bluuebunny/crossref_metadata_2025_split huggingface dataset:
Detection and pharmacokinetics of tetrahydrogestrinone in horses
Authors: MACHNIK M. , GERLACH M. , KIETZMANN M. , NIEDORF F. , THEVIS M. , SCHENK I. , GUDDAT S. , DÜE M. , SCHÄNZER W. 
Published: October 2008

The anti‐doping rules of national and international sport federations ban any use of tetrahydrogestrinone (THG) in human as well as in horse sports. Initiated by the THG doping scandals in human sports a method for the detection of 3‐keto‐4,9,11‐triene steroids in horse blood and urine was developed. The method comprises the isolation of the analytes by a combination of solid phase and liquid–liquid extraction after hydrolysis and solvolysis of the steroid conjugates. The concentrations of THG in blood and urine samples were measured by liquid chromatography‐tandem mass spectrometry (LC‐MS/MS).A THG excretion study on horses was conducted to verify the method capability for the analysis of postadministration urine samples. In addition, blood samples were collected to allow for determination of the pharmacokinetics of THG in horses. Following the administration of a single oral dose of 25 μg THG per kg bodyweight to 10 horses, samples were collected at appropriate intervals. The plasma levels of THG reached maximal concentrations of 1.5–4.8 ng/mL. Twenty‐four hours after the administration plasma levels returned to baseline. In urine, THG was detectable for 36 h. Urinary peak concentrations of total THG ranged from 16 to 206 ng/mL. For the 10 horses tested, the mean plasma clearance of THG was 2250 mL/h/kg and the plasma elimination half‐life was 1.9 h.
Opening a can of worms: unprecedented sympatric cryptic diversity within British lumbricid earthworms
Authors: KING R. ANDREW , TIBBLE AMY L. , SYMONDSON WILLIAM O. C. 
Published: October 2008

AbstractEarthworms play a major role in many aspects of soil fertility, food web ecology and ecosystem functioning, and hence are frequently the subjects of, for example, ecological and toxicological research. Our aim was to examine the genetic structure of common earthworm species, to identify cryptic lineages or species that may be distinct ecotypes or biotypes (and hence confound current research based upon morphotypes) and to try to explain the massive cryptic diversity that eventually emerged. We demonstrated that species such as Allolobophora chlorotica, Aporrectodea longa, Aporrectodea rosea and Lumbricus rubellus all comprise highly divergent lineages with species‐level divergence at the mitochondrial cytochrome oxidase I (COI) gene. In Allo. chlorotica alone, we found 55 haplotypes for COI, with 35 of these being found in pink and 20 in green morph worms. There were no cases of the two colour morphs sharing COI haplotypes. Phylogenetic analyses of mitochondrial COI and 16S genes showed the presence of five highly divergent lineages, suggesting the presence of multiple cryptic species within Allo. chlorotica. There was no clear geographical pattern to lineage distribution and many populations were polymorphic for both mitochondrial DNA lineage and colour morph. Amplified fragment length polymorphism results, based on two primer combinations, were broadly congruent with mitochondrial DNA results with one significant exception. Despite showing over 14% divergence at COI, amplified fragment length polymorphism markers showed that the two green morph lineages may be interbreeding and therefore represent a single taxon. The cryptic diversity revealed by these results has profound consequences for all areas of earthworm research.
Detection and quantification of airborne inoculum of Sclerotinia sclerotiorum using quantitative PCR
Authors: Rogers S. L. , Atkins S. D. , West J. S. 
Published: October 2008

This paper reports the development of a new specific diagnostic technique to accurately quantify airborne inoculum of Sclerotinia sclerotiorum and discusses its potential use in disease‐forecasting schemes, using examples of three contrasting epidemic seasons: 2007, when there was a severe epidemic of sclerotinia stem rot (SSR) in England and high numbers of airborne ascospores were trapped at Rothamsted, and, in contrast, 2003 and 2004, when the incidence of SSR in England was low and low numbers of airborne ascospores were trapped at Rothamsted. DNA was extracted from wax‐coated plastic tapes, such as those used in Burkard (Hirst‐type) spore traps and rotating‐arm traps. A SYBR‐green quantitative PCR (qPCR) method produced a linear relationship between ascospore numbers and S. sclerotiorum DNA (mean 0·35 pg DNA per spore) and was able to detect DNA representing as few as two ascospores. The technique was insensitive to DNA of the host plant, Brassica napus, and other plant pathogens, including Sclerotinia minor, S. trifoliorum and Botrytis cinerea, and common airborne fungal genera such as Cladosporium and Penicillium. There was no relationship between rainfall and numbers of airborne ascospores of S. sclerotiorum present at Rothamsted during the period of infection in the severe SSR season (2007).
Trichothecene mycotoxin genotypes ofFusarium graminearum sensu strictoandFusarium meridionalein wheat from southern Brazil
Authors: Scoz L. B. , Astolfi P. , Reartes D. S. , Schmale III D. G. , Moraes M. G. , Del Ponte E. M. 
Published: October 2008

