bluuebunny/crossref_metadata_2025 huggingface dataset:
TNT, a free program for phylogenetic analysis
Authors: Goloboff Pablo A. , Farris James S. , Nixon Kevin C.
Published: July 2008

The main features of the phylogeny program TNT are discussed. Windows versions have a menu interface, while Macintosh and Linux versions are command‐driven. The program can analyze data sets with discrete (additive, non‐additive, step‐matrix) as well as continuous characters (evaluated with Farris optimization). Effective analysis of large data sets can be carried out in reasonable times, and a number of methods to help identifying wildcard taxa in the case of ambiguous data sets are implemented. A variety of methods for diagnosing trees and exploring character evolution is available in TNT, and publication‐quality tree‐diagrams can be saved as metafiles. Through the use of a number of native commands and a simple but powerful scripting language, TNT allows the user an enormous flexibility in phylogenetic analyses or simulations.

© The Willi Hennig Society 2008.
Phylum Tardigrada: an “individual” approach
Authors: Sands Chester J. , McInnes Sandra J. , Marley Nigel J. , Goodall‐Copestake William P. , Convey Peter , Linse Katrin
Published: July 2008

Phylum Tardigrada consists of ∼1000 tiny, hardy metazoan species distributed throughout terrestrial, limno‐terrestrial and oceanic habitats. Their phylogenetic status has been debated, with current evidence placing them in the Ecdysozoa. Although there have been efforts to explore tardigrade phylogeny using both morphological and molecular data, limitations such as their few morphological characters and low genomic DNA concentrations have resulted in restricted taxonomic coverage. Using a protocol that allows us to identify and extract DNA from individuals, we have sequenced 18S rDNA from 343 tardigrades from across the globe. Using maximum parsimony and Bayesian analyses we have found support for dividing Order Parachela into three super‐families and further evidence that indicates the traditional taxonomic perspective of families in the class Eutardigrada are nonmonophyletic and require re‐working. It appears that conserved morphology within Tardigrada has resulted in conservative taxonomy as we have found cases of several discrete lineages grouped into single genera. Although this work substantially adds to the understanding of the evolution and taxonomy of the phylum, we highlight that inferences gained from this work are likely to be refined with the inclusion of further taxa—specifically representatives of the nine families yet to be sampled.

© The Willi Hennig Society 2008.
Accumulation of the hormone abscisic acid (ABA) at the infection site of the fungus Cercospora beticola supports the role of ABA as a repressor of plant defence in sugar beet
Authors: SCHMIDT KLAUS , PFLUGMACHER MAIKE , KLAGES SIMONE , MÄSER ANJA , MOCK ANDREA , STAHL DIETMAR J.
Published: July 2008

Inducible plant defence responses in sugar beet (Beta vulgaris L.) leaves are repressed during the early phase of infection by the fungus Cercospora beticola. In this report, we show that the concentration of the plant hormone abscisic acid (ABA) increases in sugar beet leaves during C. beticola infection. After an initial burst of ABA induced by inoculation of the fungus, elevated ABA concentrations were detected during the fungal penetration and colonization phases 3–9 days after inoculation. Fifteen days after inoculation, with visible onset of the necrotic phase of infection, the strongly elevated ABA concentrations in infected leaves were at levels similar to drought‐stressed plants. A synthetic promoter composed of four copies of the ABA‐responsive element (ABRE) A2 and the coupling element CE3 of the ABA‐inducible barley gene HVA1 was strongly induced by ABA and C. beticola infection in transgenic sugar beet leaves. Analysis of the spatial pattern of promoter activity revealed that the ABA‐inducible promoter was locally activated at the fungal infection sites. Furthermore, expression of the basic leucine zipper transcription factor AREB1 was induced by drought stress and fungal infection in the sugar beet. Application of ABA reduced the promoter activity of the phenylalanine ammonia lyase (BvPAL) gene, and this effect was observed with the –34 to +248 BvPAL promoter region. This region is equivalent to the core promoter, which is necessary for the suppression of BvPAL expression by C. beticola, as recently shown. These data indicate that ABA accumulation and activation of the ABA‐dependent signalling cascade are the primary cause of suppression of BvPAL expression during infection of sugar beet leaves.
Paroxetine and fluoxetine in pregnancy: a prospective, multicentre, controlled, observational study
Authors: Diav‐Citrin Orna , Shechtman Svetlana , Weinbaum Dafna , Wajnberg Rebecka , Avgil Meytal , Di Gianantonio Elena , Clementi Maurizio , Weber‐Schoendorfer Corinna , Schaefer Christof , Ornoy Asher
Published: July 2008

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• In recent years there has been concern regarding the possibility that selective serotonin reuptake inhibitors (SSRIs) cause an increased rate of congenital cardiovascular anomalies.

• As of today, there is still debate in the literature as to the possible effects of paroxetine and fluoxetine on the embryonic cardiovascular system.

WHAT THIS STUDY ADDS

• Based on prospective data from three Teratogen Information Services, we have demonstrated an increased rate of congenital cardiovascular anomalies among the offspring of fluoxetine‐ and paroxetine‐treated mothers.

AIMS

Recent studies have suggested a possible association between maternal use of selective serotonin reuptake inhibitors (SSRIs) in early pregnancy and cardiovascular anomalies. The aim of the present study was to evaluate the teratogenic risk of paroxetine and fluoxetine.

