Tuber rugosum, a new species from northeastern North America: Slug mycophagy aides in electron microscopy of ascospores.

Species in the genus Tuber are ascomycetous fungi that produce hypogeous fruiting bodies commonly called truffles. These fungi are ecologically relevant owing to the ectomycorrhizal symbiosis they establish with plants. One of the most speciose lineages within Tuber is the Rufum clade, which is widely distributed throughout Asia, Europe, and North America and is estimated to include more than 43 species. Most species in this clade have spiny spores, and many still have not been formally described. Here, we describe T. rugosum based on multigene phylogenetic analysis and its unique morphological characters. Tuber rugosum (previously designated in literature as Tuber sp. 69) has been collected throughout the Midwest, USA, and Quebec, Canada, and is an ectomycorrhizal symbiont of Quercus trees, as confirmed through morphological and molecular analyses of root tips presented here. We also present a novel method for preparing Tuber ascospores for scanning electron microscope imaging that includes feeding, digestion, and spore excretion by the slug Arion subfuscus. Following this method, spores become free from ascus and other mycelial debris that could obscure morphological traits during their passage through the snail gut while maintaining ornamentation. Finally, we report the fatty acid analysis, a fungicolous species association, and we provide an updated taxonomic key of the Rufum clade.

Re-examination of the Southern Hemisphere truffle genus Amylascus (Pezizaceae, Ascomycota) and characterization of the sister genus Nothoamylascus gen. nov.

Amylascus is a genus of ectomycorrhizal truffles within Pezizaceae that is known from Australia and contains only two described species, A. herbertianus and A. tasmanicus. Species of Amylascus are closely related to truffles (Pachyphlodes, Luteoamylascus) and cup fungi (Plicariella) from the Northern Hemisphere. Here we reevaluate the species diversity of Amylascus and related taxa from southern South America and Australia based on new morphological and molecular data. We identify previously undocumented diversity and morphological variability in ascospore color, ascospore ornamentation, hymenial construction, epithecium structure and the amyloid reaction of the ascus in Melzer's reagent. We redescribe two Amylascus species from Australia and describe seven new Amylascus species, five from South America and two from Australia. This is the first report of Amylascus species from South America. We also describe the new South American genus Nothoamylascus as sister lineage to the Pachyphlodes-Amylascus-Luteoamylascus clade (including Amylascus, Luteoamylascus, Pachyphlodes, and Plicariella). We obtained ITS sequences of mitotic spore mats from Nothoamylascus erubescens gen. & sp. nov. and four of the seven newly described Amylascus species, providing the first evidence of mitotic spore mats in Amylascus. Additional ITS sequences from mitotic spore mats reveal the presence of nine additional undescribed Amylascus and one Nothoamylascus species that do not correspond to any sampled ascomata. We also identify three additional undescribed Amylascus species based on environmental sequences from the feces of two grounddwelling bird species from Chile, Scelorchilus rubecula and Pteroptochos tarnii. Our results indicate that ascomata from Amylascus and Nothoamylascus species are rarely collected, but molecular data from ectomycorrhizal roots and mitotic spore mats indicate that these species are probably common and widespread in southern South America. Finally, we present a time-calibrated phylogeny that is consistent with a late Gondwanan distribution. The time since the most recent common ancestor of: 1) the family Pezizaceae had a mean of 276 Ma (217-337 HPD); 2) the Amylascus-Pachyphlodes-Nothoamylascus-Luteoamylascus clade had a mean of 79 Ma (60-100 HPD); and 3) the Amylascus-Pachyphlodes clade had a mean of 50 Ma (38-62 HPD). The crown age of Pachyphlodes had a mean of 39 Ma (25-42 HPD) and Amylascus had a mean age of 28 Ma (20-37 HPD), falling near the Eocene-Oligocene boundary and the onset of the Antarctic glaciation (c. 35 Ma). Citation: Healy RA, Truong C, Castellano MA, et al. 2023. Re-examination of the Southern Hemisphere truffle genus Amylascus (Pezizaceae, Ascomycota) and characterization of the sister genus Nothoamylascus gen. nov. Persoonia 51: 125-151. doi: 10.3767/persoonia.2023.51.03.

Two new species of Phaeohelotium (Leotiomycetes: Helotiaceae) from Chile and their putative ectomycorrhizal status.

