New details of the neural architecture of the salmonid adipose fin.

The adipose fin of salmonids, once widely regarded as vestigial and lacking in function, was shown to be important to swimming efficiency in juvenile brown trout Salmo trutta. Examination with confocal microscopy of adipose fins of S. trutta stained with various antibodies targeting the nervous system revealed several large nerves entering the fin and anastomosing throughout its length. The branching nerves form a plexus with specific patterns of fine terminal branches in the leading and trailing edges. A network of astrocyte-like cells (ALCs) that is linked through cell processes to nerves and structural collagen reacted positively with antibodies to glial cells. No other fish fins, including other adipose fins, have been shown to exhibit this type of neural architecture. Many vertebrate mechanoreceptors rely on collagen deformation to stimulate responses in afferent nerves; similarly, the adipose fin also may function as a mechanosensor, where passive mechanical deflection by water currents stimulates afferent nerves.

Neural network detected in a presumed vestigial trait: ultrastructure of the salmonid adipose fin.

A wide variety of rudimentary and apparently non-functional traits have persisted over extended evolutionary time. Recent evidence has shown that some of these traits may be maintained as a result of developmental constraints or neutral energetic cost, but for others their true function was not recognized. The adipose fin is small, fleshy, non-rayed and located between the dorsal and caudal fins on eight orders of basal teleosts and has traditionally been regarded as vestigial without clear function. We describe here the ultrastructure of the adipose fin and for the first time, to our knowledge, present evidence of extensive nervous tissue, as well as an unusual subdermal complex of interconnected astrocyte-like cells equipped with primary cilia. The fin contains neither adipose tissue nor fin rays. Many fusiform actinotrichia, comprising dense striated macrofibrils, support the free edge and connect with collagen cables that link the two sides. These results are consistent with a recent hypothesis that the adipose fin may act as a precaudal flow sensor, where its removal can be detrimental to swimming efficiency in turbulent water. Our findings provide insight to the broader themes of function versus constraints in evolutionary biology and may have significance for fisheries science, as the adipose fin is routinely removed from millions of salmonids each year.

Stimulation of insulin release by vasopressin in the clonal beta-cell line, HIT-T15: the role of protein kinase C.

We have studied the effects of vasopressin and tetradecanoyl phorbol acetate (TPA) on cytosolic free Ca2+ ([Ca2+]i) and insulin release in HIT-T15 beta-cells. Saturable binding of [3H] [Arg8]-vasopressin to HIT cell microsomes indicated a single class of receptors with a dissociation constant (Kd) of 2.5 nM and a total number of binding sites (Bmax) equal to 120 fmol/mg protein. [Arg8]-vasopressin (0.1-100 nM) elicited dose-dependent insulin release from HIT cells by up to 25-fold. This increase was dependent on the presence of extracellular glucose and was blocked by omission of extracellular Ca2+ or addition of verapamil. The stimulation was biphasic; a rapid but short-lived large increase in release was followed by a smaller sustained rise. Vasopressin also evoked a marked, concentration-dependent increase in [Ca2+]i which was also biphasic; an initial spike was followed by a sustained elevation. This increase also required glucose and was blocked by the absence of extracellular Ca2+ or the addition of verapamil. Pretreatment of the cells with TPA overnight to deplete protein kinase C activity did not affect the [Ca2+]i or insulin responses to vasopressin. However, short-term exposure to TPA markedly reduced glucose-induced steady-state [Ca2+]i, despite potentiating glucose-stimulated insulin release sevenfold, and blocked the [Ca2+]i increase induced by vasopressin. These inhibitory effects of TPA were absent in protein kinase C-depleted cells and were prevented by staurosporine. TPA had no significant effect on vasopressin-induced insulin release. Vasopressin did not modify the activity of ATP-sensitive K+ channels.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult respiratory distress syndrome in full-term newborns.