METHODS

This multicentre, prospective, controlled study evaluated the rate of major congenital anomalies after first‐trimester gestational exposure to paroxetine, fluoxetine or nonteratogens.

RESULTS

We followed up 410 paroxetine, 314 fluoxetine first‐trimester exposed pregnancies and 1467 controls. After exclusion of genetic and cytogenetic anomalies, there was a higher rate of major anomalies in the SSRI groups compared with the controls [paroxetine 18/348 (5.2%), fluoxetine 12/253 (4.7%) and controls 34/1359 (2.5%)]. The main risk applied to cardiovascular anomalies [paroxetine 7/348 (2.0%), crude odds ratio (OR) 3.47, 95% confidence interval (CI) 1.13, 10.58; fluoxetine 7/253 (2.8%), crude OR, 4.81 95% CI 1.56, 14.71; and controls 8/1359 (0.6%)]. On logistic regression analysis only cigarette smoking of ≥10 cigarettes day−1 and fluoxetine exposure were significant variables for cardiovascular anomalies. The adjusted ORs for paroxetine and fluoxetine were 2.66 (95% CI 0.80, 8.90) and 4.47 (95% CI 1.31, 15.27), respectively.

CONCLUSION

This study suggests a possible association between cardiovascular anomalies and first‐trimester exposure to fluoxetine.
Effect of sample size on cyst recovery by flotation methods: Recommendations for sample processing during EU monitoring of potato cyst nematodes (Globodera spp.)
Authors: Bellvert J. , Crombie K. , Horgan F. G.
Published: July 2008

Under EC Council Directive 2007/33/EC member states of the European Union are recommended to take soil samples ranging from 100 mL to >1500 mL to monitor populations of potato cyst nematodes [PCN] (Globodera spp.) in targeted fields. This study examines the effects of varying sample size on sample processing and cyst extraction using two widely used laboratory methods. Cyst recovery was stable using the Fenwick can from 100 mL up to the physical limits of the can. Recovery was significantly lower for low density 50 mL samples; however, this was partly due to a statistical artefact since higher numbers of cysts were lost from successively larger samples with constant cyst numbers (i.e. decreasing densities). The Schuiling centrifuge had functional limits of <100 mL and >500 mL. Outside this range, cyst recovery from low density samples was significantly reduced. Cyst recovery from samples of varying size, but with equal numbers of cysts was only constant above 100 mL. However, samples ranging from 200–500 mL were optimal for the Schuiling centrifuge. Cyst extraction efficiency was similar using both methods for samples ranging from 150 mL to 1500 mL, where larger samples were divided into <500 mL portions. However, processing times (i.e. cyst extraction and microscopic examination) and water consumption increased significantly with increasing sample size and were significantly higher when using the Fenwick can.
Assessment of the resistance of potato cultivars toSynchytrium endobioticum(Schilb.) Per. in Poland
Authors: Przetakiewicz J.
Published: July 2008

In Poland the Plant Breeding and Acclimatization Institute is responsible for officially assessing the resistance toSynchytrium endobioticumof domestic potato breeding lines and cultivars from other countries. Cultivation of potato cultivars in Poland requires confirmation of resistance to potato wart disease. The official assessment uses the modified Glynne‐Lemmerzahl method (laboratory tests) and pot tests. The full cycle of assessment of resistance to wart disease requires 52 seed potatoes per variety/breeding line. Forty two tubers are used in laboratory tests. To complete the laboratory tests the next 10 tubers are grown in pot tests (in soil with winter sporangia) during the vegetation season. The final results for domestic breeding lines of potato are available after 3 years of investigation. For cultivars from other countries the authorization of resistance toS. endobioticumtakes approximately one year. The Polish breeders (breeding lines) or the breeder's representative (cultivars from other countries) receive the certificate only for lines/cultivars with laboratory and field resistance toS. endobioticum.
Extensive damage on white variety table grapes by the Mediterranean fruit fly Ceratitis capitata (Wiedemann) in Crete
Authors: Roditakis E. , Tsagkarakou A. , Roditakis N. E.
Published: July 2008

A severe attack by the Mediterranean fruit fly Ceratitis capitata (Wiedemann) was recorded for the first time in Central‐Northern Crete (Greece) on white variety table grapes (cv. Soultanina) in August 2007. The infestation was detected in hormone (gibberellic acid) treated vineyards only and was successfully controlled using cultural measures and baited application of insecticides. This report is discussed in relation to the possible effects on the table grape export market, based on the fact that C. capitata is under quarantine regulations worldwide.
Première apparition de Sipha flava (Homoptera, Aphididae) sur la canne à sucre au Maroc
Authors: Abdelmajid Nadif
Published: July 2008

Depuis l’introduction de la canne à sucre au Maroc dans les années 70, les ravageurs rencontrés sur cette culture étaient jusqu’à présent tous considérés d’importance mineure, à l’exception de la sésamie (Sesamia nonagrioides) qui peut occasionnellement causer des dégâts par ses infestations sporadiques et irrégulières. Mais avec l’identification en Novembre 2006 d’un nouveau puceron de couleur jaunâtre connu sous le nom de Sipha flava, la situation pourrait devenir plus qu’alarmante. En effet, S. flava est particulièrement nuisible pour la canne à sucre, pouvant provoquer un jaunissement voire même un dépérissement des plants attaqués. Le puceron a déjà colonisé pratiquement toutes les variétés performantes de canne à sucre dans les deux périmètres de cultures irriguées (Gharb et Loukkos). Son identification au Maroc fait de son apparition une première sur le continent africain car il était auparavant strictement confiné au continent américain.
Occurrence of the emerald ash borer,Agrilus planipennisin Russia and its potential impact on European forestry
Authors: Baranchikov Y. , Mozolevskaya E. , Yurchenko G. , Kenis M.
Published: July 2008

