Integrin α3 negative podocytes: A gene expression study.

Integrin α3ß1 is a cell adhesion receptor widely expressed in epithelial cells. Pathogenic variants in the gene encoding the integrin α3 subunit ITGA3 lead to a syndrome including interstitial lung disease, nephrotic syndrome, and epidermolysis bullosa (ILNEB). Renal involvement mainly consists of glomerular disease caused by loss of adhesion between podocytes and the glomerular basement membrane. The aim of this study was to characterize the impact of loss of integrin α3 on human podocytes. ITGA3 was stably knocked-out in the human podocyte cell line AB8/13, designated as PodoA3-, and in human proximal tubule epithelial cell line HK2 using the targeted genome editing technique CRISPR/Cas9. Cell clones were characterized by Sanger sequencing, quantitative PCR, Western Blot and immunofluorescence staining. RNASeq of integrin α3 negative cells and controls was performed to identify differential gene expression patterns. Differentiated PodoA3- did not substantially change morphology and adhesion under standard culture conditions, but displayed significantly reduced spreading and adhesion when seed on laminin 511 in serum free medium. Gene expression studies demonstrated a distinct dysregulation of the adhesion network with downregulation of most integrin α3 interaction partners. In agreement with this, biological processes such as "extracellular matrix organization" and "cell differentiation" as well as KEGG pathways such as "ECM-receptor interaction", "focal adhesion" and the "PI3K-Akt signaling pathway" were significantly downregulated in human podocytes lacking the integrin α3 subunit.

Vasectomy and the risk of prostate cancer in a prospective US Cohort: Data from the NIH-AARP Diet and Health Study.

BACKGROUND: Several studies have linked vasectomy with the risk of prostate cancer; however, this association has been attributed to selection bias. Since vasectomy is a common and effective form of contraception, these implications are significant. Therefore, we sought to test this association in a large observational cohort. OBJECTIVE: To evaluate the potential association between prior vasectomy and the risk of developing prostate cancer. MATERIALS AND METHODS: We evaluated the relationship between vasectomy and prostate cancer in the NIH-AARP Diet and Health Study. Of the 111,914 men, prostate cancer was identified in 13,885 men and vasectomies were performed in 48,657. We used multivariate analysis to examine the relationship between prostate cancer and vasectomy. We also performed propensity score-adjusted and propensity score-matched analysis. RESULTS: Men utilizing vasectomy were more likely to be ever married, fathers, educated, white, and screened for prostate cancer. During 4,251,863 person-years of follow-up, there was a small association between vasectomy and incident prostate cancer with a hazard ratio of 1.05 (95% CI, 1.01-1.11). However, no significant association was found when looking separately at prostate cancer by grade or stage. Conclusions were similar when using propensity adjustment and matching. Importantly, a significant interaction between vasectomy and PSA screening was identified. DISCUSSION: Estimates of the association between vasectomy and prostate cancer are sensitive to analytic method underscoring the tenuous nature of the connection. Given the differences between men who do and do not utilize vasectomy, selection bias appears likely to explain any identified association between vasectomy and prostate cancer. CONCLUSIONS: With over 20 years of follow-up, no convincing relationship between vasectomy and prostate cancer of any grade was identified.

Responsiveness of stem-like human glioma cells to all-trans retinoic acid and requirement of retinoic acid receptor isotypes α, ß and γ.

Retinoic acid (RA) is required for development and homeostasis of the normal mammalian brain and may play a role in the initiation and progression of malignant brain tumors, such as the glioblastoma multiforme (GBM) and the gliosarcoma (Gsarc). The subpopulation of stem-like glioma cells (SLGCs) was shown to resist standard glioma radio-/chemotherapy and to propagate tumor regrowth. We used phenotypically distinct, self-renewing SLGC lines from six human GBMs, two Gsarcs, and two subcloned SLGC derivatives in order to investigate their responsiveness to all-trans retinoic acid (atRA) and to identify the RA-receptor (RAR) isotypes involved. In general, atRA exerted a pro-proliferative and pro-survival effect on SLGCs, though the efficacy was distinct. By means of RAR isotype-selective retinoids we disclosed that these effects were mediated by RARα and RARγ, except for one SLGC line, in which the pro-proliferative signal was induced by the RARß-selective retinoid. Only one GBM-derived cell line (T1338) and a subpopulation of another (T1389) displayed neural differentiation in response to atRA. Differentiation of T1338 was induced by RARα and RARγ isotype-selective retinoids, associated with down-regulation of Sox2, and the failure to induce orthotopic tumors in the brains of SCID mice. The differential responsiveness of the SLGC lines appeared unrelated to the expression of RARß, as (i) atRA augmented RAR isotype mRNA expression and particularly rarß mRNA in all SLGC lines, (ii) rarß promoter hypomethylation in the SLGC lines was not related to differentiation and (iii) the induction of T1338 differentiation was by RARα- and RARγ-selective ligands.

