Morbidity in California giant salamander (Dicamptodon ensatus Eschscholtz, 1833) caused by Euryhelmis sp. Poche, 1926 (Trematoda: Heterophyiidae).

In the fall of 2021, California Department of Fish and Wildlife reported larval and adult California giant salamanders (Dicamptodon ensatus Eschscholtz, 1833) with skin lesions at multiple creeks in Santa Clara and Santa Cruz Counties, California, USA. Field signs in both stages included rough, lumpy textured skin, and larvae with tails that were disproportionately long, flat, wavy, and flaccid. Presence of large-bodied larvae suggested delayed metamorphosis, with some larvae having cloudy eyes and suspected blindness. To determine the cause of the disease, three first-of-the-year salamanders from one location were collected, euthanized with 20% benzocaine, and submitted for necropsy to the U.S. Geological Survey, National Wildlife Health Center. Upon gross examination, all salamanders were emaciated with no internal fat stores, and had multiple pinpoint to 1.5-mm diameter raised nodules in the skin over the body, including the head, gills, dorsum, ventrum, all four limbs, and the tail; one also had nodules in the oral cavity and tongue. Histologically all salamanders had multiple encysted metacercariae in the dermis, subcutis, and skeletal muscles of the head, body, and tail that were often associated with granulomatous and granulocytic inflammation and edema. A small number of encysted metacercariae or empty cysts were present in the gills with minimal inflammation, and rarely in the kidney with no associated inflammation. Morphology of live metacercariae (Trematoda: Heterophyiidae), and sequencing of the 28S rRNA gene identified a species of Euryhelmis (Poche, 1926). Artificial digestion of a 1.65 g, decapitated, eviscerated carcass yielded 773 metacercariae, all of similar size and morphology as the live specimens. Based on these findings, the poor body condition of these salamanders was concluded to be due to heavy parasite burden. Environmental factors such as drought, increased temperature, and overcrowded conditions may be exacerbating parasite infections in these populations of salamander.

Automated, cryogen-free headspace-trap with gas chromatography-mass spectrometry analysis of ethylene oxide and 2-chloroethanol as residual fumigants in foods.

Ethylene oxide (EtO), although banned for use, is still being detected in foodstuffs that have been fumigated to eradicate pests during storage and transport. Residual levels over the European Union's (EU) maximum residue limit (MRL) pose severe health concerns. Recent detection of EtO and its by-product 2-chloroethanol (2-CE) at alarming levels have led to product recalls throughout the EU. Here, a simple, automated headspace (HS)-trap method for the simultaneous determination of EtO and its derivative 2-CE by gas chromatography-mass spectrometry (GC-MS) at the required MRL of ≤ 0.05 mg/kg has been implemented. Syringe-based HS combined with backflushed trapping technology provided enrichment of multiple extractions from the same sample vial (known as multi-step enrichment or MSE®) to increase sensitivity for EtO and 2-CE analysis by GC-MS using single-ion-monitoring (SIM) mode. Method detection limits (MDLs) of 0.00059 mg/kg and 0.00219 mg/kg for EtO and 2-CE, respectively, were obtained without the need for manual handling, solvent extraction or derivatization methods. Recoveries were shown to average (n = 5) at 98% and 107% for EtO and 2-CE, respectively, and the reproducibility was <10% for both compounds.

What lies beneath: White matter microstructure in pediatric myalgic encephalomyelitis/chronic fatigue syndrome using diffusion MRI.

Recent studies in adults with myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) suggest that changes in brain white matter microstructural organization may correlate with core ME/CFS symptoms, and represent a potential biomarker of disease. However, this has yet to be investigated in the pediatric ME/CFS population. We examined group differences in macrostructural and microstructural white matter properties, and their relationship with clinical measures, between adolescents recently diagnosed with ME/CFS and healthy controls. Forty-eight adolescents (25 ME/CFS, 23 controls, mean age 16 years) underwent brain diffusion MRI, and a robust multi-analytic approach was used to evaluate white and gray matter volume, regional brain volume, cortical thickness, fractional anisotropy, mean/axial/radial diffusivity, neurite dispersion and density, fiber density, and fiber cross section. From a clinical perspective, adolescents with ME/CFS showed greater fatigue and pain, poorer sleep quality, and poorer performance on cognitive measures of processing speed and sustained attention compared with controls. However, no significant group differences in white matter properties were observed, with the exception of greater white matter fiber cross section of the left inferior longitudinal fasciculus in the ME/CFS group compared with controls, which did not survive correction for intracranial volume. Overall, our findings suggest that white matter abnormalities may not be predominant in pediatric ME/CFS in the early stages following diagnosis. The discrepancy between our null findings and white matter abnormalities identified in the adult ME/CFS literature could suggest that older age and/or longer illness duration influence changes in brain structure and brain-behavior relationships that are not yet established in adolescence.

