Phylogenetic distribution of a male pheromone that may exploit a nonsexual preference in lampreys.

Pheromones are among the most important sexual signals used by organisms throughout the animal kingdom. However, few are identified in vertebrates, leaving the evolutionary mechanisms underlying vertebrate pheromones poorly understood. Pre-existing biases in receivers' perceptual systems shape visual and auditory signalling systems, but studies on how receiver biases influence the evolution of pheromone communication remain sparse. The lamprey Petromyzon marinus uses a relatively well-understood suite of pheromones and offers a unique opportunity to study the evolution of vertebrate pheromone communication. Previous studies indicate that male signalling with the mating pheromone 3-keto petromyzonol sulphate (3kPZS) may exploit a nonsexual attraction to juvenile-released 3kPZS that guides migration into productive rearing habitat. Here, we infer the distribution of male signalling with 3kPZS using a phylogenetic comparison comprising six of 10 genera and two of three families. Our results indicate that only P. marinus and Ichthyomyzon castaneus release 3kPZS at high rates. Olfactory and behavioural assays with P. marinus, I. castaneus and a subset of three other species that do not use 3kPZS as a sexual signal indicate that male signalling might have driven the evolution of female adaptations to detect 3kPZS with specific olfactory mechanisms and respond to 3kPZS with targeted attraction relevant during mate search. We postulate that 3kPZS communication evolved independently in I. castaneus and P. marinus, but cannot eliminate the alternative that other species lost 3kPZS communication. Regardless, our results represent a rare macroevolutionary investigation of a vertebrate pheromone and provide insight into the evolutionary mechanisms underlying pheromone communication.

Sex cord stromal testicular tumors: a clinical series--uniformly stage I disease.

PURPOSE: Sex cord stromal testicular tumors are rare. Historically 10% of lesions are said to be malignant but to our knowledge there are no clinical or histological features that can accurately predict potential malignant behavior. Because of this, groups at some centers have advocated prophylactic retroperitoneal lymph node dissection in patients with clinical stage I disease. We reviewed our experience with these tumors to determine whether this policy is justified. MATERIALS AND METHODS: We retrospectively reviewed the records of all 38 men older than 18 years with sex cord stromal testicular tumors who were referred to the Wessex regional cancer center for treatment or pathological review during the 25-year period of 1982 to 2006. We then compared our series with a malignant sex cord stromal testicular tumor database generated from the world literature. RESULTS: All Wessex patients were treated with excision of the primary tumor alone and metastatic disease developed in none. All remained disease-free with an overall median survival of 6.8 years (range 1.4 to 25). Features in the literature favoring malignant behavior, ie metastatic disease, included larger tumors (mean 6.43 vs 1.71 cm), a high mitotic rate, tumor necrosis, angiolymphatic invasion, infiltrative margins and extratesticular extension (each p <0.0001). The malignant group had an overall median survival of 2.3 years (range 0.02 to 17.3). CONCLUSIONS: No patient had disease progression in our study, which is to our knowledge the largest reported United Kingdom series of sex cord stromal testicular tumors. Our data suggest that malignancy is uncommon and prophylactic retroperitoneal lymph node dissection is unjustified for clinical stage I disease.

Case series: adult testicular dermoid tumours--mature teratoma or pre-pubertal teratoma?

Adult testicular dermoid tumours are rare tumours with no reported potential for recurrent or metastatic spread. Despite this they are currently classified as mature teratoma and managed as if they have equivalent malignant potential. This report describes two cases of adult mature teratoma of dermoid type and questions the classification and pathogenesis of this disease. In one of the cases there was a clear history of a testicular lump arising pre-pubertally, raising the possibility that some adult dermoid tumours may in fact be pre-pubertal teratomas that have persisted into adulthood. Classification as a mature teratoma carries with it a follow-up regimen that includes numerous radiological investigations with their attendant radiation exposure. A positive histological diagnosis and separate classification of adult dermoid tumours would allay clinical fears of recurrence and metastasis and negate the need for repeated radiological investigations.

Health related quality of life (HRQoL) after cystectomy: comparison between orthotopic neobladder and ileal conduit diversion.

