Untangling the Lactifluus clarkeae - Lf. flocktoniae(Russulaceae) species complex in Australasia.

The Lactifluus clarkeae complex is a commonly observed, generally brightly coloured, group of mushrooms that are usually associated with Nothofagus or Myrtaceous hosts in Australia and New Zealand. For this study collections labelled as 'Lactarius clarkeae', 'Russula flocktoniae' and 'Lactarius subclarkeae' were examined morphologically and molecularly. Analyses of molecular data showed a high cryptic diversity, with sequences scattered across 11 clades in three subgenera within Lactifluus, and a single collection in Russula. We select epitypes to anchor the currently accepted concepts of Lf. clarkeae s.str. and Lf. flocktoniae s.str. The name Lf. subclarkeae could not be applied to any of the collections examined, as none had a lamprotrichoderm pileipellis. Lactifluus clarkeae var. aurantioruber is raised to species level, and six new species are described, three in subg. Lactifluus: Lf. jetiae, Lf. pagodicystidiatus, and Lf. rugulostipitatus, and three in subg. Gymnocarpi: Lf. albens, Lf. psammophilus, and Lf. pseudoflocktoniae. A new collection of Lf. russulisporus provides a significant range extension for the species. Untangling this complex will enable better identification of species and increase understanding of diversity and specific habitat associations of macrofungi. Citation: Lebel T, Douch J, Tegart L, et al. 2021. Untangling the Lactifluus clarkeae - Lf. flocktoniae (Russulaceae) species complex in Australasia. Persoonia 47: 1-44. https://doi.org/10.3767/persoonia.2021.47.01.

Untangling the Lactifluus clarkeae - Lf. flocktoniae(Russulaceae) species complex in Australasia.

The Lactifluus clarkeae complex is a commonly observed, generally brightly coloured, group of mushrooms that are usually associated with Nothofagus or Myrtaceous hosts in Australia and New Zealand. For this study collections labelled as 'Lactarius clarkeae', 'Russula flocktoniae' and 'Lactarius subclarkeae' were examined morphologically and molecularly. Analyses of molecular data showed a high cryptic diversity, with sequences scattered across 11 clades in three subgenera within Lactifluus, and a single collection in Russula. We select epitypes to anchor the currently accepted concepts of Lf. clarkeae s.str. and Lf. flocktoniae s.str. The name Lf. subclarkeae could not be applied to any of the collections examined, as none had a lamprotrichoderm pileipellis. Lactifluus clarkeae var. aurantioruber is raised to species level, and six new species are described, three in subg. Lactifluus: Lf. jetiae, Lf. pagodicystidiatus, and Lf. rugulostipitatus, and three in subg. Gymnocarpi: Lf. albens, Lf. psammophilus, and Lf. pseudoflocktoniae. A new collection of Lf. russulisporus provides a significant range extension for the species. Untangling this complex will enable better identification of species and increase understanding of diversity and specific habitat associations of macrofungi. Citation: Lebel T, Douch J, Tegart L, et al. 2021. Untangling the Lactifluus clarkeae - Lf. flocktoniae (Russulaceae) species complex in Australasia. Persoonia 47: 1-44. https://doi.org/10.3767/persoonia.2021.47.01.

A multistage mixed methods study protocol to evaluate the implementation and impact of a reconfiguration of acute medicine in Ireland's hospitals.

BACKGROUND: To address deficits in the delivery of acute services in Ireland, the National Acute Medicine Programme (NAMP) was established in 2010 to optimise the management of acutely ill medical patients in the hospital setting, and to ensure their supported discharge to primary and community-based care. NAMP aims to reduce inappropriate hospital admissions, reduce length of hospital stay and ensure patients receive timely treatment in the most appropriate setting. It does so primarily via the development of Acute Medical Assessment Units (AMAUs) for the rapid assessment and management of medical patients presenting to hospitals, as well as streamlining the care of those admitted for further care. This study will examine the impact of this programme on patient care and identify the factors influencing its implementation and operation. METHODS: We will use a multistage mixed methods evaluation with an explanatory sequential design. Firstly, we will develop a logic model to describe the programme's outcomes, its components and the mechanisms of change by which it expects to achieve these outcomes. Then we will assess implementation by measuring utilisation of the Units and comparing the organisational functions implemented to that recommended by the NAMP model of care. Using comparative case study research, we will identify the factors which have influenced the programme's implementation and its operation using the Consolidated Framework for Implementation Research to guide data collection and analysis. This will be followed by an estimation of the impact of the programme on reducing overnight emergency admissions for potentially avoidable medical conditions, and reducing length of hospital stay of acute medical patients. Lastly, data from each stage will be integrated to examine how the programme's outcomes can be explained by the level of implementation. DISCUSSION: This formative evaluation will enable us to examine whether the NAMP is improving patient care and importantly draw conclusions on how it is doing so. It will identify the factors that contribute to how well the programme is being implemented in the real-world. Lessons learnt will be instrumental in sustaining this programme as well as planning, implementing, and assessing other transformative programmes, especially in the acute care setting.