Pars Plana Vitrectomy With or Without Circumferential Scleral Buckling for Primary Repair of Retinal Detachments Secondary to Retinal Dialysis.

PURPOSE: The aim of this study was to evaluate the anatomical and functional outcomes after pars plana vitrectomy with or without circumferential scleral buckling, in patients with retinal detachment secondary to retinal dialysis. DESIGN: Case series. METHODS: A retrospective consecutive case series involving 30 eyes of 30 patients with retinal detachments secondary to retinal dialysis was completed. All 30 patients underwent primary pars plana vitrectomy, with the addition of a circumferential scleral buckle in 24 patients. The main outcome measure was the primary reattachment rate at 6 months after a single surgery. RESULTS: Primary anatomical success was achieved in 90% of patients. The final anatomical success rate was 93%, with 4 of 30 eyes still having silicone oil in situ. The main indication for vitrectomy surgery was an inadequate fundal view for scleral buckle surgery alone. The macula was detached in 50% of patients and proliferative vitreoretinopathy grade C was found in 23% of cases. The mean preoperative visual acuity was 20/814 (range 20/20-light perception) and the mean postoperative visual acuity was 20/258 (range 20/20-hand movements). CONCLUSIONS: Good final anatomical success rates can be achieved with pars plans vitrectomy, plus or minus circumferential buckling, in patients with retinal detachments secondary to retinal dialysis.

Haemopedia RNA-seq: a database of gene expression during haematopoiesis in mice and humans.

During haematopoiesis, haematopoietic stem cells differentiate into restricted potential progenitors before maturing into the many lineages required for oxygen transport, wound healing and immune response. We have updated Haemopedia, a database of gene-expression profiles from a broad spectrum of haematopoietic cells, to include RNA-seq gene-expression data from both mice and humans. The Haemopedia RNA-seq data set covers a wide range of lineages and progenitors, with 57 mouse blood cell types (flow sorted populations from healthy mice) and 12 human blood cell types. This data set has been made accessible for exploration and analysis, to researchers and clinicians with limited bioinformatics experience, on our online portal Haemosphere: https://www.haemosphere.org. Haemosphere also includes nine other publicly available high-quality data sets relevant to haematopoiesis. We have added the ability to compare gene expression across data sets and species by curating data sets with shared lineage designations or to view expression gene vs gene, with all plots available for download by the user.

Transcriptional profiling of eosinophil subsets in interleukin-5 transgenic mice.

Eosinophils are important in fighting parasitic infections and are implicated in the pathogenesis of asthma and allergy. IL-5 is a critical regulator of eosinophil development, controlling proliferation, differentiation, and maturation of the lineage. Mice that constitutively express IL-5 have in excess of 10-fold more eosinophils in the hematopoietic organs than their wild type (WT) counterparts. We have identified that much of this expansion is in a population of Siglec-F high eosinophils, which are rare in WT mice. In this study, we assessed transcription in myeloid progenitors, eosinophil precursors, and Siglec-F medium and Siglec-F high eosinophils from IL-5 transgenic mice and in doing so have created a useful resource for eosinophil biologists. We have then utilized these populations to construct an eosinophil trajectory based on gene expression and to identify gene sets that are associated with eosinophil lineage progression. Cell cycle genes were significantly associated with the trajectory, and we experimentally demonstrate an increasing trend toward quiescence along the trajectory. Additionally, we found gene expression changes associated with constitutive IL-5 signaling in eosinophil progenitors, many of which were not observed in eosinophils.

MIR21 Drives Resistance to Heat Shock Protein 90 Inhibition in Cholangiocarcinoma.

