Complex Gill Disease: an Emerging Syndrome in Farmed Atlantic Salmon (Salmo salar L.).

Gill disorders have become a significant problem during the marine phase of farming Atlantic salmon (Salmo salar L.). The term complex gill disease (CGD) includes a wide range of clinical gill disease presentations generally occurring from the end of summer to early winter on marine Atlantic salmon farms. The gross and histological lesions observed are the resultant culmination of exposure to a mixture of environmental insults, pathogenic organisms and farm management practices. None of the three principal agents purportedly associated with CGD (Desmozoon lepeophtherii, salmon gill poxvirus or Candidatus Branchiomonas cysticola) have been cultured successfully in-vitro, so individual in-vivo challenge studies to identify their pathogenesis have not been possible. Studies of cohabitation of single pathogen-infected fish with naïve fish, and epidemiological investigations are required urgently to elucidate the roles of these pathogens and other factors in CGD.

Technical pre-analytical effects on the clinical biochemistry of Atlantic salmon (Salmo salar L.).

Clinical biochemistry has long been utilized in human and veterinary medicine as a vital diagnostic tool, but despite occasional studies showing its usefulness in monitoring health status in Atlantic salmon (Salmo salar L.), it has not yet been widely utilized within the aquaculture industry. This is due, in part, to a lack of an agreed protocol for collection and processing of blood prior to analysis. Moreover, while the analytical phase of clinical biochemistry is well controlled, there is a growing understanding that technical pre-analytical variables can influence analyte concentrations or activities. In addition, post-analytical interpretation of treatment effects is variable in the literature, thus making the true effect of sample treatment hard to evaluate. Therefore, a number of pre-analytical treatments have been investigated to examine their effect on analyte concentrations and activities. In addition, reference ranges for salmon plasma biochemical analytes have been established to inform veterinary practitioners and the aquaculture industry of the importance of clinical biochemistry in health and disease monitoring. Furthermore, a standardized protocol for blood collection has been proposed.

Electrophysiological function during voiding after simulated childbirth injuries.

During vaginal delivery dual injuries of the pudendal nerve and the external urethral sphincter (EUS), along with other injuries, are correlated with later development of stress urinary incontinence. It is not known how combinations of these injuries affect neuromuscular recovery of the micturition reflex. We investigated the EUS electromyogram (EMG) and the pudendal nerve motor branch potentials (PNMBP) during voiding 4 days, 3 weeks or 6 weeks after injury; including vaginal distension (VD), pudendal nerve crush (PNC), both PNC and VD (PNC+VD), and pudendal nerve transection (PNT); and in controls. Pudendal nerve and urethral specimens were excised and studied histologically. No bursting activity was recorded in the EUS EMG during voiding 4 days after all injuries, as well as 3 weeks after PNC+VD. Bursting activity demonstrated recovery 3 weeks after either VD or PNC and 6 weeks after PNC+VD, but the recovered intraburst frequency remained significantly decreased compared to controls. Bursting results of PNMBP were similar to the EMG, except bursting in PNMBP 4 days after VD and the recovered intraburst frequency was significantly increased compared to controls after PNC and PNC+VD. After PNT, neither the EUS nor the pudendal nerve recovered by 6 weeks after injury. Our findings indicate bursting discharge during voiding recovers more slowly after PNC+VD than after either PNC or VD alone. This was confirmed histologically in the urethra and the pudendal nerve and may explain why pudendal nerve dysfunction has been observed years after vaginal delivery.

Lower urogenital tract anatomical and functional phenotype in lysyl oxidase like-1 knockout mice resembles female pelvic floor dysfunction in humans.

