AXENIC CULTURE OF TWO NEW SPECIES OF BRANCHED TRICHOMYCETES

Two new endocommensal fungi belonging to the Genistellaceae (Harpellales) were isolated in pure culture from the hindguts of Diptera larvae: Smittium culesitae from the mosquito Culesita impatiens, on a 10% brain-heart infusion, and S. simulii from the black fly Simulium argus, on a potato dextrose-yeast extract medium. Both species appear to have no unusual nutritional or cultural requirements and can grow on synthetic media and a variety of extract media, as can also the other 4 existent isolates of Smittium (Rubetella). Identical isolation methods proved unsuccessful for culturing various other genera of Harpellales and Eccrinales, with the exception of an Enterobryus sp. (from the fiddler crab Uca crenulata) which survived several generations before ceasing to grow. The genus Rubetella Tuzet & Manier is discarded in favor of Smittium Poisson, for reasons discussed.     

Temporal changes in the level of infestation of Simulium ornatum Meigen (complex) (Simuliidae: Diptera) larvae by the endosymbiotic fungus Harpella melusinae Lichtwardt (Harpellales: Trichomycetes)

The fungus Harpella melusinae (Harpellales: Trichomycetes) is obligately associated with the midguts of larval Simuliidae (Diptera). The level of infestation of a population of Simulium ornatum by H. melusinae was monitored at a stream in Hampshire, England. Significant temporal changes in the level of infestation were recorded during monthly and weekly collections; a twenty-fold increase being recorded over a nine-day period. Possible mechanisms by which these changes occur are discussed.     

Secondary spore formation in <Emphasis Type="Italic">Orchesellaria mauguioi</Emphasis> (Asellariales,Trichomycetes) and its taxonomic and ecological implications

Orchesellaria (Orc.) mauguioi (Asellariales) was detected from the hindguts of Isotomurus palustris (Collembola, Hexapoda) collected in Japan. The secondary spore formation on the exuviae of its host is described. When the host molted, its exoskelton including the ectoderm-originated hindgut cuticle to which Orc. mauguioi mycelia attaches was shed. Arthrospores of Orc. mauguioi germinated and penetrated the molt skin and produced secondary spores terminally. The secondary spore is a narrowly ovoid monosporangious sporangium accompanied by a capitate-lageniform terminal cell; these fall off together as a dispersive unit. The terminal cell is sterile and bears one to three fine filamentous extensions seven times as long as the length of the cell. In contrast to the appendages produced endogenously by other harpellids, the filament is produced exogenously and is similar to those of the genus Orphella (Harpellales). The taxonomic and ecological implications of the secondary spore formation of Orc. mauguioi are discussed by comparison with those of other Trichomycetes.     

Prevalence dynamics of two endosymbiont fungi (Orphella spp. Harpellales: Kickxellomycotina) and host shift among different Leuctra (Plecoptera) species in a stream community

This study was conducted to increase the understanding of the temporal dynamics of the plecopteran-associated endosymbiont species Orphella helicospora and Orphella catalaunica (Harpellales, Zygomycota) in the middle reaches of a stream in Montseny Natural Park (Barcelona, Spain). Previous studies addressing the temporal dynamics of Harpellales have focused on Diptera-related species. Statistical analyses revealed that the temporal dynamics of Plecoptera-associated Harpellales do not respond directly to environmental factors but are more directly dependent on biotic factors, such as the particular preference and fitness of each Orphella species towards a specific Leuctra host, the relative host abundance, and its annual cycle. We report that an in situ host shift linked to Leuctra species replacement occurred during the years 2012–2013, with a resulting decline of O. catalaunica in this environment. The ecological implications of the specificity range and fitness of the host are discussed. We also provide information about the possible causes of this macroinvertebrate shift and the effects on the associated endozoic community.     