BACKGROUND & AIMS: Cholangiocarcinomas (CCA) are resistant to chemotherapy, so new therapeutic agents are needed. We performed a screen to identify small-molecule compounds that are active against CCAs. Levels of microRNA 21 (MIR21 or miRNA21) are increased in CCAs. We investigated whether miRNA21 mediates resistance of CCA cells and organoids to HSP90 inhibitors. METHODS: We performed a high-throughput screen of 484 small-molecule compounds to identify those that reduced viability of 6 human CCA cell lines. We tested the effects of HSP90 inhibitors on cells with disruption of the MIR21 gene, cells incubated with MIR21 inhibitors, and stable cell lines with inducible expression of MIR21. We obtained CCA biopsies from patients, cultured them as organoids (patient-derived organoids). We assessed their architecture, mutation and gene expression patterns, response to compounds in culture, and when grown as subcutaneous xenograft tumors in mice. RESULTS: Cells with IDH1 and PBRM1 mutations had the highest level of sensitivity to histone deacetylase inhibitors. HSP90 inhibitors were effective in all cell lines, irrespective of mutations. Sensitivity of cells to HSP90 inhibitors correlated inversely with baseline level of MIR21. Disruption of MIR21 increased cell sensitivity to HSP90 inhibitors. CCA cells that expressed transgenic MIR21 were more resistant to HSP90 inhibitors than cells transfected with control vectors; inactivation of MIR21 in these cells restored sensitivity to these agents. MIR21 was shown to target the DnaJ heat shock protein family (Hsp40) member B5 (DNAJB5). Transgenic expression of DNAJB5 in CCA cells that overexpressed MIR21 re-sensitized them to HSP90 inhibitors. Sensitivity of patient-derived organoids to HSP90 inhibitors, in culture and when grown as xenograft tumors in mice, depended on expression of miRNA21. CONCLUSIONS: miRNA21 appears to mediate resistance of CCA cells to HSP90 inhibitors by reducing levels of DNAJB5. HSP90 inhibitors might be developed for the treatment of CCA and miRNA21 might be a marker of sensitivity to these agents.

Mass Mortality Attributed to Acanthocephaliasis at a Gull-billed Tern (Gelochelidon nilotica) Colony in Coastal California, USA.

From 12 May 2013 to 29 May 2013, the Gull-billed Tern (Gelochelidon nilotica) colony at the San Diego Bay National Wildlife Refuge, California, US, experienced a mass die-off of at least 92 adults, representing 71-92% of the breeding population on the US west coast. Cause of death was determined to be peritonitis due to perforations of the intestine by a large quantity of acanthocephala (Profilicollis [=Polymorphus] altmani). This is a unique report of P. altmani infecting G. nilotica, and a report of a great impact to a tern population in southern California. Mole crabs (Emerita analoga), the intermediate host for P. altmani and a major component of the Gull-billed Tern diet in San Diego, were found in the stomachs of necropsied terns along with cystacanths, and are the presumed source of the parasite infection. The tern's dietary reliance upon mole crabs likely amplified parasite transmission and infection. We suggest additional research to determine factors that influence parasite infection of intermediate and definitive hosts, particularly mole crabs, given that they are a vital resource for migrating birds within the coastal zone.

EGFR T790M mutation testing within the osimertinib AURA Phase I study.

OBJECTIVES: Reliable epidermal growth factor receptor (EGFR) mutation testing techniques are required to identify eligible patients with EGFR mutation/T790M positive advanced non-small cell lung cancer (NSCLC), for treatment with osimertinib (AZD9291), an oral, potent, irreversible EGFR tyrosine kinase inhibitor (TKI) selective for EGFR-TKI-sensitizing and T790M resistance mutations over wild-type EGFR. There is no current consensus regarding the best method to detect EGFR T790M mutations. The aim of this study was to describe the concordance between local testing, which used a variety of methods, and central testing, using the cobas® EGFR Mutation Test, for EGFR-sensitizing mutations and the T790M resistance mutation. MATERIALS AND METHODS: Tumor samples were obtained from all patients screened for inclusion onto the osimertinib Phase I expansion component of the AURA Phase I/II study (NCT01802632). Samples underwent central laboratory testing for EGFR-sensitizing mutations and T790M resistance mutation using the cobas® EGFR Mutation Test. Results were compared with local laboratory test results, based on other testing methodologies including Sanger sequencing, therascreen®, PNAClamp™, and Sequenom MassARRAY®. RESULTS: Central laboratory testing was successful in 99% of samples passing histopathology review and testing success rates were comparable across the three central laboratories. Concordance between central and local testing for common sensitizing mutations was high (>98%) and concordance for the T790M mutation was also high (>90%). Tumor heterogeneity, along with other technical factors may have influenced this result. CONCLUSIONS: Within the osimertinib AURA Phase I study, EGFR mutation testing across three centralized laboratories using the cobas® EGFR Mutation Test was feasible and successful, with strong concordance between local and central laboratory results, including for T790M. The cobas® EGFR Mutation Test has subsequently been approved as the companion diagnostic test for osimertinib in the USA and Japan.