The emerald ash borer,Agrilus planipennis, is a beetle native to East Asia where it is considered a minor pest, preferentially attacking weakened or dying ash trees. It was first discovered in North America in 2002 and has since become one of the most serious invasive insect pests, killing millions of healthy ash trees in urban and forested settings. Similar damage is now occurring in the region of Moscow, Russia, which causes serious concern for Europe. In this paper, we review the current knowledge onA. planipennisin Asia and North America, provide new information on its occurrence in the region of Moscow and Eastern Russia and make recommendations for research and management strategies in Europe.
Native and exotic coniferous species in Europe – possible host plants for the potentially invasive Siberian moth, Dendrolimus sibiricus1 Tschtv. (Lepidoptera, Lasiocampidae)
Authors: Kirichenko N. I. , Flament J. , Baranchikov Y. N. , Grégoire J.‐C.
Published: July 2008

The performance of young larvae of the potentially invasive Siberian moth Dendrolimus sibiricus Tschtv. has been studied for the first time on the native and exotic coniferous species which are widely distributed and of considerable commercial value in Europe. Picea, Abies, the introduced Pinus species (five‐needle pine only), and species from the exotic genera Pseudotsuga and Cedrus (all Pinaceae) are found to be suitable hosts for pest development. Two‐needle pines and species from non‐native Tsuga (also Pinaceae) are poor hosts in terms of larval performance though they may support growth of neonates (most sensitive to food quality). Coniferous species from other families: Taxaceae and Cupressaceae are inedible for the pest. The fact that the Siberian moth is able to survive and develop on all the tested genera of Pinaceae and that some of them constitute two thirds of the European forests underline how harmful this defoliator could be in the case of its introduction into European countries.
Accreditation process in the National Phytosanitary laboratory in Latvia1
Authors: Bokuma G. , Lielmane I.
Published: July 2008

The National Phytosanitary laboratory is the only laboratory in Latvia that provides testing of samples for regulated pests in the phytosanitary field. In 2001 a decision was made to implement Quality Assurance systems in the laboratory and obtain accreditation. From 2001 to 2003 preparation for accreditation took place in the laboratory. During this time a client of the laboratory, Sanitary Border Inspection of Latvia (SBI), arranged three audits to assess the activities of the laboratory in accordance with the requirements of ISO 17025. After a time and labour‐consuming preparation process on 2003‐12‐29 all the necessary documents including a Quality manual, two methods and procedures were submitted to the Latvian National Accreditation Bureau (LATAK). The first audit by LATAK was carried out in March 2005. After elimination of all non‐conformities, accreditation for two methods was received in 2005 (Method for detection and identification of Clavibacter michiganensis subsp. sepedonicus in potato tubers and Method for detection and identification of Ralstonia solanacearum in potato tubers). The Quality Assurance System was implemented and applies to the whole laboratory. Every year LATAK experts carry out monitoring visits and each time imperfections are found they must be eliminated or improved.
New Zealand perspective on ISO 17025 accreditation of a plant diagnostic laboratory1
Authors: Alexander B. J. R. , Flynn A. R. , Gibbons A. M. , Clover G. R. G. , Herrera V. E.
Published: July 2008

Four plant diagnostic laboratories in New Zealand have obtained ISO 17025 accreditation since 2001: Linnaeus Laboratory; PestLab, AsureQuality Ltd; Horticulture and Food Research Institute of New Zealand Ltd; and the Ministry of Agriculture and Forestry's (MAF) Plant Health and Environment Laboratory (PHEL). The challenge of pursuing ISO 17025 accreditation for PHEL emerged in late 2003 when a review of a MAF diagnostic standard made ISO 17025 accreditation a mandatory requirement for approval. The accreditation project took three years from initiation to accreditation in 2007. The scope of PHEL's accreditation covers tests (e.g. PCR, RT‐PCR, or ELISA) for the Carlavirus group, High plains virus, Iris yellow spot virus, Pepino mosaic virus, Plum pox virus, Raspberry ringspot virus, phytoplasmas, Potato spindle tuber viroid, and Xylella fastidiosa and morphological identifications of fungi and invertebrates. This article provides a brief overview of ISO 17025 accreditation of plant diagnostic laboratories in New Zealand, describes PHEL's scope of accreditation, and discusses some of the issues and challenges PHEL faced during the process of attaining accreditation and still faces post‐accreditation.
The flexible scope of accreditation in GMO testing and its applicability to plant pathogen diagnostics1
Authors: Camloh M. , Dreo T. , Žel J. , Ravnikar M.
Published: July 2008

Accreditation, as the formal recognition of a laboratory's competence, is increasingly important in the field of plant pathogen diagnostics. As it is necessary to consider the scope of accreditation this paper presents the main differences between the fixed and flexible scope of accreditation and discusses the applicability of the flexible scope to plant pathogen diagnostics. This assessment is supported by practical experience gained during the testing of genetically modified organisms (GMOs) according to standard ISO/IEC 17025, starting with the fixed scope in 2003 and proceeding to the partially flexible scope in 2006. With increasing flexibility, more responsibility is transferred to the laboratory during this process and procedures governing validations need to be clearly specified. Our processes of planning, performing, documenting, and evaluating results of in‐house method validations are general and applicable to the validation of different methods. They are presented here with the example of real‐time PCR validation, a method that is increasingly used in plant pathogen diagnostics due to its reliability and high sensitivity.
Objective evidence for competence1
Authors: Anthoine G. , Wilson V. , Poliakoff F.
Published: July 2008