Investigations into the temporal development of epitheliocystis infections in brown trout: a histological study.

Epitheliocystis in Swiss brown trout (Salmo trutta) is a chlamydial infection, mainly caused by Candidatus Piscichlamydia salmonis and Candidatus Clavichlamydia salmonicola. To gain a better understanding of the temporal development of infections in wild brown trout, we investigated epitheliocystis infections during the course of the summer and autumn months of a single year (2015), and compared this to sampling points over the span of the years 2012-2014. The survey focused on tributaries (Venoge and Boiron) of the Rhone flowing in to Lake Geneva. When evaluated histologically, epitheliocystis infections were found throughout the period of investigation with the exception of the month of June. Fifty to 86 animals per sampling were investigated. Highest prevalence and infection intensities were seen in September. A correlation between epitheliocystis infection and water temperatures was not evident. Interyear comparison revealed consistent levels of prevalence and infection intensities in late summer. The absence of infections in June, combined with the consistent interyear results, indicates seasonal fluctuation of epitheliocystis infections in brown trout with a reservoir persisting during winter months from which infections can re-initiate each year. This could either be at levels below detection limits within the brown trout population itself or in an alternative host.

Improving acute care for adolescents and young adults on medical admission units: The interventions that matter.

It had become a familiar routine. My seventh admission with diabetic ketoacidosis (DKA) in a year. Each time I was admitted it was the same; a DKA protocol, a diabetes specialist nurse visit, and a few questions from the doctors checking if "everything is okay?" On each admission, I would be discharged home after a couple of days. We all knew I'd be back again within a month or two.

RYR3 gene polymorphisms and cardiovascular disease outcomes in the context of antihypertensive treatment.

Nearly one-third of adults in the United States have hypertension, which is associated with increased cardiovascular disease (CVD) morbidity and mortality. The goal of antihypertensive pharmacogenetic research is to enhance understanding of drug response based on the interaction of individual genetic architecture and antihypertensive therapy to improve blood pressure control and ultimately prevent CVD outcomes. In the context of the Genetics of Hypertension Associated Treatment study and using a case-only design, we examined whether single-nucleotide polymorphisms in RYR3 interact with four classes of antihypertensive drugs, particularly the calcium channel blocker amlodipine versus other classes, to modify the risk of coronary heart disease (CHD; fatal CHD and non-fatal myocardial infarction combined) and heart failure (HF) in high-risk hypertensive individuals. RYR3 mediates the mobilization of stored Ca(+2) in cardiac and skeletal muscle to initiate muscle contraction. There was suggestive evidence of pharmacogenetic effects on HF, the strongest of which was for rs877087, with the smallest P-value=0.0005 for the codominant model when comparing amlodipine versus all other treatments. There were no pharmacogenetic effects observed for CHD. The findings reported here for the case-only analysis of the antihypertensive pharmacogenetic effect of RYR3 among 3058 CHD cases and 1940 HF cases show that a hypertensive patient's genetic profile may help predict which medication(s) might better lower CVD risk.

Disseminated toxoplasmosis presenting as septic shock five weeks after renal transplantation.

We discuss a case of acute disseminated toxoplasmosis in a renal transplant recipient presenting with septic shock. Our literature review of disseminated toxoplasmosis presenting as septic shock reveals a disease process that is rapid and almost uniformly fatal. This unusual presentation warrants a high index of suspicion in transplant recipients with immediate administration of appropriate empiric antimicrobials.

Immunohistochemical expression study of proapoptotic BH3-only protein bad in canine nonneoplastic tissues and canine lymphomas.

