Rationed and satiated growth hormone transgenic Coho Salmon (Oncorhynchus kisutch) show tissue specific differences in energy stores.

Growth hormone transgenic coho salmon experience increased growth rates, driven primarily through elevated feed intake and feed conversion. However, neuropeptides that signal appetite stimulation have been shown to exhibit variable responses across fed states, suggesting a more complex system mediating growth in these fish. Studies have proposed that growth hormone may have a modulatory role on the energy reserves of fish, possibly through AMP-activated protein kinase (AMPK) activation. AMPK, an energy sensor in cells, has previously been shown to be upregulated in growth hormone transgenic salmon when compared to wild type, however, whether this effect is seen across fed states is unknown. Here, we tested the hypothesis that growth hormone induces an energetic deficit in metabolic tissues, leading to constitutive AMPK activation in growth hormone transgenic salmon. This study compared AMPK activity, ATP, and glycogen, of the liver, heart, and muscle of wild-type, and growth hormone transgenic salmon either fed to satiation or a wild-type ration. The results suggest that white muscle ATP levels in growth hormone salmon are elevated in satiation and rationed conditions. In the liver, growth hormone transgenic salmon fed a rationed wild-type diet experience reductions in ATP level and glycogen. In none of the tissues examined, did AMPK activity change. Taken together, these results indicate that growth hormone transgenic salmon experience metabolic duress when not fed to satiation.

Hardness does not affect the physiological responses of wild and domestic strains of diploid and triploid rainbow trout Oncorhynchus mykiss to short-term exposure to pH 9.5.

This study examined the effects of water hardness on the physiological responses associated with high pH exposure in multiple strains of diploid and triploid rainbow trout Oncorhynchus mykiss. To accomplish this, three wild strains and one domesticated strain of diploid and triploid O. mykiss were abruptly transferred from control soft water (City of Vancouver dechlorinated tap water; pH 6·7; [CaCO3 ] < 17·9 mg l(-1) ) to control soft water (handling control), high pH soft water (pH 9·5; [CaCO3 ] < 17·9 mg l(-1) ), or high pH hard water (pH 9·5; [CaCO3 ] = 320 mg l(-1) ) followed by sampling at 24 h for physiological measurements. There was a significant effect of ploidy on loss of equilibrium (LOE) over the 24 h exposure, with only triploid O. mykiss losing equilibrium at high pH in both soft and hard water. Furthermore, exposure to pH 9·5 resulted in significant decreases in plasma sodium and chloride, and increases in plasma and brain ammonia with no differences between soft and hard water. There was no significant effect of strain on LOE, but there were significant differences between strains in brain ammonia and plasma cortisol. Overall, there were no clear protective effects of hardness on high pH exposure in these strains of O. mykiss.

High-throughput mapping of protein occupancy identifies functional elements without the restriction of a candidate factor approach.

There are a variety of in vivo and in vitro methods to determine the genome-wide specificity of a particular trans-acting factor. However there is an inherent limitation to these candidate approaches. Most biological studies focus on the regulation of particular genes, which are bound by numerous unknown trans-acting factors. Therefore, most biological inquiries would be better addressed by a method that maps all trans-acting factors that bind particular regions rather than identifying all regions bound by a particular trans-acting factor. Here, we present a high-throughput binding assay that returns thousands of unbiased measurements of complex formation on nucleic acid. We applied this method to identify transcriptional complexes that form on DNA regions upstream of genes involved in pluripotency in embryonic stem cells (ES cells) before and after differentiation. The raw binding scores, motif analysis and expression data are used to computationally reconstruct remodeling events returning the identity of the transcription factor(s) most likely to comprise the complex. The most significant remodeling event during ES cell differentiation occurred upstream of the REST gene, a transcriptional repressor that blocks neurogenesis. We also demonstrate how this method can be used to discover RNA elements and discuss applications of screening polymorphisms for allelic differences in binding.

Modelling decisions of a multidisciplinary panel for admission to long-term care.

This paper describes a modelling study of a multidisciplinary review panel which is responsible for matching levels of long-term care to the needs of older people. The study aims to understand the decision making process of the review panel and to predict placement decisions based on an applicant's attributes. Data were collected from cases notes presented to the London Borough of Merton review panel. A model predicting placement of an individual to residential home, nursing home or long-stay nursing care was built using logistic regression. and correctly predicts 78% of placement decisions. The model can be used as a means of checking the consistency of the review panel's placement decisions.

