Isolation, characterization and molecular identification of a novel aquareovirus that infects endangered fountain darter Etheostoma fonticola.

The fountain darter Etheostoma fonticola (FOD) is a federally endangered fish listed under the US Endangered Species Act. Here, we identified and characterized a novel aquareovirus isolated from wild fountain darters inhabiting the San Marcos River. This virus was propagated in Chinook salmon embryo (CHSE)-214, rainbow trout gonad-2 and fathead minnow cells at 15°C. The epithelioma papulosum cyprini cell line was refractory at all temperatures evaluated. High throughput sequencing technologies facilitated the complete genome sequencing of this virus utilizing ribosomal RNA-depleted RNA extracted from infected CHSE-214 cells. Conventional PCR primer sets were developed for the detection and confirmation of this virus to assist diagnostic screening methods. Phylogenetic analysis suggests this virus belongs to the Aquareovirus A genus. This research provides requisite initial data critical to support hatchery and refugia biosecurity measures for this endangered species.

Disease dynamics of Montipora white syndrome within Kaneohe Bay, Oahu, Hawaii: distribution, seasonality, virulence, and transmissibility.

We report on an investigation of Montipora white syndrome (MWS), which is a coral disease reported from Hawaii, U.S.A., that results in tissue loss. Disease surveys of Montipora capitata within Kaneohe Bay (Oahu) found colonies that were affected by MWS on 9 reefs within 3 regions of Kaneohe Bay (south, central, north). Mean MWS prevalence ranged from 0.02 to 0.87% and average number of MWS cases per survey site ranged from 1 to 28 colonies. MWS prevalence and number of cases were significantly lower in the central region as compared to those in the north and south regions of Kaneohe Bay. There was a positive relationship between host abundance and MWS prevalence, and differences in host abundance between sites explained approximately 27% of the variation in MWS prevalence. Reefs in central Kaneohe Bay had lower M. capitata cover and lower MWS levels. MWS prevalence on reefs was neither significantly different between seasons (spring versus fall) nor among 57 tagged colonies that were monitored through time. MWS is a chronic and progressive disease causing M. capitata colonies to lose an average of 3.1% of live tissue mo(-1). Case fatality rate was 28% after 2 yr but recovery occurred in some colonies (32%). Manipulative experiments showed that the disease is acquired through direct contact. This is the first study to examine the dynamics of MWS within Hawaii, and our findings suggest that MWS has the potential to degrade Hawaii's reefs through time.

Coral larvae conservation: physiology and reproduction.

Coral species throughout the world's oceans are facing severe environmental pressures. We are interested in conserving coral larvae by means of cryopreservation, but little is known about their cellular physiology or cryobiology. These experiments examined cryoprotectant toxicity, dry weight, water and cryoprotectant permeability using cold and radiolabeled glycerol, spontaneous ice nucleation temperatures, chilling sensitivity, and settlement of coral larvae. Our two test species of coral larvae, Pocillopora damicornis (lace coral), and Fungia scutaria (mushroom coral) demonstrated a wide tolerance to cryoprotectants. Computer-aided morphometry determined that F. scutaria larvae were smaller than P. damicornis larvae. The average dry weight for P. damicornis was 24.5%, while that for F. scutaria was 17%, yielding osmotically inactive volumes (V(b)) of 0.22 and 0.15, respectively. The larvae from both species demonstrated radiolabeled glycerol uptake over time, suggesting they were permeable to the glycerol. Parameter fitting of the F. scutaria larvae data yielded a water permeability 2 microm/min/atm and a cryoprotectant permeability = 2.3 x 10(-4) cm/min while modeling indicated that glycerol reached 90% of final concentration in the larvae within 25 min. The spontaneous ice nucleation temperature for F. scutaria larvae in filtered seawater was -37.8+/-1.4 degrees C. However, when F. scutaria larvae were chilled from room temperature to -11 degrees C at various rates, they exhibited 100% mortality. When instantly cooled from room temperature to test temperatures, they showed damage below 10 degrees C. These data suggest that they are sensitive to both the rate of chilling and the absolute temperature, and indicate that vitrification may be the only means to successfully cryopreserve these organisms. Without prior cryopreservation, both species of coral settled under laboratory conditions.

Morphine and gastroduodenal motility.

