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Prenyltransferases (PTases) are known to play a role in embryonic development, normal tissue homeostasis and cancer by posttranslationally modifying proteins involved in these processes. They are being discussed as potential drug targets in an increasing number of diseases, ranging from Alzheimer's disease to malaria. Protein prenylation and the development of specific PTase inhibitors (PTIs) have been subject to intense research in recent decades. Recently, the FDA approved lonafarnib, a specific farnesyltransferase inhibitor that acts directly on protein prenylation; and bempedoic acid, an ATP citrate lyase inhibitor that might alter intracellular isoprenoid composition, the relative concentrations of which can exert a decisive influence on protein prenylation. Both drugs represent the first approved agent in their respective substance class. Furthermore, an overwhelming number of processes and proteins that regulate protein prenylation have been identified over the years, many of which have been proposed as molecular targets for pharmacotherapy in their own right. However, certain aspects of protein prenylation, such as the regulation of PTase gene expression or the modulation of PTase activity by phosphorylation, have attracted less attention, despite their reported influence on tumor cell proliferation. Here, we want to summarize the advances regarding our understanding of the regulation of protein prenylation and the potential implications for drug development. Additionally, we want to suggest new lines of investigation that encompass the search for regulatory elements for PTases, especially at the genetic and epigenetic levels.

Objectives: Pelvic organ prolapse is the descent of one or more reproductive organs from their normal position, causing associated negative symptoms. One conservative treatment option is pessary management. This study aimed to to investigate the cost-effectiveness of pessary self-management (SM) when compared with clinic-based care (CBC). A decision analytic model was developed to extend the economic evaluation., Methods: A randomized controlled trial with health economic evaluation. The SM group received a 30-minute SM teaching session, information leaflet, 2-week follow-up call, and a local helpline number. The CBC group received routine outpatient pessary appointments, determined by usual practice. The primary outcome for the cost-effectiveness analysis was incremental cost per quality-adjusted life year (QALY), 18 months post-randomization. Uncertainty was handled using nonparametric bootstrap analysis. In addition, a simple decision analytic model was developed using the trial data to extend the analysis over a 5-year period., Results: There was no significant difference in the mean number of QALYs gained between SM and CBC (1.241 vs 1.221), but mean cost was lower for SM (£578 vs £728). The incremental net benefit estimated at a willingness to pay of £20 000 per QALY gained was £564, with an 80.8% probability of cost-effectiveness. The modeling results were consistent with the trial analysis: the incremental net benefit was estimated as £4221, and the probability of SM being cost-effective at 5 years was 69.7%., Conclusions: Results suggest that pessary SM is likely to be cost-effective. The decision analytic model suggests that this result is likely to persist over longer durations., Competing Interests: Author Disclosures Author disclosure forms can be accessed below in the Supplemental Material section., (Copyright © 2024. Published by Elsevier Inc.)

Background: As part of a larger multi-site translational research project this study explored patient and staff experiences, perspectives and recommendations in relation to a mental health liaison nursing (MHLN) service implemented in the emergency department (ED) of two rural hospitals in New South Wales, Australia., Methods: Semi-structured interviews were conducted across the two sites with a sample of ED patients (n = 32), ED nurses (n = 14), ED medical officers (n = 11) and psychiatrists (n = 3). Data were analysed thematically., Findings: Patients highlighted the therapeutic benefits of the MHLNs, particularly being listened to and understood. Patients appreciated being seen promptly and having effective follow-up. Some negative experiences were encountered. Staff recognized that embedding the new model of care in the ED impacted positively on ED culture and practice. A nurse practitioner position as clinical lead instituted at one site made the most substantial contribution to integrating mental health care within the ED. Room for improvement was also noted., Conclusions: Findings from this study indicate that an ED-based model of MHLN care developed in a metropolitan setting was successfully translated to two rural EDs. While translating a model of care from one context to another is not without its challenges, adherence to the key principles of this model of MHLN care was associated with the most positive outcome., (Crown Copyright © 2021. Published by Elsevier Ltd. All rights reserved.)