Data requirements in a model of the natural history of Alzheimer's disease.

The "natural history" of Alzheimer's disease (AD) is discussed in terms of the data required in a general, discrete-time, non-homogeneous Markov model. The proposed model differs from similar models reported in the literature because mortality is disaggregated into AD-specific mortality and competing mortality due to other causes. Data are reviewed from the literature for AD incidence, and rates of disease progression and mortality. We conduct a preliminary sensitivity analysis using the reviewed data as base-case. The model shows that survival is sensitive to the modelling assumptions concerning mortality. This observation could have important consequences for studies that assess the cost of care following therapeutic interventions.

Analysis of bed usage and occupancy following the introduction of geriatric rehabilitative care in a hospital in Huesca, Spain.

The decision support system BOMPS (Bed Occupancy Management and Planning Software) is used to document the effects on bed usage and occupancy of policy changes in the role of the provincial hospital of Huesca, Spain. A preliminary analysis of data for 1,834 hospital discharges during the period 1987 to 1996 showed that there was an increase in hospital activity. Estimates of bed usage and occupancy during a calendar year were calculated from aggregated weekly census data. Bed usage by short-stay patients increased following the introduction of rehabilitative care. Bed usage by long-stay patients decreased immediately following the introduction of rehabilitative care, however, the time between admission and census for long-stay patients decreased only after a number of years.

Rapid direct determination of HLA-DQB1 * 0301 in the whole blood of normal individuals and cancer patients by specific polymerase chain reaction amplification.

The HLA class II DQB1 * 0301 allele is present at a higher frequency in patients with malignant melanoma than in Caucasian controls. Furthermore, HLA-DQB1 * 0301 identifies a group of melanoma patients presenting with relatively advanced disease, and independently identifies a group of melanoma patients more likely to have disease recurrence. A rapid screening test for HLA-DQB1 * 0301 may be useful in clinical research involving melanoma patients. Standard molecular oligotyping for HLA class II alleles using the polymerase chain reaction (PCR)-sequence specific oligonucleotide (SSO) method is relatively expensive, labor-intensive, and involves the use of radioisotope. We therefore developed an inexpensive, rapid, non-radioactive method using sequence-specific primers, peripheral whole blood as the substrate, and strictly defined reaction conditions in a single-step PCR to allow determination of the presence or absence of genomic HLA-DQB1 * 0301. Comparison of the single-step PCR method with standard PCR-SSO oligotyping on 63 blinded samples from Caucasian melanoma patients demonstrated complete agreement between the two methods in the detection of HLA-DQB1 * 0301. Confirmatory testing in 456 additional cancer patients and healthy controls showed a sensitivity of 98.0% and a specificity of 99.4%. Single-step PCR is accurate, rapid, inexpensive, and does not require radioisotope. These advantages make it the procedure of choice for screening melanoma patients and others for the presence of the HLA-DQB1 * 0301 allele.

Association of gastric adenocarcinoma with the HLA class II gene DQB10301.

BACKGROUND & AIMS: The HLA class II gene DQB1*0301 has been linked to several cancers. This study was designed to determine if HLA-DQB1*0301 is present at altered frequency in patients with gastric, colorectal, or pancreatic adenocarcinoma. METHODS: Oligotyping for HLA-DQB1*0301 was performed for 159 Caucasian patients with 160 gastrointestinal adenocarcinomas (52 gastric, 62 colorectal, and 46 pancreatic adenocarcinomas) and compared with 260 Caucasian noncancer controls. Patients with gastric adenocarcinoma underwent extended HLA class II region oligotyping. Immunoglobulin G to Helicobacter pylori was detected by enzyme-linked immunosorbent assay. RESULTS: HLA-DQB1*0301 was more common in patients with gastric adenocarcinoma than controls (54% vs. 27%; bonferroni-corrected chi 2 P = 0.003; odds ratio, 3.2). HLA-DQB1*0301 was not associated with colorectal or pancreatic adenocarcinoma. No other HLA-DQB1 allele and no HLA-DQA1 or transporter associated with antigen processing 2 (TAP2) allele were present at altered frequency in patients with gastric adenocarcinoma. Serological evidence for H. pylori infection was less frequent in HLA-DQB1*0301-positive patients with gastric adenocarcinoma compared with HLA-DQB1*0301-negative patients (52% vs. 88%; Fisher's Exact Test; P = 0.007). CONCLUSIONS: HLA-DQB1*0301 is more common in caucasian patients with gastric adenocarcinoma than noncancer controls. The mechanism linking HLA-DQB1*0301 with gastric adenocarcinoma is not likely through increased susceptibility to H. pylori infection.