INTRODUCTION: Radical cystectomy and urinary diversion carries a high morbidity. Quality of life and body image are important considerations for urinary diversion (UD). We wanted to conduct a systematic review of literature to see which form UD offers a better quality of life (QoL). METHODS: We searched MEDLINE, Pubmed, EMBASE, CINAHL and the Cochrane library for studies using the following key words: 'quality of life' and 'ileal conduit', 'orthotopic neobladder', 'continent diversion' and 'urinary diversion'. All English language articles on UD surgery were included in the original search from 1990 to 2014. To improve the quality of evidence, we stratified our inclusion criteria into studies that report on QoL in both forms of UD using at least one validated questionnaire. RESULTS: Twenty-one studies (2285 patients) were included in our study all of which used at least one validated tool. The most frequently used tools were the SF-36, EORTC QLQ-C30 and FACT BL (10, 8, 5 studies respectively). None of the studies were randomised and only 4 studies were prospectively designed. Sixteen studies reported no difference in QoL between the two types of urinary diversion and four studies reported a better QoL with orthotopic neobladder of which 2 studies had younger and fitter patients. On the other hand, one study reported a better QoL in ileal conduit patients. CONCLUSION: Orthotopic neobladder urinary diversion shows a marginally better quality of life scores compared to ileal conduit diversion especially when considering younger and fitter patients.

Ovarian steroid cell tumors (not otherwise specified). A clinicopathological analysis of 63 cases.

The clinical and pathological features of 63 steroid cell tumors, not otherwise specified, were reviewed. The patients ranged in age from 2 1/2 to 80 years. The most common initial manifestation was virilization (41%); four patients had estrogenic manifestations, and four had hypercortisolemia with Cushing's syndrome. The tumors, 6% of which were bilateral, ranged from 1.2 to 45 cm in greatest dimension. Microscopic examination revealed two types of cells, which had overlapping features: those with abundant eosinophilic cytoplasm and those with vacuolated cytoplasm. Fat stains were positive in 75% of the 16 cases in which they were performed. Follow-up data ranging from 1 to 19 years (average 5.2 years) in duration were available for 50 patients. In 24 cases, the tumor was designated probably benign (no evidence of spread beyond the ovary within 3 or more years postoperatively). In 18 patients, the tumor was clinically malignant. The best pathological correlates of malignant behavior were: the presence of two or more mitotic figures per 10 high power fields (92% malignant); necrosis (86% malignant); a diameter of 7 cm or greater (78% malignant); hemorrhage (77% malignant); and grade 2 or 3 nuclear atypia (64% malignant).

Determination of atrazine in water by magnetic particle immunoassay: collaborative study.

A collaborative study was performed to determine mean recovery and precision for analysis of atrazine in drinking and surface waters by immunoassay. The study design was based on the blind duplicate test plan for collaborative studies. Three blank waters (municipal drinking water, well water, and surface water) were spiked at 3 atrazine levels. Two water samples with naturally incurred atrazine loads were also spiked with atrazine at 3 levels. In the enzyme-linked immunoassay method, the water sample is mixed with a pesticide-enzyme conjugate and added to paramagnetic particles with triazine-specific antibodies attached. After separation of antibody-bound atrazine and atrazine-enzyme conjugate from free components, the bound enzyme conjugate catalyzes a reaction producing a colored end product. The color developed is inversely proportional to the original concentration of atrazine in the water sample. Fourteen laboratories participated in the collaborative study. Data were analyzed for repeatability and reproducibility, and average recoveries at the spike levels were calculated. Over the concentration range tested, the mean recovery of atrazine spiked into blank and pesticide-contaminated waters was 104%. Overall RSDR averaged about 40% for atrazine concentrations near the method detection limit (0.05 microgram/L) and about 15% at concentrations above 5 times the detection limit (0.25 microgram/L). Corresponding single-analyst RSDr values were 24 and 10%. Recovery and precision for the 3 blank water matrixes and the waters that had been naturally contaminated with atrazine showed no significant differences. The magnetic particle immunoassay for determination of atrazine in water has been adopted first action by AOAC INTERNATIONAL.

The management of coeliac disease.

One hundred children put on a gluten free diet because of suspected coeliac disease were followed for a mean period of 9.9 years. The diagnosis was eventually confirmed in 53. Under the age of two, 35 children showed subtotal villous atrophy in their initial biopsy but nine of these on subsequent gluten challenge and rebiopsy were found to tolerate gluten normally. Challenge and rebiopsy are also necessary for children over the age of two where the initial biopsy changes are less severe than subtotal villous atrophy. Such challenges can probably be carried out earlier than the recommended age of six given in the 1990 ESPGAN (European Society for Paediatric Gastroenterology and Nutrition) report with little risk of serious dental damage.

A pressing diagnosis - a compromised testicle secondary to compartment syndrome.