Evaluation of Outcomes in Adolescent Inflammatory Bowel Disease Patients Following Transfer From Pediatric to Adult Health Care Services: Case for Transition.

PURPOSE: We aimed to evaluate the impact of a transition service on clinical and developmental outcomes in adolescent Inflammatory Bowel Disease (IBD) patients on transfer to adult health care services. METHODS: We reviewed the records of IBD patients diagnosed in pediatric care following their transfer/attendance to the adult IBD service. The data on patients who attended the transition service were compared with those who did not pass through the transition service. RESULTS: Seventy-two patients were included in the study 41M and 31F. Forty-four patients went through the transition system (Group A), and 28 had no formalized transition arrangement before transfer (Group B). A significantly higher number of Group B patients needed surgery within 2 years of transfer when compared with patients in Group A (46% vs. 25%, p = .01). Sixty-one percent of patients in Group B needed at least one admission within 2 years of transfer when compared with 29% of Group A patients (p = .002). Nonattendance at clinics was higher in Group B patients with 78% having at least one nonattendance, whereas 29% of Group A failed to attend at least one appointment (p = .001). In addition, drug compliance rates were higher in the transition group when compared with Group B (89% and 46%, respectively; p = .002). A higher proportion of transitioned patients achieved their estimated maximum growth potential when completing adolescence. There was a trend toward higher dependence on opiates and smoking in Group B patients. CONCLUSIONS: In adolescent IBD patients, transition care is associated with better disease specific and developmental outcomes. Prospective studies of different models of transition care in IBD are needed.

Gefitinib treatment in EGFR mutated caucasian NSCLC: circulating-free tumor DNA as a surrogate for determination of EGFR status.

INTRODUCTION: In the phase IV, open-label, single-arm study NCT01203917, first-line gefitinib 250 mg/d was effective and well tolerated in Caucasian patients with epidermal growth factor receptor (EGFR) mutation-positive non-small-cell lung cancer (previously published). Here, we report EGFR mutation analyses of plasma-derived, circulating-free tumor DNA. METHODS: Mandatory tumor and duplicate plasma (1 and 2) baseline samples were collected (all screened patients; n = 1060). Preplanned, exploratory analyses included EGFR mutation (and subtype) status of tumor versus plasma and between plasma samples. Post hoc, exploratory analyses included efficacy by tumor and plasma EGFR mutation (and subtype) status. RESULTS: Available baseline tumor samples were 1033 of 1060 (118 positive of 859 mutation status known; mutation frequency, 13.7%). Available plasma 1 samples were 803 of 1060 (82 positive of 784 mutation status known; mutation frequency, 10.5%). Mutation status concordance between 652 matched tumor and plasma 1 samples was 94.3% (95% confidence interval [CI], 92.3-96.0) (comparable for mutation subtypes); test sensitivity was 65.7% (95% CI, 55.8-74.7); and test specificity was 99.8% (95% CI, 99.0-100.0). Twelve patients of unknown tumor mutation status were subsequently identified as plasma mutation-positive. Available plasma 2 samples were 803 of 1060 (65 positive of 224 mutation status-evaluable and -known). Mutation status concordance between 224 matched duplicate plasma 1 and 2 samples was 96.9% (95% CI, 93.7-98.7). Objective response rates are as follows: mutation-positive tumor, 70% (95% CI, 60.5-77.7); mutation-positive tumor and plasma 1, 76.9% (95% CI, 65.4-85.5); and mutation-positive tumor and mutation-negative plasma 1, 59.5% (95% CI, 43.5-73.7). Median progression-free survival (months) was 9.7 (95% CI, 8.5-11.0; 61 events) for mutation-positive tumor and 10.2 (95% CI, 8.5-12.5; 36 events) for mutation-positive tumor and plasma 1. CONCLUSION: The high concordance, specificity, and sensitivity demonstrate that EGFR mutation status can be accurately assessed using circulating-free tumor DNA. Although encouraging and suggesting that plasma is a suitable substitute for mutation analysis, tumor tissue should remain the preferred sample type when available.

Survey of helminths, ectoparasites, and chytrid fungus of an introduced population of cane toads, Rhinella marina (Anura: Bufonidae), from Grenada, West Indies.