Protection against lethal Escherichia coli bacteremia in baboons (Papio anubis) by pretreatment with a 55-kDa TNF receptor (CD120a)-Ig fusion protein, Ro 45-2081.

Fusion proteins of the human 55-kDa TNF receptor extracellular domain with hinge and C2/C3 constant domains of human IgG1 or IgG3 heavy chains were tested in a primate sepsis model. Twenty-four baboons received 4.6, or 0.2 mg/kg of TNFR5-G1,3, or placebo, before the administration of a lethal dose of live Escherichia coli. Treatment with TNFR5-G1,3 decreased 5-day mortality from 88% in the placebo group to 12% in the TNFR5-G1,3-treated animals (p < 0.01 by Fisher's exact test). Treatments with TNR5-G1 and TNFR5-G3 in doses from 0.2 to 4.6 mg/kg were efficacious. Free plasma TNF was neutralized by all treatments, but inactive TNF/TNFR5-G1,3 complexes remained in circulation for prolonged periods. TNFR5-1,3 treatments attenuated the hemodynamic disturbances, reduced fluid requirements, and decreased the systemic IL-1 beta, IL-6, and IL-8 responses. In addition, TNFR5-G1,3 treatment shortened the granulocytopenia and reduced the loss of cellular TNF receptors from granulocytes. The decrease in fibrinogen concentrations and increase in prothrombin and partial thromboplastin times were significantly attenuated by TNFR5-G1,3 treatment. TNFR5-G1,3 treatment markedly attenuated the rise in plasma lactate concentration. Histologic studies of TNFR5-G1,3 revealed dose-dependent protection against tissue injury by Escherichia coli administration.

A technique for laparoscopic repair of herniation of the anterior abdominal wall using a composite mesh prosthesis.

Improved laparoscopic techniques have engendered many new gastrointestinal and other intracavity abdominal procedures. Groin hernias have also been repaired with the assistance of the laparoscope via both transperitoneal and properitoneal approaches, but less emphasis has been placed upon repair of hernias of the anterior abdominal wall. A technique for the transperitoneal, laparoscopic repair of anterior abdominal wall hernias using a composite mesh prosthesis is presented. The technique is applicable to hernias in many locations.

Planned laparoscopic repair of a spigelian hernia using a composite prosthesis.

A planned elective repair, via the laparoscope, of a spigelian hernia is described. The repair was performed using a composite mesh prosthesis consisting of a sandwich of polyester fiber mesh and polyglactin 910 mesh, sutured together with polyglactin 910 suture at the operating table before introduction. The technique is applicable to other hernias of the anterior abdominal wall.

Anti-endotoxin therapy in primate bacteremia with HA-1A and BPI.

OBJECTIVE: The in vivo neutralizing activities of an anti-lipopolysaccharide (LPS) antibody HA-1A (Centoxin [Centocor, Malvern, PA]), a human immunoglobulin M monoclonal antibody, and of bactericidal/permeability-increasing protein (BPI), an endogenously produced human LPS-neutralizing protein, were studied in a primate model of lethal Escherichia coli bacteremia. SUMMARY BACKGROUND DATA: HA-1A has been used with variable success against LPS activity in some animal models and in a recently reported clinical trial. However, no data assessing the efficacy of this agent in subhuman primates is available. Bactericidal/permeability-increasing protein is a product of polymorphomononuclear cells (PMNs) that is stored in azurophilic granules and exhibits LPS-neutralizing activity in vitro and in some in vivo models. METHODS: Immediately after E. coli infusion and in a blinded fashion, three baboons were treated with BPI (5 mg/kg bolus infusion and 95 micrograms/kg/min infusion over 4 hr). Three animals received 3 mg/kg BW of HA-1A, whereas another three baboons received a placebo treatment. RESULTS: The BPI-treated animals demonstrated significantly (p < 0.03) lower circulating LPS-limulus amoebocyte lysate (LAL) activity compared with the control animals, but this reduction in LPS-LAL activity was not associated with improved survival. HA-1A treatment did not reduce LPS-LAL activity. However, both BPI and HA-1A treatment did attenuate the pro-inflammatory cytokine response. CONCLUSION: The current data suggests that incomplete neutralization of endotoxin activity does not alter mortality from severe bacteremia. Given the diversity of mediator production under such circumstances, a strategy of combination therapy in the form of anti-lipopolysaccharide and anticytokine treatment may be necessary to achieve optimal survival.

