Structure and dynamics of the protoplankton community in an environmentally protected urban stream / Estrutura e dinâmica da comunidade de protoplâncton em um córrego urbano ambientalmente protegido

Abstract We aimed to investigate spatial and temporal scales, abundance, and factors that structure the communities of protozoans in a tropical urban stream. Methods: Samples of water for analysis of biological communities (testate amoebae, ciliates and bacteria) and limnological variables were taken in the Mandacaru stream located in the Conservation Unit of Parque do Cinquentenário, in the city of Maringá, Paraná State, Brazil, in two hydrological periods (dry and rainy). We calculated the Water Quality Index (WQI) composed by nine parameters of water quality (dissolved oxygen, thermotolerant coliforms, pH, BOD, water temperature, total nitrogen, total phosphorous, turbidity, and total solids), Trophic State Index (TSI) for phosphorous and we used a Redundancy Analysis (RDA) to verify the influence of environmental variables in the protozoan community. Results: The WQI showed that water quality was considered good in some points and considered bad in other points. The TSI for phosphorus classified the stream as mesotrophic in the majority of sampled sites points (mean between 53.09 and 58.35). We identified 19 taxa of testate amoebae, belonging to six families, being Difflugiidae, Centropyxidae, and Arcellidae those with more species and 71 infrageneric taxa of ciliates, distributed in 12 orders being Peniculida the most representative order, followed by Euplotida. According to RDA analysis, samples of the dry period were characterized by higher values of bacteria density and concentrations of chlorophyll-a, total phosphorous, and total nitrogen. Ciliates and testate amoebae presented higher abundance values in some of the months characterized by higher precipitation and in conditions of higher system productivity. Conclusion: we may conclude that the Mandacaru stream, although strongly influenced by anthropic action, still presents an acceptable water quality. Lastly, we emphasize that protists abundance was strongly influenced by system productivity. This was evidenced by elevated protozoan densities where there was higher primary and bacterial productivity. Thus, these organisms must be considered in studies that aim at the identification of organisms that may indicate anthropic impacts and environmental quality.

Resumo O objetivo foi investigar, em escalas espacial e temporal, a abundância das comunidades de protozoários bem como os fatores intervenientes na estruturação das mesmas em um córrego urbano. Métodos: Amostras de água para análise de comunidades biológicas (amebas testáceas, ciliados e bactérias) e para análise de variáveis limnológicas foram tomadas no córrego Mandacaru localizado na Unidade de Conservação do Parque do Cinquentenário, na cidade de Maringá, Paraná State, Brasil, em dois períodos hidrológicos (seco e chuvoso). Calculamos o Índice de Qualidade de Água (IQA), o Índice de Estado Trófico (ETI) para fósforo e utilizamos uma Análise de Redundância (RDA) para verificar a influência de variáveis ambientais na comunidade de protozoários. Resultados: O IQA mostrou que a qualidade da água foi considerada boa em alguns pontos e considerada ruim em outros pontos, e o ETI para o fósforo classificou o córrego como mesotrófico na maioria dos locais amostrados (média entre 53,09 e 58,35). Foram identificados 19 táxons de amebas testáceas, pertencentes a seis famílias, sendo Difflugiidae, Centropyxidae e Arcellidae as mais especiosas, e 71 táxons de ciliados, distribuídos em 12 ordens, sendo Peniculida a ordem mais representativa, seguida por Euplotida. De acordo com a análise RDA, amostras do período seco foram caracterizadas por maiores valores de densidade bacteriana e concentrações de clorofila-a, fósforo total e nitrogênio total. As amebas testáceas e os ciliados apresentaram valores de abundância maiores em alguns dos meses caracterizados por maior precipitação e em condições de maior produtividade do sistema. Podemos concluir que o córrego Mandacaru, embora fortemente influenciado pela ação antrópica, ainda apresenta uma qualidade de água aceitável. Por fim, enfatizamos que a abundância de protistas foi fortemente influenciada pela produtividade do sistema. Isto foi evidenciado pelas elevadas densidades de protozoários onde houve maior produtividade primária e bacteriana. Assim, esses protistas devem ser considerados em estudos que visem à identificação de organismos que possam indicar impactos antrópicos e qualidade ambiental.

Structure and dynamics of the protoplankton community in an environmentally protected urban stream.

