Changes in local free-living parasite populations in response to cleaner manipulation over 12 years.

Predation on parasites is an important ecological process, but few experimental studies have examined the long-term impacts on the prey. Cleaner fish prey upon large numbers and selectively feed on the larger individuals of the ectoparasitic stage of gnathiid isopods. Removal of cleaner fish Labroides dimidiatus for 1.5-12.5 years negatively affects coral reef fishes, but the mechanism is unclear. A reduction in local parasite populations or the size of individual parasites would benefit all susceptible fishes. We tested whether cleaner presence reduces local gnathiid populations using 18 patch-reefs distributed between two sites (both at Lizard Island, Great Barrier Reef) which were maintained cleaner-free or undisturbed for 12 years. Using emergence traps (1 m2), free-living gnathiid stages were sampled before and after cleaner fish were removed during the day and night, up to 11 times over the course of the experiment. There were effects of the removal in the predicted direction, driven largely by the response at one site over the other involving 200% more gnathiids, but manifested only in the daytime sampling after 4 months. There was also a main effect (36%) for the shared sample dates at both sites after 12 years. Gnathiid size occasionally differed with cleaner presence, but in no consistent way over time. Contrary to our predictions, changes in free-living gnathiid population numbers and their size structure rarely reflected the changes in fish populations and individuals observed on cleaner-free reefs. Therefore, evidence that this predator alone regulates gnathiids remains limited, suggesting other contributing processes are involved.

Predation risk influences feeding rates but competition structures space use for a common Pacific parrotfish.

In terrestrial systems it is well known that the spatial patterns of grazing by herbivores can influence the structure of primary producer communities. On coral reefs, the consequences of varied space use by herbivores on benthic community structure are not well understood, nor are the relative influences of bottom-up (resource abundance and quality), horizontal (competition), and top-down (predation risk) factors in affecting spatial foraging behaviors of mobile herbivorous fishes. In the current study we quantified space use and feeding rates of the parrotfish, Chlorurus spilurus, across a strong gradient of food resources and predator and competitor abundance across two islands with drastically different fisheries management schemes. We found evidence that while feeding rates of this species are affected by direct interference competition and chronic predation risk, space use appears to be primarily related to exploitative competition with the surrounding herbivore community. We found no evidence that predation risk influences diurnal foraging space use in this small bodied parrotfish species. Additionally, we found the influence of chronic predation risk on feeding rates of this species to be less dramatic than the results of recent studies that used model predators to measure acute behavioral responses of other species of herbivorous fishes. Our results indicate that the non-consumptive effects of predators on the foraging behaviors of coral reef herbivores may be less dramatic than previously thought.

Detecting larval export from marine reserves.

Marine reserve theory suggests that where large, productive populations are protected within no-take marine reserves, fished areas outside reserves will benefit through the spillover of larvae produced in the reserves. However, empirical evidence for larval export has been sparse. Here we use a simple idealized coastline model to estimate the expected magnitude and spatial scale of larval export from no-take marine reserves across a range of reserve sizes and larval dispersal scales. Results suggest that, given the magnitude of increased production typically found in marine reserves, benefits from larval export are nearly always large enough to offset increased mortality outside marine reserves due to displaced fishing effort. However, the proportional increase in recruitment at sites outside reserves is typically small, particularly for species with long-distance (on the order of hundreds of kilometers) larval dispersal distances, making it very difficult to detect in field studies. Enhanced recruitment due to export may be detected by sampling several sites at an appropriate range of distances from reserves or at sites downcurrent of reserves in systems with directional dispersal. A review of existing empirical evidence confirms the model's suggestion that detecting export may be difficult without an exceptionally large differential in production, short-distance larval dispersal relative to reserve size, directional dispersal, or a sampling scheme that encompasses a broad range of distances from the reserves.

The stochastic nature of larval connectivity among nearshore marine populations.