Hydrocoeles are a common cause of scrotal swelling and discomfort in a significant proportion of men. We report a case of compartment syndrome within the tunica vaginalis. This is an unusual and previously unreported complication of a hydrocoele.

Stromal luteoma of the ovary: a clinicopathological analysis of 25 cases.

The clinical and pathological features of 25 stromal luteomas were reviewed. The patients ranged in age from 28-74 (average 58.4) years. The most common initial manifestation (60%) was abnormal vaginal bleeding; three patients (12%) were virilized. The tumors, only one of which was bilateral, ranged from 0.25-2.9 (average 1.3) cm in greatest diameter. They were all well circumscribed. Microscopic examination revealed unencapsulated nodules composed of rounded polyhedral cells that were arranged either diffusely or in nests and cords. Twenty percent showed degeneration within aggregates of cell resulting in the formation of pseudoacinar structures. The cytoplasm of the neoplastic cells was generally eosinophilic and granular, and lipochrome granules were present in over half the tumors. In 92% of the cases hyperthecosis was present in the adjacent ovarian stroma. Follow-up of 6 months to 17 years was available for 19 patients. Sixteen of them were alive and free of disease; the remaining three died of other causes. The benign course in all the cases in which follow-up data were available was expected in view of the small size of the tumors and their benign microscopic appearance.

The Pippi Salle urethral lengthening procedure; experience and outcome from three United Kingdom centres.

OBJECTIVE: To assess the outcome of the Pippi Salle (Kropp onlay) urethral lengthening procedure in the treatment of neuropathic urinary incontinence. PATIENTS AND METHODS: Twenty-eight patients (12 males and 16 females, mean age at surgery 12.6 years, range 7-32) were identified who underwent the procedure between 1993 and 1997 in the United Kingdom. Outcomes were assessed by a review of the case notes. RESULTS: Of the 28 patients, 18 (64%) were rendered continent by day, and 17 (61%) by day and night. Twelve of the 16 females were completely dry, and a further girl was dry by day, giving overall daytime continence in 13 patients; five males were rendered continent (P=0.03, chi-square test). Four patients required revision surgery; in five patients (two females and three males) the method was abandoned and they underwent an alternative procedure, and a further four are being reassessed. CONCLUSION: The Pippi Salle procedure should be considered as a first-line treatment option for neuropathic incontinence in females. Its place in the management of incontinent males is less convincing.

Confocal microscopy of idarubicin localisation in sensitive and multidrug-resistant bladder cancer cell lines.

Idarubicin is a highly lipophilic anthracycline and appears effective against tumours resistant to conventional anthracyclines. Confocal microscopy demonstrates predominantly cytoplasmic idarubicin accumulation. This distribution is unaltered by resistance status or the resistance reversing agent verapamil. Our results contrast with studies on conventional anthracyclines and suggest that nuclear accumulation may not be a prerequisite for anthracycline cytotoxicity.

Fetal heart rate and intrauterine growth.

OBJECTIVE: To assess whether fetal heart rate in early and late pregnancy relates to size at birth. DESIGN: Prospective study of fetal heart rates in early and late pregnancy. SETTING: Princess Anne Hospital, Southampton. SUBJECTS: 63 primigravid women. MAIN OUTCOME MEASURES: Anthropometric measurements made on the newborn infant. RESULTS: There were no differences in heart rate between the sexes at 18 weeks gestation but by 36 weeks the boys had rates which were 4.4 beats lower than those of the girls. Higher fetal heart rate at 18 weeks was associated with lower ponderal index, smaller head circumference and smaller mid-arm circumference. There were no trends in fetal heart rate at 36 weeks with any birth measurements. CONCLUSION: Babies born at term who have a pattern of neonatal measurements which reflect growth retardation have raised heart rates in early pregnancy. Influences which impair fetal growth appear to take effect early in gestation.

Cellular resistance to mitomycin C is associated with overexpression of MDR-1 in a urothelial cancer cell line (MGH-U1).