This study investigated the effects of intravenous morphine on gastric antral and duodenal motility in healthy volunteers. Intravenous morphine (both infusion and bolus) increased duodenal motility, typically as bursts of contractions similar to phase III of the migrating motor complex. Intravenous infusion of morphine 40 microg/kg/hr rapidly increased duodenal motility in nine of 10 subjects; in eight it was phase III-like. Intravenous infusion of naloxone (40 microg/kg/hr) blocked this effect of morphine infusion in five of six subjects. Morphine bolus injection (5-20 microg/kg) in six subjects (30-42 min following a spontaneous phase III) induced further duodenal phase III-like activity; also, morphine bolus injection (5-20 microg/kg) in five subjects (30-42 min following a liquid meal) induced duodenal phase III-like activity. Atropine (10 microg/kg intravenously) was able to prevent the action of morphine (both intravenous infusion and intravenous bolus injection) in inducing this phase III-like activity. These observations show: (1) morphine in very low dose is able to stimulate maximal duodenal contractility; (2) the motility response is typically phase III-like; and (3) morphine acts on opioid receptors to initiate this phase III-like activity, with the effect blocked by antimuscarinic drugs.

Analysis of the effects of Perkinsus marinus proteases on plasma proteins of the Eastern oyster (Crassostrea virginica) and the Pacific oyster (Crassostrea gigas).

We employed two in vitro buffer systems to determine the potential pathogenic effects of Perkinsus marinus serine proteases on the plasma proteins of the eastern oyster (Crassostrea virginica) and the Pacific oyster (Crassostrea gigas). Specifically, this study characterized the oyster plasma protein targets of P. marinus proteases. Additionally, protease-specific inhibitory activity was revealed upon comparison of artificial (PBS) and endogenous (plasma-based) diluents employed during protease digestions. It was found that a C. virginica plasma protein of approximately 35 kDa was eliminated when a standard buffer (PBS) was used as a diluent; however, this protein was preserved when a low-molecular-weight, plasma-based, diluent was used. The results strongly indicate that low-molecular-weight inhibitors of P. marinus proteases are present in oyster plasma. A control (nonparasitic) serine protease, alpha-chymotrypsin, was employed to ascertain the specificity of the protease inhibitors. Although alpha-chymotrypsin possesses ample proteolytic activity for C. virginica plasma proteins, the anti-proteases could specifically inhibit only P. marinus proteases. Such specificity of anti-protease activity is not uncommon among low-molecular-weight serine proteases. The hemolymph target protein was isolated by 2D electrophoresis and isoelectrically isolated for further characterization by N-terminal amino acid sequencing.

Successful reversal of Barrett's esophagus with multipolar electrocoagulation despite inadequate acid suppression.

BACKGROUND: Barrett's epithelium is a risk factor for esophageal cancer. In this study we attempted to reverse Barrett's epithelium in an acid-reduced environment by using multipolar electrocoagulation. METHODS: All patients had specialized columnar epithelium of at least 2 cm in length. Patients with dysplasia on successive screening examinations were excluded. Esophageal motility and 24-hour pH studies were performed before therapy and at 9 and 18 weeks. All patients received lansoprazole 30 mg twice a day regardless of pH study results. Multipolar electrocoagulation was applied to one side of the esophagus (2 to 3 cm per session) with the contralateral side serving as control. Biopsies were obtained from each 2 cm length at each endoscopy and reviewed by a single, blinded pathologist. At the 9-week evaluation of the treatment side, all patients elected to undergo multipolar electrocoagulation therapy for the control side. RESULTS: Twenty-seven patients completed the study: 21 men and 6 women, ages 33 to 81 years, length of specialized columnar epithelium 2 to 10 cm. Endoscopy at the 18-week follow-up showed normal mucosa (n = 16), residual small (< 3 mm) islands or tongue-shaped extensions of apparent specialized columnar epithelium (n = 7), untreated distal strips (< 5 mm) of apparent specialized columnar epithelium at the gastroesophageal junction (n = 3), and persistent specialized columnar epithelium (n = 1). Histologically, only 5 of 27 patients had residual specialized columnar epithelium at week 18. Of 16 patients with a normal endoscopic appearance, one still had specialized columnar epithelium on biopsy. While on lansoprazole, pH studies were normal in 15 of 26 patients at 9 weeks. Of 22 patients with histologic evidence of specialized columnar epithelium reversal, 10 had persistently abnormal 24-hour pH studies (pH < 4, 6.1% to 33.7% of total time). Four of five patients with residual specialized columnar epithelium on biopsy had persistent reflux. Side effects of multipolar electrocoagulation were transient, and treatment did not adversely alter motility patterns. CONCLUSIONS: Multipolar electrocoagulation therapy can induce re-epithelialization with normal squamous mucosa in most patients with specialized columnar epithelium and does not adversely affect esophageal motility. Adequate acid suppression is not mandatory for therapy to be effective in the short term.