Photoemission electron microscopy (PEEM) is a powerful surface technique for dynamic imaging of surface processes while PEEM studies have been performed under ultrahigh vacuum or high vacuum conditions. Here we report on a near ambient pressure PEEM (NAP-PEEM) instrument, which enables high resolution PEEM imaging in near ambient pressure (> 1 mbar) gases over a wide temperature range (150 - 1200 K). Installed with an electron gun near ambient pressure low energy electron microscopy (NAP-LEEM) can be achieved as well. The success of this new NAP-PEEM/LEEM instrument relies on the following key design concepts. First, a two-stage accelerating electric field consisting of a low field region between sample and intermediate electrode ("nozzle") and a high field between nozzle and objective lens. Second, a three-stage differential pumping system allowing a near ambient pressure atmosphere at the sample surface while ultrahigh vacuum maintained in the imaging lens systems. Third, a unique NAP cell with gas inlet/outlet, light illumination, sample cooling/heating, and precise sample positioning. The new technique will have important applications in surface catalysis, thin film growth, and energy conversion devices under nearly realistic working conditions., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Little progress has been made in the prevention of pelvic floor disorders, despite their significant health and economic impact. The identification of women who are at risk remains a key element in targeting prevention and planning health resource allocation strategies. Although events around the time of childbirth are recognized clinically as important predictors, it is difficult to counsel women and to intervene around the time of childbirth because of an inability to convey a patient's risk accurately in the presence of multiple risk factors and the long time lapse, which is often decades, between obstetric events and the onset of pelvic floor disorders later in life. Prediction models and scoring systems have been used in other areas of medicine to identify patients who are at risk for chronic diseases. Models have been developed for use before delivery that predict short-term risk of pelvic floor disorders after childbirth, but no models that predict long-term risk exist., Objective: The purpose of this study was to use variables that are known before and during childbirth to develop and validate prognostic models that will estimate the risks of these disorders 12 and 20 years after delivery., Study Design: Obstetric variables were collected from 2 cohorts: (1) women who gave birth in the United Kingdom and New Zealand (n=3763) and (2) women from the Swedish Medical Birth Register (n=4991). Pelvic floor disorders were self-reported 12 years after childbirth in the United Kingdom/New Zealand cohort and 20 years after childbirth in the Swedish Register. The cohorts were split so that data during the first half of the cohort's time period were used to fit prediction models, and validation was performed from the second half (temporal validation). Because there is currently no consensus on how to best define pelvic floor disorders from a patient's perspective, we chose to fit the data for each model using multiple outcome definitions for prolapse, urinary incontinence, fecal incontinence, ≥1 pelvic floor disorder, and ≥2 pelvic floor disorders. Model accuracy was measured in the following manner: (1) by ranking an individual's risk among all subjects in the cohort (discrimination) with the use of a concordance index and (2) by observing whether the predicted probability was too high or low (calibration) at a range of predicted probabilities with the use of visual plots., Results: Models were able to discriminate between women who experienced bothersome symptoms or received treatment at 12 and 20 years, respectively, for pelvic organ prolapse (concordance indices, 0.570, 0.627), urinary incontinence (concordance indices, 0.653, 0.689), fecal incontinence (concordance indices, 0.618, 0.676), ≥1 pelvic floor disorders (concordance indices, 0.639, 0.675), and ≥2 pelvic floor disorders (concordance indices, 0.635, 0.619). Route of delivery and family history of each pelvic floor disorder were strong predictors in most models. Urinary incontinence before and during the index pregnancy was a strong predictor for the development of all pelvic floor disorders in most models 12 years after delivery. The 12- and 20-year bothersome symptoms or treatment for prolapse models were accurate when predictions were provided for risk from 0% to approximately 15%. The 12- and 20-year primiparous model began to over predict when risk rates reached 20%. When we predicted bothersome symptoms or treatment for urinary incontinence, the 12-year models were accurate when predictions ranged from approximately 5-60%; the 20-year primiparous models were accurate from 5% and 80%. For bothersome symptoms or treatment for fecal incontinence, the 12- and 20-year models were accurate from 1-15% risk and began to over predict at rates at >15% and 20%, respectively., Conclusion: Models may provide an opportunity before birth to identify women who are at low risk of the development of pelvic floor disorders and may provide institute prevention strategies such as pelvic floor muscle training, weight control, or elective cesarean section for women who are at higher risk. Models are provided at http://riskcalc.org/UR_CHOICE/., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Background: Pelvic floor muscle training can reduce prolapse severity and symptoms in women seeking treatment. We aimed to assess whether this intervention could also be effective in secondary prevention of prolapse and the need for future treatment., Methods: We did this multicentre, parallel-group, randomised controlled trial at three centres in New Zealand and the UK. Women from a longitudinal study of pelvic floor function after childbirth were potentially eligible for inclusion. Women of any age who had stage 1-3 prolapse, but had not sought treatment, were randomly assigned (1:1), via remote computer allocation, to receive either one-to-one pelvic floor muscle training (five physiotherapy appointments over 16 weeks, and annual review) plus Pilates-based pelvic floor muscle training classes and a DVD for home use (intervention group), or a prolapse lifestyle advice leaflet (control group). Randomisation was minimised by centre, parity (three or less vs more than three deliveries), prolapse stage (above the hymen vs at or beyond the hymen), and delivery method (any vaginal vs all caesarean sections). Women and intervention physiotherapists could not be masked to group allocation, but allocation was masked from data entry researchers and from the trial statistician until after database lock. The primary outcome was self-reported prolapse symptoms (Pelvic Organ Prolapse Symptom Score [POP-SS]) at 2 years. Analysis was by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT01171846., Findings: Between Dec 21, 2008, and Feb 24, 2010, in New Zealand, and Oct 27, 2010, and Sept 5, 2011, in the UK, we randomly assigned 414 women to the intervention group (n=207) or the control group (n=207). One participant in each group was excluded after randomisation, leaving 412 women for analysis. At baseline, 399 (97%) women had prolapse above or at the level of the hymen. The mean POP-SS score at 2 years was 3·2 (SD 3·4) in the intervention group versus 4·2 (SD 4·4) in the control group (adjusted mean difference -1·01, 95% CI -1·70 to -0·33; p=0·004). The mean symptom score stayed similar across time points in the control group, but decreased in the intervention group. Three adverse events were reported, all of which were in the intervention group (one women had a fall, one woman had a pain in her tail bone, and one woman had chest pain and shortness of breath)., Interpretation: Our study shows that pelvic floor muscle training leads to a small, but probably important, reduction in prolapse symptoms. This finding will be important for women and caregivers considering preventive strategies., Funding: Wellbeing of Women charity, the New Zealand Continence Association, and the Dean's Bequest Fund of Dunedin School of Medicine., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background: The use of transvaginal mesh and biological graft material in prolapse surgery is controversial and has led to a number of enquiries into their safety and efficacy. Existing trials of these augmentations are individually too small to be conclusive. We aimed to compare the outcomes of prolapse repair involving either synthetic mesh inlays or biological grafts against standard repair in women., Methods: We did two pragmatic, parallel-group, multicentre, randomised controlled trials for our study (PROSPECT [PROlapse Surgery: Pragmatic Evaluation and randomised Controlled Trials]) in 35 centres (a mix of secondary and tertiary referral hospitals) in the UK. We recruited women undergoing primary transvaginal anterior or posterior compartment prolapse surgery by 65 gynaecological surgeons in these centres. We randomly assigned participants by a remote web-based randomisation system to one of the two trials: comparing standard (native tissue) repair alone with standard repair augmented with either synthetic mesh (the mesh trial) or biological graft (the graft trial). We assigned women (1:1:1 or 1:1) within three strata: assigned to one of the three treatment options, comparison of standard repair with mesh, and comparison of standard repair with graft. Participants, ward staff, and outcome assessors were masked to randomisation where possible; masking was obviously not possible for the surgeon. Follow-up was for 2 years after the surgery; the primary outcomes, measured at 1 year and 2 years, were participant-reported prolapse symptoms (i.e. the Pelvic Organ Prolapse Symptom Score [POP-SS]) and condition-specific (ie, prolapse-related) quality-of-life scores, analysed in the modified intention-to-treat population. This trial is registered as an International Standard Randomised Controlled Trial, number ISRCTN60695184., Findings: Between Jan 8, 2010, and Aug 30, 2013, we randomly allocated 1352 women to treatment, of whom 1348 were included in the analysis. 865 women were included in the mesh trial (430 to standard repair alone, 435 to mesh augmentation) and 735 were included in the graft trial (367 to standard repair alone, 368 to graft augmentation). Because the analyses were carried out separately for each trial (mesh trial and graft trial) some women in the standard repair arm assigned to all treatment options were included in the standard repair group of both trials. 23 of these women did not receive any surgery (15 in the mesh trial, 13 in the graft trial; five were included in both trials) and were included in the baseline analyses only. Mean POP-SS at 1 year did not differ substantially between comparisons (standard 5·4 [SD 5·5] vs mesh 5·5 [5·1], mean difference 0·00, 95% CI -0·70 to 0·71; p=0·99; standard 5·5 [SD 5·6] vs graft 5·6 [5·6]; mean difference -0·15, -0·93 to 0·63; p=0·71). Mean prolapse-related quality-of-life scores also did not differ between groups at 1 year (standard 2·0 [SD 2·7] vs mesh 2·2 [2·7], mean difference 0·13, 95% CI -0·25 to 0·51; p=0·50; standard 2·2 [SD 2·8] vs graft 2·4 [2·9]; mean difference 0·13, -0·30 to 0·56; p=0·54). Mean POP-SS at 2 years were: standard 4·9 (SD 5·1) versus mesh 5·3 (5·1), mean difference 0·32, 95% CI -0·39 to 1·03; p=0·37; standard 4·9 (SD 5·1) versus graft 5·5 (5·7); mean difference 0·32, -0·48 to 1·12; p=0·43. Prolapse-related quality-of-life scores at 2 years were: standard 1·9 (SD 2·5) versus mesh 2·2 (2·6), mean difference 0·15, 95% CI -0·23 to 0·54; p=0·44; standard 2·0 (2·5) versus graft 2·2 (2·8); mean difference 0·10, -0·33 to 0·52; p=0·66. Serious adverse events such as infection, urinary retention, or dyspareunia or other pain, excluding mesh complications, occurred with similar frequency in the groups over 1 year (mesh trial: 31/430 [7%] with standard repair vs 34/435 [8%] with mesh, risk ratio [RR] 1·08, 95% CI 0·68 to 1·72; p=0·73; graft trial: 23/367 [6%] with standard repair vs 36/368 [10%] with graft, RR 1·57, 0·95 to 2·59; p=0·08). The cumulative number of women with a mesh complication over 2 years in women actually exposed to synthetic mesh was 51 (12%) of 434., Interpretation: Augmentation of a vaginal repair with mesh or graft material did not improve women's outcomes in terms of effectiveness, quality of life, adverse effects, or any other outcome in the short term, but more than one in ten women had a mesh complication. Therefore, follow-up is vital to identify any longer-term potential benefits and serious adverse effects of mesh or graft reinforcement in vaginal prolapse surgery., Funding: UK National Institute of Health Research., (Copyright © 2017 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY license. Published by Elsevier Ltd.. All rights reserved.)