OBJECTIVE: To compare multidrug resistance (MDR)-1 and MDR-3 gene expression in a new urothelial cancer cell line (MGHU-1, with resistance to mitomycin C) against controls and the established (epirubicin-resistant) MDR clone, and to correlate MDR with cytotoxicity data. MATERIALS AND METHODS: Resistance to mitomycin C was induced by the long-term exposure of wild-type MGHU-1 cells to increasing concentrations (20-400 nmol/L) of mitomycin C. The cytotoxicity of mitomycin C or epirubicin was then compared in MGHU-1, MGHU-MMC (mitomycin C-resistant) and MGHU-1R (established MDR) cells, using the tetrazolium biomass assay. The expression of MDR-1 and -3 was investigated by the reverse transcriptase-polymerase chain reaction, using cDNA-specific primers after titration, and compared with DNA and negative controls. RESULTS: MDR-1 and -3 were significantly and equally overexpressed in MGHU-1R, and associated with a dramatic increase in the 50% inhibitory drug concentration (P < 0.001) for mitomycin C and epirubicin against controls. In MGHU-MMC, the overexpression of MDR-1 was three times greater than that of MDR-3. The cytotoxicity profile for both agents was very similar to that of MGHU-1R. Trace amounts of MDR-1, but not MDR-3, were identified in the MGHU-1 wild-type. CONCLUSIONS: Urothelial cancer cell resistance to mitomycin C is associated with cross-resistance to epirubicin and overexpression of MDR-1, suggesting that mitomycin C falls within the MDR category. Clinical application of this methodology may allow patients to be identified who are unlikely to benefit from intravesical chemotherapy.

Determination and reversal of resistance to epirubicin intravesical chemotherapy. A flow cytometric model.

OBJECTIVE: To develop a method of determining the characteristics of epirubicin resistance and to study the reversal of such resistance in the intravesical treatment of superficial bladder cancer, using sensitive and resistant derivatives of a bladder cancer cell line in vitro. MATERIALS AND METHODS: Epirubicin fluorescence and flow cytometry were used to measure the intracellular levels of epirubicin in both sensitive and resistant live cultured bladder tumour cells, with and without different doses of the resistance-reversing agent verapamil. RESULTS: There was a reliable, highly significant and consistent difference in intracellular epirubicin concentration between the resistant and sensitive bladder tumour cells. In addition, it was possible to substantially reverse the features of resistant cell subline with additional verapamil. CONCLUSION: Application of this assay to clinical specimens should allow better targeting of epirubicin intravesical chemotherapy and avoid the premature termination of such treatment in patients whose tumours remain sensitive to this agent. Furthermore, the addition of verapamil to intravesical epirubicin may permit effective treatment of those patients whose tumours have inherent or acquired resistance to epirubicin.

Determination and reversal of resistance to epirubicin intravesical chemotherapy. A confocal imaging study.

OBJECTIVE: To evaluate the use of confocal microscopy in the study of resistance to epirubicin and to determine the effect of temperature, viability and a resistance-reversing agent on the intracellular distribution of this drug in sensitive and resistant derivatives of a superficial bladder cancer cell line. MATERIALS AND METHODS: Viable and non-viable adherent cells were incubated in epirubicin solutions under various conditions. After incubation, the distribution of intracellular epirubicin fluorescence was visualized using confocal microscopy and a x50 water-immersion lens. RESULTS: There was a striking and consistent difference between resistant and sensitive cells in the intracellular distribution of the drug. In addition to having greater overall levels of epirubicin fluorescence, sensitive cells accumulated epirubicin predominantly in the nucleus. Epirubicin fluorescence in resistant cells was cytoplasmic and granular in appearance. When incubated at 0 degrees C, both cell lines showed no nuclear uptake and thus resembled resistant cells at 37 degrees C. However, dead cells rapidly acquired brightly fluorescent nuclei. The resistance-reversing agent verapamil appeared to cause reversion of the resistant to the sensitive phenotype. CONCLUSION: Confocal microscopy allows epirubicin-sensitive and resistant cultured tumour cells to be differentiated reliably and provides information about the mechanisms of action of, and resistance to, epirubicin. Applying this technique to clinical specimens should enable patients who have the resistant phenotype to be detected and the efficacy of intravesical resistance-reversing agents to be evaluated in such cases.

Estramustine reversal of resistance to intravesical epirubicin chemotherapy.

OBJECTIVE: Failure of epirubicin treatment of superficial bladder cancer implies multidrug resistance (MDR) which is common. MDR is characterised by decreased cellular levels of drug. TCC cell lines sensitive to epirubicin and resistant to both epirubicin and mitomycin C were used to investigate augmented therapy by adding the MDR reversing agent estramustine to an in vitro model. METHODS: Cells were cultured as monolayers. Fluorescence analysis was performed by flow cytometry and confocal microscopy. Cells were exposed to epirubicin 20 microg/ml for 2 h and increasing amounts of estramustine. Cytotoxicity was determined under similar exposure conditions and MTT culture (dye reduction by live cells) allowed viable biomass to be read optically. RESULTS: Resistant cells accumulated eight times less epirubicin than sensitive cells. Confocal microscopy confirmed this for nuclear uptake. Accumulation in resistant cells can be increased to near-sensitive cell levels using 40 microg/ml estramustine. Image analysis of confocal fluorescence showed a shift from cytoplasm to nucleus. This correlated with increased cytotoxicity. CONCLUSION: Estramustine plus epirubicin chemotherapy can overcome MDR and may achieve more successful tumour killing in vivo. It may also prevent emergence of resistance. Primary TCC culture examination permits detection of sensitive and resistant cells and may predict outcome allowing a more logical treatment selection.