Sterols synthesized by cultured trichomycetes

Desmosterol was synthesized by all but 2 of 14 isolates of the fungal genus Smittium (Harpellales) obtained from the hindguts of various Diptera larvae and grown axenically on a sterol-free medium. Also detected in some isolates of Smittium were cholesterol and ergosterol, and another, unidentified sterol with an apparent molecular ion of 470. Sterol content based on dry weight of fungal tissue was as high as 0.34%, but it was shown in one isolate that sterols vary both quantitatively and qualitatively during growth of the fungus. Two axenic cultures of the trichomycete Amoebidium parasiticum (Amoebidiales) isolated from the exoskeleton of a Cladocera contained cholesterol and ergosterol, but no desmosterol. Two soil fungi with possible phylogenetic affinities to Smittium, Linderina pennispora and Dipsacomyces acuminosporus (Kickxellales), produced cholesterol alone. The possible influence of the gut fungi on arthropod growth and development is discussed.Abbreviations TMS trimethylsilyl Based upon a Master of Arts thesis submitted by the senior author to the Department of Botany, University of Kansas. This research was partially supported by NSF grants BMS 72-02380-AO and DEB 77-16161 to R.W.L.     

Symbiotic Harpellales (Trichomycetes) in Tasmanian aquatic insects

Surveys for symbiotic fungi in the guts of aquatic insect larvae (Trichomycetes: Harpellales) in Tasmania, Australia, resulted in the discovery of four new species: two in Gripopterygidae (Plecoptera) nymphs, Plecopteromyces leptoperlarum and P. trinotoperlarum, and two associated with Diptera larvae, Smittium magnosporum in Thaumaleidae and Stachylina dolichospora in Chironomidae. Previously described species of Harpellales from other localities are reported and new host records summarized. A key to all Tasmanian species of Harpellales is provided.     

Trichomycetes (Zygomycota) in the digestive tract of arthropods in Amazonas, Brazil

Eight species of Harpellales and three species of Eccrinales (Zygomycota: Trichomycetes) were found associated with the digestive tract of arthropods from terrestrial and aquatic environments in the central Amazon region of Brazil. New species of Harpellales include: Harpella amazonica, Smittium brasiliense, Genistellospora tropicalis in Simuliidae larvae and Stachylina paucispora in Chironomidae larvae. Axenic cultures of S. brasiliense were obtained. Probable new species of Enterobryus (Eccrinales), Harpella, and Stachylina (Harpellales) are described but not named. Also reported are the previously known species of Eccrinales, Passalomyces compressus and Leidyomyces attenuatus in adult Coleoptera (Passalidae), and Smittium culisetae and Smittium aciculare (Harpellales) in Culicidae and Simuliidae larvae, respectively. Comments on the distribution of some of these fungi and their hosts in the Neotropics are provided.     

Prevalence of Harpellales from Chironomidae larvae in phytotelmata from punta Lara Forest, Argentina

Harpellales (Zygomycota: Trichomycetes) fungi are cosmopolitan obligate inhabitants of the gut of immature insects. A biweekly survey of gut fungi associated with chironomid (Chironomidae: Diptera) larvae living in the impounded water from Eryngium cabrerae (Apiaceae) phytotelmata from Punta Lara forest, Argentina, was done Jan 2003-Dec 2004. Two species of Harpellales were associated with chironomid larvae, Smittium phytotelmatum in the hindgut of Polypedilum sp. and Stachylina lentica in the midgut of both Polypedilum sp. and Metriocnemus eryngiotelmatus. No statistically significant differences were recorded in the prevalence of these Harpellales between seasons. Environmental variables (temperature, rainfall and relative humidity), impounded water volume, pH and chironomid larval density did not have an effect on the prevalence of the Trichomycetes.     

Molecular Identity of Holomastigotes (Spirotrichonymphea,Parabasalia) with Descriptions of Holomastigotes flavipes n. sp. and Holomastigotes tibialis n. sp.

Holomastigotes is a protist genus (Parabasalia: Spirotrichonymphea) that resides in the hindguts of “lower” termites. It can be distinguished from other parabasalids by spiral flagellar bands that run along the entire length of the cell, an anterior nucleus, a reduced or absent axostyle, the presence of spherical vesicles inside the cells, and the absence of ingested wood particles. Eight species have been described based on their morphology so far, although no molecular data were available prior to this study. We determined the 18S rRNA gene sequences of Holomastigotes from the hindguts of Hodotermopsis sjostedti, Reticulitermes flavipes, Reticulitermes lucifugus, and Reticulitermes tibialis. Phylogenetic analyses placed all sequences in an exclusive and well‐supported clade with the type species, Holomastigotes elongatum from R. lucifugus. However, the phylogenetic position of Holomastigotes within the Spirotrichonymphea was not resolved. We describe two new species, Holomastigotes flavipes n. sp. and Holomastigotes tibialis n. sp., inhabiting the hindguts of R. flavipes and R. tibialis, respectively.     