A total of 82 fungal isolates was obtained from wheat kernel samples affected by fusarium head blight collected from 20 locations in southern Brazil. Polymerase chain reaction (PCR) assays were used to characterize trichothecene mycotoxin genotypes [deoxynivalenol (DON), nivalenol (NIV) and two acetylated derivatives of DON]. To identify isolates that producing DON and NIV, portions of theTri13gene were amplified. To identify 3‐acetyl‐deoxynivalenol (3‐ADON) and 15‐acetyl‐deoxynivalenol (15‐ADON) genotypes, portions ofTri3andTri12were amplified. Nearly all of the isolates studied (76/82) were of the DON/15‐ADON genotype. Six of the isolates were of the NIV genotype. The DON/3‐ADON genotype was not observed. Portions of three genes were sequenced from representative isolates of the NIV and DON/15‐ADON genotypes and compared with sequences from curated reference isolates ofFusariumin GenBank.blastqueries for individual gene sequences and pairwise comparisons of percentage identity and percentage divergence based on 1676 bp of concatenated DNA sequence suggested that the isolates representing the DON/15‐ADON genotype wereFusarium graminearum sensu strictoand the isolates representing the NIV genotype wereFusarium meridionale. This is the first detailed report of trichothecene mycotoxin genotypes ofF. graminearumandF. meridionalein Brazil.
Tryptophan deficiency affects organ growth by retarding cell expansion in Arabidopsis
Authors: Jing Yanjun , Cui Dayong , Bao Fang , Hu Zhubing , Qin Zhixiang , Hu Yuxin 
Published: October 2008

SummaryTryptophan (Trp) is an essential amino acid required not only for protein synthesis but also for the production of many plant metabolites, including the hormone auxin. Mutations that disrupt Trp biosynthesis result in various developmental defects in plant organs, but how Trp affects organ growth and development remains unclear. Here, we identify an Arabidopsis mutant, small organ1 (smo1/trp2‐301), which exhibits a reduction in the size of its aerial organs as a result of the retardation of growth by cell expansion, rather than by the retardation of growth by cell proliferation. smo1/trp2‐301 contains a lesion in TSB1 that encodes a predominantly expressed Trp synthase β‐subunit, and is allelic with trp2 mutants. Further analyses show that in trp2 leaf cells, the nuclear endoreduplication is impaired and chloroplast development is delayed. Furthermore, cell expansion and leaf growth in trp2 can be restored by the exogenous application of Trp, but not by auxin, and the general protein synthesis is not apparently affected in trp2 mutants. Our findings suggest that the deficiency in Trp or its derivatives is a growth‐limiting factor for cell expansion during plant organogenesis.
Repression of the floral transition via histone H2B monoubiquitination
Authors: Gu Xiaofeng , Jiang Danhua , Wang Yuqi , Bachmair Andreas , He Yuehui 
Published: October 2008

SummaryThe Rad6‐Bre1 complex monoubiquitinates histone H2B in target gene chromatin, and plays an important role in positively regulating gene expression in yeast. Here, we show that the Arabidopsis relatives of the yeast Rad6, ubiquitin‐conjugating enzyme 1 (UBC1) and UBC2, redundantly mediate histone H2B monoubiquitination, and upregulate the expression of FLOWERING LOCUS C (FLC; a central flowering repressor in Arabidopsis) and FLC relatives, and also redundantly repress flowering, the developmental transition from a vegetative to a reproductive phase that is critical in the plant life cycle. Moreover, we have found that Arabidopsis relatives of the yeast Bre1, including HISTONE MONOUBIQUITINATION 1 (HUB1) and HUB2, also upregulate the expression of FLC and FLC relatives, and that HUB1 genetically interacts with UBC1 and UBC2 to repress the floral transition. These findings are consistent with a model in which HUB1 and HUB2 specifically interact with and direct UBC1 and UBC2 to monoubiquitinate H2B in developmental genes, and thus regulate developmental processes in plants.
Cyclospora spp. in herbs and water samples collected from markets and farms in Hanoi, Vietnam
Authors: Tram Nguyen Thuy , Hoang Linda M. N. , Cam Phung Dac , Chung Phan Thu , Fyfe Murray W. , Isaac‐Renton Judith L. , Ong Corinne S. L. 
Published: October 2008