Nutrient modification of inflammatory mediator production.

Recent evidence suggests that dietary manipulations may influence the production of endogenous inflammatory mediators. Hence, nutritional modulation during critical illness may affect survival through mechanisms far more complex than the maintenance of mere caloric and nitrogen balance. Provision of nutrients through parenteral routes to normal volunteers promotes an exaggerated cytokine response after an endotoxin challenge. Similarly, alterations in the specific protein, lipid, and micronutrient intake in septic animal models can dramatically influence the elaboration of inflammatory mediators generated by septic stimuli. However, unlike clinical work examining the differential outcome of parenterally and enterally fed patients, the theoretical advantages of specific amino acid or lipid modulation await confirmation by controlled human investigation and clinical trials.

Changes in polymorphonuclear leukocyte surface and plasma bactericidal/permeability-increasing protein and plasma lipopolysaccharide binding protein during endotoxemia or sepsis.

OBJECTIVE: To evaluate changes in levels of polymorphonuclear leukocyte surface bactericidal/permeability-increasing protein (BPI), plasma BPI, and plasma lipopolysaccharide (LPS) binding protein (LBP) in normal human volunteers administered Escherichia coli LPS and in patients with sepsis and gram-negative infections. DESIGN: Survey; case series. SETTING: Clinical research center and surgical intensive care unit of a medical school and an associated tertiary care hospital. PATIENTS OR OTHER PARTICIPANTS: Volunteers (n = 10) screened prior to study by history and physical examination to exclude those with underlying diseases or hematologic abnormalities. Consecutive sample of surgical intensive care unit patients (n = 10) meeting criteria for sepsis syndrome with gram-negative infection. An additional patient with systemic inflammatory response syndrome but no gram-negative infection. All patients were studied on meeting the criteria. Three of the patients with sepsis syndrome and the patient with systemic inflammatory response syndrome were evaluated on recovery (approximately 25 days after initial study). Because these studies in volunteers and patients overlapped temporally, the control values were those of volunteers evaluated prior to LPS administration. No matching was employed. MEASUREMENTS AND RESULTS: Compared with controls, LPS-challenged volunteers and patients with sepsis both exhibited significant granulocytosis (P < .01) and increased concentrations of polymorphonuclear leukocyte surface BPI (P < .01) and of plasma LBP (P < .01). Plasma BPI concentrations were increased (P < .01) in volunteers following LPS administration. There was a trend toward increased concentrations of plasma BPI in patients, but this was not significant relative to controls. Maximum concentrations of plasma LBP were approximately 250- and 3000-fold higher than plasma BPI concentrations in endotoxemic volunteers and in patients, respectively. CONCLUSIONS: Circulating polymorphonuclear leukocytes increase expression of BPI in response to LPS or gram-negative sepsis. Subsequently, concentrations of plasma BPI and LBP increase. Because both LBP and BPI bind to LPS, it is suggested that endogenously derived plasma levels of BPI are likely to be inadequate to compete for LPS binding to the much more abundant LBP in the circulation.

The metabolic effects of platelet-activating factor antagonism in endotoxemic man.