Species of the genus Phaeohelotium (Leotiomycetes: Helotiaceae) are cup fungi that grow on decaying wood, leaves, litter, and directly on soil. Northern Hemisphere species are primarily found on litter and wood, whereas in the Southern Hemisphere the genus includes a mix of saprotrophs as well as taxa that grow on soil in association with ectomycorrhizal trees. The diversity of this genus has not been fully explored in southern South America. Here we describe two species from Chile, Phaeohelotium maiusaurantium sp. nov. and Ph. pallidum sp. nov., found on soil in Patagonian Nothofagaceae-dominated forests. We present macro- and micromorphological descriptions, illustrations, and molecular phylogenetic analyses. The two new species are placed in Phaeohelotium with high support in our 15-locus phylogeny as well as phylogenetic reconstructions based on the internal transcribed spacer (ITS) region of the nuclear ribosomal RNA gene. Our ITS phylogeny places both Ph. maiusaurantium and Ph. pallidum in a well-supported subclade that includes ectomycorrhizal root tip samples from Australasia. Similar species can be separated from these new taxa based on morphological characteristics, biogeography, substrate, and sequence data. In addition, two unnamed species from Chilean Nothofagaceae forests (Phaeohelotium sp. 1 and Phaeohelotium sp. 2) are documented from scant collections and sequence data and await description until more material becomes available. Citation: Grupe II AC, Smith ME, Weier A, Healy R, Caiafa MV, Pfister DH, Haelewaters D, Quandt CA (2022). Two new species of Phaeohelotium (Leotiomycetes: Helotiaceae) from Chile and their putative ectomycorrhizal status. Fungal Systematics and Evolution 10: 231-249. doi: 10.3114/fuse.2022.10.10.

Eight new Elaphomyces species (Elaphomycetaceae, Eurotiales, Ascomycota) from eastern North America.

The hypogeous, sequestrate ascomycete genus Elaphomyces is one of the oldest known truffle-like genera. Elaphomyces has a long history of consumption by animals in Europe and was formally described by Nees von Esenbeck in 1820 from Europe. Until recently most Elaphomyces specimens in North America were assigned names of European taxa due to lack of specialists working on this group and difficulty of using pre-modern species descriptions. It has recently been discovered that North America has a rich diversity of Elaphomyces species far beyond the four Elaphomyces species described from North America prior to 2012. We describe eight new Elaphomyces species (E. dalemurphyi, E. dunlapii, E. holtsii, E. lougehrigii, E. miketroutii, E. roodyi, E. stevemilleri and E. wazhazhensis) of eastern North America that were collected in habitats from Quebec, Canada south to Florida, USA, west to Texas and Iowa. The ranges of these species vary and with continued sampling may prove to be larger than we have established. Castellano has studied authentic material of all European Elaphomyces species published through 2016 and it is interesting to note that many Elaphomyces species from eastern North America have morphological similarities but with distinct morphological differences to a number of European Elaphomyces species. Citation: Castellano MA, Crabtree CD, Mitchell D, Healy RA (2020). Eight new Elaphomyces species (Elaphomycetaceae, Eurotiales, Ascomycota) from eastern North America. Fungal Systematics and Evolution 7: 113-131. doi: 10.3114/fuse.2021.07.06.

A case of solitary kidney with duplex collecting systems and renal vascular variants in an adult male cadaver.

We describe a unique solitary kidney with duplex collecting system and vascular variation observed in an 86-year-old white male formaldehyde- and phenol-fixed cadaver during routine academic dissection. The left renal fossa was empty with an intact adrenal gland, and the right renal fossa contained a fused renal mass with apparent polarity between the superior and inferior regions and two renal pelves converging into a single ureter. There were three right renal arteries supplying the renal mass; the superior and middle arteries were noted to be postcaval and the inferior artery was precaval. There were also two right renal veins draining into the inferior vena cava and following a regional distribution with the superior vein draining the inferior portion of the renal mass. Despite generally being asymptomatic, the detection of renal anatomical variants is clinically important for appropriate patient management and surgical interventions.

Sources, variability and parameterizations of intra-city factors obtained from dispersion-normalized multi-time resolution factor analyses of PM2.5 in an urban environment.