One hundred specimens of Rhinella marina , (Anura: Bufonidae) collected in St. George's parish, Grenada, from September 2010 to August 2011, were examined for the presence of ectoparasites and helminths. Ninety-five (95%) were parasitized by 1 or more parasite species. Nine species of parasites were found: 1 digenean, 2 acanthocephalans, 4 nematodes, 1 arthropod and 1 pentastome. The endoparasites represented 98.9% of the total number of parasite specimens collected. Grenada represents a new locality record for Mesocoelium monas, Raillietiella frenatus, Pseudoacanthacephalus sp., Aplectana sp., Physocephalus sp., Acanthacephala cystacanth, and Physalopteridae larvae. The digenean M. monas occurred with the highest prevalence of 82%, contrasting many studies of R. marina where nematodes dominate the parasite infracommunity. Female toads were found to have a significantly higher prevalence of Amblyomma dissimile than male toads. Only 2 parasites exhibited a significant difference between wet and dry season with Parapharyngodon grenadensis prevalence highest in the wet season and A. dissimile prevalence highest during the dry season. Additionally, A. dissimile was significantly more abundant during the dry season.

Transitory FGF treatment results in the long-lasting suppression of the proliferative response to repeated FGF stimulation.

FGF applied as a single growth factor to quiescent mouse fibroblasts induces a round of DNA replication, however continuous stimulation results in arrest in the G1 phase of the next cell cycle. We hypothesized that FGF stimulation induces the establishment of cell memory, which prevents the proliferative response to repeated or continuous FGF application. When a 2-5 days quiescence period was introduced between primary and repeated FGF treatments, fibroblasts failed to efficiently replicate in response to secondary FGF application. The establishment of "FGF memory" during the first FGF stimulation did not require DNA synthesis, but was dependent on the activity of FGF receptors, MEK, p38 MAPK and NFκB signaling, and protein synthesis. While secondary stimulation resulted in strongly decreased replication rate, we did not observe any attenuation of morphological changes, Erk1/2 phosphorylation and cyclin D1 induction. However, secondary FGF stimulation failed to induce the expression of cyclin A, which is critical for the progression from G1 to S phase. Treatment of cells with a broad range histone deacetylase inhibitor during the primary FGF stimulation rescued the proliferative response to the secondary FGF treatment suggesting that the establishment of "FGF memory" may be based on epigenetic changes. We suggest that "FGF memory" can prevent the hyperplastic response to cell damage and inflammation, which are associated with an enhanced FGF production and secretion. "FGF memory" may present a natural obstacle to the efficient application of recombinant FGFs for the treatment of ulcers, ischemias, and wounds.

Photosynthesis of seedlings of Otoba novogranatensis (Myristicaceae) and Ruagea glabra (Meliaceae) in abandoned pasture, secondary forest and plantation habitats in Costa Rica / Fotosíntesis de plántulas de Otoba novogranatensis (Myristicaceae) y Ruagea glabra (Meliaceae) en pastizales abandonados, bosques secundarios y hábitats de plantaciones en Costa Rica

Enrichment planting in naturally recovering secondary forests or in tree plantations is increasingly being used as strategy to restore later-successional, large-seeded tropical forest trees. We seeded two tree species (Otoba novogranatensis and Ruagea glabra) in three agricultural sites in Southern Costa Rica: abandoned pastures, eight to ten year old secondary forests and three year old tree plantations (containing two N-fixing of four total tree species). We measured micrometeorological conditions, soil water content, plant water potential, leaf area, foliar C and N, and photosynthesis to better understand mechanistic responses of seedlings to conditions in the different successional habitats. Micrometeorological conditions, soil water content, and plant water potential were generally similar across habitats. Certain aspects of leaves (such as Specific Leaf Area and foliar N content), and photosynthesis (e.g. quantum yield and electron transport rate) were highest in the plantations, intermediate in the secondary forests, and lowest in abandoned pastures. Enhanced rates of photosynthetic biochemistry (such as Vcmax and Jmax) and Photosystem II efficiency (e.g. thermal energy dissipation) occurred in leaves from the plantations compared to the abandoned pastures, which may be related to higher leaf %N content. Results suggest that foliar N may be of greater importance than soil water content and micrometeorological factors in driving differences in photosynthetic processes across planting habitats. Planting seeds of these two species in plantations containing three year old trees (including two N-fixing species) enhances certain aspects of their photosynthesis and growth, compared to seedlings in abandoned pastures with non-native grasses, and thus can help increase forest recovery on abandoned agricultural lands.