Background: Pelvic organ prolapse is common and is strongly associated with childbirth and increasing age. Women with prolapse are often advised to do pelvic floor muscle exercises, but evidence supporting the benefits of such exercises is scarce. We aimed to establish the effectiveness of one-to-one individualised pelvic floor muscle training for reducing prolapse symptoms., Methods: We did a parallel-group, multicentre, randomised controlled trial at 23 centres in the UK, one in New Zealand, and one in Australia, between June 22, 2007, and April 9, 2010. Female outpatients with newly-diagnosed, symptomatic stage I, II, or III prolapse were randomly assigned (1:1), by remote computer allocation with minimsation, to receive an individualised programme of pelvic floor muscle training or a prolapse lifestyle advice leaflet and no muscle training (control group). Outcome assessors, and investigators who were gynaecologists at trial sites, were masked to group allocation; the statistician was masked until after data analysis. Our primary endpoint was participants' self-report of prolapse symptoms at 12 months. Analysis was by intention-to-treat analysis. This trial is registered, number ISRCTN35911035., Findings: 447 eligible patients were randomised to the intervention group (n=225) or the control group (n=222). 377 (84%) participants completed follow-up for questionnaires at 6 months and 295 (66%) for questionnaires at 12 months. Women in the intervention group reported fewer prolapse symptoms (ie, a significantly greater reduction in the pelvic organ prolapse symptom score [POP-SS]) at 12 months than those in the control group (mean reduction in POP-SS from baseline 3.77 [SD 5.62] vs 2.09 [5.39]; adjusted difference 1.52, 95% CI 0.46-2.59; p=0.0053). Findings were robust to missing data. Eight adverse events (six vaginal symptoms, one case of back pain, and one case of abdominal pain) and one unexpected serious adverse event, all in women from the intervention group, were regarded as unrelated to the intervention or to participation in the study., Interpretation: One-to-one pelvic floor muscle training for prolapse is effective for improvement of prolapse symptoms. Long-term benefits should be investigated, as should the effects in specific subgroups., Funding: Chief Scientist Office of the Scottish Government Health and Social Care Directorates, New Zealand Lottery Board, and National Health and Medical Research Council (Australia)., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Objectives: This study sought to identify risk factors for thrombus formation on the Amplatzer Cardiac Plug (ACP) (St. Jude Medical, St. Paul, Minnesota) after left atrial appendage occlusion., Background: Left atrial appendage occlusion with the ACP aims to reduce the risk of embolic stroke and bleeding complications associated with vitamin K antagonists in patients with atrial fibrillation., Methods: We performed transesophageal echocardiography before discharge and after 3, 6, and 12 months in 34 patients with atrial fibrillation undergoing ACP implantation and receiving dual antiplatelet therapy. Clinical, echocardiographic, and hemostaseological parameters were retrospectively analyzed to identify risk factors for thrombus formation., Results: Three patients had thrombi before discharge, 3 more at the 3-month follow-up. No differences were found in left atrial volume, left atrial appendage velocity, spontaneous echo contrast, transmitral gradient, or mitral regurgitation between patients without or with thrombi. CHADS2 (Congestion, Hypertension, Age, Diabetes, and Stroke) score (2.0 ± 1.1 vs. 4.3 ± 1.0), CHA2DS2-VASc (CHADS2 plus Vascular Disease and Sex Category) score (5.2 ± 1.3 vs. 6.8 ± 0.8), and pre-interventional platelet count (215.9 ± 63.9/nl vs. 282.5 ± 84.4/nl) were higher and ejection fraction (50.6 ± 11.4% vs. 39.7 ± 10.6%) lower in those with thrombi. Factor 2, factor 5, or methylenetetrahydrofolate reductase mutations and genetic variants associated with reduced clopidogrel activity were not more frequent in patients with thrombi., Conclusions: Transesophageal echocardiography identified 17.6% of patients with thrombus formation on the ACP despite dual antiplatelet therapy. CHADS2 and CHA2DS2-VASc scores, platelet count, and ejection fraction are risk factors for such thrombus formation., (Published by Elsevier Inc.)