The BH3-only protein Bad is a proapoptotic Bcl-2 family member that acts as a sensitizer in intrinsic apoptosis by inactivating antiapoptotic members through heterodimer formation. Bad has been shown to contribute to tumorigenesis, including lymphoma formation in humans and mice, through alteration in expression or functional status. Here, its immunohistochemical expression was analyzed in canine nonneoplastic and lymphoma tissues using tissue microarrays. Bad was expressed in the cytoplasm of a wide range of nonneoplastic tissues, especially epithelial cells. Nonneoplastic lymph nodes displayed weak immunostaining in the follicular germinal centers only. Immunoblotting supported these observations but also revealed presence of nonspecific labeling in some organs. Of 81 lymphomas, 29 (35.8%) displayed moderate to strong immunohistochemical Bad labeling, and a significant expression increase was found in lymphomas (especially B cell and double negative) compared to nonneoplastic lymph nodes. These findings warrant further investigations of the functional status, the involvement of partner proteins, and a possible impact of Bad on prognosis in canine lymphoma.

Novel Chlamydiales associated with epitheliocystis in a leopard shark Triakis semifasciata.

The Chlamydiales is a diverse order of obligate intracellular gram-negative bacteria that are known to cause a wide range of diseases in terrestrial animals, including humans. Molecular analyses have revealed that these organisms are also associated with epitheliocystis in teleost fish species, highlighting the suspected deep evolutionary origin of members of this bacterial order. However, our knowledge of their fish host range and of the diversity of the bacteria themselves is still very limited. In this study, we provide molecular evidence for a novel member of the Order Chlamydiales in a nonteleost species, the leopard shark Triakis semifasciata. Based on phylogenetic analysis of the 16S rRNA gene, this novel organism appears to represent a unique lineage in the Order Chlamydiales despite appearing histologically similar to epitheliocystis-causing organisms in other fish species. A greater understanding of the genetic diversity of marine Chlamydiales will assist our attempts to manage and control epitheliocystis outbreaks and to understand the evolution of this unique obligate intracellular pathogen.

Cartilage contains mixed fibrils of collagen types II, IX, and XI.

The distribution of collagen XI in fibril fragments from 17-d chick embryo sternal cartilage was determined by immunoelectron microscopy using specific polyclonal antibodies. The protein was distributed throughout the fibril fragments but was antigenically masked due to the tight packing of collagen molecules and could be identified only at sites where the fibril structure was partially disrupted. Collagens II and IX were also distributed uniformly along fibrils but, in contrast to collagen XI, were accessible to the antibodies in intact fibrils. Therefore, cartilage fibrils are heterotypically assembled from collagens II, IX, and XI. This implies that collagen XI is an integral component of the cartilage fibrillar network and homogeneously distributed throughout the tissue. This was confirmed by immunofluorescence.

J1/tenascin in substrate-bound and soluble form displays contrary effects on neurite outgrowth.

The influence of J1/tenascin adsorbed to polyornithine-conditioned plastic (substrate-bound J1/tenascin) and J1/tenascin present in the culture medium (soluble J1/tenascin) on neurite outgrowth was studied with cultured single cells from hippocampus and mesencephalon of embryonic rats. Neurons at low density grew well on J1/tenascin substrates and extended neurites that were approximately 40% longer than on the polyornithine control substrate after 24 h in vitro. The neurite outgrowth promoting effect of substrate bound J1/tenascin was largely abolished in the presence of mAb J1/tn2, but not by mAb J1/tn1. In contrast to the neurite growth-promoting effects of substrate bound J1/tenascin, neurite outgrowth on polyornithine, laminin, fibronectin, or J1/tenascin as substrates was inhibited by addition of soluble J1/tenascin to the cultures. Neither of the two mAbs neutralized the neurite outgrowth-inhibitory properties of soluble J1/tenascin. In contrast to their opposite effects on neurite outgrowth, both substrate-bound and soluble J1/tenascin reduced spreading of the neuronal cell bodies, suggesting that the neurite outgrowth-promoting and antispreading effects are mediated by two different sites on the molecule. This was further supported by the inability of the mAb J1/tn2 to neutralize the antispreading effect. The J1/tn2 epitope localizes to a fibronectin type III homology domain that is presumably distinct from the putative Tn68 cell-binding domain of chicken tenascin for fibroblasts, as shown by electronmicroscopic localization of antibody binding sites. We infer from these experiments that J1/tenascin contains a neurite outgrowth promoting domain that is distinguishable from the cell-binding site and presumably not involved in the inhibition of neurite outgrowth or cell spreading. Our observations support the notion that J1/tenascin is a multifunctional extracellular matrix molecule.

Protein tyrosine phosphatase alpha (PTPalpha) and contactin form a novel neuronal receptor complex linked to the intracellular tyrosine kinase fyn.