Near real time detection of hazardous airborne substances.

A fast near real-time monitoring system for hazardous airborne substances, such as chemical warfare agents (CWA) is presented and limits of detection (LOD) for five CW simulants are determined. A tandem thermal desorber (TTD) continuously collects and pre-concentrates air. The pre-concentrated samples are then separated in a fast gas chromatographic (GC) run of 6.9min. and detected by a time-of-flight mass spectrometer (TOFMS). The GC-TOFMS signals are evaluated using chemometric methods for deconvolution and target identification. The high toxicity of nerve agents requires extremely low detection limits; for some as low as 100 ng/m(3) (10 ppt). The combination of TTD, TOFMS and chemometric data evaluation methods enables the system to fulfill this requirement. Calibration measurements for five different CWA simulants show lower limits of detection in the range of 10 ng/m(3)-60 ng/m(3) (1-11 ppt). In addition, the ability to detect trace concentrations of real CWA is demonstrated with a measurement of 30 pg Sarin on column. Several other real CWA measurements are shown, like sulfur mustard in diesel, lewisite under humid conditions and VX. As part of this work the influence of stationary film thickness on peak tailing of organophosphates is investigated for peak shape optimization.

Maltreatment, MAOA, and delinquency: sex differences in gene-environment interaction in a large population-based cohort of adolescents.

The present study investigated a possible interaction between a functional polymorphism in the MAOA gene promoter (MAOA-VNTR) and childhood maltreatment in the prediction of adolescent male and female delinquency. A cohort of 1,825 high school students, 17-18 years old, completed an anonymous questionnaire during class hours which included questions on childhood maltreatment, sexual abuse, and delinquency. Saliva samples were collected for DNA isolation, and analyzed for the MAOA-VNTR polymorphism. Self-reported maltreatment was a strong risk factor for adolescent delinquent behavior. The MAOA genotype also showed a significant main effect when controlled for maltreatment. Boys with a short variant and girls with one or two long variants of the polymorphism showed a higher risk for delinquency when exposed to maltreatment. Our results confirm previous findings of an interaction between the MAOA-VNTR polymorphism and self-reported maltreatment. Results for boys and girls differ according to MAOA-VNTR genotype and direction of phenotypic expression.

Experience with 750 consecutive laparoscopic donor nephrectomies--is it time to use a standardized classification of complications?

PURPOSE: Laparoscopic living donor nephrectomy offers patients the benefits of decreased morbidity and improved cosmesis, while maintaining equivalent graft outcomes and complication rates similar to those of open donor surgery. With expressed concern for donor safety, using a standardized complication scale would allow combining data in a donor registry so potential donors could be adequately followed and counseled. We present the largest series to our knowledge of laparoscopic living donor nephrectomy by a single surgeon. MATERIALS AND METHODS: The institution's initial 750 laparoscopic living donor nephrectomies were included in the study, and a retrospective and prospective chart and database analysis was performed. RESULTS: Mean donor age was 40.5 years and average body mass index was 25.7 kg/m(2). There were 175 patients (23%) with 2 or more renal arteries while 161 (21.5%) had early arterial bifurcations. There were 3 open conversions (0.4%) and the overall complication rate was 5.46%. Median hospital stay was 1 day and the readmission rate was 1.2%. There were 5 reoperations (0.67%), none of which was for the control of bleeding. No patients required a blood transfusion and there were no mortalities. Using a modified Clavien classification of complications for living donor nephrectomy 65.8% were grade 1, 31.7% grade 2 (12.2% grade 2a, 14.6% grade 2b, 4.9% grade 2c) and 2.4% grade 3. There were no grade 4 complications. CONCLUSIONS: With appropriate patient selection and operative experience, laparoscopic living donor nephrectomy is a safe procedure associated with low morbidity. The use of a standardized complication system specific for this procedure is encouraged and could aid in counseling potential donors in the future.