In the area of plant pest diagnostics, increasing numbers of laboratories apply for accreditation according to the standard ISO 17025 General requirements for the competence of testing and calibration laboratories, which guarantees the compliance with standard operating procedures and the technical competence of the staff involved in the tests. In this standard, each accredited laboratory is required to supervise their staff's skills. This article explains how different French accredited diagnostic laboratories interpret and apply this accreditation requirement.
Results and experiences from the first EU proficiency test for the detection of Phytophthora ramorum1
Authors: Inghelbrecht S. , Heungens K. , De Wael L. , Maes M.
Published: July 2008

The Crop Protection unit of the Institute for Agricultural and Fisheries Research offers plant disease diagnostic services through its ‘Diagnostic Centre for Plants’ (ILVO‐DCP). ILVO‐DCP has requested accreditation (ISO 17025) for a number of diagnostic detection methods in bacteriology, mycology, entomology and nematology. Accreditation forms an essential part of quality control programs such as ISO17025, which can in part be realized by proving the laboratory's competence in inter‐laboratory proficiency or ring tests. In 2006, ILVO‐DCP organized such a proficiency test for the detection of Phytophthora ramorum. The protocol was developed using the standard ILAC‐G13:2000, and defined rules for participation, sample preparation and transport, communication, fraud prevention and reporting. Eight European diagnostic centres participated in the proficiency test including ILVO‐DCP. Each participant received one set of 10 coded samples, each sample consisting of either leaves or stems that were artificially inoculated with either the target or an alternative organism. Participants could use one or more methods listed in the EPPO diagnostic protocol PM 7/66. They had to report within a specific timeframe and received a detailed report of their performance. The success rate of the proficiency test was 100%. This paper lists some of the experiences gained from organising this type of proficiency test.
The Nordic Baltic forum for plant health diagnostic laboratories and its use of ‘on‐site’ proficiency testing to improve laboratory testing1
Authors: Thrane C. , Boa E.
Published: July 2008

During an Estonia‐UK‐Denmark twinning project, a new forum for Plant Health Diagnostic Laboratories in the Nordic‐Baltic region emerged. At the first meeting held in Estonia in July 2006, a new approach to proficiency testing was evaluated. Proficiency tests are key elements assessing the ability of a laboratory to perform a certain test in comparison with other laboratories. ‘On‐site’ proficiency testing means that experts are meeting to look at exactly the same samples under the same conditions on the same day. This is especially relevant for tests based on morphological interpretation as this minimizes the inherent differences in samples due to natural variation within a given species. The benefits of ‘on‐site’ proficiency testing outweigh the additional costs of organising a regional meeting which might include discussion of related scientific topics.

Diagnostic capacity is an essential part of European Plant Health policies but staff may be spread out in different laboratories and countries. ‘On‐site’ proficiency testing helps create a stronger group identity for diagnosticians as well as maintaining quality control. In the plant health field there are also several difficulties in organising conventional proficiency tests. This is further discussed in the paper.
Development of National Reference Laboratories in the European Union1
Authors: Müller P.
Published: July 2008

Since 2004 discussions on the need for greater international co‐operation and coordination between diagnostic laboratories have taken place at several meetings of the EU Heads of National Plant Protection Organisations (NPPOs) and of the European and Mediterranean Plant Protection Organization (EPPO). Technical discussions on criteria for, and tasks and responsibilities of, National Reference Laboratories (NRLs) took place in a Commission Expert Working Group in September 2007. The meeting was attended by representatives from 13 Member States. The experts discussed the issues broadly and very actively and it was possible to elaborate an agreed Guidance Paper containing the main elements for the establishment of NRLs, including a priority list of individual pests for which such laboratories are needed. At their meeting in December 2007, the EU Heads of the NPPOs supported the establishment of NRLs as far as possible and where necessary. It was agreed that the Guidance Paper is a suitable common basis for the establishment of NRLs in the Member States and that it should be reviewed on the basis of the experience gained at national level.
Recent developments in vulvovaginal pathology
Authors: McCluggage W G
Published: July 2008