El enriquecimiento de bosques secundarios o plantaciones forestales en proceso de regeneración natural por medio de la siembra de plántulas es una práctica cada vez más utilizada para restaurar bosques tropicales en estado de sucesión tardía. Sembramos dos especies de árboles (Otoba novogranatensis y Ruagea glabra) en pastizales abandonados, bosques secundarios de ocho a diez años de edad y plantaciones forestales de tres años de edad (con dos especies fijadoras de Nitrógeno de un total de cuatro especies) en tres sitios agrícolas en el Sur de Costa Rica. Medimos condiciones micrometeorológicas, contenido de agua del suelo, potencial hídrico de las plantas, área foliar, C y N foliar, y fotosíntesis para entender de una mejor manera las respuestas funcionales de las plántulas ante condiciones de distintos estadíos sucesionales. Las condiciones micrometeorológicas, contenido hídrico del suelo y el potencial hídrico de las plantas fueron mayoritariamente similares entre hábitats. Algunos aspectos de las hojas (como Área Foliar Específica y contenido de N foliar) y fotosíntesis (ej.: rendimiento cuántico y tasa de transporte de electrones) presentaron valores mayores en las plantaciones, intermedios en los bosques secundarios y menores en los pastizales abandonados. Se obtuvo un aumento en las tasas fotosintéticas bioquímicas (como Vcmax, Jmax) y la eficiencia del Fotosistema II (ej.: disipación de energía térmica) en hojas provenientes de las plantaciones comparado a las de los pastizales, posiblemente relacionado a un mayor %N foliar. Los resultados sugieren que el N foliar puede ser más importante que el contenido de agua del suelo y que los factores micrometeorológicos para marcar diferencias en los procesos fotosintéticos entre hábitats. Las plántulas de estas dos especies en las plantaciones con árboles de tres años de edad (incluyendo dos fijadoras de N) incrementaron ciertos aspectos de su fotosíntesis y crecimiento comparado a las plántulas en los pastizales abandonados de especies exóticas, por lo tanto, esta práctica puede ayudar a incrementar la recuperación de los bosques en áreas agrícolas abandonadas.

Restauración ecológica de bosques tropicales en Costa Rica: efecto de varios modelos en la producción, acumulación y descomposición de hojarasca

Tropical forest restoration in Costa Rica: the effect of several strategies on litter production, accumulation and decomposition. Tropical forest restoration strategies have the potential to accelerate the recovery of the nutrient cycles in degraded lands. Litter production and its decomposition represent the main transfer of organic material and nutrients into the soil substrate. We evaluated litter production, accumulation on the forest floor, and its decomposition under three restoration strategies: plantation (entire area planted with trees), island (trees planted in patches of three different sizes) and control (natural regeneration) plots. We also compared restoration strategies to young secondary forest (7-9yr). Restoration treatments were established in 50x50m plots in June 2004 at six sites in Southern Costa Rica. Planted tree species included two native timber species (Terminalia amazonia and Vochysia guatemalensis) interplanted with two N fixers (Erythrina poeppigiana and Inga edulis). Litter was collected every 15 days between September 2008 and August 2009 in 12 0.25m² litter traps distributed within each plot; litter that accumulated on the soil surface was collected at four locations (0.25m² quadrats) within each plot in February and May 2009. Total litter production in plantation (6.3Mg/ha) and secondary forest (7.3Mg/ha) did not differ, but were greater than in islands (3.5Mg/ha) and control (1.4Mg/ha). Plantation had greatest accumulation of litter on the soil surface (10.6Mg/ha) as compared to the other treatments (SF=7.2; I=6.7; C=4.9). Secondary forest was the only treatment with a greater annual production of litter than litter accumulation on the soil surface. Carbon storage in litter was similar between plantation and secondary forest, and significantly greater than the other treatments. No differences were found for carbon concentration and storage in the soil among treatments. There was also high variability in the production and accumulation of litter and carbon among sites. Active restoration treatments accelerated the production of litter and carbon storage in comparison to areas under natural recovery. However, the nutrient cycle has not necessarily been restored under these conditions, as high litter accumulation on the soil surface indicates a low decomposition rate, which slows nutrient return to the soil. Rev. Biol. Trop. 59 (3): 1323-1336. Epub 2011 September 01.