Diagnostic route is associated with care satisfaction independently of tumour stage: Evidence from linked English Cancer Patient Experience Survey and cancer registration data

BackgroundWhether diagnostic route (e.g. emergency presentation) is associated with cancer care experience independently of tumour stage is unknown.MethodsWe analysed data on 18 590 patients with breast, prostate, colon, lung, and rectal cancers who responded to the 2014 English Cancer Patient Experience Survey, linked to cancer registration data on diagnostic route and tumour stage at diagnosis. We estimated odds ratios (OR) of reporting a negative experience of overall cancer care by tumour stage and diagnostic route (crude and adjusted for patient characteristic and cancer site variables) and examined their interactions with cancer site.ResultsAfter adjustment, the likelihood of reporting a negative experience was highest for emergency presenters and lowest for screening-detected patients with breast, colon, and rectal cancers (OR versus two-week-wait 1.51, 95% confidence interval [CI] 1.24–1.83; 0.88, 95% CI 0.75–1.03, respectively). Patients with the most advanced stage were more likely to report a negative experience (OR stage IV versus I 1.37, 95% CI 1.15–1.62) with little confounding between stage and route, and no evidence for cancer-stage or cancer-route interactions.ConclusionsThough the extent of disease is strongly associated with ratings of overall cancer care, diagnostic route (particularly emergency presentation or screening detection) exerts important independent effects.     

Does Multifocal Papillary Thyroid Microcarcinoma With a Total Tumor Diameter >1 cm Indicate Poor Biological Behavior? The Evidence is Insufficient

ObjectiveMultifocal cancer is common in papillary thyroid microcarcinoma (PTMC). Our aim was to investigate the correlation between multifocal PTMC, total tumor diameter (TTD), and clinicopathologic features.MethodsIn total, 206 patients were included and grouped as stage cT1a or cT1b. The primary tumor diameter and TTD (the sum of the maximal diameter of each focus) were calculated. These patients were further subgrouped as TTD ≤1 cm or 1 cm < TTD ≤ 2 cm. The relationships of clinicopathological features between these groups were analyzed.ResultsMultifocal cancer was more likely to occur with stage cT1a than stage cT1b (P = .028). Stage cT1b papillary thyroid carcinoma was more prone to central lymph node metastasis (CLNM) and capsular invasion than stage cT1a. There was no difference in clinicopathological factors, such as sex, age, CLNM, number of CLNMs, capsular invasion, BRAF mutation, or recurrence between the multifocal PTMC and TTD >1 cm and primary tumor diameter + TTD ≤1 cm groups. Comparing stage cT1a and cT1b tumors with a 1 cm < TTD ≤ 2 cm using a multivariate analysis, stage cT1b tumors were more prone to capsular invasion than stage cT1a tumors, with an odds ratio of 19.013 (95% confidence interval, 2.295-157.478), but there was no significant correlation with CLNM.ConclusionStage cT1b tumors are more prone to capsular invasion and CLNM than than stage cT1a tumors. For multifocal PTMC, calculating the TTD to evaluate adverse biological behavior is insufficient and limited, and further research is needed.     

Termite pathogens: Effects of ingested Metarhizium,beauveria, and Gliocladium conidia on worker termites (Reticulitermes sp.)

Separate groups of subterranean termites (Reticulitermes sp.) were exposed to whole cultures of Beauveria bassiana, Gliocladium virens, or Metarhizium anisopliae. Individuals were removed after varying time intervals and hindgut contents were plated onto potato dextrose agar. Viable spores first appeared in the hindguts within 8 hr of exposure. Fungi reisolated from the hindguts of diseased termites were pathogenic of healthy termites. Histological examination showed that invasion of the hemocoel by M. anisopliae occurred exclusively through direct invasion of the integument ca. 24 hr after death. B. bassiana invaded, primarily through the alimentary tract, ca. 12 hr prior to termite death.     