This review discusses recent developments in vulvovaginal pathology. A variety of morphologically bland mesenchymal lesions occur at this site with considerable histological and immunohistochemical overlap. Aggressive angiomyxoma exhibits HMGA2 immunoreactivity in approximately 50% of cases, and this nuclear transcription factor is emerging as a useful and relatively specific marker for aggressive angiomyxoma, although occasional vulvovaginal smooth muscle neoplasms are positive. HMGA2 is useful in the diagnosis of aggressive angiomyxoma and its distinction from mimics, in the evaluation of resection margins and in the assessment of the presence or absence of residual disease in re‐excisions. Aggressive angiomyxoma is almost invariably positive with oestrogen and progesterone receptors, and there have been several reports of a dramatic reduction in size following gonadotropin releasing hormone agonist therapy. Recent series of the relatively newly described entities cellular angiofibroma and superficial myofibroblastoma of the lower female genital tract have expanded upon the morphological spectrum of these neoplasms. Recently described mesenchymal lesions at this site include massive oedema and prepubertal vulval fibroma. Gastrointestinal stromal tumours have been described as primary neoplasms in the vagina, and rectovaginal septum and extragastrointestinal stromal tumour should be added to the differential diagnosis of a vulvovaginal mesenchymal lesion. Many mesenchymal lesions in the vulvovaginal region exhibit immunoreactivity with both CD34 and desmin, a somewhat unusual immunophenotype in mesenchymal lesions at other sites. It is now established that there are two distinct types of vulval intraepithelial neoplasia (VIN), most commonly termed classic and differentiated VIN, the former associated with human papillomavirus (HPV) infection. There are two corresponding types of vulval squamous carcinoma with HPV‐associated and non‐HPV‐associated variants, the latter often arising in a vulval dystrophy and associated with p53 mutation. However, in some cases there is clinicopathological overlap between HPV‐associated and non‐HPV‐associated squamous carcinomas, and immunohistochemistry with p16 is more reliable than morphology in predicting the presence of HPV. There have been new developments regarding Paget’s disease of the vulva with the identification of markers that are useful in diagnosis and evidence that the neoplastic cells represent a proliferation of adnexal stem cells residing in sebaceous units. The newly described entity vaginal tubulo‐squamous polyp typically exhibits immunopositivity with prostatic markers, possibly indicating derivation from displaced periurethral Skene’s glands.
Is hypospadias a genetic, endocrine or environmental disease, or still an unexplained malformation?
Authors: Kalfa Nicolas , Philibert Pascal , Sultan Charles
Published: July 2008

Hypospadias is one of the most frequent genital malformations in the male newborn and results from an abnormal penile and urethral development. This process requires a correct genetic programme, time‐ and space‐adapted cellular differentiation, complex tissue interactions, and hormonal mediation through enzymatic activities and hormonal transduction signals. Any disturbance in these regulations may induce a defect in the virilization of the external genitalia and hypospadias. This malformation thus appears to be at the crossroads of various mechanisms implicating genetic and environmental factors. The genes of penile development (HOX, FGF, Shh) and testicular determination (WT1, SRY) and those regulating the synthesis [luteinizing hormone (LH) receptor] and action of androgen (5α reductase, androgen receptor) can cause hypospadias if altered. Several chromosomal abnormalities and malformative syndromes include hypospadias, from anterior to penoscrotal forms. More recently, CXorf6 and ATF3 have been reported to be involved. Besides these genomic and hormonal factors, multiple substances found in the environment can also potentially interfere with male genital development because of their similarity to hormones. The proportion of hypospadias cases for which an aetiology is detected varies with the authors but it nevertheless remains low, especially for less severe cases. An interaction between genetic background and environment is likely.
The effects of 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin on foetal male rat steroidogenesis
Authors: Adamsson A. , Simanainen U. , Viluksela M. , Paranko J. , Toppari J.
Published: July 2008

2,3,7,8‐Tetrachlorodibenzo‐p‐dioxin (TCDD) is the most toxic and widely investigated dioxin congener. In utero and lactational exposure to TCDD results in developmental and reproductive defects that are the most sensitive endpoints for TCDD toxicity. TCDD has a potential to interfere with steroid metabolism, but the mechanisms by which this occurs are not well understood. In this study, we investigated the effects of TCDD on prenatal rat steroidogenesis. Pregnant Sprague–Dawley female rats were treated once with TCDD (0, 0.3 or 1 μg/kg) by gavage on embryonic day (ED) 11 and the expression levels of androgen (AR) and estrogen receptors (ER), steroidogenic enzymes (P450scc and 3β‐HSD) and four regulatory factors (StAR, SF‐1, GATA‐4 and Insl‐3) involved in foetal Leydig cell and adrenal function were analysed on ED 19.5. Hormonal status of male foetuses was determined by measuring testicular testosterone (T) levels, plasma luteinizing hormone (LH) and corticosterone concentrations. In utero exposure to TCDD reduced intratesticular T of foetal males (significant at 0.3 μg/kg TCDD) and tended to reduce the protein expression of ERα and AR of foetal male rat testis. Foetal male rat plasma LH levels were significantly reduced at the dose of 1 μg/kg TCDD, while corticosterone levels tended to be increased possibly because of the TCDD‐induced stress. Only minor alterations in steroidogenesis were observed in rat adrenal. mRNA expression of developmental regulatory factors was not influenced by foetal TCDD exposure, except for significantly reduced adrenal SF‐1. The results demonstrate that maternal exposure to TCDD suppressed testicular steroidogenesis of 19.5‐day‐old foetal male Sprague–Dawley rat. The highest dose of TCDD (1 μg/kg) had also an effect on pituitary LH secretion. Our data implicate that TCDD has direct testicular and pituitary effects on foetal male rat but with different dose–responses. These changes can lead to impaired steroidogenesis and it may result in the maldevelopment of the testis and weaken masculinization.
Epigenetic regulation and downstream targets of the Rhox5 homeobox gene
Authors: Shanker S. , Hu Z. , Wilkinson M. F.
Published: July 2008