Estrategias de restauración tienen el potencial de acelerar el restablecimiento del ciclo de nutrientes en áreas degradadas. En este estudio, se evaluó la producción de hojarasca, su acumulación y descomposición bajo tres tratamientos: plantación (toda la superficie plantada); islas (árboles sembrados en parches de tres tamaños) y testigo (regeneración natural). También se compararon bosques secundarios jóvenes (7-9 años). Los tratamientos fueron establecidos en parcelas de 50x50m en junio 2004 en seis sitios en el sur de Costa Rica. Las especies introducidas fueron dos maderables (Terminalia amazonia y Vochysia guatemalensis) intercaladas con dos fijadoras de nitrógeno (Erythrina poeppigiana e Inga edulis). La producción total de hojarasca en la plantación y bosque secundario no difirió significativamente, fue mayor que en las islas y el testigo. La plantación presentó mayor acumulación de hojarasca sobre el suelo. Los resultados indican una gran variabilidad entre los sitios. Ambas estrategias de restauración activas aceleraron la producción y acumulación de hojarasca en comparación con la regeneración. No obstante, eso no implica la restauración del ciclo de nutrientes. La elevada acumulación de hojarasca sobre el suelo indica baja tasa de descomposición y de retorno de nutrientes al suelo.

Noradrenergic neurons of the area postrema mediate amylin's hypophagic action.

Circulating amylin inhibits food intake via activation of the area postrema (AP). The aim of this study was to identify the neurochemical phenotype of the neurons mediating amylin's hypophagic action by immunohistochemical and feeding studies in rats. Expression of c-Fos protein was used as a marker for neuronal activation and dopamine-beta-hydroxylase (DBH), the enzyme-catalyzing noradrenaline synthesis, as a marker for noradrenergic neurons. We found that approximately 50% of amylin-activated AP neurons are noradrenergic. To clarify the functional role of these neurons in amylin's effect on eating, noradrenaline-containing neurons in the AP were lesioned using a saporin conjugated to an antibody against DBH. Amylin (5 or 20 microg/kg s.c.)-induced anorexia was observed in sham-lesioned rats with both amylin doses. Rats with a lesion of > 50% of the noradrenaline neurons were unresponsive to the low dose of amylin (5 microg/kg) and only displayed a reduction in food intake 60 min after injection of the high amylin dose (20 microg/kg). In a terminal experiment, the same rats received amylin (20 microg/kg) or saline. The AP and nucleus of the solitary tract (NTS) were stained for DBH to assess noradrenaline lesion success and for c-Fos expression to evaluate amylin-induced neuronal activation. In contrast to sham-lesioned animals, noradrenaline-lesioned rats did not show a significant increase in amylin-induced c-Fos expression in the AP and NTS. We conclude that the noradrenergic neurons in the AP mediate at least part of amylin's hypophagic effect.

Amylin-mediated restoration of leptin responsiveness in diet-induced obesity: magnitude and mechanisms.

Previously, we reported that combination treatment with rat amylin (100 microg/kg.d) and murine leptin (500 microg/kg.d) elicited greater inhibition of food intake and greater body weight loss in diet-induced obese rats than predicted by the sum of the monotherapy conditions, a finding consistent with amylin-induced restoration of leptin responsiveness. In the present study, a 3 x 4 factorial design was used to formally test for a synergistic interaction, using lower dose ranges of amylin (0, 10, and 50 microg/kg.d) and leptin (0, 5, 25, and 125 microg/kg.d), on food intake and body weight after 4 wk continuous infusion. Response surface methodology analysis revealed significant synergistic anorexigenic (P < 0.05) and body weight-lowering (P < 0.05) effects of amylin/leptin combination treatment, with up to 15% weight loss at doses considerably lower than previously reported. Pair-feeding (PF) experiments demonstrated that reduction of food intake was the predominant mechanism for amylin/leptin-mediated weight loss. However, fat loss was 2-fold greater in amylin/leptin-treated rats than PF controls. Furthermore, amylin/leptin-mediated weight loss was not accompanied by the counterregulatory decrease in energy expenditure and chronic shift toward carbohydrate (rather than fat) utilization observed with PF. Hepatic gene expression analyses revealed that 28 d treatment with amylin/leptin (but not PF) was associated with reduced expression of genes involved in hepatic lipogenesis (Scd1 and Fasn mRNA) and increased expression of genes involved in lipid utilization (Pck1 mRNA). We conclude that amylin/leptin interact synergistically to reduce body weight and adiposity in diet-induced obese rodents through a number of anorexigenic and metabolic effects.