Prolonged recovery stay and unplanned admission of the pediatric surgical outpatient: an observational study.

STUDY OBJECTIVE: To determine the incidence of and reasons for prolonged length of stay in the postanesthesia care unit and unplanned hospital admissions of children scheduled for outpatient surgery. DESIGN: Prospective, observational cohort study. SETTING: C. S. Mott Children's Hospital, a tertiary care setting. PATIENTS: 168 ASA physical status I, II, and III children (birth to 18 years), 130 of whom experienced a prolonged length of stay and 61 who had an unplanned hospital admissions. MEASUREMENTS AND MAIN RESULTS: 3.9% of annual outpatient population experienced a prolonged length of stay, and 1.9% had an unplanned outpatient admission. Prolonged length of stay was most commonly due to postoperative nausea and vomiting (19%) or respiratory complications (16%), whereas unplanned hospital admissions were primarily for respiratory or surgical reasons (32% and 30%, respectively). Higher ASA status had a significant direct relationship with the incidence of unplanned outpatient admission and respiratory complications. Although most families were satisfied with the length of their child's care, 28% of parents whose children were sent home after a prolonged length of stay would have preferred a short hospital admission, and 16% of parents of children with an unplanned hospital admission would have preferred a longer stay in recovery and discharge home. CONCLUSION: Prolonged length of stay and unplanned hospital admissions were uncommon outcomes following pediatric outpatient surgery. However, the impact of such outcomes on hospital staffing and family convenience may have implications related to cost containment and patient satisfaction.

Evaluation of simethicone for the treatment of postoperative abdominal discomfort in infants.

STUDY OBJECTIVE: To determine whether abdominal discomfort is a cause for distress symptoms in infants following administration of inhalational anesthesia, and to evaluate the effectiveness of simethicone in treating this discomfort. DESIGN: Randomized, double-blinded study. SETTING: Large tertiary care, university-based medical center. PATIENTS: 175 ASA physical status I and II infants under 28 months of age who underwent an inhalational anesthetic for a variety of procedures that were expected to cause relatively little pain. INTERVENTIONS: Children were assessed for the presence of postoperative abdominal discomfort, and, if evident, were randomly given either simethicone or placebo in a double-blinded fashion. MEASUREMENTS AND MAIN RESULTS: Abdominal discomfort was measured using the Faces Legs Activity Cry and Consolability (FLACC) Behavioral Pain Scale. Scores were recorded pre-drug; at 10, 20, and 30 minutes following drug administration; and at discharge. If discomfort had not resolved within 15 minutes after the drug was given, routine analgesics or other medications were administered. Abdominal girth was measured preoperatively, on admission into the postanesthesia care unit (PACU), and at discharge. 21% of infants exhibited symptoms of abdominal discomfort postoperatively. Younger infants were at greater risk for this condition. 36 infants were given either placebo or simethicone, and of these, infants who received simethicone were comfortable earlier and required fewer rescue medications compared with placebo. There were no differences in ability to tolerate oral fluids prior to discharge or in the length of stay in the PACU. CONCLUSIONS: Simethicone is a safe and inexpensive medication that may provide anesthesiologists with an effective treatment choice for suspected postoperative abdominal discomfort in infants.

Orthotopic cardiac transplantation: evaluation with CT.

As cardiac transplantation has become widely available, computed tomography (CT) of the chest has played a useful role in the examination of patients after heart transplantation. To determine anatomic features related to the procedure, the authors evaluated 59 scans in 46 patients who had undergone orthotopic cardiac transplantation. Aortic anastomosis (seen in 98% of scans) and altered spacing between the great vessels (83%) proved to be the most common and most reliable findings. Other features including atrial anastomosis, high main pulmonary artery segment, remnant superior vena cava, and cardiac reorientation were also seen. Accurate interpretation of adenopathy, mediastinal abscess, and pericardial effusion will be enhanced in these patients through a better understanding of the cardiovascular-pericardial complex, which is afforded by CT.

Needle embolus: a unique complication of intravenous drug abuse.

We report a unique complication of intravenous drug abuse, central embolization of a needle from a peripheral site. A heroin addict dislodged a needle into the soft tissues of the forearm during self injection. Immediate surgical attempts to visualize and remove the needle were unsuccessful. On a subsequent admission, chest radiograph showed the needle in the right mid-lung field. No attempt at removal was made.

Gastroduodenal motility in a case of dystrophia myotonica.