We aimed to investigate spatial and temporal scales, abundance, and factors that structure the communities of protozoans in a tropical urban stream. METHODS: Samples of water for analysis of biological communities (testate amoebae, ciliates and bacteria) and limnological variables were taken in the Mandacaru stream located in the Conservation Unit of Parque do Cinquentenário, in the city of Maringá, Paraná State, Brazil, in two hydrological periods (dry and rainy). We calculated the Water Quality Index (WQI) composed by nine parameters of water quality (dissolved oxygen, thermotolerant coliforms, pH, BOD, water temperature, total nitrogen, total phosphorous, turbidity, and total solids), Trophic State Index (TSI) for phosphorous and we used a Redundancy Analysis (RDA) to verify the influence of environmental variables in the protozoan community. RESULTS: The WQI showed that water quality was considered good in some points and considered bad in other points. The TSI for phosphorus classified the stream as mesotrophic in the majority of sampled sites points (mean between 53.09 and 58.35). We identified 19 taxa of testate amoebae, belonging to six families, being Difflugiidae, Centropyxidae, and Arcellidae those with more species and 71 infrageneric taxa of ciliates, distributed in 12 orders being Peniculida the most representative order, followed by Euplotida. According to RDA analysis, samples of the dry period were characterized by higher values of bacteria density and concentrations of chlorophyll-a, total phosphorous, and total nitrogen. Ciliates and testate amoebae presented higher abundance values in some of the months characterized by higher precipitation and in conditions of higher system productivity. CONCLUSION: we may conclude that the Mandacaru stream, although strongly influenced by anthropic action, still presents an acceptable water quality. Lastly, we emphasize that protists abundance was strongly influenced by system productivity. This was evidenced by elevated protozoan densities where there was higher primary and bacterial productivity. Thus, these organisms must be considered in studies that aim at the identification of organisms that may indicate anthropic impacts and environmental quality.

The Hospital at Home program: no place like home.

BACKGROUND: The treatment of children with cancer is associated with significant burden for the entire family. Frequent clinic visits and extended hospital stays can negatively affect quality of life for children and their families. METHODS: Here, we describe the development of a Hospital at Home program (H@H) that delivers therapy to pediatric hematology, oncology, and blood and marrow transplant (bmt) patients in their homes. The services provided include short infusions of chemotherapy, supportive-care interventions, antibiotics, post-chemotherapy hydration, and teaching. RESULTS: From 2013 to 2015, the H@H program served 136 patients, making 1701 home visits, for patients mainly between the ages of 1 and 4 years. Referrals came from oncology in 82% of cases, from hematology in 11%, and from bmt in 7%. Since inception of the program, no adverse events have been reported. Family surveys suggested less disruption in daily routines and appreciation of specialized care by hematology and oncology nurses. Staff surveys highlighted a perceived benefit of H@H in contributing to early discharge of patients by supporting out-of-hospital monitoring and teaching. CONCLUSIONS: The development of a H@H program dedicated to the pediatric hematology, oncology, or bmt patient appears feasible. Our pilot program offers a potential contribution to improvement in patient quality of life and in cost-benefit for parents and the health care system.

Structure and dynamics of the protoplankton community in an environmentally protected urban stream

Abstract We aimed to investigate spatial and temporal scales, abundance, and factors that structure the communities of protozoans in a tropical urban stream. Methods: Samples of water for analysis of biological communities (testate amoebae, ciliates and bacteria) and limnological variables were taken in the Mandacaru stream located in the Conservation Unit of Parque do Cinquentenário, in the city of Maringá, Paraná State, Brazil, in two hydrological periods (dry and rainy). We calculated the Water Quality Index (WQI) composed by nine parameters of water quality (dissolved oxygen, thermotolerant coliforms, pH, BOD, water temperature, total nitrogen, total phosphorous, turbidity, and total solids), Trophic State Index (TSI) for phosphorous and we used a Redundancy Analysis (RDA) to verify the influence of environmental variables in the protozoan community. Results: The WQI showed that water quality was considered good in some points and considered bad in other points. The TSI for phosphorus classified the stream as mesotrophic in the majority of sampled sites points (mean between 53.09 and 58.35). We identified 19 taxa of testate amoebae, belonging to six families, being Difflugiidae, Centropyxidae, and Arcellidae those with more species and 71 infrageneric taxa of ciliates, distributed in 12 orders being Peniculida the most representative order, followed by Euplotida. According to RDA analysis, samples of the dry period were characterized by higher values of bacteria density and concentrations of chlorophyll-a, total phosphorous, and total nitrogen. Ciliates and testate amoebae presented higher abundance values in some of the months characterized by higher precipitation and in conditions of higher system productivity. Conclusion: we may conclude that the Mandacaru stream, although strongly influenced by anthropic action, still presents an acceptable water quality. Lastly, we emphasize that protists abundance was strongly influenced by system productivity. This was evidenced by elevated protozoan densities where there was higher primary and bacterial productivity. Thus, these organisms must be considered in studies that aim at the identification of organisms that may indicate anthropic impacts and environmental quality.