Leptin responsiveness restored by amylin agonism in diet-induced obesity: evidence from nonclinical and clinical studies.

Body weight is regulated by complex neurohormonal interactions between endocrine signals of long-term adiposity (e.g., leptin, a hypothalamic signal) and short-term satiety (e.g., amylin, a hindbrain signal). We report that concurrent peripheral administration of amylin and leptin elicits synergistic, fat-specific weight loss in leptin-resistant, diet-induced obese rats. Weight loss synergy was specific to amylin treatment, compared with other anorexigenic peptides, and dissociable from amylin's effect on food intake. The addition of leptin after amylin pretreatment elicited further weight loss, compared with either monotherapy condition. In a 24-week randomized, double-blind, clinical proof-of-concept study in overweight/obese subjects, coadministration of recombinant human leptin and the amylin analog pramlintide elicited 12.7% mean weight loss, significantly more than was observed with either treatment alone (P < 0.01). In obese rats, amylin pretreatment partially restored hypothalamic leptin signaling (pSTAT3 immunoreactivity) within the ventromedial, but not the arcuate nucleus and up-regulated basal and leptin-stimulated signaling in the hindbrain area postrema. These findings provide both nonclinical and clinical evidence that amylin agonism restored leptin responsiveness in diet-induced obesity, suggesting that integrated neurohormonal approaches to obesity pharmacotherapy may facilitate greater weight loss by harnessing naturally occurring synergies.

Anti-MuSK patient antibodies disrupt the mouse neuromuscular junction.

OBJECTIVE: A subset of myasthenia gravis patients that are seronegative for anti-acetylcholine receptor (anti-AChR) antibodies are instead seropositive for antibodies against the muscle-specific kinase (anti-MuSK-positive). Here, we test whether transfer of IgG from anti-MuSK-positive patients to mice confers impairment of the neuromuscular junction and muscle weakness. METHODS: IgG from anti-MuSK-positive myasthenia gravis patients or control IgG (seronegative for AChR and MuSK) was injected intraperitoneally (45 mg daily for 14 days) into 6-week-old female FVB/NJ and C57BL/6J mice. Changes at neuromuscular junctions in the tibialis anterior and diaphragm muscles were assessed by confocal fluorescent imaging of AChRs stained with fluorescent-alpha-bungarotoxin. Loss of function was assessed by electromyography. RESULTS: In experimental mice injected with anti-MuSK-positive patient IgG, postsynaptic AChR staining was reduced to as little as 22% of that seen in control mice. Experimental mice showed reduced apposition of the nerve terminal (labeled with antibodies against synaptophysin and neurofilament) and the postsynaptic AChR cluster (labeled with fluorescent-alpha-bungarotoxin). Mice injected with IgG from two of three anti-MuSK-positive patients lost weight and developed muscle weakness associated with a decremental electromyographic trace on repetitive nerve stimulation. INTERPRETATION: IgG from anti-MuSK-positive patients can cause myasthenia gravis when injected into mice. This may be explained by a progressive reduction in the density of postsynaptic AChR combined with changes in the nerve terminal and its relation to the postsynaptic structure.

Mortality in Laysan ducks (Anas laysanensis) by emaciation complicated by Echinuria uncinata on Laysan Island, Hawaii, 1993.

In November 1993, unusual mortality occurred among endangered Laysan ducks on Laysan Island, one of the remote refugia of the Northwestern Hawaiian Islands National Wildlife Refuge (USA). Ten live ducks were emaciated, and blood samples documented anemia, heterophilia, and eosinophilia. Pathology in 13 duck carcasses revealed emaciation, marked thickening of the proventricular wall, abundant mucus, and nodules in the gastrointestinal tract. Histology revealed granulomata associated with nematodes in the proventriculus, small intestines, and body walls of nine of 10 ducks examined on histology. We suspect that low rainfall and low food abundance that year contributed to enhanced pathogenicity of parasite infection, either through increased exposure or decreased host resistance. Because the Laysan duck is found only on Laysan island and is critically endangered, translocation of this species to other islands is being considered. Given that we have not seen pathology associated with Echinuria spp. in native waterfowl on other Hawaiian Islands and given the parasite's potential to cause significant lesions in Laysan ducks, it will be important to prevent the translocation of Echinuria spp.