A 53-year-old man with dystrophia myotonica developed symptoms of a chronic intestinal motility disorder. Incoordinate small bowel contractions as well as dilated segments were observed on x-ray examination. Gastroduodenal manometry in the fasted state showed contractions of reduced amplitude. A migrating motor complex occurred in the basal state. Pentagastrin and edrophonium injections increased motility; a liquid meal was followed by a normal motor response. The maximum gastric contraction rate (after pentagastrin injection) was 3.8/min. The maximum duodenal contraction rate observed at each duodenal recording site varied between 11.0 and 18.5/min. The lowest rate at each recording site was 11.1, 11.5, 11.0, and 11.2/min, while the fastest rate was 17.6, 16.6, 18.5, and 17.6/min. These maximum rates occurred independently of the rates at adjacent sites and of the drug infusions. The gastroduodenal motor abnormality in this patient thus predominantly affects smooth muscle, with the intrinsic neurons able to release acetylcholine and the muscle capable of responding to it. The results may indicate that electrical control activity in the duodenum intermittently oscillates more rapidly than normal, and when this occurs, phase-locking is absent. We conclude that in dystrophia myotonica there may be a defect in the cell membrane, which can cause more frequent electrical oscillations of the cell; alternatively, there may be a defect in cell-to-cell coupling.

Initiation of migrating myoelectric complexes in human subjects: role of duodenal acidification and plasma motilin.

The hypothesis that acid, emptied intermittently from the stomach during fasting, might initiate the duodenal phase of the migrating motor complex was tested in normal human subjects. In addition, the relationship between plasma motilin concentrations and the initiation of migrating motor complexes was examined. Migrating complexes occurred spontaneously in the absence of acid in the duodenal bulb and in the presence of duodenal bulb neutralization with sodium bicarbonate. Thus duodenal bulb acidification is not necessary for initiation of the duodenal phase of the migrating motor complexes. Furthermore, cyclical increases in plasma motilin concentrations were not closely correlated with the initiation of the gastric phase of maximal activity of the migrating motor complexes. However, motilin concentrations were decreased significantly following onset of the duodenal phase III. We conclude that neither duodenal acidification nor increases in motilin concentration are necessary to initiate migrating motor complexes in man.

Changes in plasma motilin concentration in response to manipulation of intragastric and intraduoduenal contents in man.

Immunoreactive plasma motilin concentrations were studied following a variety of stimuli in 24 healthy fasting subjects. Plasma motilin was measured by a radioimmunoassay using antibody GP 71 (J. C. Brown) and natural porcine motilin as standard. Basal motilin levels ranged from undetectable to 365 pg/mL. Antral and intraduodenal infusion of 50 mL o.1 N HCl (pH 1.2) at 5 mL/min failed to alter significantly plasma motilin levels but duodenal acid infusions at 17 mL/min caused a significant increase (70.8 +/- 29.5 pg/mL, mean +/- SEM; n = 6), maximal at 40 min. Duodenal alkalinization with 50 mL 0.3 M Tris buffer (pH 8.0) infused at 5 mL/min produced no change in plasma motilin. A mixed meal did not affect plasma motilin levels. Ingestion of 60 g fat significantly increased plasma motilin (n = 13; maximal increase 150.3 +/- 43.3 pg/mL at 30 min) but duodenal infusions of fat failed to increase plasma motilin levels. These results suggest that motilin secretion induced by fat requires that the fat be present initially within the stomach for secretion to occur. We conclude that ingested fat is a potent stimulus of motilin release. As duodenal acidification (50 mL 0.1 N HCl over 10 min) induces duodenal activity resembling migrating motor complexes but does not release motilin, our data argue against the release of motilin following duodenal acidification as a trigger for the initiation of these complexes in man.

Disorganised electrical activity in a child with idiopathic intestinal pseudo-obstruction.

This report presents the findings of investigation of a child with idiopathic intestinal pseudo-obstruction (IIP). Functional abnormalities of the smooth muscle of the gastrointestinal tract were disclosed by electrical recordings from the gut obtained after laparotomy. In vitro analysis of tissue and ultrastructure were undertaken and a possible aetiology of the disorder in this patient based on these findings is presented.

Small-bowel malabsorption and gastrointestinal malignancy.

In addition to lymphoma, there is an increased incidence of gastrointestinal carcinoma in patients with malabsorption due to celiac disease. This is frequently manifested by a loss of response to gluten withdrawal. Four such cases are described: one patient had lymphoma and the other three had cancer of the esophagus, jejunum, and pancreas, respectively. The literature indicates that carcinoma of the esophagus and small bowel is particularly common in patients with celiac disease. These findings suggest that celiac disease should be considered a premalignant condition and that such patients should undergo a regular radiographic survey to detect early cancer.