OBJECTIVE: To determine if the inflammatory phospholipid platelet-activating factor (PAF) participated in the symptomatologic, metabolic, and counterregulatory hormonal responses of human endotoxemia. DESIGN: In a double-blind, placebo-controlled study, five subjects received 10 mg of the PAF antagonist Ro 24-4736 orally, while five control subjects received a placebo. Eighteen hours later, all subjects were administered 4 ng/kg of endotoxin (lipopolysaccharide) intravenously. SETTING: The Clinical Research Center of The New York Hospital-Cornell Medical Center. PARTICIPANTS: Healthy male volunteers. MAIN OUTCOME MEASURES: Repeated measurements of vital signs, symptoms, cytokine and hormone levels, resting energy expenditure, platelet aggregation, and bleeding times were performed during a 24-hour period. RESULTS: Subjects who were pretreated with the PAF antagonist experienced fewer symptoms, including rigors at 1 hour (P < .05) and myalgias at 1 through 4 hours (P < .05) after administration of lipopolysaccharide. This was in concert with a diminished peak cortisol level (668 +/- 107 vs 959 +/- 159 nmol/L in controls; P < .05), epinephrine secretion (1057 +/- 165 vs 2029 +/- 431 nmol/L in controls; P < .05), and almost complete inhibition of PAF-induced platelet aggregation ex vivo. CONCLUSIONS: These findings in the face of unaltered circulating cytokines tumor necrosis factor alpha, interleukin 1 beta, and interleukin 6, as well as the tumor necrosis factor receptor-I s, suggest that PAF may influence some endotoxin-induced, counterregulatory hormonal responses and symptoms through cytokine-independent mechanisms. This study further supports the role of PAF antagonists as an adjunct to cytokine blockade in the treatment of gram-negative sepsis.

Interleukin-1 receptor blockade improves survival and hemodynamic performance in Escherichia coli septic shock, but fails to alter host responses to sublethal endotoxemia.

The present study was undertaken to evaluate the extent to which an endogenous interleukin-1 (IL-1) response contributes to the hemodynamic and metabolic consequences of sublethal endotoxemia or lethal Gram-negative septic shock. Young, healthy baboons received either a sublethal dose of lipopolysaccharide (LPS) or an LD100 of live Escherichia coli bacteria, and one half of the animals in each group were continuously infused with IL-1 receptor antagonist (IL-1ra). Plasma IL-1 beta was not detected in this model of endotoxemia. Administration of IL-1ra had only minimal effects on the modest hemodynamic and metabolic responses to sublethal endotoxemia, and did not attenuate the plasma cytokine response. In contrast, high circulating levels of IL-1 beta (range 300-800 pg/ml) were seen during lethal E. coli septic shock. IL-1ra treatment significantly attenuated the decrease in mean arterial blood pressure (MAP) (from -72 +/- 8 to -43 +/- 6 mm Hg; P less than 0.05) and cardiac output (from -0.81 +/- 0.17 to -0.48 +/- 0.15 liter/min; P less than 0.05), and significantly improved survival from 43 to 100% at 24 h (P less than 0.05). The plasma IL-1 beta and IL-6 responses to lethal E. coli septic shock were also significantly diminished by IL-1ra treatment (P less than 0.05), whereas tumor necrosis factor-alpha (TNF alpha) concentrations were unaffected. We conclude that an exaggerated systemic IL-1 beta response is characteristic of lethal E. coli septic shock, and contributes significantly to the hemodynamic and metabolic consequences of E. coli septic shock. IL-1ra can significantly attenuate the cytokine cascade and improve survival.

Pyrazoloquinoline benzodiazepine receptor ligands: effects on schedule-controlled behavior in dogs.

The effects of diazepam and the pyrazoloquinoline benzodiazepine receptor ligands CGS8216, CGS9896, and CGS9895 on schedule-controlled responding were studied in dogs. Responding was maintained under a multiple fixed-interval (FI) 5-min fixed-ratio (FR) 30 response schedule of food presentation. Diazepam (PO) produced dose-related decreases in response rates under FR component. Under the FI, rates first increased and then decreased with increasing doses of diazepam. Diazepam also produced a dose-related disruption of the temporal pattern of responding under the FI as measured by decreases in quarter-life values. CGS8216 IV produced dose-related decreases in response rates under both components. The highest oral dose of CGS8216 also decreased rates in both components. CGS8216 was approximately 100 times more potent by the IV route as compared to the oral route. CGS9896 IV had no significant effect on responding under either component of the multiple schedule. However, with increasing doses of CGS9896 PO, response rates under both components first decreased and then returned to control values. CGS9895 PO was without significant effect on responding. When CGS8216 was administered concomitantly with graded doses of diazepam, the former drug blocked the rate-decreasing effects of diazepam under the FR component, but not the rate-increasing effects of diazepam under the FI. The present results demonstrate that although these three pyrazoloquinolines are benzodiazepine receptor ligands, they do not exhibit diazepam-like effects on schedule-controlled behavior.