Female pelvic floor dysfunction (FPFD) is a complex group of conditions that include urinary incontinence and pelvic organ prolapse (POP). In humans, elastin homeostasis has been implicated in the pathophysiology of FPFD. Lysyl oxidase-like 1 knockout (LOXL1-KO) mice demonstrate abnormal elastic fiber homeostasis and develop FPFD after parturition. We compared the lower urogenital tract (LUT) anatomy and function in LOXL1-KO mice with and without POP. LUT anatomy was assessed in LOXL1-KO mice over 28 wk. Pelvic visceral anatomy in LOXL1-KO was evaluated with a 7-Tesla magnetic resonance imaging (MRI) scanner. LUT function was assessed using conscious cystometry and leak point pressure (LPP) testing. Quantitative histological analysis of elastic fibers was performed on external urethral sphincter (EUS) cross sections. By 25 wk of age, 50% of parous LOXL1-KO mice developed POP. LOXL1-KO mice with POP had greater variability in the size and location of the bladder on MRI compared with mice without POP. Parity and POP were associated with lower LPP. Elastin clusters were significantly increased in the EUS of LOXL1-KO mice with POP. Because parity triggers POP in LOXL1-KO mice, LOXL1-KO mice with POP have variable internal pelvic anatomy, and both parity and POP are associated with a decrease in LPP, we conclude that LOXL1 LUT anatomical and functional phenotype resembles FPFD in humans. The increase in elastin clusters in the urethra of LOXL1-KO mice with POP suggests that elastin disorganization may lead to functional abnormalities. We conclude that LOXL1 warrants further investigation in the pathphysiology of FPFD.

Efficacy of a standardized technique of trans-vaginal entry into the peritoneal cavity for post-hysterectomy vaginal vault suspension.

The aim of this paper is to describe the efficacy and safety of a standardized technique of trans-vaginal entry into the peritoneal cavity for post-hysterectomy vault prolapse and determine how well the pre-operative pelvic examination predicted successful peritoneal entry. A retrospective review of patients undergoing post-hysterectomy trans-vaginal apical suspensions using a standardized technique of peritoneal entry was employed in this study. A subset of patients underwent a standardized pre-operative evaluation to predict what organ was behind the vaginal cuff and posterior cul-de-sac. Peritoneal entry was attempted in 280 patients, and successfully achieved in 223(80%). One cystotomy and one proctotomy (0.3%) occurred during attempted entry. Of those who underwent the standardized pre-operative evaluation, peritoneal entry was successful 86% of the time when small bowel was predicted to lie behind the vaginal cuff or posterior cul-de-sac. Peritoneal entry can be safely achieved in the majority of post-hysterectomy prolapse patients. Pre-operative examination assists in predicting successful peritoneal entry.

Bowel symptoms 1 year after surgery for prolapse: further analysis of a randomized trial of rectocele repair.

OBJECTIVE: The purpose of this study was to analyze change in bowel function and its relationship to vaginal anatomy 1 year after rectocele repair and pelvic reconstruction in a randomized trial of 3 techniques of rectocele repair. STUDY DESIGN: The study is an ancillary analysis of data from a randomized trial of 3 techniques of rectocele repair: posterior colporrhaphy, site-specific repair, and site-specific repair with Fortagen graft augmentation. Pelvic examination and validated questionnaires were obtained at baseline, 6 months, and 1 year after surgery. Bowel symptoms included straining, splinting, incomplete emptying, painful defecation, fecal urgency, and fecal incontinence. Logistic regression was used to identify predictors of bothersome postoperative symptoms. RESULTS: One hundred six women with Stage > or = 2 POP, which included a rectocele, were enrolled in the study. Ninety-nine underwent prolapse surgery that included a rectocele repair and completed at least 1 follow-up visit. Ninety-six percent of subjects underwent concomitant prolapse surgery. No differences in change in bowel symptoms were noted between treatment groups. On average, all bowel symptoms evaluated were significantly improved 1 year after surgery. The development of new "bothersome" bowel symptoms after surgery was uncommon (11%). After controlling for age, treatment group, comorbidities, and preoperative bowel symptoms, corrected postoperative vaginal support (Stage 0/1) was associated with a reduced risk of postoperative straining (adj. OR 0.17 95% CI 0.03 to 0.9) and feeling of incomplete emptying (adj. OR 0.1 95% CI 0.01 to 0.52). Normal support of the posterior vaginal wall (Bp < or = -2) was associated with a reduced risk of bothersome incomplete emptying (OR 0.08 95% CI 0.004 to 0.58) but not with other symptoms. CONCLUSION: Resolution or improvement in bowel symptoms can be expected in the majority of women after rectocele repair and pelvic reconstruction. While all symptoms improved after surgery, a reduction in bothersome postoperative straining and incomplete emptying were specifically associated with cure of posterior vaginal wall prolapse.

