems to have an additive effect on the GPalpha subunit gene transcription. These results suggest that in tilapia the expression of GPalpha subunit is regulated by GnRH mainly via PKC and PKA pathways. Furthermore, PACAP and NPY can elevate the GnRH-stimulated GPalpha subunit transcription and can directly affect the subunit mRNA levels, via the same transduction pathways.

Psychiatric illness in family practice.

One hundred twenty patients from each of 6 family practices were surveyed to determine the prevalence of symptom-based psychiatric illness (emotional distress) in family practice. Assessments of presence or absence of emotional distress and the severity of this distress were made using ratings made by the treating physician and by the patient. Nineteen percent of the surveyed patients were designated by their physicians as having "high distress," and 21% by the patient-completed abbreviated Hopkins Symptom Checklist (HSCL-25). High concordance (86.7%) existed between the "case" assessment by the physician and the patient's own rating of distress. For the majority of "high distress" patients, pharmacotherapy was considered the treatment of choice by physicians.

Diphenhydramine in insomniac family practice patients: a double-blind study.

One hundred eleven mildly to moderately insomniac patients participated in a double-blind, placebo-controlled study to establish the efficacy of diphenhydramine as an over-the-counter (OTC) sleep aid. A two-week crossover design was employed in which all patients received both diphenhydramine and placebo for one week each. The daily diphenhydramine dose was 50 mg at bedtime. Results obtained indicate that diphenhydramine improved various sleep parameters, including sleep latency, to a significantly higher degree than did placebo. In addition, patients on diphenhydramine reported feeling more restful the following morning and patients preferred the diphenhydramine drug to placebo despite experiencing more side effects. This study thus supports the use of 50 mg diphenhydramine as an OTC sleep aid in the treatment of temporary mild to moderate insomnia.

Primary repair of aortopulmonary septal defect, interrupted aortic arch, and anomalous origin of the right pulmonary artery.

Aortopulmonary septal defect, interruption of the aortic arch, and anomalous origin of the right pulmonary artery from the aorta comprised a challenging constellation of defects in a 13-day-old neonate, who underwent complete repair without homograft tissue or synthetic graft. After the aorta was separated from the pulmonary artery, the descending aorta was anastomosed to the septal defect in the ascending aorta. The right pulmonary artery was anastomosed to the septal defect in the main pulmonary artery, anterior to the aorta. The postoperative course was uncomplicated, and the potential for growth of both great vessels was optimized.

Pulmonary artery translocation: a surgical option for complex anomalous coronary artery anatomy.

Rather than perform a difficult and potentially high risk coronary reimplantation in a patient with an aberrant right coronary artery coursing between the aorta and pulmonary artery, the main pulmonary artery was translocated toward the left pulmonary hilum to create additional space between the aortic and pulmonic trunks.