Resumo O objetivo foi investigar, em escalas espacial e temporal, a abundância das comunidades de protozoários bem como os fatores intervenientes na estruturação das mesmas em um córrego urbano. Métodos: Amostras de água para análise de comunidades biológicas (amebas testáceas, ciliados e bactérias) e para análise de variáveis limnológicas foram tomadas no córrego Mandacaru localizado na Unidade de Conservação do Parque do Cinquentenário, na cidade de Maringá, Paraná State, Brasil, em dois períodos hidrológicos (seco e chuvoso). Calculamos o Índice de Qualidade de Água (IQA), o Índice de Estado Trófico (ETI) para fósforo e utilizamos uma Análise de Redundância (RDA) para verificar a influência de variáveis ambientais na comunidade de protozoários. Resultados: O IQA mostrou que a qualidade da água foi considerada boa em alguns pontos e considerada ruim em outros pontos, e o ETI para o fósforo classificou o córrego como mesotrófico na maioria dos locais amostrados (média entre 53,09 e 58,35). Foram identificados 19 táxons de amebas testáceas, pertencentes a seis famílias, sendo Difflugiidae, Centropyxidae e Arcellidae as mais especiosas, e 71 táxons de ciliados, distribuídos em 12 ordens, sendo Peniculida a ordem mais representativa, seguida por Euplotida. De acordo com a análise RDA, amostras do período seco foram caracterizadas por maiores valores de densidade bacteriana e concentrações de clorofila-a, fósforo total e nitrogênio total. As amebas testáceas e os ciliados apresentaram valores de abundância maiores em alguns dos meses caracterizados por maior precipitação e em condições de maior produtividade do sistema. Podemos concluir que o córrego Mandacaru, embora fortemente influenciado pela ação antrópica, ainda apresenta uma qualidade de água aceitável. Por fim, enfatizamos que a abundância de protistas foi fortemente influenciada pela produtividade do sistema. Isto foi evidenciado pelas elevadas densidades de protozoários onde houve maior produtividade primária e bacteriana. Assim, esses protistas devem ser considerados em estudos que visem à identificação de organismos que possam indicar impactos antrópicos e qualidade ambiental.

ALG6-CDG in South Africa: Genotype-Phenotype Description of Five Novel Patients.

ALG6-CDG (formerly named CDG-Ic) (phenotype OMIM 603147, genotype OMIM 604566), is caused by defective endoplasmic reticulum α-1,3-glucosyltransferase (E.C 2.4.1.267) in the N-glycan assembly pathway (Grünewald et al. 2000). It is the second most frequent N-glycosylation disorder after PMM2-CDG; some 37 patients have been reported with 21 different ALG6 gene mutations (Haeuptle & Hennet 2009; Al-Owain 2010). We report on the clinical and biochemical findings of five novel Caucasian South African patients. The first patient had a severe neuro-gastrointestinal presentation. He was compound heterozygous for the known c.998C>T (p.A333V) mutation and the novel c.1338dupA (p.V447SfsX44) mutation. Four more patients, presenting with classical neurological involvement were identified and were compound heterozygous for the known c.257 + 5G>A splice mutation and the c.680G>A (p.G227E) missense mutation. The patients belong to a semi-isolated Caucasian community that may have originated from European pioneers who colonized South Africa in the seventeenth/eighteenth centuries.

Screening of subfertile men for testicular carcinoma in situ by an automated image analysis-based cytological test of the ejaculate.