De novo rectal prolapse after obliterative and reconstructive vaginal surgery for urogenital prolapse.

OBJECTIVE: The purpose of this study was to compare the incidence of de novo rectal prolapse after obliterative vaginal surgery with the incidence that was seen after reconstructive vaginal surgery for urogenital prolapse. STUDY DESIGN: A chart review was performed on subjects who underwent vaginal surgery for urogenital prolapse from Jan. 1, 2001, through Dec. 31, 2004, at the Cleveland Clinic. Diagnosis of postoperative rectal prolapse was identified with ICD-9 code 569.1. RESULTS: Nine hundred sixteen women underwent vaginal surgery for urogenital prolapse. Ninety-two percent of the women (n = 840) underwent reconstructive surgery, and 8% of the women (n = 76) underwent obliterative surgery. The incidence of postoperative full-thickness rectal prolapse in women who were > or = 65 years old who underwent obliterative surgery was 3 of 74 (4.1%; 95% CI, 1.4-11), with an estimated odds ratio of 22 (95% CI, 2.3-196; P < .002) compared with women who were > or = 65 years old who underwent reconstructive surgery. CONCLUSION: Obliterative surgery is associated with a substantially greater risk of de novo rectal prolapse than reconstructive vaginal surgery for urogenital prolapse.

Treatment of urinary tract endometriosis.

Endometriosis involving the urinary tract, although infrequent, can have significant impact on patients' symptoms, response to treatment, and urologic function. The purpose of this article is to review the epidemiology, pathophysiology, diagnosis, and management of endometriosis that affects the urinary tract.

The incidence of ureteral obstruction and the value of intraoperative cystoscopy during vaginal surgery for pelvic organ prolapse.

OBJECTIVE: The objective of the study was to determine the incidence of ureteral obstruction during vaginal surgery for pelvic organ prolapse and the accuracy and efficacy of intraoperative cystoscopy. STUDY DESIGN: The study was a retrospective review of 700 consecutive patients who underwent vaginal surgery for anterior and/or apical pelvic organ prolapse with universal intraoperative cystoscopy. RESULTS: Thirty-seven patients (5.3%) had no spillage of dye from 1 or both ureters intraoperatively. The false-positive and negative cystoscopy rates were 0.4% and 0.3%, respectively. Thus, the true incidence of intraoperative ureteral obstruction was 5.1%. Intraoperative cystoscopy was accurate in 99.3% of cases, with a sensitivity and specificity of 94.4% and 99.5%, respectively. Suture removal relieved ureteral obstruction in 88% of cases. Six subjects (0.9%) had true ureteral injuries. CONCLUSION: Vaginal surgery for anterior and/or apical pelvic organ prolapse is associated with an intraoperative ureteral obstruction rate of 5.1%. Intraoperative cystoscopy accurately detects ureteral obstruction and allows for relief of obstruction in the majority of cases.

Cloning and expression studies during vegetative and sexual development of Pbs1, a septin gene homologue from Pyrenopeziza brassicae.