Glycosyl phosphatidylinositol (GPI)-linked receptors and receptor protein tyrosine phosphatases (RPTPs), both play key roles in nervous system development, although the molecular mechanisms are largely unknown. Despite lacking a transmembrane domain, GPI receptors can recruit intracellular src family tyrosine kinases to receptor complexes. Few ligands for the extracellular regions of RPTPs are known, relegating most to the status of orphan receptors. We demonstrate that PTPalpha, an RPTP that dephosphorylates and activates src family kinases, forms a novel membrane-spanning complex with the neuronal GPI-anchored receptor contactin. PTPalpha and contactin associate in a lateral (cis) complex mediated through the extracellular region of PTPalpha. This complex is stable to isolation from brain lysates or transfected cells through immunoprecipitation and to antibody-induced coclustering of PTPalpha and contactin within cells. This is the first demonstration of a receptor PTP in a cis configuration with another cell surface receptor, suggesting an additional mode for regulation of a PTP. The transmembrane and catalytic nature of PTPalpha indicate that it likely forms the transducing element of the complex, and we postulate that the role of contactin is to assemble a phosphorylation-competent system at the cell surface, conferring a dynamic signal transduction capability to the recognition element.

D-periodic distribution of collagen type IX along cartilage fibrils.

It has recently become apparent that collagen fibrils may be composed of more than one kind of macromolecule. To explore this possibility, we developed a procedure to purify fibril fragments from 17-d embryonic chicken sternal cartilage. The fibril population obtained shows, after negative staining, a uniformity in the banding pattern and diameter similar to the fibrils in situ. Pepsin digestion of this fibril preparation releases collagen types II, IX, and XI in the proportion of 8:1:1. Rotary shadowing of the fibrils reveals a d-periodic distribution of 35-40-nm long projections, each capped with a globular domain, which resemble in form and dimensions the aminoterminal globular and collagenous domains, NC4 and COL3, of type IX collagen. The monoclonal antibody (4D6) specific for an epitope close to the amino terminal of the COL3 domain of type IX collagen bound to these projections, thus confirming their identity. Type IX collagen is therefore distributed in a regular d-periodic arrangement along cartilage fibrils, with the chondroitin sulfate chain of type IX collagen in intimate contact with the fibril.

Neuronal cell adhesion molecule contactin/F11 binds to tenascin via its immunoglobulin-like domains.

Adhesive interactions between neurons and extracellular matrix (ECM) play a key role in neuronal pattern formation. The prominent role played by the extracellular matrix protein tenascin/cytotactin in the development of the nervous system, tied to its abundance, led us to speculate that brain may contain yet unidentified tenascin receptors. Here we show that the neuronal cell adhesion molecule contactin/F11, a member of the immunoglobulin(Ig)-superfamily, is a cell surface ligand for tenascin in the nervous system. Through affinity chromatography of membrane glycoproteins from chick brain on tenascin-Sepharose, we isolated a major cell surface ligand of 135 kD which we identified as contactin/F11 by NH2-terminal sequencing. The binding specificity between contactin/F11 and tenascin was demonstrated in solid-phase assays. Binding of immunopurified 125I-labeled contactin/F11 to immobilized tenascin is completely inhibited by the addition of soluble tenascin or contactin/F11, but not by fibronectin. When the fractionated isoforms of tenascin were used as substrates, contactin/F11 bound preferentially to the 190-kD isoform. This isoform differs in having no alternatively spliced fibronectin type III domains. Our results imply that the introduction of these additional domains in some way disrupts the contactin/F11 binding site on tenascin. To localize the binding site on contactin/F11, proteolytic fragments were generated and characterized by NH2-terminal sequencing. The smallest contactin/F11 fragment which binds tenascin is 45 kD and also begins with the contactin/F11 NH2-terminal sequence. This implies that contactin/F11 binds to tenascin through a site within the first three Ig-domains.

Molecular evidence for chlamydial infections in the eyes of sheep.

Ocular infections by chlamydiae are associated with ocular disease manifestations such as conjunctivitis and keratitis in humans and animals. Limited evidence exists that members of the order Chlamydiales can also cause ocular disease in sheep. In the current study, the prevalence of chlamydiae in the eyes of sheep was investigated by using PCR methods. Data obtained in sheep by broad-range 16S rRNA order Chlamydiales-specific PCR were compared to the prevalence of antibodies against chlamydiae detected by a competitive enzyme-linked immunosorbent assay (cELISA). Flocks tested included a clinically healthy flock and two flocks suffering from ocular disease and with histories of Ovine Enzootic Abortion (OEA). PCR detected DNA of Chlamydophila (Cp.) abortus and Cp. pecorum in the eyes of both healthy and sick animals but also identified Chlamydia (C.) suis and a variety of uncultured chlamydia-like organisms. Good correlation was found between the presence of Cp. abortus DNA in sheep conjunctival samples and seropositivity detected by cELISA. Despite these findings, no association was found between the presence of chlamydial DNA in the sheep conjunctival samples and the onset of clinical disease. These results suggest that the biodiversity of chlamydiae in the eyes of sheep is greater than that previously thought. Further investigations are needed to determine whether a causal relationship between infection by chlamydiae and ocular disease exists in these animals.