Objective: To assess preliminary effects of a program of transcutaneous posterior tibial nerve stimulation (TPTNS) on lower urinary tract symptoms and number of episodes of urinary and fecal incontinence in older adults in residential care homes and the feasibility of a full-scale randomized trial., Design: Pilot randomized single-blind, placebo-controlled trial., Setting: Seven residential care homes and 3 sheltered accommodation complexes in the United Kingdom., Participants: Thirty care home residents aged 65 and older with urinary or bowel symptoms and/or incontinence., Interventions: Twelve 30-minute sessions of TPTNS or sham stimulation (placebo)., Measurements: Lower urinary tract symptoms using American Urological Society Symptom Index, urinary incontinence using International Consultation on Incontinence Questionnaire-Short Form (ICIQ-SF), postvoid residual urine volumes using portable bladder scanning, bowel symptoms and fecal incontinence using selected ICIQ questions., Results: Total American Urological Society Symptom Index scores improved, showing a median reduction of 7 (interquartile range [IQR] -8 to -3) in the TPTNS group and a median increase in the sham stimulation (placebo) group of 1 (IQR -1 to 4) (Mann-Whitney U 16.5000, Z -3.742, P < .001). Total ICIQ-SF scores improved by a median of 2 (IQR -6 to 0) in the TPTNS group and 0 points (IQR -3 to 3) in the sham stimulation group (Mann-Whitney U 65.000, Z -1.508, P = .132). Significant reduction was found in postvoid residual urine of 55 mL in the TPTNS group (t = -2.215, df 11.338, P = .048). Bowel urgency improved in 27% of the TPTNS group compared with 8% of the sham group (χ(2) 2.395, df 2, P > .302), fecal leakage improved in 47% of the TPTNS group compared with 23% of the sham group (χ(2) 4.480, df 2, P > .106); however, these differences were not significant. No adverse effects were reported by older adults or care staff., Conclusion: TPTNS is safe and acceptable with evidence of potential benefit for bladder and bowel dysfunction in older male and female residents of care homes. Data support the feasibility of a substantive trial of TPTNS in this population., (Copyright © 2013. Published by Elsevier Inc.)

A set of 13 dinucleotide STR loci (G1A, G10B, G1D, G10L, MU05, MU09, MU10, MU15, MU23, MU26, MU50, MU51, MU59) were selected as candidate markers for a DNA forensic profiling system for Northern European brown bear (Ursus arctos). We present results from validation of the markers with respect to their sensitivity, species specificity and performance (precision, heterozygote balance and stutter ratios). All STRs were amplified with 0.6ng template input, and there were no false bear genotypes in the cross-species amplification tests. The validation experiments showed that stutter ratios and heterozygote balance was more pronounced than in the tetranucleotide loci used in human forensics. The elevated ratios of stutter and heterozygote balance at the loci validated indicate that these dinucleotide STRs are not well suited for interpretation of individual genotypes in mixtures. Based on the results from the experimental validations we discuss the challenges related to genotyping dinucleotide STRs in single source samples. Sequence studies of common alleles showed that, in general, the size variation of alleles corresponded with the variation in number of repeats. The samples characterized by sequence analysis may serve as standard DNA samples for inter laboratory calibration. A total of 479 individuals from eight Northern European brown bear populations were analyzed in the 13 candidate STRs. Locus MU26 was excluded as a putative forensic marker after revealing large deviations from expected heterozygosity likely to be caused by null-alleles at this locus. The remaining STRs did not reveal significant deviations from Hardy-Weinberg equilibrium expectations except for loci G10B and MU10 that showed significant deviations in one population each, respectively. There were 9 pairwise locus comparisons that showed significant deviation from linkage equilibrium in one or two out of the eight populations. Substantial genetic differentiation was detected in some of the pairwise population comparisons and the average estimate of population substructure (F(ST)) was 0.09. The average estimate of inbreeding (F(IS)) was 0.005. Accounting for population substructure and inbreeding the total average probability of identity in each of the eight populations was lower than 1.1×10(-9) and the total average probability of sibling identity was lower than 1.3×10(-4). The magnitude of these measurements indicates that if applying these twelve STRs in a DNA profiling system this would provide individual specific evidence., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

The B cell receptor (BCR)-elicited calcium flux results in activation of mature B cells. We have recently shown that the adaptor protein Swiprosin-1/EFhd2 (EFhd2) amplifies the BCR-induced calcium flux in B cell lines. EFhd2 is a calcium binding adaptor protein with two predicted EF-hands. Here we asked whether these domains are functional and control its function. Using a blot-overlay assay with radioactive calcium we show that both EF-hands of EFhd2 have an intrinsic capacity to bind calcium. Equilibrium centrifugation confirmed that EFhd2 binds 2 calcium ions, with an apparent Kd of 110 μM. Point mutations revealed that the conserved residues E116 and E152, which reside in the canonical calcium binding loop in EF-hands 1 and 2, are essential for calcium binding by EFhd2. These mutations as well as deletion of the EF-hands, in particular EF-hand 1, abolished the ability of EFhd2 to restore BCR-induced calcium signaling in EFhd2-deficient WEHI231 cells. N-terminal deletions, but not C-terminal deletions, acted similarly. Thus, the N-terminal part of EFhd2 as well as calcium binding to its EF-hands control the intracellular calcium concentration in response to BCR stimulation in WEHI231 cells. Hence, EFhd2 regulates the BCR-elicited calcium flux through a calcium-dependent positive feedback mechanism in WEHI231 cells., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Background: Urinary incontinence is common immediately after prostate surgery. Men are often advised to do pelvic-floor exercises, but evidence to support this is inconclusive. Our aim was to establish if formal one-to-one pelvic floor muscle training reduces incontinence., Methods: We undertook two randomised trials in men in the UK who were incontinent 6 weeks after radical prostatectomy (trial 1) or transurethral resection of the prostate (TURP; trial 2) to compare four sessions with a therapist over 3 months with standard care and lifestyle advice only. Randomisation was by remote computer allocation. Our primary endpoints, collected via postal questionnaires, were participants' reports of urinary incontinence and incremental cost per quality-adjusted life year (QALY) after 12 months. Group assignment was masked from outcome assessors, but this masking was not possible for participants or caregivers. We used intention-to-treat analyses to compare the primary outcome at 12 months. This study is registered, number ISRCTN87696430., Findings: In the intervention group in trial 1, the rate of urinary incontinence at 12 months (148 [76%] of 196) was not significantly different from the control group (151 [77%] of 195; absolute risk difference [RD] -1·9%, 95% CI -10 to 6). In trial 2, the difference in the rate of urinary incontinence at 12 months (126 [65%] of 194) from the control group was not significant (125 [62%] of 203; RD 3·4%, 95% CI -6 to 13). Adjusting for minimisation factors or doing treatment-received analyses did not change these results in either trial. No adverse effects were reported. In both trials, the intervention resulted in higher mean costs per patient (£180 and £209 respectively) but we did not identify evidence of an economically important difference in QALYs (0·002 [95% CI -0·027 to 0·023] and -0·00003 [-0·026 to 0·026])., Interpretation: In settings where information about pelvic-floor exercise is widely available, one-to-one conservative physical therapy for men who are incontinent after prostate surgery is unlikely to be effective or cost effective. The high rates of persisting incontinence after 12 months suggest a substantial unrecognised and unmet need for management in these men., Funding: National Institute of Health Research, Health Technology Assessment (NIHR HTA) Programme., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Purpose: To evaluate the repeatability and reproducibility of retinal thickness measurements in exudative age-related macular degeneration (AMD) using Stratus optical coherence tomography (OCT) (Carl Zeiss Meditec, Inc., Dublin, CA)., Design: Prospective, observational case series., Participants: A total of 200 eyes of 200 subjects with exudative AMD., Methods: Macular thickness and fast macular thickness programs of Stratus OCT were performed twice by the same examiners or 2 different examiners. The sequence of examiners was randomized 1:1:1:1. The variability of 1-mm subfield central retinal thickness (CRT), center point thickness (CPT), and retinal volume (RV) was calculated., Main Outcome Measures: Interobserver and intraobserver variability of retinal thickness measurements., Results: Ninety-nine patients/eyes were enrolled in study arm 1 (repeated by the same examiner), and 101 patients/eyes were enrolled in study arm 2 (repeated by different examiners). Values of CPT, CRT, and RV were well correlated (interclass correlation coefficient, 0.71-0.93) in both study arms, revealing better results for the macular thickness program than for the fast macular thickness program. Threshold algorithm line failures were significantly correlated to the absolute differences of 2 repeated measurements for CPT, CRT, and RV but not with manually corrected maximum retinal thickness (MRT). Maximum retinal thickness was significantly influenced by the examiner performing the measurement. Age, lesion composition, examiner performing OCT examination, and sequence of examination had no significant influence., Conclusions: The repeatability and reproducibility of retinal thickness measurements were high, presenting better results for CRT and RV versus CPT, and for the macular thickness program versus the fast macular thickness program. The reliability of retinal thickness measurement was most frequently affected by algorithm line failures and fixation problems. A possible solution may be manually corrected measurement, such as MRT., (Copyright 2010 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