Since 1984, 11 newborns with severe respiratory distress have been treated whose clinical characteristics appear distinctive. Characteristics of these neonates were as follows: (1) they were full term by obstetric and neonatal criteria, (2) they had diffuse bilateral alveolar opacification on chest radiographs during the acute illness, (3) each had an acute perinatal triggering insult, (4) the neonates required continuous positive pressure ventilation for at least 48 hours with FiO2 greater than 0.50 for at least 12 hours, (5) they needed positive end-expiratory pressure of 6 cm of H2O or greater within three days of the triggering event, (6) there were no other known causes of these clinical conditions. Ten (91%) neonates had evidence of other organ dysfunction in addition to the lungs. Trials of hyperventilation in nine and tolazoline in five failed to improve oxygenation. Ten infants who underwent trials of increased positive end-expiratory pressure greater than or equal to 6 cm of H2O without other concurrent changes in ventilator settings responded with prompt increases in PaO2 (median increase 84 mm Hg, range 22 to 196 mm Hg). All 11 babies survived but required prolonged mechanical ventilation and supplemental oxygen. We suggest that adult respiratory distress syndrome can and does occur in newborns. A trial of positive end-expiratory pressure greater than or equal to 6 cm of H2O should be considered in full-term infants with severe respiratory distress in whom other causes can be excluded.

The effects of dietary n - 3 fatty acid in animal models of type 1 and type 2 diabetes.

We studied the incorporation of dietary n - 3 fatty acids from marine oils into red cell membranes, using animal models of type 1 diabetes (streptozotocin-treated Wistar rats) and type 2 diabetes (gold-thioglucose-injected CBA/T6 mice). In type 1 diabetic rats, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were higher following marine oil supplementation, and the increase in EPA was significantly greater than in non-diabetic animals (3.4 +/- 1.4% vs. 0.8 +/- 1.6%). Marine oil supplementation also resulted in higher levels of EPA and DHA in mice, but the increases were quantitatively similar with and without type 2 diabetes. Improvement in glycosylated haemoglobin following n - 3 fatty acid feeding was only found in type 2 diabetes (6.5 +/- 2.9% vs. 9.5 +/- 1.2%). This was associated with a higher plasma insulin concentration (170 +/- 40 vs. 136 +/- 41 mU/ml). The theory that n - 3 fatty acids improve insulin sensitivity would have predicted a decrease in plasma insulin levels. Our results suggest that n - 3 fatty acids may modify insulin secretion.

Exosurf rescue surfactant improves high ventilation-perfusion mismatch in respiratory distress syndrome.

OBJECTIVE: To assess ventilation/perfusion (VA/Q) mismatch of the high type, following rescue surfactant therapy for respiratory distress syndrome. HYPOTHESIS: Surfactant therapy reduces such mismatch. DESIGN: Randomized, double-blind, placebo-controlled study, assessing VA/Q with the arterial-alveolar difference of CO2 tension (P(a-A)CO2). This difference was determined with capnometry and arterial blood gases, using the equation: P(a-A)CO2 equals arterial CO2 minus alveolar CO2 partial pressure. SETTING: A level III nursery. PATIENTS: Ten intubated infants with respiratory distress syndrome. INTERVENTION: Infants were randomized to each receive two doses of surfactant or two doses of air placebo. RESULTS: P(a-A)CO2 improved after surfactant and worsened after placebo (P = 0.0021), comparing slopes of 12-hr regression lines. A similar pattern occurred with oxygenation. These changes in P(a-A)CO2 and in oxygenation were minimally correlated within the surfactant group. CONCLUSION: Exosurf rescue surfactant reduced VA/Q mismatch of the high type, over several hours.

Proximal high-frequency jet ventilation of the newborn.

During a 14-month period 11 preterm infants were treated with high-frequency jet ventilation for severe respiratory failure that had been unresponsive to conventional mechanical ventilation. Primary indications included intractable air leaks (pulmonary interstitial emphysema, pneumothorax, or both), congenital diaphragmatic hernia, and progressive pneumonia. The Sechrist 990 HFV respirator, a solenoid-driven, pulse-generated high frequency jet ventilator, was used. This device delivers a volume of gas at a controlled pressure to a jet located in the patient connector proximal to a standard single lumen endotracheal tube. The ventilator was operated in tandem with a Sechrist IV-100B ventilator to provide a sigh effect and to improve humidification. Although only 3 of 11 patients survived, 9 displayed short-term benefits from HFJV with significant reduction in mean airway pressure and improvement in mean arterial blood pressure, and no concomitant adverse effects on oxygenation or ventilation. Preliminary results suggest short-term benefits of HFJV in infants who have severe respiratory failure that is unresponsive to conventional mechanical ventilation.

A comparison of proximal and distal high-frequency jet ventilation in an experimental animal model.