The discovery of the Rhox homeobox gene cluster on the X chromosome opens up new vistas in the regulation of reproductive processes in mammals. In mice, this cluster comprises more than 30 genes that are selectively expressed in reproductive tissues. A subset of Rhox genes are androgen and AR regulated in postnatal and adult Sertoli cells, making them candidates to mediate androgen‐dependent steps during spermatogenesis. The best characterized of these androgen/AR‐regulated genes is Rhox5 (Pem), the founding member of the Rhox gene cluster. Targeted deletion of Rhox5 in mice causes male subfertility marked by increased germ‐cell apoptosis and decreased sperm count and motility. Microarray analyses identified a wide variety of genes regulated by Rhox5 in Sertoli cells. One of them is the tumour suppressor UNC5C, a pro‐apoptotic molecule previously only known to be involved in brain development. Targeted deletion of Unc5c causes decreased germ‐cell apoptosis in postnatal and adult testes, indicating that it also has a role in spermatogenesis and supporting a model in which Rhox5 promotes germ‐cell survival by downregulating Unc5c. Rhox5 has two independently regulated promoters that have distinct expression patterns. The unique tissue‐specific and developmentally regulated transcription pattern of these two promoters appear to be controlled by DNA methylation. Both promoters are methylated in tissues in which they are not expressed, suggesting that DNA methylation serves to repress Rhox5 expression in inappropriate cell types and tissues. In summary, the Rhox gene cluster is an epigenetically regulated set of genes encoding a large number of transcription factors that are strong candidates to regulate gametogenesis and other aspects of reproduction.
Evaluation of MENT on primary cell cultures from benign prostatic hyperplasia and prostate carcinoma
Authors: Mendoza Patricia , Sánchez Catherine , Contreras Héctor R. , Vergara Jorge , Acevedo Cristian , Cabezas Juan , Huidobro Christian , Noé Gabriela , Castellón Enrique A.
Published: July 2008

7‐alpha‐Methyl‐19‐Nortestosterone (MENT) is a synthetic androgen more potent than testosterone (T) and cannot be reduced at 5‐alpha position. No important effects of MENT on prostate growth have been reported. However, little is known about the effect of MENT on benign prostatic hyperplasia (BPH) or prostate carcinoma (CaP). We evaluate the effect of MENT, T and dihydrotestosterone (DHT) on secretion, proliferation and gene expression of primary cell cultures from human BPH and CaP. Moreover, the effect of these androgens was examined in the presence of finasteride to determine the influence of the 5‐alpha reductase (5‐AR) activity on the androgenic potency. BPH and CaP primary cultures were treated with 0, 1, 10 and 100 nmof T, MENT or DHT during 24 and 48 h. Prostate‐specific antigen (PSA) was measured by micro particles immunoassay and proliferation rate by spectrophotometric assay (MTT) and by the immunochemical detection of the proliferation marker Ki‐67. Gene expression of FGF8b (androgen sensitive gene) was evaluated by semi‐quantitative RT‐PCR. Results showed that MENT treatments increased PSA secretion and proliferation rate with a potency ranged between T and DHT. Similar effects of MENT were observed in both BPH and CaP cultures. The studies with finasteride showed that in BPH and CaP cells, the conversion of T into DHT significantly contributes to its effect on the proliferation and PSA secretion, and corroborated the resistance of MENT to the 5‐AR. The effect of MENT on the gene expression of FGF8b in CaP cells was similar to T and lower than DHT. It is concluded that MENT increases proliferative and secretory activities and gene expression on pathological prostate cells although in less extent than the active metabolite DHT. Furthermore, the fall of endogenous concentration of T during MENT treatment anticipates that this androgen will be of low impact for the prostate.
Gonadotrophin‐Inhibitory Hormone Receptor Expression in the Chicken Pituitary Gland: Potential Influence of Sexual Maturation and Ovarian Steroids
Authors: Maddineni S. , Ocón‐Grove O. M. , Krzysik‐Walker S. M. , Hendricks III G. L. , Proudman J. A. , Ramachandran R.
Published: July 2008

Gonadotrophin‐inhibitory hormone (GnIH), a hypothalamic RFamide, has been found to inhibit gonadotrophin secretion from the anterior pituitary gland originally in birds and, subsequently, in mammalian species. The gene encoding a transmembrane receptor for GnIH (GnIHR) was recently identified in the brain, pituitary gland and gonads of song bird, chicken and Japanese quail. The objectives of the present study are to characterise the expression of GnIHR mRNA and protein in the chicken pituitary gland, and to determine whether sexual maturation and gonadal steroids influence pituitary GnIHR mRNA abundance. GnIHR mRNA quantity was found to be significantly higher in diencephalon compared to either anterior pituitary gland or ovaries. GnIHR mRNA quantity was significantly higher in the pituitaries of sexually immature chickens relative to sexually mature chickens. Oestradiol or a combination of oestradiol and progesterone treatment caused a significant decrease in pituitary GnIHR mRNA quantity relative to vehicle controls. GnIHR‐immunoreactive (ir) cells were identified in the chicken pituitary gland cephalic and caudal lobes. Furthermore, GnIHR‐ir cells were found to be colocalised with luteinising hormone (LH)β mRNA‐, or follicle‐stimulating hormone (FSH)β mRNA‐containing cells. GnIH treatment significantly decreased LH release from anterior pituitary gland slices collected from sexually immature, but not from sexually mature chickens. Taken together, GnIHR gene expression is possibly down regulated in response to a surge in circulating oestradiol and progesterone levels as the chicken undergoes sexual maturation to allow gonadotrophin secretion. Furthermore, GnIHR protein expressed in FSHβ or LHβ mRNA‐containing cells is likely to mediate the inhibitory effect of GnIH on LH and FSH secretion.
Staphylococcus aureus haem oxygenases are differentially regulated by iron and haem
Authors: Reniere Michelle L. , Skaar Eric P.
Published: July 2008