Sense of coherence and psychological well-being: improvement with age.

BACKGROUND: Psychological well-being is important for individuals, communities and health services throughout the world because of the costs associated with psychological ill-health and the loss of quality of life for those affected and their relatives. Following a salutogenic approach, there is a link between health-promoting resources, such as generalised resistance resources and a positive state of health. Generalised resistance resources have been proposed to relate to an individual's sense of coherence (SOC). The objectives of the present study were (i) to investigate SOC in relation to age and sex, (ii) to investigate psychological well-being in relation to age and sex, and (iii) to investigate the relationship between generalised resistance resources and psychological well-being. METHODS: A random sample of 43 598 respondents (54% female) aged 18-85 years participated in the present study via a postal survey questionnaire. SOC was measured by the SOC-13 and well-being by the General Health Questionnaire-12 questionnaire. RESULTS: Males had both stronger SOC and well-being compared to females. There was a relationship between SOC and age, with stronger SOC in the older age groups. There was a larger proportion of individuals who experienced well-being as a function of age. In addition, an increase in SOC was related to a decrease in psychological well-being, that is, a stronger SOC corresponded to higher well-being. CONCLUSION: Males showed a stronger SOC and more well-being than females. Moreover, SOC and well-being increased with age in both sexes. Our findings suggest that SOC may develop over a entire lifetime.

Do psychosocial factors predict disability and health at a 3-year follow-up for patients with non-acute musculoskeletal pain? A validation of the Orebro Musculoskeletal Pain Screening Questionnaire.

PURPOSE: Early identification and intervention with those that run the risk of developing long-term disability would offer a great opportunity for reducing costs and personal suffering associated with long-term work absenteeism. The Orebro Musculoskeletal Pain Screening Questionnaire (OMPSQ) has been used and validated in several studies for participants with mainly acute pain problems. The aim of this study was to validate the OMPSQ for patients with non-acute pain problems (e.g. 1-6 months sick leave) and compare to other relevant questionnaires. METHOD: One hundred and fifty-eight patients with musculoskeletal pain and disability recruited to a multidisciplinary rehabilitation project completed a battery of questionnaires at baseline and at 3-year follow-up visits. The main analysis involved the relationship between risk levels in the questionnaire and sick leave and perceived health after 3 years. RESULTS: The OMSPQ predicted future sick leave and health and was found to have six factors. The function and pain factors were the best predictors of sick leave after 3 years, while the distress factor was the best predictor of perceived mental health and return to work-expectancy was borderline significant. Perceived physical health at 3 years was best predicted by the function and pain factors with the fear-avoidance factor being marginally significant. CONCLUSION: The results demonstrate that psychosocial factors as measured by OMPSQ are related to work disability and perceived health even 3 years after treatment for patients with non-acute pain problems. The OMSPQ was a good predictor of outcome.

Complications of laparoscopic renal surgery.

Initial excitement for laparoscopy's potential to decrease patient morbidity and convalescence by avoiding a flank incision was initially tempered by concerns for increased operative time, technical complexity and the suitability of laparoscopy approaches to oncologic surgery. With experience, the benefits of laparoscopic approaches to renal surgery have become clear. As laparoscopic techniques are mastered and the indications expanded, it is important to remember that minimally invasive surgery remains associated with significant risks and potential complications. Several complications are theoretically more difficult to control laparoscopically than with open exposure. Control of bleeding, identification of injury to solid organs, and positive margins due to the lack of haptic feedback have been of special concern during the rapid advancement of laparoscopic surgical technique between 1991 and today. It is critical for the urologist to be familiar with these complications in order to maximize patients' clinical outcomes through appropriate patient selection and intraoperative planning. Know-ledge of the complications of laparoscopic renal surgery will also aid in providing patients with true informed consent and realistic surgical expectations. The purpose of this manuscript is to review the complications associated with laparoscopic renal surgery in general, with specific attention paid to laparoscopic radical, partial, pediatric, and donor nephrectomy.