High-frequency jet ventilation using either a proximal or a distal endotracheal injection site through a triple-lumen endotracheal tube was studied in 10 adult cats. The comparative effects on pulmonary gas exchange, tracheal pressure, heart rate, and blood pressure were examined for each injection site at both high (8-12 pounds per square inch [PSI] and low (5-8 PSI) jet-driving pressures in normal and lung-injured cats. Lung injury was created by modification of a surfactant washout technique previously demonstrated in rabbits. Alveolar ventilation (PaCO2) was found to be significantly better with distal than with proximal jet injection under all experimental conditions. At high jet-driving pressures, peak inspiratory pressure was higher in both normal (p = 0.03) and lung-injured cats (p = 0.002) with distal high-frequency jet ventilation. In addition, lung-injured animals were observed to have higher distal mean airway pressures at high jet-driving pressures (p less than 0.01). No differences in oxygenation were found in any circumstances. The results of this animal study suggest that distal high-frequency jet ventilation may be more effective in those situations in which improvement in alveolar ventilation is the major goal and that during proximal high-frequency jet ventilation airway pressures should be monitored as far distally as possible.

A water-sealed indirect calorimeter for measurement of oxygen consumption (VO2), carbon dioxide production (VCO2), and energy expenditure in infants.

We have developed a water-sealed infant calorimeter (IC) system which uses the techniques of closed-circuit spirometry to measure oxygen consumption (VO2) in premature and full-term infants. Carbon dioxide production (VCO2) is simultaneously calculated from the effluent mixed expired CO2 and the circulating flowrate. Respiratory Quotient (RQ) and Energy Expenditure (EE) are then calculated from the primary data. Measurement of VO2, VCO2, and calculation of RQ were +/- 5.0% of predicted values determined by burning ethyl alcohol or volume extraction and CO2 infusion in our bench model. Measurement in 11 premature infants produced mean values for VO2 and VCO2 of 8.5 +/- 2.5 ml/min/kg and 8.5 +/- 2.4 ml/min/kg, respectively. This system is noninvasive, does not interfere with infant tube feedings or iv infusions, and permits safe, long-term monitoring of the infant's metabolic activity. It allows a more exact matching or oral or intravenous feedings to the actual energy expenditure of the infants, and offers potential advantages for the nutritional management of sick infants.

Flow-synchronized ventilation of preterm infants with respiratory distress syndrome.

Asynchrony of delivered and spontaneous breaths in mechanically ventilated infants may impair gas exchange and prolong the need for assisted ventilation. We conducted a randomized, controlled trial of a patient-triggered, flow-synchronized ventilator on 30 preterm infants with respiratory distress syndrome who weighed between 1100 and 1500 gm at birth. Entry criteria included radiographic evidence of respiratory distress syndrome and the need for mechanical ventilation and surfactant replacement therapy. Patients were assigned to either conventional time-cycled, pressure-limited ventilation or patient-triggered, flow-synchronized ventilation in an assist/control mode. Otherwise clinical management was identical. Time to extubation was the primary outcome measure. Patients treated with flow-synchronized ventilation were weaned more rapidly and had a significantly shorter mean time to extubation than those treated with time-cycled, pressure-limited ventilation, 119 versus 271 hours, p = 0.0152. In addition, there was no difference in the rate of complications between the two groups. There were, however, considerable reductions in patient charges of $4344 per patient in the flow-synchronized ventilation group.

Tracheobronchial and pulmonary histopathology following conventional and high-frequency jet ventilation.

The advent of high-frequency jet ventilation (HFJV) as an alternative method of respiratory support for newborns has been implicated as a causative agent of necrotizing tracheobronchitis (NTB). We conducted a controlled trial of prolonged HFJV and conventional mechanical ventilation (CMV) in adult cats to determine effects on airway injury related to mode of ventilation and placement of the jet injector (proximal vs distal trachea). Fifteen cats were randomly assigned to either high-frequency positive pressure ventilation, proximal injection jet ventilation, or distal injection jet ventilation. The animals were ventilated for more than 33 hours. Post mortem, the trachea and respiratory tree were removed en bloc and fixed in formalin. A pathologist, unaware of the mode of ventilation, examined tracheobronchial histology and assigned scores using a technique previously described. Lung parenchymal tissue was also assessed using a similar grading system. Statistical analysis (Kruskal-Wallis analysis of variance) demonstrated no significant differences between tracheobronchial or lung parenchymal histopathology regardless of the mode of ventilation. We conclude that (1) the adult cat serves as a useful model for evaluating histopathologic effects of prolonged ventilation, (2) the etiology and pathogenesis of airway injury appears to be multifactorial, and the mode of ventilation is only one of many contributing variables, and (3) previously demonstrated differences between CMV and HFJV may be related to a much shorter duration of ventilation.

