Phylogenetic placement of lichenicolous Phoma species in the Phaeosphaeriaceae (Pleosporales, Dothideomycetes)

More than twenty species of lichenicolous fungi have been described in Phoma, a large anamorphic genus of primarily plant-associated pathogens with broad geographic distributions. We obtained nuclear and mitochondrial rDNA sequences from 19 fungal cultures isolated from specimens representing four described and two undescribed lichenicolous species in the genus. Our multilocus phylogeny indicates that lichenicolous Phoma species represent at least two phylogenetically distinct clades in the Phaeosphaeriaceae, one including a new species, Phoma puncteliae, isolated from a specimen of Punctelia rudecta collected in Maryland, USA, and another group of primarily lichenicolous species. This latter group includes four described lichenicolous Phoma species, an unidentified melanized rock fungus, and a new lichenicolous Phoma species isolated from Xanthomendoza species collected in Canada that we are naming P. xanthomendozae. Some specimens in this clade collected from different lichen genera and species were found to be very similar genetically, which calls into question the recent practice of recognizing lichenicolous Phoma species mainly by differences in host preference.     

New insights into relationships of lichen-forming Dothideomycetes

Here we continue to investigate the phylogenetic relationships of taxa ascribed to the primarily lichen-forming families Trypetheliaceae, Monoblastiaceae and Arthopyreniaceae. We demonstrate that the genera Julella and Arthopyrenia do not form monophyletic groups with taxa from these genera instead being placed both in Pleosporales and Trypetheliales. Within Dothideomycetes, lichen-forming species with brown ascospores are generally placed in the genera Mycomicrothelia, Architrypethelium, and Aptrootia in the family Trypetheliaceae. We tested the taxonomic placement of Anisomeridium phaeospermum, in Monoblastiaceae. This species produces brown-spores with wall ornamentation and therefore appears morphologically similar to Mycomicrothelia. Despite these morphological similarities, molecular data confirmed its placement in Anisomeridium. Consequently, the distinction between these two genera is in need of clarification and ascus characters are identified as the principal discriminating feature. Finally, we identify the non-lichenized taxa Heleiosa barbatula and Funbolia dimorpha as being part of the otherwise lichen-forming family Monoblastiaceae. This is confirmed by the perithecial anatomy of the sexually reproducing Heleiosa barbatula.     

Phylogenetic relationships of five genera of Xylariales and Rosasphaeria gen. nov. (Hypocreales)

Eight inconspicuous non-stromatic perithecial fungi immersed in plant tissue are assessed with respect to their morphology, ecology and phylogenetic position. Emphasis is laid on two genera now and then placed in the family Hyponectriaceae, Xylariales: Leiosphaerella with its type species L. praeclara occurring on Vaccinium, and Pseudomassaria with its type species P. chondrospora occurring on Tilia. In molecular phylogenetic analyses of LSU and ITS sequences, the generic types of these genera are closely related, but their familial affiliation within Xylariales remains unresolved. Pseudomassaria sepincoliformis clusters with P. chondrospora, whereas P. fallax is not congeneric with Pseudomassaria and P. lycopodina is combined in Leiosphaerella despite its apiospores. Three species thought to belong to Leiosphaerella are re-assessed: L. moravica that occurs on Rosa, is disposed in the new genus Rosasphaeria, which is close to Eucasphaeria in the Niessliaceae (Hypocreales), according to multi-gene phylogenetic analyses (ITS, LSU, rpb2 and tef1). For L. vexata the genus Pseudomassariella is revived. A Leiosphaerella-like fungus on Lycopodium is described as the new species Monographella lycopodina. In addition the phylogenetic relationships of two fungi forming superficial stromata are here clarified: Collodiscula japonica belongs to the Xylariaceae, while Melogramma campylosporum may currently be interpreted as representing a family of its own, the Melogrammataceae.     