Behavior maintained under fixed-interval and second-order schedules by intravenous injections of endogenous noncatecholic phenylethylamines in dogs.

The effectiveness of i.v. injections of the endogenously occurring amines beta-phenylethylamine (PEA), N-methyl phenylethylamine (NMPEA) and phenylethanolamine in maintaining schedule-controlled behavior was investigated in dogs. Behavior was maintained under either a fixed-interval (FI) 5-min schedule of i.v. drug injection or a second-order FI 5-min schedule where every fifth response (FR 5) resulted in a 2-sec visual stimulus and the first FR 5 completed after the interval elapsed resulted in both the visual stimulus and i.v. drug injection [FI 5-min (FR 5:S)]. Experimental sessions, with 10 intervals per session, were conducted 5 days/week. Each drug injection was followed by a 5-min timeout period to minimize the direct effects of the drugs on responding. As the dose per injection increased, rates of responding maintained under both schedules by PEA and NMPEA first increased and then decreased. When saline was substituted for drug, responding occurred at very low rates. PEA and NMPEA were approximately equieffective and equipotent in maintaining responding under the FI 5-min schedule. PEA maintained somewhat higher rates under the FI 5-min (FR 5:S) schedule; rates maintained by NMPEA under the second-order schedule were comparable to those maintained under the simple FI schedule. Phenylethanolamine failed to consistently maintain responding under either schedule. Injections of PEA and NMPEA controlled overall patterns of positively accelerated responding under both schedules, whereas the local pattern of responding under the second-order schedule was under the control of both the brief stimulus presentations as well as drug delivery.(ABSTRACT TRUNCATED AT 250 WORDS)

Computed tomography of the gastroesophageal junction.

Computed tomography (CT) of the chest and abdomen has proved to be helpful in the preoperative staging of both esophageal and gastric carcinoma. The gastroesophageal junction however, is a difficult area to evaluate as variations in normal anatomy may mimic pathological processes. Pseudomasses at the gastroesophageal junction can be confused with neoplasm. The CT appearance of the GE junction was evaluated in 150 normal patients. CT scans were also performed on 15 patients with carcinoma involving the GE junction. Twenty cases of benign diseases of the GE junction were also studied by CT. Anatomy--The normal anatomy of the gastroesophageal junction will be illustrated with both line diagrams and CT images. The hepatogastric ligament and the caudate lobe of the liver will be demonstrated and their use in locating the GE junction will be shown. Technique--A short segment describing the appropriate technique for CT of the gastroesophageal junction will follow. The use of oral and intravenous contrast will be discussed. The need for distension of the stomach with effervescent agents and oral contrast as well as the use of decubitus and prone positioning will be emphasized when a mass-like density is seen at the GE junction. Examples will be provided. A pseudomass at the GE junction on a supine CT will be shown that disappears with distension and decubitus scanning. This will be used to lead into the next section on neoplasm in which the first example will have an identical appearance on supine CT images. Neoplasm--The relative incidence of gastric adenocarcinoma and esophageal squamous cell carcinoma at the GE junction will be briefly reviewed. The similar CT appearance of the neoplasms will be described and liberally illustrated. Metastatic involvement of lymph nodes adjacent to the GE junction will also be shown. The staging classification for CT evaluation of GE neoplasms will be reviewed. The utility of preoperative staging of esophageal and gastric neoplasms will be briefly reviewed and applied to the GE junction. Our series of patients with cancer of the GE junction will be discussed. The importance of the CT detection of criteria of inoperability will be demonstrated with examples of metastatic involvement of the liver and lymph nodes as well as direct invasion of adjacent organs. Benign Disease--Examples of benign stricture, hiatal hernia, and achalasia will be illustrated. Our cases where CT scans helped rule out a malignant process that had been suggested on barium studies will be reviewed. Summary and Conclusions--Important points of technique, normal anatomy, benign and malignant disease will be briefly reviewed.