Distension of painful structures in the treatment for chronic pelvic pain in women.

BACKGROUND: There is a lack of established treatment for Chronic pelvic pain (CPP), defined as acyclic pain of at least six months duration. We decided to study the pain-alleviating effects of stretching on defined structures in women with CPP, and the treatment's impact on quality of life variables. DESIGN OF STUDY: An open, randomized study. SETTING: Primary Health Care Centre, Kolbäck, Sweden. METHODS: Fifty women, median age 33 years (range 19-54), complaining of CPP for a median duration of 25.5 months (range 6-264) were randomly assigned to either a treatment or a control group. A short questionnaire containing 17 questions was administered before randomization and two to three weeks after a second treatment of distension of pelvic structures. Visual analog scales were used for questions concerning intensity of pain and quality of life. Five-point scales were used for questions dealing with duration and frequency of pain. RESULTS: Intensity, frequency and duration of pelvic pain, painful intercourse, lower back pain, sleep disturbance, sleep quality, mental fatigue, depression, mood and anger improved significantly more in the treatment group than in the control group. Treatment proved more effective than counseling as reflected by self-rating scales: pain intensity (OR 18.37, 95% CI 3.39-99.64) and pain during intercourse (OR 8.59, 95% CI 1.57-46.68). CONCLUSION: In this open, randomized study, distension of painful pelvic structures in women with CPP resulted in significant relief of pain and improvement in quality of life measures.

Time-domain and spectral-domain optical coherence tomography in the analysis of brain tumor tissue.

INTRODUCTION: Detection of residual tumor during resection of glial brain tumors remains a challenge because of a low inherent contrast of adjacent edematous brain, the surrounding infiltration zone, and the solid tumor. Therefore, new technologies that may facilitate an intraoperative analysis of the tissue at the resection edge are of great interest to neurosurgeons. MATERIALS AND METHODS: For ex vivo imaging of gliomas in a mouse model and human biopsy specimens of brain tumors and nervous system tissue we have used a time-domain Sirius 713 Tomograph with a central wavelength of 1,310 nm and a coherence length of 15 microm equipped with a mono mode fiber and a modified optical coherence tomography (OCT) adapter containing a lens system for imaging at a working distance of 2.5 cm. A spectral-domain tomograph using 840 nm and 930 nm superluminescence diodes (SLD) with a central wavelength of 900 nm was used as a second imaging modelity. RESULTS: Both time-domain and spectral-domain coherence tomography delineated normal brain, the infiltration zone and solid tumor in murine intracerebral gliomas. Histological evaluation of H&E sections parallel to the optical plain demonstrated that tumor areas of less than a millimeter could be detected and that not only solid tumor, but also brain invaded by a low-density single tumor cells produced an OCT signal different from normal brain. Spectral-domain OCT (SD-OCT) demonstrated a significantly more detailed microstructure of tumor and normal brain up to a tissue depth of 1.5-2.0 mm, whereas the interpretation of time-domain OCT (TD-OCT) was difficult at a tissue depth >1.0 mm. Because of rapid scanning times SD-OCT data could be acquired as 3D data maps, which allowed a multi-planar analysis of the tumor to brain interface. Similar to our findings in experimental gliomas, images of human nervous system tissue acquired using SD-OCT showed a characteristic signal of normal brain tissue and a detailed microstructure of tumor parenchyma. CONCLUSION: Spectral-domain OCT of experimental gliomas and human brain tumor specimens differentiates solid tumor, diffusely invaded brain tissue, and adjacent normal brain based on microstructure and B-scan signal characteristics. In conjunction with the rapid image acquisition rates of SD-OCT, this technology carries the potential of a novel intraoperative imaging tool for the detection of residual tumor and guidance of neurosurgical tumor resections.

Quality of life and maintenance of improvements after early multimodal rehabilitation: a 5-year follow-up.