Association of plasma neutrophil gelatinase-associated lipocalin with acute kidney injury and clinical outcome in cardiac arrest survivors depends on the time of measurement

Purpose: The optimal timing for measurement of neutrophil gelatinase-associated lipocalin (NGAL) level to predict acute kidney injury (AKI) and prognosis in cardiac arrest (CA) survivors has not been elucidated. We aimed to compare the diagnostic and prognostic performance of NGAL levels after return of spontaneous circulation (ROSC) and at 48?h after CA.

Methods: We included 231 adult cardiac arrest survivors who underwent targeted temperature management between May 2013 and December 2016. The primary outcome was stage 2 and 3 AKI (high stage AKI), and the secondary outcomes were in-hospital mortality and neurologic outcome. Sixty-one (26.4%) developed high stage AKI, 50 (21.6%) died, and 152 (65.8%) had a poor neurologic outcome.

Results: NGAL level at 48?h (0.876; 95% confidence interval [CI], 0.826–0.916) had a higher area under receiver operating characteristic curve than NGAL level after ROSC (0.694; 95% CI, 0.631–0.753). Both NGAL levels were independently associated with high stage AKI. NGAL level at 48?h (1.001; 95% CI, 1.000–1.002) remained a significant predictor for in-hospital mortality, while neither of the NGAL levels were independently associated with neurologic outcome.

Conclusions: NGAL at 48?h after CA seems to be a robust predictor for high stage AKI and in-hospital mortality.     

Surgical resection as a predictor of cancer-specific survival by stage at diagnosis and cancer type,United States, 2006–2015

BackgroundWhen solid tumors are amenable to definitive resection, clinical outcomes are generally superior to when those tumors are inoperable. However, the population-level cancer survival benefit of eligibility for surgery by cancer stage has not yet been quantified.MethodsUsing Surveillance, Epidemiology and End Results data allowing us to identify patients who were deemed eligible for and received surgical resection, we examined the stage-specific association of surgical resection with 12-year cancer-specific survival. The 12-year endpoint was selected to maximize follow-up time and thereby minimize the influence of lead time bias.ResultsAcross a variety of solid tumor types, earlier stage at diagnosis allowed for surgical intervention at a much higher rate than later-stage diagnosis. At every stage, surgical intervention was associated with a substantially higher rate of 12-year cancer-specific survival, with absolute differences of up to 51% for stage I, 51% for stage II, and 44% for stage III cancer, and stage-specific mortality relative risks of 3.6, 2.4, and 1.7, respectively.ConclusionsDiagnosis of solid cancers in early stages often enables surgical resection, which reduces the risk of death from cancer. Receipt of surgical resection is an informative endpoint that is strongly associated with long-term cancer-specific survival at every stage.     

Relationship between a host plant compound,myrcene and pheromone production in the bark beetle, IPS paraconfusus

The pheromonal components, ipsenol and ipsdienol were found in increasing quantities in hindguts of only the male sex of Ips paraconfusus following exposure of both sexes to a series of increasing concentrations of myrcene vapour. Hindguts of female and male beetles contained similar quantities of myrcene and other volatile compounds associated with myrcene exposure. Unexposed beetles of both sexes did not contain detectable amounts of any volatile compound. This indicates that myrcene induces or is a precursor for sex-specific pheromone biosynthesis.     

Occurrence of comorbidity with colorectal cancer and variations by age and stage at diagnosis

