Liverpool Ultrasound Pictorial Chart: the development of a new method of documenting anal sphincter injury diagnosed by endoanal ultrasound

Objective  To develop and validate a pictorial chart that documents ultrasound examination of the anal sphincter.
Design  A new pictorial chart (Liverpool Ultrasound Pictorial Chart [LUPIC]) depicting the normal anatomy of the anal sphincter was developed.
Methods  To validate LUPIC, two observers documented the findings of 296 endoanal scans. Reliability was assessed between observers using kappa agreement for presence and position of sphincter defects. To validate the use of LUPIC by different observers, a video of ten endoanal ultrasound scans was reviewed by our local expert (gold standard). Seven clinicians underwent test-retest analysis. Kappa agreement was calculated to assess intra-observer and gold standard versus observer agreement for the overall presence of sphincter defects and compared with the gold standard. Complete agreement for the position and level of sphincter defects was assessed for the five abnormal scans.
Main outcome measures  Excellent agreement between the two observers was found for the presence (kappa 0.99), position and level of external anal sphincter defects documented using LUPIC. The intra-observer and gold standard versus observer kappa values of experienced clinicians (A–E) showed good agreement for the overall presence of sphincter defects. Complete agreement for the position and level of sphincter defects was found in 23 of 35 (66%) observations.
Conclusions  LUPIC is designed and validated method of documenting anal sphincter injury diagnosed by endoanal ultrasound. Standardisation of endoanal ultrasound findings by using LUPIC may help correlate the degree of damage with patient symptoms.     

Multifocal urothelial cancers with the mutator phenotype are of monoclonal origin and require panurothelial treatment for tumor clearance

PURPOSE: UC is a disease of the entire urothelium, characterized by multiplicity and multifocality. The clonal relationship among multiple UCs has implications regarding adjuvant chemotherapy. It has been investigated in studies of chromosomal alteration and single gene mutation. However, these genetic changes can occur in unrelated tumors under similar carcinogenic selection pressures. Tumors with high MSI have numerous DNA mutations, of which many provide no selection benefit. While these tumors represent an ideal model for studying UC clonality, their low frequency has prevented their previous investigation. MATERIALS AND METHODS: We investigated 32 upper and lower urinary tract UCs with high MSI and 4 nonUC primary cancers in 9 patients. We used the high frequency and specificity of individual DNA mutations in these tumors (MSI at 17 loci) and the early timing of epigenetic events (methylation of 7 gene promoters) to investigate tumor clonality. RESULTS: Molecular alterations varied among tumors from different primary organs but they appeared related in the UCs of all 9 patients. While 7 patients had a high degree of concordance among UCs, in 2 the UCs shared only a few similar alterations. Genetic and epigenetic abnormalities were frequently found in normal urothelial samples. CONCLUSIONS: Multiple UCs in each patient appeared to arise from a single clone. The molecular order of tumor development varied from the timing of clinical presentation and suggested that residual malignant cells persist in the urinary tract despite apparent curative surgery. These cells lead to subsequent tumor relapse and new methods are required to detect and eradicate them.     

ATM is required for the cellular response to thymidine induced replication fork stress

Genetically distinct checkpoints, activated as a consequence of either DNA replication arrest or ionizing radiation-induced DNA damage, integrate DNA repair responses into the cell cycle programme. The ataxia-telangiectasia mutated (ATM) protein kinase blocks cell cycle progression in response to DNA double strand breaks, whereas the related ATR is important in maintaining the integrity of the DNA replication apparatus. Here, we show that thymidine, which slows the progression of replication forks by depleting cellular pools of dCTP, induces a novel DNA damage response that, uniquely, depends on both ATM and ATR. Thymidine induces ATM-mediated phosphorylation of Chk2 and NBS1 and an ATM-independent phosphorylation of Chk1 and SMC1. AT cells exposed to thymidine showed decreased viability and failed to induce homologous recombination repair (HRR). Taken together, our results implicate ATM in the HRR-mediated rescue of replication forks impaired by thymidine treatment.     

Assessment of an immunoassay for interferon-alpha in cerebrospinal fluid as a diagnostic aid in infections of the central nervous system

A highly sensitive and specific immunoradiometric assay, based upon a monoclonal antibody, was used to measure interferon-alpha (IFN-alpha) in the cerebrospinal fluid (CSF) of patients with central nervous system infections and in controls with non-infectious neurological disorders. IFN-alpha was detected in all 21 patients with viral meningitis but in only one of four patients with non-viral aseptic meningitis. It was also present in the CSF of three of four patients with herpes encephalitis and five of seven patients with acute bacterial meningitis. By contrast, IFN-alpha was present in the CSF in low concentrations in only five (7%) of 71 neurological controls. This rapid test is positive in viral meningitis and may help in distinguishing viral infection from other causes of aseptic meningitis. It is usually negative in non-infective disorders but will not distinguish between viral and bacterial infections.     

Immunoreactive IFN-gamma in CSF in neurological disorders.

Interferon gamma (IFN-gamma) was measured in the CSF of neurological patients using a highly specific and sensitive immunoradiometric assay. It was detected in 52% of patients presenting as suspected meningitis or encephalitis and in 83% with proven viral meningitis. In contrast IFN-gamma was detected in only 26% of patients who did not have an acute infection at the time of presentation. Only 15% of patients with multiple sclerosis had detectable CSF IFN-gamma. The presence of IFN-gamma in CSF in response to acute viral infections of the central nervous system may be of importance in relation to the pathophysiology of immunologically mediated neurological disorders.     