A taxonomic revision and phylogenetic reconstruction of the Jahnulales (Dothideomycetes), and the new family Manglicolaceae

Genera assigned to the Jahnulales are morphologically diverse, especially in ascospores equipped with or without appendages, sheaths or apical caps. They are predominantly freshwater fungi occurring on woody substrata, with Manglicola guatemalensis, Xylomyces chlamydosporus and X. rhizophorae the only species known from marine habitats. The order Jahnulales with 4 teleomorphic genera: Jahnula (15 species), Aliquandostipite (5), Megalohypha (1), Manglicola (2) and the anamorphic genera Brachiosphaera (2), Speiropsis (9), Xylomyces (8), amounting to a total of 42 species, is reviewed and nomenclatural changes are proposed. Twenty species are treated at the molecular level, with 94 sequences, 13 of which are newly generated for this review. Three species are rejected (Speiropsis irregularis, Xylomyces aquaticus, X. elegans) while the phylogenetic placement of 6 Xylomyces, 7 Speiropsis, 1 Brachiosphaera and 1 Manglicola require molecular data to confirm their placement in the order. Sequences are derived from ex-holotype isolates and new collections made in Thailand. Most taxa are included in the family Aliquandostipitaceae and a new family Manglicolaceae is erected for the marine ascomycete Manglicola guatemalensis with its large ascomata (1,100–1,750?×?290–640 μm), wide ostioles and ascospores that are fusiform, unequally one-septate with the apical cell larger than the turbinate basal cell and bear apical gelatinous appendages. The genus Jahnula is polyphyletic grouping in three clades with J. aquatica, J. granulosa, J. rostrata, J. potamophila and Megalohypha aqua-dulces in the Jahnula sensu stricto clade. No taxonomical changes are proposed for Jahnula species not grouping in the Jahnula sensu stricto clade, until further species are isolated and sequenced.     

Phylogenetic relationships of snake venom toxic proteins

Two kinds of distance matrices have been formed from minimum mutational distances and absolute hydrophobicity differences obtained by comparison of aligned homologous sequences of 56 toxins from venom of snakes belonging to 7 genera. Phylogenetic trees were constructed from these distance matrices, employing the unweighted pair-group method using arithmetic averages (UPGMA). The Pearson product-moment correlation coefficient has been used to estimate the agreement between the original distance matrix and that obtained directly from the dendrogram. For all these procedures the set of computer programs PHYTREE (written in BASIC for micro-computer, and available from the author) has been used.     

A molecular, morphological and ecological re-appraisal of Venturiales―a new order of Dothideomycetes

The Venturiaceae was traditionally assigned to Pleosporales although its diagnostic characters readily distinguish it from other pleosporalean families. These include a parasitic or saprobic lifestyle, occurring on leaves or stems of dicotyledons; small to medium-sized ascomata, often with setae; deliquescing pseudoparaphyses; 8-spored, broadly cylindrical to obclavate asci; 1-septate, yellowish, greenish or pale brown to brown ascospores; and hyphomycetous anamorphs. Phylogenetically, core genera of Venturiaceae form a monophyletic clade within Dothideomycetes, and represent a separate sister lineage from current orders, thus a new order—Venturiales is introduced. A new family, Sympoventuriaceae, is introduced to accommodate taxa of a well-supported subclade within Venturiales, which contains Sympoventuria, Veronaeopsis simplex and Fusicladium-like species. Based on morphology and DNA sequence analysis, eight genera are included in Venturiaceae, viz. Acantharia, Apiosporina (including Dibotryon), Caproventuria, Coleroa, Pseudoparodiella, Metacoleroa, Tyrannosorus and Venturia. Molecular phylogenetic information is lacking for seven genera previously included in Venturiales, namely Arkoola, Atopospora, Botryostroma, Lasiobotrys, Trichodothella, Trichodothis and Rhizogenee and these are discussed, but their inclusion in Venturiaceae is doubtful. Crotone, Gibbera, Lineostroma, Phaeocryptopus, Phragmogibbera, Platychora, Polyrhizon, Rosenscheldiella, Uleodothis and Xenomeris are excluded from Venturiales, and their ordinal placement needs further investigation. Zeuctomorpha is treated as a synonym of Acantharia.     