Testicular cancer (TC) is usually diagnosed after manifestation of an overt tumour. Tumour formation is preceded by a pre-invasive and asymptomatic stage, carcinoma in situ (CIS) testis, except for very rare subtypes. The CIS cells are located within seminiferous tubules but can be exfoliated and detected in ejaculates with specific CIS markers. We have built a high throughput framework involving automated immunocytochemical staining, scanning microscopy and in silico image analysis allowing automated detection and grading of CIS-like stained objects in semen samples. In this study, 1175 ejaculates from 765 subfertile men were tested using this framework. In 5/765 (0.65%) cases, CIS-like cells were identified in the ejaculate. Three of these had bilateral testicular biopsies performed and CIS was histologically confirmed in two. In total, 63 bilateral testicular biopsy were performed in conjunction with analysis of the ejaculates because of infertility work-up. Histological analysis of the biopsies for the presence of CIS yielded a test sensitivity of 0.67 and a specificity of 0.98. In addition, ejaculates from 45 patients with clinical signs of an overt TC were investigated and yielded a slightly lower sensitivity (0.51), possibly because of obstruction. We conclude that this novel non-invasive test combining automated immunocytochemistry and advanced image analysis allows identification of TC at the CIS stage with a high specificity, but a negative test does not completely exclude CIS. On the basis of the results, we propose that the assay could be offered to subfertile men and other patients who are at increased risk of TC.

Trans-cranial placement of an amplatzer device to control intractable cardiac failure in an infant with a vein of galen anomaly. A case report.

Neonates presenting with intractable cardiac failure due to vein of Galen aneurysmal malformations (VGAMs) rapidly progress to multisystem organ failure and death if left untreated. Currently the only viable treatment option is endovascular embolization. Although intracranial embolization of a neonate is a high-risk procedure, successful treatment can reverse cardiac failure and prevent neurological complications associated with VGAMs. Embolization via the arterial route is thought to have a better outcome than embolization via the venous system. However, multiple transarterial embolizations in different sessions may well be contraindicated in neonates, because repeat access via the femoral artery, carries a risk of arterial trauma which, in turn, can jeopardize lower limbs. With this case study we show that after repeat failure of arterial embolization, the transcranial placement of an Amplatzer PFO occluder (AGA Medical, Plymouth, USA) in the aneurysm can effectively reduce intrafistular pressure and venous outflow velocity. We also propose a mathematical model that can be used to calculate flow velocity through the aneurysm, which, in turn, could be used to aid clinical decision-making. Unlike some conventional techniques, the placement of an Amplatzer occluder does not pose the risk of completely obstructing venous drainage and therefore does not increase the risk of venous breakthrough hemorrhage. We propose this endovascular technique as a treatment option for high risk neonates in need of emergency embolization of VGAMs, where multiple arterial embolizations failed to control the condition sufficiently.

Distal truncation of KCC3 in non-French Canadian HMSN/ACC families.

BACKGROUND: Hereditary motor and sensory neuropathy with agenesis of the corpus callosum (HMSN/ACC) is a severe and progressive autosomal recessive polyneuropathy. Mutations in the potassium-chloride cotransporter 3 gene (KCC3) were identified as responsible for HMSN/ACC in the French Canadian (FC) population. In the present study, the authors were interested in finding new mutations in non-FC populations, assessing the activity of mutant proteins and refining genotype-phenotype correlations. METHODS: The authors screened KCC3 for mutations using direct sequencing in six non-FC HMSN/ACC families. They then assessed the functionality of the most common mutant protein using a flux assay in Xenopus laevis oocytes. RESULTS: The authors identified mutations in exon 22 of KCC3: a novel mutation (del + 2994-3003; E1015X) in one family, as well as a known mutation (3031C-->T; R1011X) found in five unrelated families and associated with two different haplotypes. The function of the cotransporter was abolished, although a limited amount of mutant proteins were correctly localized at the membrane. CONCLUSIONS: KCC3 mutations in exon 22 constitute a recurrent mutation site for hereditary motor and sensory neuropathy with agenesis of the corpus callosum (HMSN/ACC), regardless of ethnic origin, and are the most common cause of HMSN/ACC in the non-French Canadian (FC) families analyzed so far. Therefore, for genetic analysis, exon 22 screening should be prioritized in non-FC populations. Finally, the R1011X mutation leads to the abrogation of KCC3's function in Xenopus laevis oocytes, likely due to impaired transit of the cotransporter.