Ambient fine particulate matter (PM2.5) data of similar continuously monitored species at two air monitoring sites with different characteristics within the City of Toronto were used to gauge the intra-city variations in the PM composition over a largely concurrent period spanning two years. One location was <8 m from the side of a major highway while the other was an urban background location. For the first time, multi-time resolution factor analysis was applied to dispersion-normalized concentrations to identify and quantify source contributions while reducing the influence of local meteorology. These factors were particulate sulphate (pSO4), particulate nitrate (pNO3), secondary organic aerosols (SOA), crustal matter (CrM) that were common to both sites, a hydrocarbon-like organic matter (HOM) exclusive to the urban background site, three black carbon related factors (BC, BC-HOM at the highway site, and a brown carbon rich factor (BC-BrC) at the urban background site), biomass burning organic matter (BBOM) and brake dust (BD) factors exclusive to the highway site. The PM2.5 composition was different between these two locations, over only a 10 km distance. The sum of SOA, pSO4 and pNO3 at the urban background site averaged 57% of the PM2.5 mass while the same species represented 43% of the average PM2.5 mass at the highway site. Local or site-specific factors may be of greater interest for control policy design. Thus, regression analyses with potential explanatory, site-specific variables were performed for results from the highway site. Three model approaches were explored: multiple linear regression (MLR), regression with a generalized reduced gradient (GRG) algorithm, and a generalized additive model (GAM). GAM gave the largest fraction of variance for the locally-found factors at the highway site. Heavy-duty vehicles were most important for explaining the black carbon (BC and BC-HOM) factors. Light-duty vehicles were dominant for the brake dust (BD) factor. The auxiliary modelling for the local factors showed that the traffic-related factors likely originated along the main roadways at their respective sites while the more regional factors, - pSO4, pNO3, SOA, - had sources that were both regional and local in origin and with contributions that varied seasonally. These results will be useful in understanding ambient particulate matter sources on a city scale that will support air quality management planning.

Resurrecting the genus Geomorium: Systematic study of fungi in the genera Underwoodia and Gymnohydnotrya (Pezizales) with the description of three new South American species.

Molecular phylogenetic analyses have addressed the systematic position of several major Northern Hemisphere lineages of Pezizales but the taxa of the Southern Hemisphere remain understudied. This study focuses on the molecular systematics and taxonomy of Southern Hemisphere species currently treated in the genera Underwoodia and Gymnohydnotrya. Species in these genera have been identified as the monophyletic /gymnohydnotrya lineage, but no further research has been conducted to determine the evolutionary origin of this lineage or its relationship with other Pezizales lineages. Here, we present a phylogenetic study of fungal species previously described in Underwoodia and Gymnohydnotrya, with sampling of all but one described species. We revise the taxonomy of this lineage and describe three new species from the Patagonian region of South America. Our results show that none of these Southern Hemisphere species are closely related to Underwoodia columnaris, the type species of the genus Underwoodia. Accordingly, we recognize the genus Geomorium described by Spegazzini in 1922 for G. fuegianum. We propose the new family, Geomoriaceae fam. nov., to accommodate this phylogenetically and morphologically unique Southern Hemisphere lineage. Molecular dating estimated that Geomoriaceae started to diverge from its sister clade Tuberaceae c. 112 MYA, with a crown age for the family in the late Cretaceous (c. 67 MYA). This scenario fits well with a Gondwanan origin of the family before the split of Australia and South America from Antarctica during the Paleocene-Eocene boundary (c. 50 MYA).

Awareness of surgical expenditure amongst UK trainees and consultants: A questionnaire study.