PURPOSE: There is a paucity of long-term evaluations on rehabilitation of musculoskeletal disorders, e.g., neck, shoulder or back pain. The aim of this study was to assess quality of life and the effect of early multimodal rehabilitation on 91 patients with musculoskeletal pain and disability at a 5-year follow-up. METHOD: The follow-up assessment, which included questions on pain, function, quality of life, perceived health, sick leave and psychosomatic symptoms, was performed 5 years after the assessment of baseline status. RESULTS: Improvements in pain, perceived health and psychosomatic symptoms were maintained at the 5-year follow-up. In addition, improvements in function, quality of life, and level of acceptable pain were significant in comparison to baseline. At the time of the baseline assessment all patients were on sick leave (13% were on partial sick leave). At the 5-year follow-up, 58% of the patients were at work part or full time. The results show that those working differed significantly from those not working at the 5-year follow-up on almost all variables, indicating that those working enjoy better health. The most salient prognostic factors for return to work were perceived health and educational level at the time of the baseline evaluation. CONCLUSIONS: These results show that treatment improved quality of life and the effects were basically maintained at 5 years. Work capacity as reflected in return to work increased greatly (81%) at a 1-year follow-up and was substantial (58%) at the 5-year follow-up. Moreover, perceived health and educational levels were important prognostic factors. Finally, the fact that patients working reported better health underscores the probable importance of return to work. Our results imply that it may be feasible to obtain long-term benefits from such a primary care-based intervention.

Analysis of mouse brain using a clinical 1.5 T scanner and a standard small loop surface coil.

With increasing numbers of in vivo experiments in the field of neuroscience, the interest in methods for in vivo imaging of animal brains as small as those of mice has increased. Because highly specialized small bore scanners with high field strengths are not commonly available, clinical magnetic resonance imaging (cMRI) scanners have been used in the past to image rat and more recently also mouse brains in combination with specifically developed RF coils. These studies have demonstrated that imaging of small animal brains is feasible, and that tumor volumes measured by cMRI correlate well with histological tumor volume analysis. This protocol describes the cMRI settings at 1.5 T for imaging of mouse brain with resolutions up to 120 x 120 microm using an inexpensive, commercially available small loop surface coil. This allows easy establishment of a small animal MRI facility without the need for cost intensive dedicated small animal scanners or special custom made coils. In this study, we demonstrate high-resolution imaging of intracranial xenografts in a mouse glioma model and monitor the treatment effect of external field irradiation by cMRI.

Basal glucometabolic status has an impact on long-term prognosis following an acute myocardial infarction in non-diabetic patients.

OBJECTIVES: Patients with diabetes are known to have a worse prognosis after an acute myocardial infarction (AMI) compared with non-diabetic patients. The primary aim of this study was to investigate the effect of glucometabolic status on long-term prognosis in non-diabetic patients with an AMI. The second aim was to evaluate the extent to which blood glucose levels at admission depended on acute stress, assessed as serum cortisol, previous glucometabolic status, measured as haemoglobin A1c (HbA1c), or both. DESIGN: In a prospective study of patients with an AMI, blood glucose, HbA1c and cortisol were measured at admission. Fasting blood glucose was determined before discharge and also afterwards, if necessary, for classification. Patients were followed-up for 5.5 years. SUBJECTS: Of the 305 consecutive patients 24% were diagnosed as diabetic and 76% as non-diabetic. MAIN OUTCOME MEASURES: Death or non-fatal myocardial re-infarction. RESULTS: In non-diabetic patients, a Cox regression model was used. With death or re-infarction as endpoint, the following prognostic factors had an impact on event-free survival: age (P<0.001), HbA1c (P=0.002), cortisol (P<0.001) and thrombolytic treatment (P=0.001). There was a correlation between cortisol and blood glucose at admission (r=0.44, P<0.001). Fasting blood glucose day 5 showed no association with event-free survival. CONCLUSIONS: In non-diabetic patients with AMI, admission HbA1c and cortisol were predictors for 5.5-year survival without recurrent non-fatal myocardial infarction. The glucometabolic status of importance for prognosis was detected by HbA1c but not by fasting blood glucose or admission blood glucose, of which the latter was influenced by cortisol.