Relative ventricular volume measurements by intracardiac impedance.

The purpose of this study was to investigate whether changes of the right ventricular end-diastolic and end-systolic volumes can be detected by the DC-coupled electrical intracardiac impedance signal. Measurements were conducted in 13 patients with an implanted or external pacemaker, testing various measurement configurations. Volume changes were induced by incremental overpacing and by postural changes. The pulse contour systolic integral from the peripheral blood pressure was used as a marker of stroke volume changes. The results show, that DC-coupled impedance can detect volume changes of the right ventricle. The quadrupolar impedance measurement configurations offer better amplitude resolution, but the tripolar and intraventricular bipolar configurations are better hemodynamic markers.

Closed-loop stimulation using intracardiac impedance as a sensor principle: correlation of right ventricular dP/dtmax and intracardiac impedance during dobutamine stress test.

Changes of the unipolar right ventricular impedance during the cardiac cycle are related to the changing content of blood (low impedance) and tissue (high impedance) around the tip of the pacing electrode. During myocardial contraction, the impedance continuously increases reaching its maximum in late systole. This impedance increase is thought to correlate with right ventricular contractility, and thus, with the inotropic state of the heart. In the new Inos2 DDDR pacemaker, integrated information from the changing ventricular impedance (VIMP) is used for closed-loop regulation of the rate response. The aim of this study was to analyze the effect of increasing dobutamine challenge on RV contractility and the measured impedance signals. In 12 patients (10 men, 68 +/- 12 years) undergoing implantation of an Inos2 DDDR pacemaker (Biotronik), a right ventricular pigtail catheter was inserted for continuous measurements of RV-dP/dtmax and simultaneous VIMP signals during intrinsic and ventricular paced rhythm. Then, a stress test with a stepwise increase of intravenous dobutamine (5-20 micrograms/kg per min) was performed. To assess the relationship between RV contractility and measured sensor signals, normalized values of dP/dtmax and VIMP were compared by linear regression. There was a strong and highly significant correlation between dP/dtmax and VIMP for ventricular paced (r2 = 0.93) and intrinsic rhythm (r2 = 0.92), although the morphologies of the original impedance curves differed quite substantially between paced and intrinsic rhythm in the same patient. Furthermore, VIMP correlated well with sinus rate (r2 = 0.82), although there were at least four patients with documented chronotropic incompetence. We conclude, that for intrinsic and ventricular paced rhythms sensor signals derived from right ventricular unipolar impedance curves closely correlate with dP/dtmax, and thus, with a surrogate of right ventricular contractility during dobutamine stress testing. Our results suggest that "inotropy-sensing" via measurement of intracardiac impedance is highly accurate and seems to be a promising sensor principle for physiological rate adaptation in a closed-loop pacing system.

Men's attitudes regarding genetic testing for hereditary prostate cancer risk.

OBJECTIVES: Little is known about the attitudes of men unselected for a family history for prostate cancer concerning genetic testing for prostate cancer risk or genetic testing for inherited cancer predisposition. To explore this, we examined the interest in molecular testing for hereditary prostate cancer (HPC) predisposition among a self-selected cohort of 342 men presenting for prostate cancer screening. METHODS: Participants were surveyed concerning their attitudes about DNA testing for HPC predisposition and knowledge of prostate cancer-associated risk factors, including heredity. RESULTS: Of the participants completing the survey, 92% expressed interest in learning about DNA testing, and 89% stated that they would undergo DNA analysis for HPC predisposition, if available. Twenty-eight percent of respondents failed to demonstrate an adequate understanding of the concept of "inherited tendency." The demonstrated level of understanding of this concept did not differ by the respondent's family history, although it varied by race. An interest in learning about or undergoing testing did not vary by race, family history, or demonstrated understanding of the concept of inherited risk. CONCLUSIONS: Among men presenting for routine prostate cancer screening, interest in learning about testing for HPC predisposition and in having such testing performed may be high. The data also provide evidence that, in a population of men unselected for family history, interest in molecular testing for this common, male-specific cancer may parallel the high interest level demonstrated among women in DNA testing for inherited breast and ovarian cancer risk.