SummaryObjective  To determine the prevalence of Cyclospora spp. oocysts in herb and water samples as well as in fecal specimens of clinical cases of diarrhoea in Hanoi, Vietnam.Method  From November 2004 to October 2005, water and herb samples collected from markets and farms in Hanoi were examined for the presence of Cyclospora spp. oocysts in concentrated sediments and washings using UV epifluorescence examination of a wet mount. In addition, hospital based surveillance studies were carried out using a structured questionnaire which focused on potential risk factors for cyclosporiasis. Stool specimens were collected from individuals with diarrhoea attending primary healthcare facilities and examined for Cyclospora spp. oocysts by modified acid fast smear and wet mount examination using both light and UV epifluorescence microscopy.Results  Cyclospora spp. were found in 34/288 (11.8%) market water and herb samples, and in 24/287 (8.4%) farm samples. All varieties of herbs sold at the market and grown in farms were contaminated with Cyclospora spp. oocysts. A marked seasonal increase in Cyclospora spp. contamination was observed before the rainy season (39/288) from November to April compared to the rainy season (19/268) from May to October (χ2 = 7.593, P = 0.006). However, Cyclospora spp. was not found in any stool samples collected in hospital‐based surveillance studies.Conclusions  These results confirm the presence of Cyclospora spp. which varies seasonally in environmental samples (water and herbs collected from farms and markets) within the Hanoi metropolitan area.
Allotyping human complement factor B in Asian Indian type 1 diabetic patients
Authors: Kumar N. , Kaur G. , Tandon N. , Mehra N. K. 
Published: October 2008

AbstractHuman complement factor B (BF) is an essential component of the alternate complement pathway and therefore important in innate immune and autoimmune responses. The BF gene is located in the central region of major histocompatibility complex (MHC) and is known to encode more than 30 protein variants that can be resolved by isoelectric focusing and gel electrophoresis. There are three BF alleles – BF*S, BF*FB and BF*FA – that differ in codon 7 at nucleotide positions 94 and 95. These alleles have CGG, TGG or CAG triplets at their codon 7, respectively, that code for Arg, Trp or Gln residues. We have developed a novel polymerase chain reaction using sequence‐specific primers‐based allotyping assay that can identify nucleotide substitutions in codon 7 in all the three BF alleles. The assay was validated by sequencing and amplified fragment length polymorphism. Using this SSP assay, we report the BF alleles located on the multiple human leukocyte antigen (HLA)‐DR3 haplotypes that are unique in the Indian population and are associated with autoimmunity. The common type 1 diabetes (T1D)‐favoring Caucasian haplotype HLA‐A1‐B8‐DR3 (ancestral haplotype AH8.1) carries BF*S. However, in the North Indian T1D patients, the most common haplotype is HLA‐A26‐B8‐DR3 (AH8.2) and this carried BF*FB. Because of its association with AH8.2, the BF*FB was overrepresented in the patients (51.03%) compared with healthy controls (32.7%, OR  = 2.148, 95% CI  = 1.34–3.44, P = 0.002). Similar studies on allotyping BF alleles in different haplotypes in various populations could have important implications in understanding mechanisms of MHC haplotypic diversifications and disease associations and designing future therapeutic approaches.
No easy solution for the fractionation of faecal nitrogen in captive wild herbivores: results of a pilot study
Authors: Schwarm A. , Schweigert M. , Ortmann S. , Hummel J. , Janssens G. P. J. , Streich W. J. , Clauss M. 
Published: October 2008