Expanded taxon sampling disentangles evolutionary relationships and reveals a new family in Peltigerales (Lecanoromycetidae, Ascomycota)

Lichens that incorporate cyanobacterial symbionts (cyanolichens) are an ecologically key group of species used as biomonitors at all latitudes. Cyanolichen evolution is however based on intense studies of few keystone species and the bulk of species diversity, especially of small species in cold climates, has yet to be accounted for in phylogenetic studies. We assembled an expanded data set including members of all nine currently accepted Peltigeralean families as well as hitherto undersampled representatives of small, radially symmetrical, placodioid cyanolichen genera from the Northern and Southern Hemispheres. Bayesian and maximum likelihood consensus trees from our multilocus analyses (nuSSU, nuLSU and mtSSU) recovered the genera Koerberia, Vestergrenopsis and Steinera as a new, fully supported, family-level clade within the Peltigerales. This clade is further supported by a posteriori morphological analysis and we describe it here as the new family Koerberiaceae. The recently described and physiognomically similar genus Steineropsis, by contrast, is recovered as sister to Protopannaria in the Pannariaceae (Collematineae). Previous analyses have recovered strong monophyletic groups around Pannariaceae, Lobariaceae and Peltigeraceae. We discuss in detail the phylogenetic relationships of all these taxa, provide a pan-Peltigeralean overview of phenotypic characteristics and illustrate all major ascus apical structures. Our topology provides strong backbone support for the sister relationship of Peltigerineae to Collematineae as well as for most currently recognized families of the Peltigerales. The following new combinations are made: Steinera symptychia (Tuck.) T. Sprib. & Muggia, and Vestergrenopsis sonomensis (Tuck.) T. Sprib. & Muggia.     

Structure-activity relationships (SAR) of hydroxyapatite-binding molecules

Summary Carboxyl-containing molecules can bind to HA with sufficient affinity to prevent the uptake of AZ by this mineral. Structural features that favor binding can be discerned. Isolated carboxyls are inactive; adjacent dicarboxylic functions are marginally active. Crowding of oxygens in the form of vicinal carboxyls or hydroxyls contributes the property of HA binding ability. This property exhibits gradation in magnitude and the variation can be correlated with structural features of the carboxylcontaining molecule. Phospho functions are generally more powerful contributors to HA binding property than are carboxyls.     

Temporal and Spatial Analysis of the New World Screwworm (Cochliomyia hominivorax) in Darien and Embera,Panama (2001–2011)

Larvae (maggots) of Cochliomyia hominivorax, the New World Screwworm fly, are voracious consumers of living flesh that have a negative economic impact by decreasing productivity, predisposing to other pathogens, and, in severe cases, causing death of domestic livestock. Screwworm caused extensive financial losses to the livestock industry in North America prior to its eradication. Sterile insect technique (SIT) was used to eradicate screwworm throughout North and Central America and continues to be the main tool to control it in eastern Panama. The goal of this study was to evaluate the temporal and spatial trends of screwworm myiasis cases reported in the Province of Darien and Comarca Embera (border with Colombia), Panama, from 2001 to 2011. We hypothesized that screwworm cases would vary seasonally and be spatially clustered near Colombia as a result of effective eradication strategies in Panama and the presence of an autochthonous population of flies in western Colombia. Temporal and spatial data were retrieved from COPEG‐USDA records (Panama) and analysed by anova , Ripley's K function, discrete Poisson spatial statistic scan and Getis‐Ord Gi*. No significant temporal trend was found, but cases were spatially distributed in four clusters. One cluster of cases occurred from 2001 to 2003 and was considered a focal temporal and spatial cluster. One cluster occurred in 2001 and 2007 indicating more rare outbreaks in an area with fewer cattle. The two remaining clusters contained cases from 2004 to 2011 and 2001 to 2011 suggesting regular breaks in the control barrier due to occasional failures of the SIT programme, difficulties implementing border quarantine strategies, livestock smuggling or the movement of infested wildlife.     

Refractory disease,new horizons and patient-physician relationships

Understanding the role of chemotherapy for prostate cancer has advanced along two axes, better definition of endpoints, and biologic understanding of disease targets. Use of PSA as a surrogate endpoint makes possible the more rational design of phase III trials which will include survival and disease free intervals as an endpoint. Pain control has emerged as an important treatment endpoint through which more cost-effective care can be envisioned. Discovery of growth factor interactions with prostate cancer cells and the elucidation of apoptotic pathways have provided numerous new targets for biologic and chemotherapy of advanced disease.     