Many nearshore fish and invertebrate populations are overexploited even when apparently coherent management structures are in place. One potential cause of mismanagement may be a poor understanding and accounting of stochasticity, particularly for stock recruitment. Many of the fishes and invertebrates that comprise nearshore fisheries are relatively sedentary as adults but have an obligate larval pelagic stage that is dispersed by ocean currents. Here, we demonstrate that larval connectivity is inherently an intermittent and heterogeneous process on annual time scales. This stochasticity arises from the advection of pelagic larvae by chaotic coastal circulations. This result departs from typical assumptions where larvae simply diffuse from one site to another or where complex connectivity patterns are created by transport within spatially complicated environments. We derive a statistical model for the expected variability in larval settlement patterns and demonstrate how larval connectivity varies as a function of different biological and physical processes. The stochastic nature of larval connectivity creates an unavoidable uncertainty in the assessment of fish recruitment and the resulting forecasts of sustainable yields.

Carcinoid case presentation and discussion: the American perspective.

The rationale underlying an aggressive approach in the management of some carcinoid patients is explained and illustrated by the presented case of a middle-aged man with advanced classic typical midgut carcinoid. The patient exhibited somatostatin receptor scintigraphy-positive massive liver metastases, carcinoid syndrome, severe tricuspid and pulmonic cardiac valve disease with congestive heart failure, ascites and malnutrition. He had been treated for several years with supportive medications and biotherapy including octreotide and alpha interferon but his tumor eventually progressed and his overall condition was markedly deteriorated when he first sought more aggressive treatment. This consisted of prompt replacement of both tricuspid and pulmonic valves, followed by hepatic artery chemoembolus (HACE) injection and then surgical tumor debulking including excision of the primary tumor in the small intestine. In addition, radiofrequency ablation was utilized to reduce the volume of metastases in the liver. Prophylactic cholecystectomy was also performed and a biopsy of tumor was submitted for cell culture drug resistance testing. This was followed by systemic chemotherapy utilizing the drug (docetaxel) which the in vitro studies suggested as most likely to be effective. His excellent response to this succession of treatments exemplifies the successful application of aggressive sequential multi-modality therapy.

Dandruff has an altered stratum corneum ultrastructure that is improved with zinc pyrithione shampoo.

Transmission electron microscopy of scalp tape strips indicates that dandruff scalp possesses abnormal stratum corneum (SC) ultrastructure that is normalized by treatment with small-particle zinc pyrithione (ZPT). Similar abnormalities occur throughout the scalp of those with dandruff, even where no flaking is present. SC abnormalities are consistent with hyperproliferation, including parakeratosis, lipid droplets within corneocytes, few desmosomes, corneocyte membrane interdigitation, and excessive disorganized intercellular lipid. Reversal of SC abnormalities would require treatment of the cause(s) of dandruff, not merely flake removal. A protocol was developed to quantify scalp structural abnormalities by scoring cells from scalp tape strips for yeast number, amount of intercellular lipid, normal intercellular lipid structures, prevalence of intracellular lipid droplets, parakeratotic corneocytes, and corneocyte interdigitation. This protocol was used to compare dandruff and normal SC to dandruff SC treated with either commercial ZPT-containing shampoo or a placebo. Treatment with commercial ZPT shampoo significantly returned SC ultrastructure to normal, suggesting control of the cause of dandruff.

Historical overfishing and the recent collapse of coastal ecosystems.

Ecological extinction caused by overfishing precedes all other pervasive human disturbance to coastal ecosystems, including pollution, degradation of water quality, and anthropogenic climate change. Historical abundances of large consumer species were fantastically large in comparison with recent observations. Paleoecological, archaeological, and historical data show that time lags of decades to centuries occurred between the onset of overfishing and consequent changes in ecological communities, because unfished species of similar trophic level assumed the ecological roles of overfished species until they too were overfished or died of epidemic diseases related to overcrowding. Retrospective data not only help to clarify underlying causes and rates of ecological change, but they also demonstrate achievable goals for restoration and management of coastal ecosystems that could not even be contemplated based on the limited perspective of recent observations alone.

Characterization of vernix caseosa: water content, morphology, and elemental analysis.