Purpose: To assess prognostic factors in epiretinal membrane (ERM) surgery using spectral-domain (SD) optical coherence tomography (OCT)., Design: Prospective, interventional case series., Participants: Forty-one patients., Methods: Patients with a diagnosis of ERM were examined with spectral-domain and time-domain (TD) OCT before and after surgery., Main Outcome Measures: Main outcome measures were functional results and predefined OCT patterns. Cofactors analyzed were the type of ERM, the duration and grading of subjective metamorphopsia, simultaneous cataract surgery, the type of dyes used, the duration of surgery, gender, and age. A multivariate regression analysis was performed., Results: Thirty-nine patients (95%) showed an improved or stable best-corrected distance visual acuity (DVA) at 3 months, whereas 36 patients (88%) showed an improved or stable best-corrected near visual acuity (NVA) at 3 months. Significant correlations (P>0.4) were found between the course of central retinal thickness (CRT) assessed using SD OCT and TD OCT (P<0.02). No clinically relevant correlations (P<0.3) were seen between CRT and VA (P>0.1). Baseline DVA and NVA were found to be significant prognostic values for the postoperative decrease in CRT in both OCT systems (P<0.04) as well as for the visual outcomes (P<0.007) at 3 months. In addition, the integrity of the junction between the photoreceptor inner segment and outer segment (IS/OS) significantly influenced the visual outcomes at 3 months (P<0.038). The baseline profile of the internal limiting membrane (ILM) significantly influenced the NVA at 3 months (P<0.009), whereas the postoperative foveal contour significantly influenced the DVA at 3 months (P<0.025). The type of ERM, subjective metamorphopsia, simultaneous cataract surgery, the type of dyes used, the duration of surgery, gender, or age had no significant influence on patient outcome (P>0.05). Compared with TD OCT, SD OCT allowed for a more precise differentiation between the ERM and the retinal surface and for a better evaluation of the IS/OS line., Conclusions: Besides the baseline VA, the integrity of the IS/OS line, better visualized by SD OCT, can be used to predict the functional outcomes after surgery. Additionally, analyzing the ILM profile and the foveal contour may help to understand limited visual outcomes after surgery., Financial Disclosure(s): The author(s) have no proprietary or commercial interest in any materials discussed in this article., (Copyright 2010 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.)

The deep inferior epigastric perforator (DIEP) flap is the gold standard for breast reconstruction using abdominal tissue. Unlike the transverse rectus abdominis myocutaneous (TRAM) flap, no rectus abdominis muscle is removed with the flap, but intra-muscular scarring can still cause post-operative complications. Strong abdominal muscles have been advocated as a prerequisite for surgery, but without any evidence as to the potential benefits. This study aimed to investigate the effect of pre-operative abdominal exercises on inpatient pain levels, length of hospital stay, post-operative abdominal muscle strength and function following a DIEP flap.Ninety-three women undergoing delayed breast reconstruction with a DIEP flap between October 1999 and November 2000 were randomly allocated to either a control or exercise group. The exercise group performed pre-operative exercises using the Abdotrim abdominal exerciser. Pre-operatively, outcome measures included trunk muscle strength measured on an isokinetic dynamometer, SF-36, rectus muscle thickness measured using ultrasound, and submaximal fitness. Post-operative pain and length of hospital stay were recorded. Subjects were reassessed using the same outcome measures 1 year post-operatively. There was a statistically significant increase in static (isometric) muscle strength and thickness pre-operatively for the exercise group. One year following surgery, there was a significant decrease in dynamic (concentric and eccentric) flexion strength for both groups, although the clinical significance of this is questionable as the majority of women had returned to pre-operative fitness and the surgery had no impact on functional activities. The static flexion strength of the control group was reduced at 1 year, whereas it was maintained in the exercise group, although this was not statistically significant. One third of women in the control group complained of functional problems or abdominal pain post-operatively compared to one fifth of the exercise group. Overall, the DIEP flap had no major impact on abdominal muscle strength for either group, demonstrating its superiority over the TRAM flap. There was no statistically significant benefit to the exercise group of the pre-operative exercises 1 year following surgery. However, there was a subjective benefit, albeit statistically nonsignificant, in terms of reduced functional problems post-operatively and improved well-being prior to surgery.