Evaluating linkage disequilibrium and recombination provides a haplotype-tagging SNP panel of the major histocompatibility complex in African Americans.

The major histocompatibility complex (MHC) (Chromosome 6p21.3) is a dynamic, immune gene-rich region that is associated with multiple diseases. Haplotype-tagging single-nucleotide polymorphism (htSNP) panels for the MHC can aid association studies but have only been reported for African, Asian and Caucasian populations to date. We genotyped 2154 SNPs spanning a 3.8-Mb region of the classical MHC in 94 healthy African Americans using Illumina BeadArray technology. We describe the haplotype structure of the MHC in African Americans, calculate the recombination rate (0.35 cM Mb(-1)) across the region, identify recombination hot spots and develop a panel of htSNPs for future genetic association studies in this population. We conclude that while patterns of LD and recombination are similar within the MHC to that reported in other populations, differences in minor allele frequency at specific markers necessitates an htSNP panel unique to African Americans, which we provide here for use in future genetic association studies.

Dynamical implications of Viral Tiling Theory.

The Caspar-Klug classification of viruses whose protein shell, called viral capsid, exhibits icosahedral symmetry, has recently been extended to incorporate viruses whose capsid proteins are exclusively organised in pentamers. The approach, named 'Viral Tiling Theory', is inspired by the theory of quasicrystals, where aperiodic Penrose tilings enjoy 5-fold and 10-fold local symmetries. This paper analyses the extent to which this classification approach informs dynamical properties of the viral capsids, in particular the pattern of Raman active modes of vibrations, which can be observed experimentally.

Comparison of the Immulite and RIA assay methods for measuring peripheral blood progesterone levels in Greyhound bitches.

Determination of optimal breeding time in bitches earmarked for single insemination only is based on measurement of peripheral blood serum or plasma progesterone concentration. In this paper a comparison is made between radioimmune assay (RIA) and chemoluminescent assay (Immulite) for determination of P4 concentrations in the bitch. The Immulite assay is shown to be an accurate and reliable method for serum or plasma P4 measurement. It compares favourably with other methods in terms of turn-around time, cost and accessibility for veterinarians in practice.

Breeding soundness evaluation of bulls by semen analysis, testicular fine needle aspiration cytology and trans-scrotal ultrasonography.

The aim of this study was to evaluate the usefulness of trans-scrotal ultrasonography and testicular fine needle aspiration cytology in assessing bulls for breeding suitability. These two techniques were also compared with semen analysis. Bulls presented for breeding soundness evaluation were assessed using all three techniques. The findings of each technique were compared. There was agreement in classification of fertile bulls using all three techniques, suggesting that the combined use of these techniques enhances routine breeding soundness examination. Use of the three techniques also enhances detailed investigation of suspected sub-fertile bulls while accurately identifying testicular cause(s) of sire sub-fertility.

Correspondence-free determination of the affine fundamental matrix.

Fundamental matrix estimation is a central problem in computer vision and forms the basis of tasks such as stereo imaging and structure from motion. Existing algorithms typically analyze the relative geometries of matched feature points identified in both projected views. Automated feature matching is itself a challenging problem. Results typically have a large number of false matches. Traditional fundamental matrix estimation methods are very sensitive to matching errors, which led naturally to the application of robust statistical estimation techniques to the problem. In this work, an entirely novel approach is proposed to the fundamental matrix estimation problem. Instead of analyzing the geometry of matched feature points, the problem is recast in the frequency domain through the use of Integral Projection, showing how this is a reasonable model for orthographic cameras. The problem now reduces to one of identifying matching lines in the frequency domain which, most importantly, requires no feature matching or correspondence information. Experimental results on both real and synthetic data are presented that demonstrate the algorithm is a practical technique for fundamental matrix estimation. The behavior of the proposed algorithm is additionally characterized with respect to input noise, feature counts, and other parameters of interest.