Recent studies have prompted interest in the use of epidermal barrier creams as protective biofilms for very low birthweight preterm infants. The key to understanding the role of epidermal barrier films is an elucidation of their interaction with water and a basic knowledge of their composition. In this study, we investigated the morphologic properties and elemental composition of the naturally occurring biofilm, vernix caseosa. This biofilm is typically lacking in preterm infants and its production coincides in utero with terminal differentiation of the epidermis and formation of the stratum corneum. Significantly, vernix (80.5+/-1.0% H2O) had a much higher water content than other barrier creams (Eucerin: 17.1+/-0.6%, Aquaphor: 0.33+/-0.03%, Ilex: 0.19+/-0.02%, petrolatum: 0.03+/-0.01%; all p<0.05). Phase contrast microscopy of vernix showed multiple cellular elements with nucleic "ghosts" embedded in a putative lipid matrix. Transmission electron microscopy revealed flattened structures approximately 1-2 microm in thickness with distinct cellular envelopes indicative of differentiated corneocytes. Compared with mature corneocytes in adult stratum corneum, vernix corneocytes appeared swollen, the density of the keratin filaments was less, and there was a relative lack of tonofilament orientation. Cryofractured specimens were examined by cryoscanning electron microscopy with subsequent elemental localization by X-ray beam analysis. The findings indicate the high water content of vernix is largely compartmentalized within fetal corneocytes. These results are consistent with the novel view of vernix as a "fluid phase" stratum corneum consisting of a hydrophobic lipid matrix with embedded fetal corneocytes possessing unique biomechanical and water-binding properties.

Relative fitness components measured with competitive PCR.

In mating systems with sperm competition, paternity is frequently established with modern DNA techniques. These methods are often expensive and cumbersome, and can be especially difficult for highly fecund species. An additional objective of many paternity studies is to discover the relationship between sperm number and paternity. We present here a competitive polymerase chain reaction (PCR) protocol, coupled with the use of an automated sequencer, that has two functions: (i) to measure directly relative sperm output of males in sperm competition; and (ii) to estimate paternity distributions of large numbers of offspring simultaneously. Our technique was calibrated using a microsatellite locus of the bluehead wrasse, Thalassoma bifasciatum, with the result that product ratio after competitive PCR accurately reflected the initial template proportions of known mixtures of DNA. When we applied our technique to multiple larvae of separate mating events we found that paternity distributions estimated with the competitive PCR technique closely matched the estimates derived from the traditional method of pooling paternity data from individual larvae. Finally, we compared paternity of these spawns with relative sperm contribution estimates. This comparison suggests that ejaculate size alone does not predict a male's proportion of paternity within the group.

Hypothalamic arginine vasotocin mRNA abundance variation across sexes and with sex change in a coral reef fish.

Gonadal hormones are important mediators of sexual and aggressive behavior in vertebrates. Recent evidence suggests that the peptide hormones arginine vasotocin (AVT) and its mammalian homologue arginine vasopressin (AVP) often critically mediate these gonadal hormone effects on behavior and have direct influences on behavioral variation. Behavioral differences between sexes, across reproductive states, and even among closely related species are correlated with differences in central AVT/AVP systems in many species. We report differences in hypothalamic AVT mRNA levels between distinct alternate male phenotypes and with female-to-male sex change in the bluehead wrasse (Thalassoma bifasciatum), a teleost fish. The aggressively dominant and strongly courting male phenotype has greater numbers of AVT mRNA producing cells in the magnocellular preoptic area of the hypothalamus than females. Levels of AVT mRNA within these cells in dominant males are also approximately three times female levels whereas the non-aggressive male phenotype has AVT mRNA levels approximately twice female levels. Behavioral sex change is very rapid in this species and is not dependent on the presence of gonads. Conversely, rapid increases in sexual and aggressive behavior during sex change are closely paralleled by approximate fourfold increases in hypothalamic AVT-mRNA levels. The behavioral plasticity shown by bluehead wrasses in response to social environment might be mediated in part by a neuropeptide, AVT, with changes in the gonads and gonadal hormones as the result rather than the cause of behavioral dominance.

Water disrupts stratum corneum lipid lamellae: damage is similar to surfactants.