We have measured the effect of temperature and denaturant concentration on the rate of intrachain diffusion in an unfolded protein. After photodissociating a ligand from the heme iron of unfolded horse cytochrome c, we use transient optical absorption spectroscopy to measure the time scale of the diffusive motions that bring the heme, located at His18, into contact with its native ligand, Met80. Measuring the rate at which this 62 residue intrachain loop forms under both folding and unfolding conditions, we find a significant effect of denaturant on the chain dynamics. The diffusion of the chain accelerates as denaturant concentration decreases, with the contact formation rate approaching a value near approximately 6x10(5) s(-1) in the absence of denaturant. This result agrees well with an extrapolation from recent loop formation measurements in short synthetic peptides. The temperature dependence of the rate of contact formation indicates an Arrhenius activation barrier, Ea approximately 20 kJ/mol, at high denaturant concentrations, comparable to what is expected from solvent viscosity effects alone. Although Ea increases by several kBT as denaturant concentration decreases, the overall rate of diffusion nevertheless increases. These results indicate that inter-residue energetic interactions do not control conformational diffusion in unfolded states, even under folding conditions.

Abdominal weakness is a known potential complication of breast reconstruction with a pedicled or free TRAM flap. It has been presumed that the DIEP flap, which involves no muscle resection, does not compromise abdominal muscle strength but little objective research exists to substantiate this. The aims of this retrospective study were to compare abdominal muscle strength following free TRAM flap and DIEP flap, to compare both groups with a control group and to establish the effect of both procedures on functional activities. Fifty women (23 with a DIEP flap, 27 with a free TRAM flap) plus 32 non-operated controls underwent assessment of their abdominal and back extensor muscle strength on a KIN COM isokinetic dynamometer. Two questionnaires were used to establish the impact on function. The TRAM flap group had significant weakness of the abdominal and back extensor muscles compared with the DIEP flap group and the control group. The trend was for the DIEP flap group to have weaker abdominal muscles than the control group. There was a higher level of abdominal pain and a greater number of reported functional difficulties in the TRAM flap group than in the DIEP flap group. This study demonstrates that whilst the DIEP flap can reduce the strength deficit caused by the free TRAM flap, abdominal weakness can still result from the DIEP flap. A randomised controlled trial is currently underway to investigate the effect of preoperative abdominal exercises in preventing/minimising postoperative abdominal muscle weakness in this group., (Copyright 2000 The British Association of Plastic Surgeons.)

The initial phase of folding for many proteins is presumed to be the collapse of the polypeptide chain from expanded to compact, but still denatured, conformations. Theory and simulations suggest that this collapse may be a two-state transition, characterized by barrier-crossing kinetics, while the collapse of homopolymers is continuous and multi-phasic. We have used a laser temperature-jump with fluorescence spectroscopy to measure the complete time-course of the collapse of denatured cytochrome c with nanosecond time resolution. We find the process to be exponential in time and thermally activated, with an apparent activation energy approximately 9 k(B)T (after correction for solvent viscosity). These results indicate that polypeptide collapse is kinetically a two-state transition. Because of the observed free energy barrier, the time scale of polypeptide collapse is dramatically slower than is predicted by Langevin models for homopolymer collapse.

A natural infection with Helicobacter pylori (H. pylori) in domestic cats (Felis cattus) less than 2 years of age has been well described in a closed colony of animals. Six cats from this colony that were serially evaluated by culture, polymerase chain reaction, and light and electron microscopy for a period of 3 years demonstrated persistent gastric colonization with a single cag(-) vac(+) strain of H. pylori. In these cats, as well as five other 5- to 6-year-old cats that were examined, a long-term infection resulted in chronic diffuse lymphofollicular atrophic gastritis with areas of mucosal dysplasia in the antrum and predominantly midsuperficial gastritis in the body and cardia. Topographically, the distribution of lesions was similar in both young and older cats and closely resembled that found in humans, with the most severe changes occurring in the gastric antrum. Few granulocytes and no significant elevation in mast cells were seen in older H. pylori-infected cats compared with uninfected controls; however, marked increases in interepithelial globule leukocytes and numerous active mucosal lymphoid follicles were present in infected animals. Indices of gastritis were significantly greater in older infected cats when compared with uninfected controls and younger cats (P < 0.05). The antral cell proliferation index of infected older cats was significantly (P = 0.021) greater than that of uninfected controls. Apoptotic indices of the gastric antrum and body of infected cats were significantly (P = 0.01) increased versus controls. Chronic infection with H. pylori in cats shares many features of long-term H. pylori infection in humans, including the development of preneoplastic processes. This similarity provides useful, comparative insights into host-pathogen interactions.

The standardization of natural allergenic extracts and the characterization of recombinant allergens ensures a continuing requirement for highly purified natural allergens. The extraction and purification methods have to be reproducible and also preserve the biological and immunological activity of the allergen. A simple two-step purification system has been established in order to provide milligram amounts of purified natural Phl p 1 and Phl p 2/3. Both major allergens were separated from other proteins of timothy grass pollen extract in one step by hydrophobic interaction chromatography (HIC) under mild conditions. The allergens elute in the flow-through fraction while the rest of the proteins remain bound to the column. The very different molecular weights of Phl p 1 and Phl p 2/3 permitted separation of the allergens by a second step using gel filtration.

The CC chemokine RANTES is synthesized, stored, and upregulated in response to interferon-gamma (IFN-gamma) in human peripheral blood eosinophils. In this report, we propose that RANTES is rapidly mobilized from eosinophil crystalloid granules during agonist-induced degranulation. We stimulated purified eosinophils (>99%) from atopic asthmatics with 500 U/mL IFN-gamma to analyze the kinetics of mobilization and release of RANTES (0 to 240 minutes). We used subcellular fractionation, immunogold analysis, two-color confocal laser scanning microscopy (CLSM), and enzyme-linked immunosorbent assay (ELISA) to trace the movement of eosinophil-derived RANTES from intracellular stores to release. RANTES was rapidly mobilized (10 minutes) and released after 120 minutes of stimulation (80 +/- 15 pg/mL per 2 x 10(6) cells). RANTES appeared to be stored in at least two intracellular compartments: the matrix of crystalloid granules, detected by major basic protein and eosinophil peroxidase activities, and a specialized small secretory vesicle present in light membrane fractions. The extragranular RANTES was mobilized more rapidly than that of crystalloid granules during IFN-gamma stimulation. This effect was not observed in eosinophils treated with IFN-alpha, interleukin-3 (IL-3), IL-5, granulocyte-macrophage colony-stimulating factor (GM-CSF), or genistein followed by IFN-gamma. Our findings suggest that RANTES may be mobilized and released by piecemeal degranulation upon stimulation, involving transport through a putative pool of small secretory vesicles.