Alternative medicine use in patients with localized prostate carcinoma treated with curative intent.

BACKGROUND: Alternative medicine therapies are estimated to be used by 7-64% of cancer patients but up to 72% do not inform their physician. To the authors' knowledge little useful information is available regarding the prevalence of alternative medicine use by patients with prostate carcinoma. Thus, the authors attempted to evaluate the prevalence of alternative medicine use by prostate carcinoma patients treated with curative intent by either radical prostatectomy, brachytherapy alone, or a combination of brachytherapy and external radiation therapy. METHODS: Between January 1997 and May 1998, 234 men with clinically localized prostate carcinoma underwent radical prostatectomy (54 patients) or brachytherapy (180 patients) with (74 patients) or without (106 patients) external beam radiation therapy. In July 1998 a questionnaire was mailed to all patients comprised of multiple questions regarding alternative medicine use to which 190 patients (81%) responded. The overall prevalence and types of alternative medicine therapies used were assessed and the relation between age at procedure, pretreatment prostate specific antigen level, clinical stage, pretreatment Gleason score, and type of treatment with the use of alternative medicine therapies was evaluated using univariate and multivariate analysis. RESULTS: The prevalence of alternative medicine use by prostate carcinoma patients responding to the survey was 43% (81 of 190 patients). No significant differences in alternative medicine use were observed between the patients who received brachytherapy alone (38%), those who underwent radical prostatectomy (40%), and those treated with combined brachytherapy and external beam radiation therapy (51%). Vitamins, prayer or other religious practices, and herbal medicines were the most commonly used therapies in these patients. Higher pretreatment Gleason scores were associated with a greater use of alternative medicine therapies on both multivariate and univariate analyses. Finally, using multivariate analysis, younger prostate carcinoma patients were more likely to use alternative medicine therapies than older patients. CONCLUSIONS: Alternative medicine use is very prevalent among patients treated for localized prostate carcinoma. Because some of these treatments may have a potential biologic impact on tumor behavior and, consequently, on definitive or surrogate therapeutic endpoints, patients should be questioned carefully regarding alternative medicine use during routine tumor follow-up.

Breast cancer screening and family history among rural women in Wisconsin.

This study examines the relationship between family history of breast cancer and current compliance with mammography screening guidelines. A random telephone survey of women age 40 or older living in rural Wisconsin (N = 2398) was conducted to determine mammography screening knowledge, family history of breast cancer, attitudes, intentions, physician recommendation, and compliance with screening guidelines. Compared with women without a family history of breast cancer, women with a family history were significantly more likely to demonstrate correct knowledge (p = 0. 01); express intentions in compliance with recommended screening guidelines (p < 0.001); report having been advised by a physician to obtain a mammogram (p < 0.001); and be in current compliance with mammography screening guidelines (p < 0.001). Results of simultaneous and individual logistic regression suggest that the effects of family history on compliance with screening guidelines are directly mediated through the combination of women's knowledge, women's intentions, and physician recommendation. Thus, programs to increase compliance with mammography screening guidelines should address both women and providers.

Memory effects in the action of ozone on conifers.

Conifers are known to possess relative ozone tolerance in short-term experiments. A scenario for ozone damage of conifers is now derived from the first exposure experiments in which both the initial biochemical response phase and delayed visible symptom development were studied. A number of early biochemical ozone responses could be detected in Norway spruce (Picea abies [L.] Karst.) and Scots pine (Pinus sylvestris L.). The stress metabolite catechin persisted over several months. In the year following ozone treatment of spruce, decreases in pigment content and photosynthetic capacity, as well as development of visible symptoms (chlorosis, banding), were determined in the needle age classes previously exposed to an accumulated hourly ozone dose above 40 ppb (AOT40) of >/=60-80 ppm small middle doth. The visible symptoms developed during spring emergence of the new flush. In the case of Scots pine, an ozone dose (AOT40) of >/=30 ppm small middle doth caused the premature shedding of needles 9 months after treatment. The delayed symptoms of both spruce and pine occurred during known phases of endogenous stress. The symptoms appeared to reflect an ozone "memory" imprinted by the induced early stress reactions. Ambient AOT40 ozone doses in Central Europe are in the range 4 and 50 ppm small middle doth per growing season. Ozone is proposed to potentially damage conifers through memory effects ("abiotic" pathway) or through predisposition for pathogen attack ("biotic" pathway).