INTRODUCTION: Limited knowledge of surgical device and service costs restricts a surgeon's ability to make cost efficient choices and contribute to the efficiency savings required by the NHS to reduce the financial deficit. This study aims to assess how aware surgeons are of surgical equipment and regularly used services. METHODS: A single sided hard copy questionnaire asking for the estimate cost of 24 surgical devices/services was handed out to individuals at two separate UK annual conferences. Items and services which are regularly used and/or clinically significant were selected and, where possible, alternatives to those items were included for comparison. Participants were also asked for their grade and specialty. An estimate was deemed correct if it was within 20% of the actual cost. Planned subgroup analyses for grade and specialty were performed. RESULTS: The 143 participants consisted of 23 (16%) consultants, 39 (27%) registrars, 33 (23%) SHOs and 48 (34%) foundation doctors. Of the 95 participants who were SHO grade or more senior, 67 (71%) work within general surgery. Across all items, only 9.6% of estimates were correct. There was no statistically significant difference between training levels (consultant 11.5%, registrar 10.1%, SHO 8.6%, foundation 8.9%; p = 0.253). Participants were significantly less successful in correctly estimating the cost of high value (>£150 [USD $198; EUR €175]) items (8.5% vs. 11.1%); p = 0.011, and the cost of devices as compared to the cost of services (7.4% vs. 15.0%); p = 0.001. CONCLUSION: Surgeons across all grades and specialties have poor knowledge of device and service costs. It is important that this improves in order to allow surgeons to make a meaningful contribution to NHS efficiency savings by making informed decisions about their use of devices and services.

Increased non-melanoma skin cancer risk in young patients with inflammatory bowel disease on immunomodulatory therapy: a retrospective single-centre cohort study.

BACKGROUND: Recent studies report an increased risk of non-melanoma skin cancer (NMSC) in immunosuppressed patients with inflammatory bowel disease (IBD). Concurrently, paediatric IBD incidence is rising, with more patients now exposed to immunomodulators from a younger age. OBJECTIVES: To investigate NMSC incidence and to examine the risk associated with immunomodulators in the development of NMSC in patients with IBD. METHODS: This was a retrospective single-centre cohort study. Patients with IBD attending a tertiary adult hospital from 1994 to 2013 were included. Skin cancer incidence was compared with population data from the National Cancer Registry of Ireland (NCRI) to calculate standardized incidence ratio (SIR). Logistic regression was utilized for risk factor analysis. RESULTS: Two thousand and fifty-three patients with IBD were studied. The SIR for NMSC in patients with IBD taking immunomodulators overall was 1.8 (95% CI: 1.0-2.7) with age-specific rates significantly elevated across certain age categories. Exposure to thiopurines (OR: 5.26, 95% CI: 2.15-12.93, P < 0.001) and in particular thiopurines and/or tumour necrosis factor alpha (TNF-α) inhibitors (OR: 6.45, 95% CI: 2.69-15.95, P < 0.001) was significantly associated with NMSC. The majority (82%) of those exposed to a TNF-α inhibitor also had thiopurine exposure. CONCLUSIONS: Compliance with skin cancer preventative measures should be highlighted to all patients with IBD. There should be a low threshold for dermatology referral for immunosuppressed patients, particularly those with a history of exposure to dual immunomodulators from a young age.

Scanning electron microscopy-energy dispersive X-ray spectrometry (SEM-EDX) and aerosol time-of-flight mass spectrometry (ATOFMS) single particle analysis of metallurgy plant emissions.

The chemical composition of single particles deposited on industrial filters located in three different chimneys of an iron-manganese (Fe-Mn) alloy manufacturing plant have been compared using aerosol time-of-flight mass spectrometry (ATOFMS) and scanning electron microscopy-energy dispersive X-ray spectrometry (SEM-EDX). Very similar types of particles were observed using both analytical techniques. Calcium-containing particles dominated in the firing area of the sintering unit, Mn and/or Al-bearing particles were observed at the cooling area of the sintering unit, while Mn-containing particles were dominant at the smelting unit. SEM-EDX analysis of particles collected downstream of the industrial filters showed that the composition of the particles emitted from the chimneys is very similar to those collected on the filters. ATOFMS analysis of ore samples was also performed to identify particulate emissions that could be generated by wind erosion and manual activities. Specific particle types have been identified for each emission source (chimneys and ore piles) and can be used as tracers for source apportionment of ambient PM measured in the vicinity of the industrial site.

Prevalence and incidence of blindness and other degrees of sight impairment in patients treated for neovascular age-related macular degeneration in a well-defined region of the United Kingdom.