Comparison of two methods of surfactant administration and the effect on dosing-associated hypoxemia.

OBJECTIVE: The purpose of our study was to test the hypothesis that surfactant dosing through a proximal sideport adapter on an endotracheal tube leads to more dosing-associated hypoxemia compared with a method of dosing that uses a double-lumen endotracheal tube. STUDY DESIGN: Using adequate sample size to compare significant changes in O2 saturation (power > 0.8, alpha < 0.05) we enrolled 36 infants with respiratory distress syndrome in this randomized trial. A 10% change in O2 saturation was considered clinically significant. Nineteen infants received 38 doses of surfactant through the sideport adapter. Seventeen infants received 31 doses of surfactant through the nonventilation lumen of a double-lumen endotracheal tube. Two main outcome measures were assessed: time-averaged O2 saturation values 30 minutes after dosing and the largest absolute fall in O2 saturation for each patient. RESULTS AND CONCLUSIONS: Time-averaged O2 saturation measures were higher in the proximal sideport group (p = 0.02), but the magnitude of difference was probably not clinically significant. No significant difference was detected between groups when we compared largest absolute drop in O2 saturation. Secondary analyses found no effect of birth weight or dose number (second vs third dose) on either outcome measure.

Fertilization in Callochiton castaneus (Mollusca).

A fine-structural study of fertilization in Callochiton castaneus has revealed that the mechanism of sperm penetration into the egg is intermediate between the primitive condition found in members of the order Lepidopleurida and the more derived condition found in the Chitonida. C. castaneus sperm have the long needlelike nuclear filament and reduced acrosome that characterizes all Chitonida, but they have retained several plesiomorphic features such as an unspecialized mid-piece and a lack of flagellar reinforcement. As in some Lepidopleurida but unlike any Chitonida, the egg hull in this species comprises a thick, smooth jelly coat permeated by pores that permit sperm rapid access to the vitelline layer. The jelly coat is delicate and quickly dissolves when a sperm concentrate is used, suggesting that excess acrosomal enzymes may be responsible. Once the sperm have penetrated the vitelline layer, the long nuclear filament bridges the gap to cups in the egg membrane. However, once the fertilization membrane is raised, the perivitelline space exceeds the length of the nuclear filament, preventing other sperm from penetrating the egg. A fertilization cone forms around the nuclear filament of the penetrating sperm, but it does not appear to engulf the body of the sperm. Rather, the nuclear chromatin is injected into the egg as a long thread. The remaining sperm organelles are apparently abandoned on the egg surface. If this is the case, it would be a significant departure from fertilization in other molluscs and many other metazoans, in which sperm organelles, such as centrioles and mitochondria, enter the egg. New sperm and egg characters, as well as significant differences in fertilization, indicate that Callochitonidae are basal to all other members of the order Chitonida and may warrant separation as the sister taxon to the suborders Chitonina and Acanthochitonina.

Multiple paternity in Littorina obtusata (Gastropoda, Littorinidae) revealed by microsatellite analyses.

Parental identity for juvenile Littorina obtusata was determined from three egg masses by means of microsatellite DNA markers. Results confirm that the attendant adult female in each case was the dam of the offspring and that at least 4-6 males contributed to each brood. This correlates with our behavioral observations that indicated multiple copulations between the female and several males in each experimental aquarium. A significant number of offspring from each brood were sired by non-sampled males (males that had copulated with females before capture) whose sperm had been stored by the female. This is the first direct evidence of multiple paternity in the Littorinidae. Results are discussed in reference to current theories of sperm competition, male precedence, and cryptic female choice.

Hull Cupules of Chiton Eggs: Parachute Structures and Sperm Focusing Devices?

The extracellular hull of chiton eggs is often elaborated into cupules or spines that may be open or closed to the external environment. Scanning electron microscopy was used to examine the location of fertilizing sperm in eggs that had been exposed to a dilute sperm suspension to create natural fertilization or to a sperm concentrate to induce polyspermic egg penetration. The effect of cupules on sinking rates was tested in cupulous (free-spawning) and non-cupulous (brooding) species, by timing descent of eggs over a fixed distance in a large container of seawater. Densities of eggs were compared on Percoll gradients and found to be similar. It was found that hull cupules focus the sperm to specific regions of the egg surface in both brooding and free-spawning species. Furthermore, protruding cupules act as parachute structures that can significantly reduce sinking rates.