A septin gene homologue designated Pyrenopeziza brassicae septin 1 (Pbs1) has been identified and cloned from the plant pathogenic fungus Pyrenopeziza brassicae and its expression analysed. Pbs1 is present in both mating types and in a single copy within each genome and is transcribed in proportionate levels during both vegetative and sexual growth.

Intra-specific and inter-specific conservation of mating-type genes from the discomycete plant-pathogenic fungi Pyrenopeziza brassicae and Tapesia yallundae.

In previous work, four genes involved in mating-type determination were cloned from reference strains of Pyrenopeziza brassicae; three genes, PAD1, PMT1, and PHB1 (re-named henceforth as MAT-1-1, MAT-1-4, and MAT1-3, respectively), are encoded by the MAT-1 idiomorph, and one gene, PHB2 (re-named MAT-2), by the corresponding MAT-2 idiomorph. To assess MAT gene organisation within field-populations of P. brassicae, 30 field-isolates of both mating-types from different geographical locations were analysed by PCR using primers designed for the MAT genes of P. brassicae. The results indicate that mating-type gene structure and organisation within these isolates is conserved and is consistent with the mating-type designations established by crossing experiments. The four P. brassicae MAT genes were then used as probes against gel blots of the genomic DNA of a discomycete Tapesia yallundae from the same family (Dermateaceae, order Helotiales) and one, Ascobolus stercorarius, from a distantly related family (Ascobolaceae, order Pezizales), in order to determine whether P. brassicae MAT-gene homologs were present. MAT-specific hybridisation signals were obtained with T. yallundae using all four probes. In particular, MAT-1 DNA of T. yallundae gave a strongly hybridising signal with MAT-1-4 (PMT1), a putative metallothionein gene found in the P. brassicae MAT-1 idiomorph but not in any other MAT idiomorph examined to-date. No MAT-specific hybridisation was obtained with A. stercorarius. A fragment of the MAT-2 gene of T. yallundae was obtained by PCR using degenerate primers designed to amplify the high-mobility group (HMG) domain present in other ascomycete MAT genes. Sequencing of this PCR product revealed similarities to MAT HMG domains from other ascomycetes with the greatest degree of similarity exhibited with P. brassicae. The T. yallundae HMG-DNA sequence was shown to co-segregate with mating type (MAT-2) in progeny from a sexual cross.

Cloning of the mating type loci from Pyrenopeziza brassicae reveals the presence of a novel mating type gene within a discomycete MAT 1-2 locus encoding a putative metallothionein-like protein.

The mating type loci were cloned from Pyrenopeziza brassicae by chromosome walking from a mating type-linked polymerase chain reaction (PCR) fragment and shown to be idiomorphic by sequence analysis. The MAT 1-1 locus is approximately 3.2 kb and contains a single gene encoding a putative high-mobility group (HMG) domain protein. The MAT 1-2 locus is approximately 3.9 kb with three open reading frames (ORFs) encoding a putative HMG domain, an alpha-1 domain and metallothionein-like proteins. The putative alpha-1 domain ORF on MAT 1-2 is transcribed in the opposite orientation to the other two transcripts and extends into non-idiomorphic sequence. This is the first report of sequence analysis of the mating type loci from a discomycete fungus, which has revealed an interesting mating type infrastructure within the MAT 1-2 locus. Although metallothionein-like proteins have been implicated in a number of processes in animals and plants, they have not to date been implicated in the mating process of filamentous fungi. Possible roles for metallothionein-like proteins in the mating process are discussed.

Plasma and erythrocyte profiles of nonesterified polyunsaturated fatty acids during normal pregnancy and labor.