SummaryFaecal nitrogen (FN) concentration is used as a marker for habitat quality and digestive efficiency in free‐ranging herbivores. In herbivores, FN can be separated into undigested plant N (analysed as the N concentration of the neutral detergent residue) and metabolic faecal N (MFN). It has been suggested that by differential analysis of the faecal fibre‐bound N, the MFN fraction can be further split into a bacterial N and an endogenous N fraction [Hesta et al., Br. J. Nutr. 90 (2003) 1007]. We applied these methods to 96 faecal samples of 48 mammalian herbivore species from zoos. Species were grouped into coprophageous and non‐coprophageous hindgut fermenters and ruminating and non‐ruminating foregut fermenters. Diet was not controlled. The FN decreased with body mass, possibly reflecting higher proportions of concentrates in diets of smaller animals. The proportion of MFN increased with FN, indicating that higher quality food might enhance the gastrointestinal bacterial flora. The only outlier to this pattern was the lesser panda (Ailurus fulgens), confirming the low relevance of fermentative digestion in this herbivorous ‘carnivore’. No relevant differences between the four digestion types were noted. The proportion of endogenous faecal N (32–80% of FN) was always higher than that of bacterial faecal N (7–30%), which contradicts basal understanding of herbivore digestive physiology. Thus, the method of Hesta et al. (2003) does not appear applicable to herbivores. While the results do not exclude the possibility that detailed differences might occur between digestion types, they indicate a high degree of similarity between herbivores that rely on bacterial fermentation, regardless of their digestion type, with respect to metabolic faecal losses.
Identification of a SCAR marker linked to a recessive male sterile gene (Tems) and its application in breeding of marigold (Tagetes erecta)
Authors: He Y. H. , Ning G. G. , Sun Y. L. , Qi Y. C. , Bao M. Z. 
Published: October 2008

AbstractIn marigold, an F2 segregation population of 167 plants was constructed from a cross of a line (M525A) carrying the male sterility trait × an inbred line (f53f). In line M525A, the male sterility trait was controlled by the recessive gene, Tems. The intersimple sequence repeat (ISSR) and sequence‐related amplified polymorphism (SRAP) techniques combined with bulked segregant analysis were used to develop markers linked to the trait. From a survey of the 38 ISSR primers and 170 SRAP primer combinations, only one SRAP marker that was closely linked to the target trait was identified and successfully converted into sequence characterized amplified region (SCAR) marker that was located within 2.4 cM from Tems locus. The marker was validated with five other two‐type lines and in each case the male fertile plants were reliably identified. This SCAR marker therefore permits the efficient marker‐assisted selection of male sterile individuals in breeding programmes of marigold and will greatly facilitate the breeding of F1 cultivars.
A Study of Endometritis Causing Repeat Breeding of Cycling Iraqi Buffalo Cows
Authors: Azawi OI , Omran SN , Hadad JJ 
Published: October 2008

ContentsThe objectives of this study were to determine the non‐specific aerobic and anaerobic bacterial causes of endometritis causing repeat breeding of cycling Iraqi buffalo cows at Nineveh province, validate diagnostic criteria for endometritis and to evaluate the treatment efficiency of using systemic or intra‐uterine infusion of antibiotics for the treatment of endometritis. Data were collected from 60 buffalo cows with history of repeat breeding in different herds. All buffaloes were subjected to detailed clinical examination including external inspection, vaginoscopy and transrectal palpation of the cervix, uterus and ovaries. Swabs for bacteriology and biopsies for histopathology were collected from the uterine lumen from each cow. Character, odour and estimation of polymorphonuclear cells (PMN) of the vaginal mucus were scored. Blood samples were collected from cows for creatine kinase (CK) and aspartate aminotransferase (AST) measurement. Treatment conducted using oxytetracycline with tylosin in local intrauterine infusion or systemically with hormonal treatment. The most pre‐disposing factor for uterine infection was retained placenta (13.3%). The most prevalent bacteria in uterine lumen were E. coli (23%), Archanobacterium pyogenes (13%) and Staphylococcus aureus (10%) were mostly isolated from buffaloes with repeat breeding. Vaginal mucus character score was associated with the bacterial growth density score. The difference in PMN was highly significant (p < 0.01) in animals with repeat breeding than control groups. In addition, PMNs was significantly (p < 0.01) correlated r = 0.894 with the character of vaginal discharge. High level of PMNs observed in buffaloes infected with A. pyogenes. Buffalo cows with endometritis had higher CK (321.47 ± 39.06 vs 162.01 ± 16.41 U/l) and AST (133.93 ± 12.43 vs 97.01 ± 6.86 U/l) activities (p < 0.05) than control‐heifers, but no significant difference was observed between buffalo cows with endometritis in CK (321.47 ± 39.06 vs 208.33 ± 5.84) and AST (133.93 ± 12.43 vs 156.17 ± 9.65) activities than control‐pluriparious. It could be concluded that A. pyogenes was the only non‐specific uterine pathogen directly associated with severe endometrial lesions. Vaginoscopy examination combined with palpation of uterus increase the accuracy of diagnosing endometritis and cytogenic examination of uterine discharge is more reliable method of establishing the presence or absence of uterine inflammation in buffalo cows. Animals with repeat breeding (endometritis) showed clinical cure and improved pregnancy in all treatment groups with no significant difference. The use of oestradiol in repeat breeder cases has no effect in improving neither clinical cure rate nor pregnancy rate.
Expression of Occludin in Testis and Epididymis of Wild Rabbits, Lepus sinensis coreanus
Authors: Yoon SI , Park CJ , Nah WH , Gye MC 
Published: October 2008