Multidrug resistance evaluation by confocal microscopy in primary urothelial cancer explant colonies.

Assessing functional multidrug resistance (MDR) status in clinical biopsy material using drug autofluorescence has potential applications to clinical management. The small size of many cystoscopy specimens has led us to develop, as an alternative to flow cytometry, a protocol for studying epirubicin accumulation in adherent colonies of primary bladder cancer cells viewed live and in situ by confocal microscopy. The limitations to quantitation inherent in this technique are compensated for by preservation of cellular organisation and the elimination of non-malignant cells. Biopsy material is disaggregated and explanted into culture-grade petri dishes. After incubation for three to seven days plaques of epithelial cells have developed. Classical patterns of sensitive and resistant drug distribution are observed. Cells of the rolled edges of the colony accumulate more drug than those of the inner epithelial monolayer. Some central areas of larger colonies give the appearance of drug arrested at the intercellular junctions to give a fenestrated pattern. These observations contribute to the understanding of mechanisms in MDR as well as forming the basis for a clinical urological MDR evaluation protocol.

Identification and characterization of elevated microbial counts in bulk tank raw milk.

The bacterial composition of bulk tank milk from 13 farms was examined over a 2-wk period to characterize sudden elevations in the total bacterial count referred to as "spikes." Bulk tank milk samples collected at each pick-up were analyzed for standard plate count, Petrifilm aerobic count, somatic cell count, gram-negative organisms, and streptococci. Twenty standard plate count spikes were observed: 12 associated with streptococci, 4 associated with gram-negative organisms, 2 associated with streptococci and gram-negative organisms, and 2 that were not definitively characterized. Spikes ranged from 14,000 to 600,000 cfu/ml. Streptococcus uberis was isolated as the predominant organism from 11 spikes, and Escherichia coli was isolated from 4 spikes. Statistical analysis of total bacterial counts indicated a high correlation (r = 0.94) between standard plate counts and Petrifilm aerobic count. Regression analysis of standard plate counts and Petrifilm aerobic counts yielded the equation log10 (standard plate count) = 0.73 + 0.85log10 (Petrifilm aerobic count), indicating that the correlation, although strong, is not one to one. In a related pilot study, triplicate bulk tank milk samples were collected and analyzed for total bacterial count and presumptive streptococcus, gram-negative, and staphylococcus counts. Two-way ANOVA of these triplicate data indicated a lack of significant variation among the triplicate samples, suggesting that one sample can reliably gauge the microbial status of the entire bulk tank.

Is gamma-linolenic acid an effective intravesical agent for superficial bladder cancer? In vitro cytotoxicity and in vivo tolerance studies.

The essential fatty acid gamma-linolenic acid (GLA) is an effective cytotoxic agent when applied topically and for prolonged periods to tumour cells. Topical application, by intravesical therapy, is firmly established in the treatment of superficial bladder cancer. However, this form of therapy is limited to a maximum duration of 2 h. At such a short drug exposure time, does GLA retain its cytotoxicity? We have examined this question by exposing the superficial bladder cancer cell lines MGH-U1 and RT112 to meglumine-GLA (MeGLA) for time intervals ranging from 30 min to 2 h, at drug concentrations ranging from 1000 to 1.95 microg/ml. The MTT viable biomass assay was used to assess cell kill. Greater than 90% inhibition was observed at a concentration of 125 microg/ml (IC > 90), at 2 h drug exposure. At shorter drug exposure times, higher drug concentrations were needed to induce the same effect. At 1 h drug exposure, the IC > 90 was recorded at 500 microg/ml. In vivo intravesical tolerance studies were conducted in rats. Rats exposed to 2.5 mg/ml MeGLA intravesically for 2 h or less remained well and bladder histology showed minimal changes. This study confirms that GLA retains its cytotoxicity at short drug exposure times and is well tolerated by normal bladder mucosa in vivo. Bladder mucosa tolerated > 10x the concentration required for the IC > 90 in vitro. MeGLA is therefore a feasible intravesical agent for superficial bladder cancer.