AIMS: This study aimed to evaluate the incidence and prevalence of blindness, sight impairment, and other visual acuity (VA) states in patients receiving ranibizumab for neovascular age-related macular degeneration (nAMD) in Gloucestershire. METHODS: Serial VA and injection data for all treatment-naive patients receiving their first intravitreal injections of ranibizumab for nAMD in the Gloucestershire National Health Service Ophthalmology department between 2008 and 2010 were extracted from an electronic medical record system. RESULTS: The prevalence of blindness (VA in the better-seeing eye ≤25 Early Treatment Diabetic Retinopathy Study (ETDRS) letters) at the time of first intravitreal injection was 0.8%, increasing to 3.5% after 3 years. The prevalence of sight impairment (VA in the better-seeing eye 26-39 ETDRS letters) increased from 4.1% at baseline to 5.5% after 3 years. The incidence of initiating ranibizumab treatment for nAMD in people aged ≥50 years in Gloucestershire was 111 people per 100 000 population in 2009, and 97 people in 2010. The incidence of patients meeting the visual criteria for blindness and sight impairment registration from treated nAMD in people aged ≥50 years in Gloucestershire was 3.5 and 9.7 people, respectively per 100 000 population in 2010. CONCLUSION: This is the first real-world study on the incidence and prevalence of eligibility for blindness and sight impairment registration in treated nAMD in the UK based on VA data. The incidence and prevalence of eligibility for certification of blindness or sight impairment in patients treated with ranibizumab for nAMD is low in Gloucestershire, with only 3.6% of the incident population progressing to blindness in 2010.

The impact of animal introductions during herd restrictions on future herd-level bovine tuberculosis risk.

In Ireland new cases of bovine tuberculosis (bTB) are detected using both field (with the single intradermal comparative tuberculin test (SICTT)) and abattoir surveillance. Once a new case has been detected, herd restrictions, including restrictions on animal movements into and out of the herd, are implemented until the herd has passed two consecutive clear tests. While a herd is restricted, there may be several reasons why it may be desirable to introduce new stock, such as enabling routine management practices to continue 'as near to normal'. In Ireland, introduction of animals during a bTB episode is permitted under specific conditions, with permission from the local veterinary office. The objectives of this study were (1) to provide an overview of movement events associated with each bTB episode, (2) to determine whether introduction of animals during a bTB episode is associated with increased future bTB risk and (3) to identify the practices relating to the introduction of animals that are the most risky. All herds that were not restricted at the start of 2006, but experienced a bTB episode during 2006 with 2 or more SICTT standard reactors (the eligible bTB episode) were included in the study. We calculated the number of extended eligible bTB episodes and subsequent bTB episodes that could be directly attributed to introduced animals. The main outcome of interest was the time from de-restriction of the eligible bTB episode to the start of a subsequent bTB episode or the date of the last test prior to the end of the study (31 December 2010). Cox proportional-hazard models were developed, each using a different introduction variable: introduced animals during an episode (yes/no), introduced animals prior to the first retest/first clear test, time from start of episode until first animals introduced and number of animals introduced during the episode. Only a small proportion of subsequent bTB episodes (1.8%) or extended eligible bTB episodes (2.7%) could be directly attributed to introduced animals. The results highlight an increased risk of a subsequent bTB episode among only a subset of herds that introduced animals during the eligible bTB episode. Specifically, herds that introduced animals early during the eligible bTB episode were at significantly greater future bTB risk than herds where animals were only introduced later. To illustrate, herds that introduced animals after the first retest did not have a significantly different risk compared to herds that did not introduce animals at all. In contrast, herds that did introduce animals prior to the first retest had 1.5 times higher risk of a subsequent bTB episode. Future practices concerning the introduction of animals during an episode now need to be reviewed.

High diversity and widespread occurrence of mitotic spore mats in ectomycorrhizal Pezizales.

Fungal mitospores may function as dispersal units and/ or spermatia and thus play a role in distribution and/or mating of species that produce them. Mitospore production in ectomycorrhizal (EcM) Pezizales is rarely reported, but here we document mitospore production by a high diversity of EcM Pezizales on three continents, in both hemispheres. We sequenced the internal transcribed spacer (ITS) and partial large subunit (LSU) nuclear rDNA from 292 spore mats (visible mitospore clumps) collected in Argentina, Chile, China, Mexico and the USA between 2009 and 2012. We collated spore mat ITS sequences with 105 fruit body and 47 EcM root sequences to generate operational taxonomic units (OTUs). Phylogenetic inferences were made through analyses of both molecular data sets. A total of 48 OTUs from spore mats represented six independent EcM Pezizales lineages and included truffles and cup fungi. Three clades of seven OTUs have no known meiospore stage. Mitospores failed to germinate on sterile media, or form ectomycorrhizas on Quercus, Pinus and Populus seedlings, consistent with a hypothesized role of spermatia. The broad geographic range, high frequency and phylogenetic diversity of spore mats produced by EcM Pezizales suggests that a mitospore stage is important for many species in this group in terms of mating, reproduction and/or dispersal.