Porous poly(para-divinylbenzene) and poly(meta-divinylbenzene) particles were synthesised from para-divinylbenzene and meta-divinylbenzene monomers with toluene and 2-ethylhexanoic acid as porogens. The residual vinyl groups in the particles were thereafter reacted using aluminium chloride with dichlorobenzene as a catalyst. The conversion of vinyl groups was followed by analysing polymer particles taken from the reaction mixture at different time intervals. Infrared spectroscopy both in the mid and near infrared region was used as the analytical technique. The intensity changes in the overtone absorption at 1628 nm due to the vinyl bonds were used as the basis for the quantification of the vinyl group consumption. Infrared spectra of the particles in the mid IR were also measured to understand changes taking place in the polymer matrix during the reaction. The results indicated that residual vinyl groups in these polymer particles were consumed during the reaction with aluminium chloride. The reaction of aluminium chloride with the polymer matrix was explained by proposing mechanisms for the formation of different products during the reaction. The complex formed between aluminium chloride and the residual vinyl groups seemed to induce addition of HCl to the vinyl group or leads to crosslinking and/or cyclisation in the case poly(para-DVB) particles. The reaction of aluminium chloride with poly(meta-DVB) takes place to a lesser extent.

The Newark Bay Complex includes the Newark Bay, tidal portions of the Hackensack River, Passaic River, Arthur Kill, and Kill van Kull. It is a highly industrialized urban area including five counties and more than 20 local governments with a large racially-mixed population of more than 3 million people. In 1982, research conducted by the New Jersey Department of Environmental Protection (NJDEP) showed elevated levels of chemical contaminants in five species of fish and one type of crab in the Newark Bay Complex. Subsequently, the State of New Jersey adopted advisories to guide citizens on safe consumption practices for fish and crabs. Since then, fish consumption advisories have been issued primarily through the Fish and Game Digest, a publication distributed by the state to licensed anglers. However, anglers in the Complex are not required to have a fishing license because the waters are marine. Therefore, most anglers in this area do not receive advisory information. To gain greater insight into the information sources and risk perceptions of urban anglers, a survey was conducted of 300 anglers at 26 fishing and crabbing locations in the Newark Bay Complex during the summer and early fall of 1995. The objectives of the study were to learn anglers': (1) knowledge of fish consumption advisories; (2) belief in the advisories; (3) perception of how safe fish are to eat; (4) sources for information about fish and fishing; and (5) sources for information on fish consumption advisories. The study concluded that while 60% had heard about advisories, they either did not believe or were unconcerned about health effects from eating contaminated species. In addition, the most used source for information about fish and fishing was other fishermen, while newspapers were selected as a source for information about community news, health, and food safety.

Eosinophils, prominent cells in asthmatic inflammation, have been shown to synthesize, store, and release an array of up to 18 cytokines and growth factors, including interleukin-6 (IL-6). In this report, we show that IL-6 immunofluorescence localizes to the matrix of the crystalloid granule in peripheral blood eosinophils from atopic asthmatics using confocal laser scanning microscopy (CLSM). Granule localization of IL-6 was confirmed using dot-blot analysis and enzyme-linked immunosorbent assay (ELISA) on subcellular fractions of highly purified eosinophils produced from density centrifugation across a 0% to 45% Nycodenz gradient. IL-6 was found to coelute with eosinophil crystalloid granule marker proteins, including eosinophil peroxidase (EPO), major basic protein (MBP), arylsulfatase B, and beta-hexosaminidase. Immunoreactivity to IL-6 colocalized with granule-associated IL-2 and IL-5 in subfractionated eosinophils. We also made the novel and compelling observation that interferon gamma (IFNgamma), a Th1-type cytokine, stimulated an early elevation in eosinophil IL-6 immunoreactivity. A 2.5-fold enhancement of IL-6 immunoreactivity in eosinophil granules was observed within 10 minutes of IFNgamma treatment (500 U/mL), as determined by subcellular fractionation and CLSM. These findings suggest that IFNgamma has short-term effects on human eosinophil function and imply that a physiologic role exists for Th1-type cytokine modulation of Th2-type responses in these cells.

MPL immunostimulant, a 4'-monophosphoryl lipid A (MLA) preparation obtained from the lipopolysaccharide of Salmonella minnesota R595, is being developed for several clinical indications. MPL comprises a mixture of MLA congeners that contain 4, 5, and 6 fatty acids. In this paper, we report a new high-performance liquid chromatography (HPLC) method for analyzing the congener composition and purity of MPL. MPL is first derivatized with dinitrobenzyloxyamine (DNBA), resulting in incorporation of the dinitrobenzyl chromophore at the reducing end of all MLA congeners. DNBA-MPL is then analyzed by reversed-phase HPLC on a Waters NovaPak C18, 4 microns particle size, 300 mm x 3.9 mm column. Optimal separation of DNBA-MLA species is obtained using a linear gradient of 10% to 80% isopropanol-water (95:5, v/v), 5 mM tetrabutylammonium dihydrogenphosphate (TBAP), in acetonitrile-water (95:5, v/v), 5 mM.TBAP, over 45 min. A synthetic compound, corresponding to a hexaacyl MLA congener, is used for determination of the detector response factor, allowing the MLA content of MPL (i.e., purity) to be determined. Overall, this method provides better separation, higher sensitivity, and is faster and saler than previous methods used for the analysis of MPL.

Clinicians often do not agree whether a treatment is given with a palliative or curative intent. A common clear definition does not exist. This study has assessed the usefulness of dividing the goal of treatment into three distinct categories: curative treatment; palliative, symptom preventive treatment; and palliative, symptom relieving treatment. In a cross-sectional study among all cancer centres in Norway, a total of 629 patients were included into the study. Of these patients, 60% received palliative treatment, with an equal distribution between symptom preventive and symptom relieving. The definitions were found easy to use by the physicians. It gave important information of differences between cancer diagnosis with respect to the number of patients receiving palliative treatment. In order to refine the classification system, the authors will, in future studies, include a fourth category, life prolonging treatment, which is located between curative and palliative treatment with respect to treatment intensity.

Physical-chemical and biological studies of hepatic bile suggest that biliary phospholipid molecules are secreted as unilamellar vesicles. Systematic ultrastructural studies of bile canaliculi were undertaken to visualize this event. Liver tissue was obtained from normal adult male rats (control), from bile salt-depleted rats (by overnight biliary diversion), and from depleted rats infused intravenously with a hydrophilic-hydrophobic congener series of common taurine-conjugated bile salts. Livers were fixed in situ either by modified chemical methods or by ultrarapid cryofixation. In control rats, chemical fixation revealed unilamellar vesicles 63 +/- 17 (+/- SD) nm in diameter, mostly free within canalicular lumena. Vesicles were infrequent in canaliculi of bile salt-depleted rats, but were present in canaliculi of rats infused with taurocholate. In cryofixed liver tissue, vesicles 67 +/- 13 nm in diameter were observed in canaliculi of control rats and bile-salt depleted rats infused with common bile salts. The majority of these vesicles were affixed to the luminal side of the canalicular membrane. The average number of vesicles per bile canaliculus was in agreement with that estimated on the basis of biliary phospholipid secretion rates, mean vesicle size, and area of close-packed phosphatidylcholine molecules. By immunoelectron microscopy, canalicular vesicles were free of actin and of a 100 kDa canalicular membrane protein. We conclude that biliary phospholipid molecules are secreted from hepatocytes into bile canalicular lumena as unilamellar vesicles approximately 63-67 nm in average diameter. We postulate that this secretion mechanism involves lumenal bile salt-induced vesiculation of lipid microdomains in the exoplasmic hemileaflet of the canalicular membrane.

