Generation, Annotation and Analysis of First Large-Scale Expressed Sequence Tags from Developing Fiber of Gossypium barbadense L

BACKGROUND: Cotton fiber is the world's leading natural fiber used in the manufacture of textiles. Gossypium is also the model plant in the study of polyploidization, evolution, cell elongation, cell wall development, and cellulose biosynthesis. G. barbadense L. is an ideal candidate for providing new genetic variations useful to improve fiber quality for its superior properties. However, little is known about fiber development mechanisms of G. barbadense and only a few molecular resources are available in GenBank. METHODOLOGY AND PRINCIPAL FINDINGS: In total, 10,979 high-quality expressed sequence tags (ESTs) were generated from a normalized fiber cDNA library of G. barbadense. The ESTs were clustered and assembled into 5852 unigenes, consisting of 1492 contigs and 4360 singletons. The blastx result showed 2165 unigenes with significant similarity to known genes and 2687 unigenes with significant similarity to genes of predicted proteins. Functional classification revealed that unigenes were abundant in the functions of binding, catalytic activity, and metabolic pathways of carbohydrate, amino acid, energy, and lipids. The function motif/domain-related cytoskeleton and redox homeostasis were enriched. Among the 5852 unigenes, 282 and 736 unigenes were identified as potential cell wall biosynthesis and transcription factors, respectively. Furthermore, the relationships among cotton species or between cotton and other model plant systems were analyzed. Some putative species-specific unigenes of G. barbadense were highlighted. CONCLUSIONS/SIGNIFICANCE: The ESTs generated in this study are from the first large-scale EST project for G. barbadense and significantly enhance the number of G. barbadense ESTs in public databases. This knowledge will contribute to cotton improvements by studying fiber development mechanisms of G. barbadense, establishing a breeding program using marker-assisted selection, and discovering candidate genes related to important agronomic traits of cotton through oligonucleotide array. Our work will also provide important resources for comparative genomics, polyploidization, and genome evolution among Gossypium species

Distinct colonization patterns and cDNA-AFLP transcriptome profiles in compatible and incompatible interactions between melon and different races of Fusarium oxysporum f. sp. melonis

Background: Fusarium oxysporum f. sp. melonis Snyd. & Hans. (FOM) causes Fusarium wilt, the most important infectious disease of melon (Cucumis melo L.). The four known races of this pathogen can be distinguished only by infection on appropriate cultivars. No molecular tools are available that can discriminate among the races, and the molecular basis of compatibility and disease progression are poorly understood. Resistance to races 1 and 2 is controlled by a single dominant gene, whereas only partial polygenic resistance to race 1,2 has been described. We carried out a large-scale cDNA-AFLP analysis to identify host genes potentially related to resistance and susceptibility as well as fungal genes associated with the infection process. At the same time, a systematic reisolation procedure on infected stems allowed us to monitor fungal colonization in compatible and incompatible host-pathogen combinations. Results: Melon plants (cv. Charentais Fom-2), which are susceptible to race 1,2 and resistant to race 1, were artificially infected with a race 1 strain of FOM or one of two race 1,2 w strains. Host colonization of stems was assessed at 1, 2, 4, 8, 14, 16, 18 and 21 days post inoculation (dpi), and the fungus was reisolated from infected plants. Markedly different colonization patterns were observed in compatible and incompatible host-pathogen combinations. Five time points from the symptomless early stage (2 dpi) to obvious wilting symptoms (21 dpi) were considered for cDNA-AFLP analysis. After successful sequencing of 627 transcript-derived fragments (TDFs) differentially expressed in infected plants, homology searching retrieved 305 melon transcripts, 195 FOM transcripts expressed in planta and 127 orphan TDFs. RNA samples from FOM colonies of the three strains grown in vitro were also included in the analysis to facilitate the detection of in planta-specific transcripts and to identify TDFs differentially expressed among races/strains. Conclusion: Our data suggest that resistance against FOM in melon involves only limited transcriptional changes, and that wilting symptoms could derive, at least partially, from an active plant response. We discuss the pathogen-derived transcripts expressed in planta during the infection process and potentially related to virulence functions, as well as transcripts that are differentially expressed between the two FOM races grown in vitro. These transcripts provide candidate sequences that can be further tested for their ability to distinguish between races. Sequence data from this article have been deposited in GenBank, Accession Numbers: HO867279-HO867981