Acquired perforating dermatosis associated with pregnancy.

Acquired perforating collagenosis (APC) is a rare condition in which altered collagen and elastin fibres undergo transepidermal elimination. The pathogenesis is unknown but it is usually associated with underlying systemic disease, most commonly diabetes mellitus and renal impairment. We describe a case of an acquired perforating disorder occurring in pregnancy. APC is a diagnosis that can easily be missed, especially in atypical cases such as ours, and clinicians and histopathologists need to be aware of this condition to avoid unnecessary patient morbidity.

Amyloplast to chromoplast conversion in developing ornamental tobacco floral nectaries provides sugar for nectar and antioxidants for protection.

Tobacco floral nectaries undergo changes in form and function. As nectaries change from green to orange, a new pigment is expressed. Analysis demonstrated that it is ß-carotene. Plastids undergo dramatic changes. Early in nectary development, they divide and by stage 9 (S9) they are engorged with starch. About S9, nectaries shift from quiescent anabolism to active catabolism resulting in starch breakdown and production of nectar sugars. Starch is replaced by osmiophilic bodies, which contain needle-like carotenoid crystals. Between S9 and S12, amyloplasts are converted to chromoplasts. Changes in carotenoids and ascorbate were assayed and are expressed at low levels early in development; however, following S9 metabolic shift, syntheses of ß-carotene and ascorbate greatly increase in advance of expression of nectar redox cycle. Transcript analysis for carotenoid and ascorbate biosynthetic pathways showed that these genes are significantly expressed at S6, prior to the S9 metabolic shift. Thus, formation of antioxidants ß-carotene and ascorbate after the metabolic shift is independent of transcriptional regulation. We propose that biosynthesis of these antioxidants is governed by availability of substrate molecules that arise from starch breakdown. These processes and events may be amenable to molecular manipulation to provide a better system for insect attraction, cross pollination, and hybridization.

Mattirolomyces tiffanyae, a new truffle from Iowa, with ultrastructural evidence for its classification in the Pezizaceae.

A new species of hypogeous Pezizales, Mattirolomyces tiffanyae, is described and illustrated. Its asci are typically three-spored, an unusually small number in the non-Tuber Pezizales. Ascus septal pore ultrastructure consists of a uni- or bi-convex band, which suggests an affinity with the Pezizaceae. Secondary spore-wall development is similar to that of Peziza, and several species of hypogeous Pezizaceae.

The significance of suicide notes in the elderly.

Suicide notes are traditionally considered as markers of the severity of the suicide attempt and are said to provide valuable insight into the thinking of suicide victims before the fatal act. Few studies have described the characteristics of elderly suicide note-writers and their final thoughts. This study is a retrospective view of suicide notes obtained from coroners' records of all elderly suicides in Cheshire over a period of 10 years, 1989-1998. Out of 125 suicides, 54 (43%) had suicide notes, which were reported in the coroner's records. Of these there were 31 (57%) male and 23 (43%) female subjects. Elderly suicide note-leavers were more likely to be unknown to psychiatric services (p < 0.01) and to have used a non-violent method of suicide (p < 0.01). Sex, marital status, social isolation, mental or physical morbidity did not appear to be linked with leaving a suicide note. More cases that took an overdose, used plastic bags, electrocuted themselves, or used car exhaust also left suicide notes. Those who died by more violent means such as hanging, drowning, jumping from height, immolation or wounding appeared less likely to have left a suicide note. No statistically significant difference in the content of the suicide notes was observed in relation to sex or age. Many elderly are isolated and may have no one to write a note to, while others have lost the ability to express themselves. Therefore, it is not possible to say that the different incidence of note leaving observed for suicide was due to differing levels of suicidal intent. Although only a proportion of elderly suicide victims leave suicide notes, the absence of a suicide note must not be considered an indicator of a less serious attempt.