Rad51 supports triple negative breast cancer metastasis

In contrast to extensive studies on familial breast cancer, it is currently unclear whether defects in DNA double strand break (DSB) repair genes play a role in sporadic breast cancer development and progression. We performed analysis of immunohistochemistry in an independent cohort of 235 were sporadic breast tumours. This analysis suggested that RAD51 expression is increased during breast cancer progression and metastasis and an oncogenic role for RAD51 when deregulated. Subsequent knockdown of RAD51 repressed cancer cell migration in vitro and reduced primary tumor growth in a syngeneic mouse model in vivo. Loss of RAD51 also inhibited associated metastasis not only in syngeneic mice but human xenografts and changed the metastatic gene expression profile of cancer cells, consistent with inhibition of distant metastasis. This demonstrates for the first time a new function of RAD51 that may underlie the proclivity of patients with RAD51 overexpression to develop distant metastasis. RAD51 is a potential biomarker and attractive drug target for metastatic triple negative breast cancer, with the capability to extend the survival of patients, which is less than 6 months.     

A urinary control device for management of female stress incontinence

OBJECTIVE: To examine the performance of a silicon urinary control device for nonsurgical management of women with genuine stress incontinence. METHODS: A 3-month prospective study involved 41 women with genuine stress incontinence. They completed urinary diaries of voiding, incontinence, and severity of incontinence on a 4-point scale over a week. Subjects were taught how to apply the device and used it as required from the second week. Visual analogue scales were used to record aspects of use (such as acceptability, comfort, and ease of application), and 2-hour perineal pad tests were completed at recruitment, after 2 weeks, and after 3 months. Data were compared by Mann-Whitney U test, or Wilcoxon test. RESULTS: Ten women (24.4%) declined to participate and six (14.6%) withdrew before 2 weeks. Ten (24.4%) failed to attend for 2-week follow-up and 11 (26.8%) did not continue for 3 months. Two (4.9%) did not attend 3-month follow-up. Only two women (4.9%) completed the study. There was no difference in pad test results or in results from voiding diaries. CONCLUSION: The urinary incontinence device had low acceptability and was ineffective, and we cannot recommend it for nonsurgical management of genuine stress incontinence.     

A tumour-derived mutant allele of XRCC2 preferentially suppresses homologous recombination at DNA replication forks

Homologous recombination repair (HRR) is required for both the repair of DNA double strand breaks (DSBs) and the maintenance of the integrity of DNA replication forks. To determine the effect of a mutant allele of the RAD51 paralog XRCC2 (342delT) found in an HRR-defective tumour cell line, 342delT was introduced into HRR proficient cells containing a recombination reporter substrate. In one set of transfectants, expression of 342delT conferred sensitivity to thymidine and mitomycin C and suppressed HRR induced at the recombination reporter by thymidine but not by DSBs. In a second set of transfectants, the expression of 342delT was accompanied by a decreased level of the full-length XRCC2. These cells were defective in the induction of HRR by either thymidine or DSBs. Thus 342delT suppresses recombination induced by thymidine in a dominant negative manner while recombination induced by DSBs appears to depend upon the level of XRCC2 as well as the expression of the mutant XRCC2 allele. These results suggest that HRR pathways responding to stalled replication forks or DSBs are genetically distinguishable. They further suggest a critical role for XRCC2 in HRR at replication forks, possibly in the loading of RAD51 onto gapped DNA.     

Effect of a new guideline on outcome following third-degree perineal tears: results of a 3-year audit

The aim of this study was to assess the impact of a new guideline on the outcome of repair of obstetric anal sphincter tears by examining adherence to the guideline and the effect upon the incidence of symptoms of anal incontinence. An audit of third-degree perineal tears was conducted in 1997. A reaudit was completed in 1998 and 1999 after the introduction of a new guideline. The audits were conducted in a tertiary obstetric unit with 5000 deliveries per annum. Over the 3-year period between 1997 and 1999 124 women with a third-degree tear were identified; 14 381 women who had delivered vaginally without third-degree tears were used as controls. The main outcome measure was the number of cases with adherence to the protocol, and the number of patients with ongoing symptoms. Cases were identified from the hospital database, and case notes were reviewed to obtain clinical data. The incidence of third-degree perineal tears was 0.81% over the 3-year period. Following the introduction of a new guideline there was a significant increase in the number of repairs performed in theatre (70% vs 82% vs 97%, P<0.05), using Prolene (64% vs 76% vs 93%, P< 0.05), with adequate anesthesia (70% vs 82% vs 97%, P<0.05). At follow-up there was a transient improvement in defecatory symptoms in the first year of the protocol only (45% vs 32% vs 50%, P<0.01). More patients had complete follow-up data after introduction of the protocol: 66% vs 86% vs 80% (P<0.001). There were more cases of Prolene suture migration (7% vs 34% vs 16%, P<0.01). We concluded that the introduction of a new guideline was followed by improved performance of appropriate repair. There was no sustained improvement in fecal symptoms at follow-up. Editorial Comment: This study highlights to important issues regarding primary repair of obstetric anal sphincter lacerations. First, the need for adequate training and supervision of residents is required for optimal repair of obstetric anal sphincter lacerations. Junior residents often perform these repairs with inadequate training, lack of supervision, and in adverse operative conditions. The authors devised a protocol primary repair of obstetric anal sphincter lacerations in order to improve surgical outcome and resident training. Second, the optimal method of repairing obstetric anal sphincter lacerations has not been fully determined. The feasibility of overlap versus end-to-end techniques has been examined in two randomized clinical trials, each with significantly different results. A multicenter randomized study comparing overlap versus end-to-end repair is required.