Surgical aspects and the outcomes of kidney transplants with multiple renal arteries

Introduction: The effect of multiple arterial anastomosis and different reconstruction techniques on the outcome of renal transplant recipients is not clearly defined. In this retrospective study, we report our experience of living donor kidney transplantation using allografts with multiple renal arteries, regarding the patient and graft survival, as well as surgical complications. Materials and Methods: Patients were divided into two groups according to the arterial anatomy of the renal allograft. While group I consisted of patients with single renal artery grafts, group II included patients with multiple renal artery grafts. Results were compared between single and multiple artery groups and further analyzed between the groups, which were constituted according to vascular reconstruction technique. Results: Forty-five patients (15.7%) had grafts with multiple renal arteries. The mean total ischemia time was longer in multiple renal artery group (90.8±17.3 vs. 71.9±16.6 minutes, p0.001). The mean serum creatinine level at posttransplant one year was significantly higher in multiple artery group than in single renal artery group (1.74±0.45 vs.1.46±44 mg/dl), but it was similar at fifth year. There was no significant difference between the groups with regard to mean systolic blood pressure, the incidence of acute tubular necrosis and acute rejection, vascular and urological complications, graft and patient survival rates. Similar findings were obtained in patients with multiple arteries whose anastomosis were performed with different reconstruction techniques. Conclusion: Our data shows that transplantation of the kidney with multiple arteries is safe. Any reconstruction technique can be chosen according to the vascular anatomy of the graft and the recipient

Prognostic parameters for the prediction of acute gangrenous cholecystitis

Background/Purpose: The aim of this study was to identify preoperative prognostic parameters for gangrenous cholecystitis to differentiate this subgroup of patients with acute cholecystitis in order to provide immediate surgical therapy. Methods: The medical records of patients who had an emergency cholecystectomy with the diagnosis of acute cholecystitis between January 2002 and June 2005 were reviewed retrospectively. Univariate and multivariate analysis were performed on the data. Results: Out of 203 individuals with the clinical diagnosis of acute cholecystitis, 21 (10.3%) patients had a histological diagnosis of gangrenous cholecystitis. Multivariate analysis demonstrated an independent association of male sex, diabetes mellitus and white blood cell (WBC) count with the development of acute gangrenous cholecystitis. Conclusions: The risk for gangrenous cholecystitis is increased in male patients who have diabetes and a greater WBC count than 14900/mm3. Urgent surgical intervention should be considered for these patients because of the high morbidity and mortality rate of the condition. © Springer-Verlag Tokyo 2006

Viral transmission during surgery: What do we know?

Surgeons are frequently exposed to blood and body fluids that have the potential to transmit disease. During surgery, intact gloves act as a protective barrier against bloodborne pathogens. In this study, we investigated the knowledge of the surgeons and residents at our hospital about the transmission risk of the three bloodborne viruses most commonly involved in the health care setting: human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV). We also investigated their immunization against the HBV, gloving attidues and puncture rates during surgery. A survey form was filled reciprocally with physicians who accepted to answer. From surgical specialties, a number of 127 physicians consisting of 78 resident (%61) and 49 staff surgeon (%39) participated to our questionnaire. The rate of immunized, naturally immunized and those who were checked their anti-HbS titration at least once among the respondents were %79, %12 and %52 respectively. While the estimation ratio of the transmisson risk for HBV and HCV was both %25, only %10 of the respondents estimated the transmission risk for HIV correctly. The needle puncture rates in major risk vs minor risk specialties (p=0.002) and in residents vs staff surgeons (p=0.0001) were significantly higher. However, the rate of double gloving was only %4.7 in all series. The proportion of double glovers were significantly higher among staff surgeons (p=0.004)