ContentsTight junctions (TJs) in inter‐Sertoli junctional areas and epididymal epithelia build up the blood–testis barrier (BTB) and the blood–epididymal barrier (BEB), respectively. In this study, the expression of occludin, an integral member of the TJs, was examined in testis and different regions of epididymis of Lepus sinensis coreanus, an Korean wild rabbit species. In testis, intense occludin immunoreactivity was found in the basally located inter‐Sertoli junctional area together with diffused immunoreactivity of occludin in the cytoplasm of Sertoli cells. It can be suggested that occludin is one of the robust elements of BTB in seminiferous tubules of rabbit testis. In proximal and distal caput epididymis, occludin immunoreactivity was found in the lateral as well as apical contacts of epithelial cells. In corpus epididymis, intense occludin immunoreactivity was found in the basolateral as well as apical contacts of epithelial cells together with cytoplasmic signal. In cauda epididymis, occludin immunoreactivity in luminal epithelia was relatively strong but largely found in the cytoplasm. This suggests that intriguing regulatory mechanisms differentially recruit occludin to the TJ in the different regions of epididymal epithelia. The differences in the subcellular localization as well as expression levels of occludin among the epididymal segments may reflect differential paracellular permeability of epithelia along the epididymal tubules and be correlated with sperm maturation in rabbit. In Western blot, a major form of occludin was MW 62 kDa together with small fragments of MW 34–39 kDa in testis and epididymis, suggesting the peptide cleavage of occludin. This is the first report on the molecular nature of TJs in a wild rabbit testis and epididymis.
Effects of Plasma Urea Nitrogen Levels on the Bovine Oocyte Ability to Develop After In vitro Fertilization
Authors: Santos P , Marques A , Antunes G , Chaveiro A , Andrade M , Borba A , Da Silva F Moreira 
Published: October 2008

ContentsThe overall aim of the present study was to evaluate in vitro development ability of oocytes recovered from 56 Holstein Frisian heifers with low [group 1 (G1): <13 mg /dl], moderate [group 2 (G2): 13–16 mg /dl] and high [group 3 (G3): >16 mg /dl] plasma urea nitrogen (PUN) concentrations, to determine whether PUN concentrations affect the competence of oocytes to progress to blastocysts after in vitro fertilization. In vitro oocyte and embryo development was assessed by blastocyst rates, embryo total cell numbers and apoptosis. Blood samples for the determination of PUN were collected 24 h prior to collection of the ovaries at the slaughter. A total of 112 ovaries were collected at a local abattoir and oocytes (n = 697) were aspirated, in vitro matured and fertilized. On day 8, blastocysts were assigned to the terminal dUTP nick end labelling assay. Cleavage rates were significantly higher (p < 0.001) for groups 1 and 2 than for group 3 (i.e. 72.5% and 72.2% vs 61.7%, respectively). The proportion of fertilized oocytes that developed into blastocysts was higher (p < 0.05) for group 1 than for group 3 (34.0% vs 23.0%, respectively). Day 8 blastocysts showed higher total cell counts (p < 0.05) for group 1 than for group 3 (123.7 vs 76.3), and a higher (p < 0.05) total apoptotic cell rate was found in group 3 (25.9 and 19.0 vs 43.2 for G1, G2 and G3, respectively). In conclusion, the ability of oocytes from heifers with increased levels of PUN to develop to the blastocyst stage was significantly reduced when standard routines for in vitro maturation, fertilization and culture were followed. These detrimental effects can be mediated in part through direct effect of urea and/or by the metabolic products on the process of follicle‐enclosed oocyte nuclear and cytoplasmic development.