Iron acquisition is a vital process for most pathogenic bacteria, as iron is a limiting nutrient during infection. Staphylococcus aureus, an increasingly important pathogen, acquires iron from host haem via elaboration of the iron‐regulated surface determinant system (Isd). IsdG and IsdI are haem oxygenases that have been proposed to degrade exogenous haem in the bacterial cytoplasm as a mechanism to liberate free iron for use as a nutrient source. Herein, we report that IsdG and IsdI are both important for S. aureus growth on haemin as a sole iron source and are necessary for full S. aureus pathogenesis. Investigations into the regulation of these enzymes revealed that IsdG and IsdI are differentially regulated by iron and haem through both transcriptional and post‐transcriptional mechanisms. Additionally, IsdI was found to be expressed in infected tissues at the sites of abscess formation, suggesting that abscesses are iron‐starved microenvironments inside the host. These findings suggest that S. aureus differentially regulates IsdG and IsdI in response to alterations in iron and haem availability during infection.
Novel mechanism of outer membrane targeting of proteins in Gram‐negative bacteria
Authors: Ferrandez Yann , Condemine Guy
Published: July 2008

In Gram‐negative bacteria, all the proteins destined for the outer membrane are synthesized with a signal sequence that is cleaved, either by the signal peptidase LepB for integral outer membrane proteins or by LspA for lipoproteins, when they cross the cytoplasmic membrane. The Dickeya dadantii protein PnlH does not possess a cleavable signal sequence but is anchored in the outer membrane by an N‐terminal targeting signal. Addition of the 41 N‐terminal amino acids of PnlH is sufficient for anchoring various hybrid proteins in the outer membrane. This targeting signal presents some of the characteristics of a Tat (twin arginine translocation) signal sequence but without an obvious cleavage site. We found that the Tat translocation pathway is required for the targeting process. This new mechanism of outer membrane protein targeting is probably widespread as PnlH was also addressed to the outer membrane when expressed in Escherichia coli. As PnlH was not detected as a substrate by Tat signal sequence prediction programmes, this would suggest that there may be many other unknown Tat‐dependent outer membrane proteins.
Nitrites in induced sputum as a simple and cheap non‐invasive marker of airway inflammation for asthmatic schoolchildren
Authors: Recabarren Arturo , Apaza Carlos , Castro‐Rodríguez José A.
Published: July 2008

To determine if there are differences in the nitric oxide metabolites (nitrites) in sputum of patients with persistent asthma and healthy schoolchildren, we performed a case‐control study in a tertiary care hospital in Arequipa, Perú. Nitrites in induced sputum samples were measured using the Griess assay in 30 persistent asthmatics (mean age of 10.1 yr) and 30 controls (mean age of 11.9 yr). The mean ± s.d. of nitrites among asthmatics was significantly higher than the controls (16.30 ± 8.6 vs. 10.25 ± 4.68 nmol/ml, respectively, p = 0.001). Moreover, the nitrite level in the sputum in children with severe persistent asthma was higher than in the level found in the moderate and mild asthmatics (32.83 ± 9.48 vs. 18.10 ± 1.96 vs. 11.84 ± 4.73 nmol/ml, respectively, p < 0.01 for linear trend). This study showed for the first time in children that asthmatics have significantly higher levels of nitrites in induced sputum than healthy controls and that the level of nitrite correlates with the severity of the asthma. Nitrite levels in sputum, a simple and cheap, non‐invasive method, may be a good alternative to measure the severity of inflammation in asthmatic children.
The effect of digestion and pH on the allergenicity of kiwifruit proteins
Authors: Lucas Jane S. A. , Cochrane Stella A. , Warner John O. , Hourihane Jonathan O’.B.
Published: July 2008

It is suggested that patients with oral allergy syndrome (OAS) respond to pepsin‐sensitive allergens, and systemic reactors identify pepsin‐resistant allergens. We sought to assess the digestibility of kiwifruit proteins in simulated gastric fluid (SGF), and to compare the immunogenicity of the digests in patients with isolated oral and systemic reactions to kiwifruit. In addition, the effect of pH on digestibility of kiwifruit proteins was investigated. The in vitro resistance of kiwifruit proteins to digestion was determined using SGF. G‐immunoglobulin (IgE) binding to digested proteins was investigated by Western blotting using sera from children and adults (aged 5–72 yr) with systemic reactions and patients with isolated oral symptoms. To determine whether pH conditions influence digestion of kiwifruit extracts, digestion at pHs 1.5–7 were compared by SDS‐PAGE. Patients with systemic reactions showed IgE binding to digestion‐resistant allergens, but patients with oral symptoms reacted only to digestion‐labile allergens. An increase in pH from 1.5 to 2.5 significantly reduced pepsin breakdown of kiwifruit allergens. Immunoreactive digested protein fragments were detectable by immunoblot but not Coomassie stain. This study confirms a difference in the lability of food allergens recognized by patients with systemic reactions and those with OAS. Pepsin digestion of kiwifruit proteins was impaired by hypoacidic conditions suggesting that patients with hypoacidic gastric conditions are at increased risk of systemic absorption of allergens. The data indicate that commonly used methods for predicting allergenicity of novel proteins using Coomassie stains may be flawed.
Sexual selection, sexual size dimorphism and Rensch’s rule in Odonata
Authors: SERRANO‐MENESES M. A. , CÓRDOBA‐AGUILAR A. , AZPILICUETA‐AMORÍN M. , GONZÁLEZ‐SORIANO E. , SZÉKELY T.
Published: July 2008