Calcium absorption from a new calcium delivery system (CCM)

Absorption of calcium from a highly soluble form of calcium, a mixed calcium citrate-malate^* salt (CCM), was tested against calcium carbonate and milk in both rats and humans. The rat method estimated absorption from the 6-day retention of an oral tracer, and the human method employed the standard double-isotope procedure. CCM was given both as a dry powder and in an organe juice beverage. In two experiments in rats calcium from CCM was absorbed at least as well as, if not better than from calcium carbonate or milk. In two separate experiments in humans, calcium from CCM was absorbed significantly better than from calcium carbonate or milk. We conclude that CCM exhibits excellent bioavailability and that this formulation is a useful addition to the forms of calcium now available either for direct supplementation or for food fortification.     

Phylogenetic conservation of gp96-mediated antigen-specific cellular immunity: new evidence from adoptive cell transfer in xenopus

BACKGROUND: In vertebrates from man to frogs, the heat shock protein (hsp) gp96 elicits T-cell responses against antigenic peptides that it chaperones. In Xenopus, immunization with gp96 purified from normal tissues accelerates rejection of MHC identical, minor histocompatibility (H) antigen-disparate skin grafts in vivo and induces MHC-restricted CTL responses in vitro. Also in Xenopus, gp96 derived from MHC class I-negative tumors elicits peptide-specific responses against these tumors in vivo and MHC-unrestricted CD8 killing in vitro. We have developed an adoptive cell transfer protocol to further characterize these gp96-stimulated Xenopus effectors in vivo. METHODS AND RESULTS: Carboxyfluorescein diacetate succinimidyl ester (CFSE)-stained splenocytes from cloned LG-6 donor frogs immunized with gp96 purified from minor H-antigen-disparate LG-15 tissues were transferred into LG-6 recipients bearing a LG-15 minor H antigen (ag)-disparate skin graft. Primed anti-LG-15 but not naive CFSE T cells accumulated and divided in the spleen of allografted recipients to a greater extent than in those of autografted recipients. Similar accumulation and division occurred when CD8 T cells primed by 15/0 tumor-derived gp96 were transferred to an isogeneic recipient bearing the same MHC class I-negative tumor. Furthermore, the transfer of such primed antitumor splenocytes into naive recipients before tumor challenge delayed the appearance of tumors. CONCLUSIONS: These data provide new in vivo evidence that in frogs as in mammals, gp96 can prime CD8 T cells against antigens they chaperone. In addition, at least in Xenopus, gp96 can prime CD8(+) T-cell effectors that are not MHC restricted.     

Phylogenetic and chemotaxonomic resolution of the genus <Emphasis Type="Italic">Annulohypoxylon</Emphasis> (Xylariaceae) including four new species

Fourty-three species of microfungi from bamboo are treated, including one new family, Occultibambusaceae, three new genera, Neoanthostomella, Occultibambusa and Seriascoma, 27 new species, one renamed species and 15 re-described or re-illustrated species, and four designated reference specimens are treated in this paper, the majority of which are saprobic on dead culms. To determine species identification, separate phylogenetical analyses for each group are carried out, based on molecular data from this study and sequences downloaded from GenBank. Morphologically similar species and phylogenetically close taxa are compared and discussed. In addition a list of bambusicolous fungi published since Hyde and colleagues in 2002 is provided.     

Establishment and characterization of a new cell line from human bladder cancer (JMSU1)

A new human bladder cancer cell line designated JMSU1 has been established from malignant ascitic fluid of a 75-year-old Japanese man with bladder cancer, and maintained in culture for more than 7 years and over 240 passages. Inverted phase-contrast microscopy revealed that JMSU1 was composed of morphologically distinct cells (polygonal to spindle-shaped cells), showing morphological heterogeneity in vitro. Histological examination of xenografts showed poorly differentiated transitional cell carcinoma, resembling the original tumor. Immunohistochemical staining for cytokeratin and electron microscopic examination suggested that JMSU1 was of epithelial origin. Chromosome analysis gave a modal number of 69 with no Y chromosome. Isozyme analysis (LDH, G6PD, and NP) showed the mobility pattern of human type B. DNA fingerprint analysis demonstrated that there was no cross-culture contamination of JMSU1 during the passages. In conclusion, a newly established and well-characterized cell line, JMSU1, offers promising material for the investigation of the biological properties of bladder cancer.     