Mycolic acids from Mycobacterium phlei and M. bovis cell wall skeletons (CWSs) were analyzed by HPLC. After saponifying lyophilized CWSs in methanolic KOH, the mycolic acids were quantitatively extracted into chloroform. Aliquots of the CWS mycolic acid extracts were then derivatized prior to HPLC analysis with a UV reagent, p-bromophenacylbromide (PBPB), and three fluorescent reagents, 4-bromomethyl-6,7-dimethoxycoumarin, 4-bromomethyl-7-acetoxycoumarin and 3-bromomethyl-7-methoxy-1,4-benzoxazin-2-one. A synthetic alpha-branched carboxylic acid was derivatized with the same reagents and used as an internal standard along with the mycolic acids. The derivatized samples were analyzed by reversed-phase HPLC on a Waters Novapak C18, 4 microns particle size, 150 mm x 3.9 mm stainless-steel column. Two solvent systems were used: (1) methanol and methylene chloride with the column at 30 degrees C, and (2) methanol and isopropanol with the column at 50 degrees C. Detection sensitivity with the fluorescent reagents was 16-50 times greater than the sensitivity observed with PBPB-derivatized samples. Unique mycolic acid elution profiles for the two mycobacterial species could be achieved with each of the solvent systems and derivatization reagents tested. Thus, the HPLC analysis of pre-column derivatized mycolic acids was useful as a means of rapidly identifying mycobacterial species. Replacement of methylene chloride with isopropanol and PBPB with a fluorescent derivatizing reagent could increase the safety and sensitivity of the assay, and make it more useful for the clinical identification of mycobacterial infections.

Entactin/nidogen (E/N) was isolated from bovine renal tubular basement membrane. Apparent molecular weight, amino acid composition, and molecular configuration by electron microscopy rotary shadowing were similar to that of nidogen from EHS mouse tumor. The identity of bovine E/N was confirmed using a thrombin derived peptide, the sequence of which corresponded to a region within mouse and human E/N. Monoclonal and polyclonal anti-E/N antibodies were used to determine the distribution of E/N in human kidney by immunofluorescent and immunoelectron microscopy. E/N was present in all renal basement membranes and was distributed through the full width of the glomerular basement membrane (GBM) with accentuation along its epithelial aspects. E/N distribution was similar to that of novel collagen chain alpha 3(IV) NC domain in the GBM. In the mesangium, E/N was distributed mainly in the peripheral mesangial region that is bounded by the GBM, while classical collagen chain alpha 1(IV) NC as present diffusely throughout the mesangium. In the developing nephron, E/N was present in basement membranes of the ureteric bud, primitive vesicle and S-form. In all instances, E/N co-localized with laminin B2 chain. Prominent E/N detection within the mesangium was observed in diseases where mesangial expansion was present. This process was also seen in early diabetic nephropathy, but disappeared with disease progression. However, all thickened diabetic renal basement membranes showed an increase in E/N which was also present in Kimmelstiel-Wilson lesions. E/N was observed in the GBM "spikes" of membranous glomerulonephritis and in epithelial crescents associated with various disorders. The association between E/N, laminin and type IV collagen chains observed in the normal kidney were maintained in disorders with altered E/N distribution. We could not detect any changes in the distribution of E/N in other acquired and hereditary kidney diseases. These observations reflect the involvement of E/N in the structure and disease alteration of renal basement membranes and mesangial matrix.

MHC class II antigen-associated invariant (Ii) chain mRNA appears in mouse small intestine during postnatal development. Ii chain cDNA hybridizes to RNA from epithelial sheets dissociated from the lamina propria with EDTA. Of several mouse organs tested, only bone marrow and spleen contain higher levels of Ii chain mRNA than small bowel. Ii chain mRNA is not detected in stomach, colon, duodenum, testis, liver, submandibular gland, or L-cell RNA; brain contains a cross-reactive but uncharacterized sequence. cDNA amplification using primers specific for both Ii31 and Ii41 chain mRNAs showed that both forms occur in small intestine. These results support the conclusion that regulation of the class II Ii chain gene is associated with the ontogeny of intestinal immunity.

Cyclic adenosine 3',5'-monophosphate is known to modulate smooth muscle contractility. Because both relaxin and progesterone have been demonstrated to affect myometrial cyclic adenosine monophosphate activity, we questioned whether the previously observed synergism of these two hormones in inhibiting uterine contractility is mediated via cyclic adenosine monophosphate. Immature rats were treated with estradiol benzoate (n = 7) or a combination of estradiol benzoate and progesterone (n = 7). Uterine horns were isolated, each horn was divided into two segments, and these horn segments were incubated in Ringer-Locke solution, either alone (control) or with 3-isobutyl-1-methylxanthine (MIX) 0.5 mM, MIX 0.5 mM + relaxin 10 ng/ml, or MIX 0.5 mM + relaxin 50 ng/ml. When compared with uterine segments incubated in MIX alone, treatment with MIX + relaxin 50 ng/ml significantly increased cyclic adenosine monophosphate levels in animals treated with estradiol benzoate alone or in combination with progesterone. Relaxin 10 ng/ml was sufficient to significantly elevate mean (+/- SEM) uterine cyclic adenosine monophosphate levels above that of control MIX-treated uteri in animals receiving both estradiol benzoate and progesterone (2.49 +/- 0.39 pm/micrograms deoxyribonucleic acid [DNA] versus 1.08 +/- 0.16 pm/microgram DNA, p less than 0.05) but not in animals receiving estradiol benzoate alone (2.08 +/- 0.32 pm/micrograms DNA versus 1.28 +/- 0.16 pm/micrograms DNA, NS). Compared with treatment with MIX only, MIX + relaxin 10 ng/ml and MIX + relaxin 50 ng/ml produced greater increases in uterine cyclic adenosine monophosphate in the steroid combination group than in the estradiol benzoate controls (144.8% and 233.7% versus 71.7% and 156.6%, respectively). These results suggest that the synergism of relaxin and progesterone in inhibiting uterine contractility may be mediated by intracellular cyclic adenosine monophosphate.