Odonata (dragonflies and damselflies) exhibit a range of sexual size dimorphism (SSD) that includes species with male‐biased (males > females) or female‐biased SSD (males < females) and species exhibiting nonterritorial or territorial mating strategies. Here, we use phylogenetic comparative analyses to investigate the influence of sexual selection on SSD in both suborders: dragonflies (Anisoptera) and damselflies (Zygoptera). First, we show that damselflies have male‐biased SSD, and exhibit an allometric relationship between body size and SSD, that is consistent with Rensch’s rule. Second, SSD of dragonflies is not different from unit, and this suborder does not exhibit Rensch’s rule. Third, we test the influence of sexual selection on SSD using proxy variables of territorial mating strategy and male agility. Using generalized least squares to account for phylogenetic relationships between species, we show that male‐biased SSD increases with territoriality in damselflies, but not in dragonflies. Finally, we show that nonagile territorial odonates exhibit male‐biased SSD, whereas male agility is not related to SSD in nonterritorial odonates. These results suggest that sexual selection acting on male sizes influences SSD in Odonata. Taken together, our results, along with avian studies (bustards and shorebirds), suggest that male agility influences SSD, although this influence is modulated by territorial mating strategy and thus the likely advantage of being large. Other evolutionary processes, such as fecundity selection and viability selection, however, need further investigation.
Frequency of independent origins of viviparity among caecilians (Gymnophiona): evidence from the first ‘live‐bearing’ Asian amphibian
Authors: GOWER D. J. , GIRI V. , DHARNE M. S. , SHOUCHE Y. S.
Published: July 2008

Viviparity is reported for Gegeneophis seshachari (Gymnophiona: Caeciliidae) from a gravid female containing four oviductal foetuses. The oviducts are highly vascularized and contain patches of thickened, layered tissue similar to foetal gut contents. Gegeneophis seshachari probably resemble other viviparous caecilians in having foetuses that ingest thickened oviduct lining using specialized deciduous teeth. This is the first report of viviparity in Asian amphibians and Indo‐Seychellean caeciliids. Gegeneophis is the only caecilian genus known to include oviparous and viviparous species, and G. seshachari is the smallest known viviparous caecilian. Phylogenetic analysis of mitochondrial DNA sequences supports assignment of G. seshachari to a monophyletic Gegeneophis. Character optimization indicates that viviparity has evolved independently at least four times within Gymnophiona – a rate of incidence relative to the number of extant species that is higher than for other vertebrate groups except squamate reptiles. Our findings strengthen the proposal that caecilian reproduction demands further attention.
Spatial and temporal prokaryotic variability in the northern Adriatic Sea
Authors: Del Negro Paola , Celussi Mauro , Crevatin Erica , Paoli Alessandro , Bernardi Aubry Fabrizio , Pugnetti Alessandra
Published: July 2008

The prokaryotic community, both in terms of abundance and activity (exoenzymatic hydrolysis of proteins, polysaccharides and phosphorylated molecules and leucine uptake), was investigated seasonally for a 3‐year period (2004–2006) in the Gulf of Venice (northern Adriatic Sea). By focusing on spatial and temporal variability, the prokaryote dynamics showed significant variations on a horizontal and seasonal scale, but no substantial differences were observed among years. The basin‐scale variability was mainly influenced by allochthonous inputs from the Po river and the Venice Lagoon, which were the main source of nutrients, acting as a ‘bottom up’ control on prokaryotes. On a seasonal scale, all the microbial parameters (except the polysaccharide degradation) showed significant variations following the temperature fluctuations. The annual rate of change was very low for all the chemical, physical and biological parameters and only the abundance and phosphatase activity of the prokaryotes differed significantly among years.
Influence of hospital and surgeon volumes on operative time, blood loss and perioperative complications in radical nephrectomy
Authors: Yasunaga Hideo , Yanaihara Hitoshi , Fuji Kohzo , Matsuyama Yutaka , Deguchi Nobuhiro , Ohe Kazuhiko
Published: July 2008

Objectives:  We conducted a nationwide multi‐center survey using medical record‐based data to investigate the relationship between hospital/surgeon volumes and various outcomes, including operative time, volume of blood loss, and incidence of perioperative complications, in radical nephrectomy for renal cell carcinoma.

Methods:  We investigated a total of 1704 patients who underwent radical nephrectomy at 461 hospitals in Japan between November 2006 and February 2007. The association between hospital/surgeon volumes and operative time, volume of blood loss, and incidence of perioperative complications were independently analyzed using multivariate regression analysis against age, gender, operation site, cancer stage, serum creatinine levels, comorbid conditions, and surgical technique (open surgery or minimally invasive surgery).

Results:  Neither hospital volume nor surgeon volume was a significant predictor of operative time or volume of blood loss. We did not identify any association between hospital volume and perioperative complications. High‐volume (> 100) surgeons were unlikely to have perioperative complications compared to low‐volume (< 20) surgeons, but the relation was not significant (odds ratio, 0.50; 95% confidence intervals, 0.14–1.87; P = 0.30).

Conclusions:  Volume–outcome relationships were not confirmed for radical nephrectomy. According to our study, the justification of regionalizing radical nephrectomy to centers of excellence is not yet clear.