BackgroundWhile age and stage at diagnosis are known to affect treatment choices and survival from colorectal cancer (CRC), few studies have investigated the extent to which these effects are influenced by comorbidity. In this study, we describe the occurrence of comorbidity in CRC cases in South Australia and associations of comorbidity with age, stage and the age-stage relationship. Furthermore, we report on the association of individual comorbidities with age and stage at diagnosis.MethodsThe South Australian Cancer Registry (SACR) provided CRC data (C18-C20, ICD-10) for 2004–2013 diagnoses. CRC data were linked with comorbidity data drawn from hospital records and health insurance claims. Logistic regression was used to model associations of comorbidity with age and stage.ResultsFor the 8462 CRC cases in this study, diabetes, peptic ulcer disease, and previous cancers were the most commonly recorded co-existing conditions. Most comorbidities were associated with older age, although some presented more frequently in younger people. Patients at both ends of the age spectrum (<50 and 80 + years) had an increased likelihood of CRC diagnosis at an advanced stage compared with other ages (50–79 years old). Adjusting for comorbidities moderated the association of older age with advanced stage. Conditions associated with advanced stage included dementia (OR = 1.25 (1.01–1.55)), severe liver disease (OR = 1.68 (1.04–2.70)), and a previous cancer (OR = 1.18 (1.08–1.28)).ConclusionComorbidities are prevalent with CRC, especially in older people. These comorbidities differ in their associations with age at diagnosis and stage. Dementia and chronic heart failure were associated with older age whereas inflammatory bowel disease and alcohol access were associated with younger onset of the disease. Severe liver disease and dementia were associated with more advanced stage and rheumatic disease with less advanced stage. Comorbidities also interact with age at diagnosis and appear to vary the likelihood of advanced-stage disease. CRC patient have different association of age with stage depending on their comorbidity status.     

New species and summary of Iberian Harpellales

The occurrence of three new species of Harpellales in the Iberian Peninsula is reported. Capniomyces celatus was found in Nemouridae (Plecoptera) nymphs, Stipella latispora in Simuliidae (Diptera) larvae and Legeriomyces dolabrae with Baetidae (Ephemeroptera) nymphs. These species are differentiated from others by thalli and/or reproductive structures. Geographic range extensions for other species of Harpellales also are reported; these are Graminella bulbosa, Pennella angustispora, Spartiella barbata, Stachylina euthena, St. grandispora, St. pedifer, St. penetralis, St. prolifica, St. robusta and Stipella vigilans. All are compared with related taxa and are illustrated with photographs. Finally a summary of the known Harpellales occurring in the Iberian territory is provided with data on their distribution and ecology.     

The Prevalence of Dysglycemia-Based Chronic Disease in a European Population – a New Paradigm to Address Diabetes Burden: A Kardiovize Study

ObjectiveTo determine the prevalence rate and associated risk factors for each stage of the Dysglycemia-Based Chronic Disease (DBCD) model, which 4 distinct stages and prompts early prevention to avert Diabetes and cardiometabolic complications.MethodsSubjects between 25 and 64 years old from a random population-based sample were evaluated in Czechia from 2013 to 2014 using a cross-sectional design. DBCD stages were: stage 1 “insulin resistance” (inferred risk from abdominal obesity or a family history of diabetes); stage 2 “prediabetes”(fasting glucose between 5.6 and 6.9 mmol/L); stage 3 “type 2 diabetes (T2D)” (self-report of T2D or fasting glucose ≥7 mmol/L); and stage 4 “vascular complications” (T2D with cardiovascular disease).ResultsA total of 2147 subjects were included (57.8% women) with a median age of 48 years. The prevalence of each DBCD stage were as follows: 54.2% (stage 1); 10.3% (stage 2), 3.7% (stage 3); and 1.2% (stage 4). Stages 2 to 4 were more frequent in men and stage 1 in women (P < .001). Using binary logistic regression analysis adjusting by age/sex, all DBCD stages were strongly associated with abnormal adiposity, hypertension, dyslipidemia, and smoking status. Subjects with lower educational levels and lower income were more likely to present DBCD.ConclusionUsing the new DBCD framework and available metrics, 69.4% of the population had DBCD, identifying far more people at risk than a simple prevalence rate for T2D (9.2% in Czechia, 2013-2014). All stages were associated with traditional cardiometabolic risk factors, implicating common pathophysiologic mechanisms and a potential for early preventive care. The social determinants of health were related with all DBCD stages in alarming proportions and will need to be further studied.     

Bojamyces repens (Harpellales) from exuviae of mayfly,a new record from Japan

Bojamyces repens (Harpellales, Legeiomycetaceae) was discovered from exuviae of Paraleptophrebia sp. (Ephemeroptera: Leptophlebiidae). Trichosporogenesis required at least 2 days from the accumulation of cytoplasm in generative cells to mature trichospores at 5 °C in exuviae, which were kept in a drop of water on glass slides. Each mature trichospore possessed a structure with different refraction than the cytoplasm and was considered as a spore body, including a holdfast substance for the next infection. This is the first record of this genus in Japan.