Using electron microscopy, we investigated the effect of (i) a dilute surfactant and of water alone on the ultrastructure of stratum corneum lipids in pig skin exposed in vitro at 46 degrees C, and (ii) of water alone on human skin exposed in vivo at ambient temperature. For pig skin, the surfactant sodium dodecyl sulfate disrupts stratum corneum intercellular lamellar bilayers, leading to bilayer delamination and "roll-up" in a water milieu after 1 h, extensive bilayer disruption after 6 h, and nearly complete dissociation of corneocytes after 24 h. Corneodesmosomes show progressive degradation with exposure time. Water alone also disrupts the stratum corneum, but with a slower onset. Alterations in intercellular lamellar bilayers, but not intercellular lamellar bilayer roll-up, are detected after 2 h. Intercellular lamellar bilayer roll-up occurs after 6 h. Extensive dissociation of corneocytes occurs after 24 h of water exposure. Unlike sodium dodecyl sulfate, water exposure results in the formation of amorphous intercellular lipid. Corneodesmosome degradation parallels intercellular lamellar bilayer disruption; calcium appears to offer some protection. Similar disruption of intercellular lamellar bilayers occurs in human skin in vivo at ambient temperature. Our studies show that water can directly disrupt the barrier lipids and are consistent with surfactant-induced intercellular lamellar bilayer disruption being due at least in part to the deleterious action of water. Intercellular lamellar bilayer disruption by water would be expected to enhance permeability and susceptibility to irritants; accordingly, increased attention should be given to the potential dangers of prolonged water contact. For common in vitro procedures, such as skin permeation studies or isolation of stratum corneum sheets, exposure to water should also be minimized.

Bismuth-mediated disruption of the glycocalyx-cell wall of Helicobacter pylori: ultrastructural evidence for a mechanism of action for bismuth salts.

The mechanism of bismuth's bactericidal activity against Helicobacter pylori was investigated using transmission electron microscopy (TEM) and analytical electron microscopy (AEM); time-kill kinetic methods evaluated the effect of excess divalent cations. TEM analysis of untreated H. pylori revealed a normal morphology. In contrast, H. pylori exposed to bismuth salts had swollen, distorted cells with membrane-cell wall blebbing and a cytoplasm containing electron-dense, sometimes crystalline aggregates. By AEM, swollen cells contained bismuth at the cell periphery, whereas bacillary forms contained cytoplasmic bismuth localizations. Time-kill studies showed that the bactericidal activity of bismuth could be prevented by pretreatment with divalent cations. The effects of bismuth salts on the glycocalyces-cell walls of H. pylori with reversal of bactericidal activity by divalent cations are identical to those produced by other polycationic agents on various gram-negative bacilli. We conclude that disruption of the glycocalyces-cell walls of H. pylori is one mechanism of action for bismuth salts.

Behavioural sex change in the absence of gonads in a coral reef fish.

It is an axiom of vertebrate behavioural endocrinology that full expression of a male behavioural phenotype depends on testicular influences during development, in adulthood, or both. Sex change in fishes challenges this necessity: behavioural changes are often rapid and greatly precede gonadal changes. However, steroid hormones can have fast actions on the nervous system, so gonadal influences on behavioural sex change cannot be excluded based solely on the speed of these changes. We report that surgical gonad removal does not prevent or discernibly alter female-to-male behavioural sex change in a protogynous coral reef fish. Male behaviour assumption is instead purely dependent on attaining social dominance. This is the first example of a vertebrate fully expressing a male behavioural phenotype without current or previous exposure to a functioning testis or testicular products.

An 18-year follow-up of primary hepatic carcinoid with carcinoid syndrome.

Primary hepatic carcinoid is extremely rare. Although one of the 18 previously reported cases was accompanied by clinical features of carcinoid syndrome, no patient manifested these features as the presenting complaint, as was true in our case. During the 18 years this patient has been followed, she has been treated with most of the major therapeutic methods, including systemic chemotherapy, hepatic artery chemoembolus injection, extended right hepatic lobectomy, and, eventually, more systemic chemotherapy and octreotide. She continues to be nearly asymptomatic and is still working. We present the results of extensive chemical and hormonal assays, briefly summarize the primary hepatic carcinoids reported previously, and review therapy of this disease.

Sexual conflict: males with highest mating success convey the lowest fertilization benefits to females.