Lichenicolous fungi of the genus Abrothallus (Dothideomycetes: Abrothallales ordo nov.) are sister to the predominantly aquatic Janhulales

This study provides new insights on the phylogenetic position of the lichenicolous fungal genus Abrothallus based on six molecular markers (nuSSU, nuLSU, mtSSU, RPB1, RPB2 and TEF-α). In a broad-scale analysis, we detected high support for inclusion of the genus within Dothideomycetes. A further analysis provided support for Abrothallus as a member of the subclass Pleosporomycetidae as a sister group of Jahnulales, an order of aquatic Dothideomycetes. Given the exclusive characters of this group of apotheciate fungi within the Dothidiomycetes, a new monotypic order Abrothallales is here introduced together with the new family Abrothallaceae. In a multi-locus analysis (based on the six loci indicated above plus ITS) restricted to 12 putative Abrothallus species, two clearly separated clades were observed: one comprising species growing on lichens of the families Parmeliaceae and Ramalinaceae, and the second including species that live on lichens of the order Peltigerales and the family Cladoniaceae.     

日间手术病历书写规范专家共识（2019年）

随着国家医改层面促进日间手术的发展,日间手术占择期手术比例逐步提高。由于日间手术住院时间短、病种多、周转快,病历书写工作量较大,普通病历模板很难适用于日间手术发展的临床需要。目前国家层面尚缺乏统一的日间手术病历书写规范和标准。为促进和规范日间手术的发展,国家老年疾病临床医学研究中心(湘雅)组织专家,依据原卫生部《病历书写基本规范》(2010年版)和国家卫生计生委医疗管理服务指导中心《日间手术管理导则》(2016版征求意见稿),在《中南大学湘雅医院日间手术病历书写规范(2018)》的基础上制订《日间手术病历书写规范专家共识》。     

中心静脉穿刺新进针点的可行性研究

Objective To evaluate the feasibility of a new approach to internal jugular vein (IJV) catheterization. Methods Sixty patients undergoing cardiac surgery were randomly divided into two groups (n=30). In group Ⅰ, a new method was used. In group Ⅱ, IJV catheterization from a high medial approach. The overall succcess rate of puncture, the success rate of puncture at first attempt, the success rate of cannulation and complication were recorded. Chest X-ray was taken for confirmation of the position of catheter in subclavian vein. Results The overall success rate of puncture was 100% and the success rate of puncture at first attempt was 93.33% in group Ⅰ, which is significantly higher than those in group Ⅱ(P<0.05). No complication occurred in group Ⅰ including arterial puncture, pneumothorax, hematothorax and difficult catheter placement. In group II there were 3 arterial punctures. Con-clusion This new method is a reliable and safe technique in patienst as compared with the high medial approach.     

中心静脉穿刺新进针点的可行性研究

Objective To evaluate the feasibility of a new approach to internal jugular vein (IJV) catheterization. Methods Sixty patients undergoing cardiac surgery were randomly divided into two groups (n=30). In group Ⅰ, a new method was used. In group Ⅱ, IJV catheterization from a high medial approach. The overall succcess rate of puncture, the success rate of puncture at first attempt, the success rate of cannulation and complication were recorded. Chest X-ray was taken for confirmation of the position of catheter in subclavian vein. Results The overall success rate of puncture was 100% and the success rate of puncture at first attempt was 93.33% in group Ⅰ, which is significantly higher than those in group Ⅱ(P<0.05). No complication occurred in group Ⅰ including arterial puncture, pneumothorax, hematothorax and difficult catheter placement. In group II there were 3 arterial punctures. Con-clusion This new method is a reliable and safe technique in patienst as compared with the high medial approach.