This study was undertaken to determine the plasma and erythrocyte profiles of metabolically important nonesterified polyunsaturated fatty acids (PUFA), that is, free fatty acids, in each trimester of pregnancy and in labor. Blood was drawn from patients in the first, second, and third trimester and in labor. Nonesterified polyunsaturated fatty acids were extracted from erythrocytes with chloroform and methanol. The PUFAs from erythrocytes and plasma were then measured in their methylester form using high performance liquid chromatography (HPLC). Plasma levels of all PUFAs were similar in each trimester of pregnancy but levels of linoleic and linolenic acids were higher in laboring patients. Plasma levels of linoleic and arachidonic acid in the n-6 pathway (range 40 to 162 mg/L plasma) were higher than linolenic, eicosapentaenoic, and docosahexaenoic acids measured in the n-3 pathway (range 2.1 to 12.8 mg/L plasma). PUFA levels in erythrocytes were generally higher in the second trimester (range 2.6 to 79.7 mg/100-microL spun erythrocytes). In these erythrocytes, docosahexaenoic acid in the n-3 pathway and linoleic and arachidonic acids in the n-6 pathways were present in the highest amounts. Polyunsaturated fatty acids appear to be absorbed and mobilized in increasing amounts in plasma and erythrocytes with advancing gestational age and labor. This activity appears to be most pronounced in the second trimester. Further investigations into PUFA metabolism and the mechanisms which govern it could lead to a better understanding of the role of these important substances in normal and abnormal pregnancies as well as in the initiation of labor.

Ti plasmid-specified chemotaxis of Agrobacterium tumefaciens C58C1 toward vir-inducing phenolic compounds and soluble factors from monocotyledonous and dicotyledonous plants.

Twelve phenolic compounds with related structures were analyzed for their ability to act as chemoattractants for Agrobacterium tumefaciens C58C1 and as inducers of the Ti plasmid virulence operons. The results divided the phenolic compounds into three groups: compounds that act as strong vir inducers and are chemoattractants for A. tumefaciens C58C1 harboring the nopaline Ti plasmid pDUB1003 delta 31, but not the isogenic cured strain; compounds that are at best weak vir inducers and are weak chemoattractants for Ti plasmid-harboring and cured A. tumefaciens C58C1; and compounds that are vir noninducers and are also nonattractants. A strong correlation between vir-inducing ability and Ti plasmid requirement for chemotaxis is thus established. In addition, chemical structure rules for vir induction and chemotaxis are outlined. Positive chemotaxis toward root and shoot homogenates from monocotyledonous and dicotyledonous plants was observed. At low extract concentrations, chemotaxis was enhanced by the presence of Ti plasmid. The chemoattractants do not derive from intact cell walls. Lack of attraction is not responsible for the apparent block to monocot transformation by A. tumefaciens.

virA and virG are the Ti-plasmid functions required for chemotaxis of Agrobacterium tumefaciens towards acetosyringone.

Octopine and nopaline Ti-plasmids confer upon Agrobacterium tumefaciens C58C1 the ability to respond chemotactically to the vir-inducing phenolic wound exudate, acetosyringone. A. tumefaciens C58C1 containing Ti-plasmids with Tn5 insertions in virB, C, D or E exhibited marked chemotaxis towards acetosyringone. However, Ti-plasmids with mutations in virA or virG were unable to confer the responsive phenotype. Of the cosmid clones pVK219 (virAB) pVK221 (virBGC) pVK225 (virGCDE) and pVK257 (virABGC) mobilized to cured A. tumefaciens C58C1, only pVK257 bestowed acetosyringone chemotaxis. virA and virG are thus required for chemotaxis of A. tumefaciens towards acetosyringone. This suggests a multifunctional role for virA and virG: at low concentrations of acetosyringone they mediate chemotaxis and at higher concentrations they effect vir-induction.

Developmental study of short-term memory characteristics for kinesthetic movement information.

The present experiment was designed to examine age-related characteristics of short-term retention of kinesthetic movement information. 20 children from each age group (6, 8, 10 yr.) were tested for recall of kinesthetic end-location. The results indicate that 6- and 8-yr.-old children became less accurate and more variable in recall performance following a delay interval. However, 10-yr.-old children were capable of maintaining performance accuracy during a delay interval.