In natural populations of a coral reef fish (the bluehead wrasse, Thalassoma bifasciatum), males with the highest daily mating success produce the fewest sperm per mating, and this is reflected in significantly lower fertilization rates. The average amount released by males in pair-mating was 3.3 x 10(6) spermatozoa, resulting in a fertilization rate of 96%. Sperm released per spawn declined with increasing mating success, so that females mating with the most successful males had less than 93% of their eggs fertilized. It is unlikely that high mating-success males are physiologically incapable of increasing sperm production, because younger males with different mating strategies have absolutely larger testes and higher daily sperm output. Feeding experiments suggest that high-success males are diverting energy from gamete production to other fitness-enhancing activities such as mate guarding. Females incur the cost of low sperm release by having fewer of their eggs fertilized. There are no obvious compensatory benefits to females from mating with high-success males.

Corneocytes undergo systematic changes in element concentrations across the human inner stratum corneum.

Using analytical electron microscopy of freeze-dried cryosections, physiologic elements were visualized within individual cells across the human inner stratum corneum. Human corneocytes undergo systematic changes in element composition as they advance through this region. Phosphorus is largely excluded from the stratum corneum, undergoing a precipitous drop in concentration at the granular/stratum corneum interface. The cellular potassium concentration has a profile similar to that of phosphorus but with a slower decline, thus migrating further into the stratum corneum. In contrast, the cellular chloride concentration increases in the innermost corneocyte layer, increases further in the subsequent layer or two (as potassium declines), and then decreases to values comparable to those in the innermost corneocyte. The cellular sodium concentration (per unit volume of tissue) is relatively unaltered in transit across the inner stratum corneum. The initial potassium and chloride movements are oppositely directed and have the appearance of creating an electrical charge imbalance. The position-dependent alterations in corneocyte elemental composition may reflect sequential stages of chemical maturation occurring intracellularly during stratum corneum transit, an example of which is the breakdown of filaggrin that occurs over this same region of the inner stratum corneum.

Octreotide treatment of carcinoid hypertensive crisis.

In the severe crisis of carcinoid syndrome the flush is usually accompanied by hypotension and occasionally shock. Injection of octreotide, the long-acting analog of somatostatin, usually prevents or aborts this vasomotor reaction. A small minority of carcinoid syndrome patients manifest hypertension during their crises and little has been reported in the literature on their management. We present the first case reports of the response of patients with hypertensive carcinoid crisis to treatment with octreotide. The world literature contains reports of 20 prior cases of hypertensive carcinoid crises occurring in association with the stress of surgery and anesthesia. Review of these cases reveals no common feature, other than hypertension, that might clearly distinguish them from the typical hypotensive carcinoid syndrome patient. It is hypothesized that the mechanism of action of octreotide correcting the blood pressure changes in all carcinoid crises is via its known inhibition of vasomotor product release from the tumor and blocking receptors for these substances. We suggest that hypertensive as well as hypotensive carcinoid crises respond to octreotide and that this agent should be considered for prophylactic and emergency use in all carcinoid syndrome patients prior to and during anesthesia and surgery.

Analytical electron microscopy of chlorhexidine-induced tooth stain in humans: direct evidence for metal-induced stain.

Using analytical electron microscopy, we directly tested the hypothesis that transition metals and sulfur are the cause of chlorhexidine-induced tooth stain. Plaque scraped from the teeth of water-treated individuals or chlorhexidine-treated "non-stainers" was non- or lightly-stained and contained low concentrations of sulfur and transition metals. Tooth scrapings from heavily-staining chlorhexidine-treated individuals consisted of distinct unstained and stained regions. The stained regions were organic but were in close proximity to mineralized areas. Enhanced levels of sulfur and transition metals, particularly iron, were found in stained regions, whereas unstained regions contained low sulfur and metal levels similar to the water-treated or non-staining individuals. Excluding decreased mineralization, the major elemental change in heavily-stained plaque was an increase in sulfur, and to a lesser extent, iron and other transition metals. Sulphur and iron levels were directly correlated. Following chlorhexidine treatment with iron supplementation, staining was enhanced, the Fe/S ratio increased, and sulfur and iron remained correlated. These data support the hypothesis that transition metals and sulfur are the cause of chlorhexidine-induced tooth stain. The data are consistent with chlorhexidine treatment altering the incorporation into plaque of a natural sulfur-containing organic component of saliva or bacteria. This natural component appears to readily interact with transition metals, particularly iron, producing stained material.