Increased transcription decreases the spontaneous mutation rate at the thymidine kinase locus in human cells.

Transcription increases DNA repair efficiency and modulates the distribution of certain types of DNA damage. Furthermore, increased transcription level stimulates spontaneous mutation rate in yeast. We explored whether transcription level affects spontaneous mutation rate in human cells. We first developed two thymidine kinase (tk) inducible human cell lines using the Gal4-Estrogen receptor system. In our TK6i-G3 and G9 tk heterozygous cell lines, the active tk allele is linked to an inducible promoter element. Tk mRNA is induced following treatment with estrogen. Spontaneous mutation rate was significantly decreased in human cell lines after induction in contrast to the report in yeast. Thus, humans may have evolved different or additional mechanisms to deal with transcription related spontaneous mutagenesis.

Early prostate-specific antigen failure following radical perineal versus retropubic prostatectomy: the importance of seminal vesicle excision.

OBJECTIVES: Because of renewed interest in the radical perineal prostatectomy, we chose to evaluate factors influencing differences in biochemical failure as measured by prostate-specific antigen (PSA) between radical perineal and the radical retropubic prostatectomies. METHODS: We undertook a retrospective review of 87 men with clinically localized prostate cancer who underwent radical retropubic (64%) or radical perineal (36%) prostatectomy, noting age, race, preoperative PSA, Gleason score, clinical stage, capsular penetration, surgical approach, and completeness of seminal vesicle (SV) excision. The two groups were comparable with respect to tumor factors such as preoperative PSA, Gleason score, clinical stage, and capsular penetration. Time to postoperative PSA failure (0.2 ng/mL or greater) was evaluated with univariate and multivariate analysis of multiple contributing factors. RESULTS: Twenty-eight percent of patients had a PSA level rising to 0.2 ng/mL or greater in the follow-up period. Patients who underwent perineal prostatectomy had a higher PSA failure rate (45%) than those treated by the retropubic approach (18%) and patients with incomplete SV excision had a higher failure rate (69%) than patients with bilateral SV excision (20%). When time to PSA failure was examined by multivariate analysis, completeness of SV excision, clinical stage, and Gleason score had a statistically significant impact on this outcome. In perineal prostatectomy patients, bilateral SV excision had a significantly longer time to PSA failure than in patients with incomplete excision. There was no significant difference in time to PSA failure between patients who underwent radical retropubic prostatectomy and the patients who underwent perineal prostatectomy with bilateral SV excision. CONCLUSIONS: Incomplete excision of SVs during a radical perineal prostatectomy contributes to an earlier postoperative biochemical recurrence as measured by a rising PSA, and may explain the higher disease recurrence rate for radical perineal prostatectomies as opposed to radical retropubic prostatectomies in this study.

The Hautmann neobladder with a chimney: a versatile modification.

PURPOSE: We describe a modification to the popular Hautmann ileal neobladder comprised of a chimney for the ureterointestinal anastomosis in order to enhance its flexibility, reduce its pitfalls and hence make more patients candidates for this type of diversion. MATERIALS AND METHODS: The Hautmann ileal neobladder is prepared in the standard fashion except that an additional 5 to 10 cm. (or more if needed in the case of a short ureter) of small bowel are selected to be added for either a left or right "chimney" (depending on which side the ureter may be shorter or if there is a solitary kidney). The V segment of the W which will be anastomosed to the urethra is made longer than the contralateral V segment. A neobladder neck is created by sewing together the cut edges of a 5 cm. lip in the lowermost W configuration of the neobladder, allowing an end-to-end urethrointestinal anastomosis. RESULTS: Between March and October 1996, 3 patients have undergone this procedure at the University of Virginia. Two had right chimneys and 1 had a left chimney. Followup is short, but there have been no complications except for a transient postoperative partial small bowel obstruction in 1 patient. Two patients have no urinary incontinence, the third has only occasional nocturnal incontinence. CONCLUSIONS: This modification of the Hautmann ileal neobladder provides greater versatility for short ureters, minimizes tension from the mesentery on the urethrointestinal anastomosis, simplifies the ureterointestinal anastomosis and provides easy postoperative access for ureterointestinal anastomosis revision or resection in case of recurrent disease.