Sexual problems in MS: Sex differences and their impact on quality of life.

BACKGROUND: Low sexual function and satisfaction are common problems among people with multiple sclerosis (PwMS), but the literature on which patient variables are associated with these issues is inconsistent. OBJECTIVE: To investigate the associations between sexual function and satisfaction in PwMS with clinical, demographic, and patient-reported quality of life (QOL) measures and determine if sex differences exist. METHODS: This analysis includes PwMS enrolled in the Comprehensive Longitudinal Investigation of Multiple Sclerosis at the Brigham and Women's Hospital (CLIMB), who completed patient-reported outcome measures: Multiple Sclerosis Quality of Life-54 (MSQOL-54), Modified Fatigue Impact Scale (MFIS), and Center for Epidemiologic Studies Depression Scale (CES-D). Regression models were used to analyze associations between patient variables and function and satisfaction. Results were stratified by sex. Cross-sectional and longitudinal data were used. RESULTS: 702 PwMS (526 females,176 males, mean age 42.2 +/-11.1, median EDSS 1.5) were included in the cross-sectional analysis. Data from 341 PwMS were used in the three-year longitudinal analysis. Increasing age, disease duration, and disability were associated with reduced sexual function and satisfaction to the same degree in males and females. However, sex differences existed in the strength of associations with QOL variables. There was no significant longitudinal change in females or males. CONCLUSIONS: Age and disease duration were associated with reduced sexual function and satisfaction in males and females. In females, function was significantly associated with disability and satisfaction with fatigue. Males had stronger associations with sexual function in domains related to emotional well-being, health perceptions, and overall QOL. Males had stronger associations with satisfaction in emotional and social functioning and physical health domains. These findings can help better understand the multidimensional problems of sexual function and satisfaction in PwMS and better guide patient care.

Response to Comment on "Sensitivity of seafloor bathymetry to climate-driven fluctuations in mid-ocean ridge magma supply".

Tolstoy reports the existence of a characteristic 100 thousand year (ky) period in the bathymetry of fast-spreading seafloor but does not argue that sea level change is a first-order control on seafloor morphology worldwide. Upon evaluating the overlap between tectonic and Milankovitch periodicities across spreading rates, we reemphasize that fast-spreading ridges are the best potential recorders of a sea level signature in seafloor bathymetry.

Response to Comment on "Sensitivity of seafloor bathymetry to climate-driven fluctuations in mid-ocean ridge magma supply".

Huybers et al present new bathymetric spectra from an intermediate-spreading ridge as evidence for a primary contribution of sea level cycles to the morphology of the seafloor. Although we acknowledge the possibility that sea level-modulated magmatic constructions may be superimposed on a first-order tectonic fabric, we emphasize the difficulty of deciphering these different contributions in the frequency domain alone.

Sensitivity of seafloor bathymetry to climate-driven fluctuations in mid-ocean ridge magma supply.

Recent studies have proposed that the bathymetric fabric of the seafloor formed at mid-ocean ridges records rapid (23,000 to 100,000 years) fluctuations in ridge magma supply caused by sealevel changes that modulate melt production in the underlying mantle. Using quantitative models of faulting and magma emplacement, we demonstrate that, in fact, seafloor-shaping processes act as a low-pass filter on variations in magma supply, strongly damping fluctuations shorter than about 100,000 years. We show that the systematic decrease in dominant seafloor wavelengths with increasing spreading rate is best explained by a model of fault growth and abandonment under a steady magma input. This provides a robust framework for deciphering the footprint of mantle melting in the fabric of abyssal hills, the most common topographic feature on Earth.

[Thoracic actinomycosis: diagnostic pitfalls and therapeutic considerations]. / Thorakale Aktinomykose: diagnostische Schwierigkeiten und therapeutische Uberlegungen.

We report two patients admitted to our hospital suspected to suffer from cancer in the lung or mediastinum, respectively. Both patients had a diagnosis of thoracic actinomycosis. A 76 year old man revealed pulmonary and endobronchial actinomycosis associated with broncholithiasis. Diagnosis was achieved by bronchoscopy. Therapy with ampicillin/sulbactam was successful. A 36 year old patient presented with bilateral pleural effusions, extended pericardial, mediastinal and pulmonary actinomycosis with pericarditis constrictiva and superior vena cava syndrome. Diagnosis was finally made by cardiac surgery with therapeutic pericardectomy. Prolonged therapy with ampicillin/sulbactam was administered with satisfactory result. Here we discuss the importance to include actinomycosis in the differential diagnosis of pulmonary affections and mediastinal masses in order to avoid diagnostic errors and to limit invasive procedures to the necessary amount. We illustrate the need of an individualized treatment approach.

[Bullectomy as emergency intervention in a mechanically ventilated patient with refractory type II respiratory failure due to chronic obstructive lung disease (COPD)]. / Erfolgreiche Notfall-Bullektomie bei einer invasiv beatmeten Patientin mit refraktärer ventilatorischer Insuffizienz im Rahmen einer akuten Exazerbation einer chronisch-obstruktiven Lungenerkrankung (COPD).

A 44-year-old female patient presented with an extensive exacerbation of severe chronic obstructive lung disease (COPD) and bullous emphysema. Because of a severe type II respiratory failure, the patient was intubated and mechanically ventilated. Respiratory failure was refractory despite appropriate ventilation regimes and pCO2 values ranged from 110 mm Hg to 180 mm Hg. Chest radiography revealed hyperinflation of two giant bullae with mediastinal shifting to the left lung. We describe a successful rescue bullectomy.

Differential frequencies of p16(INK4a) promoter hypermethylation, p53 mutation, and K-ras mutation in exfoliative material mark the development of lung cancer in symptomatic chronic smokers.

PURPOSE: The aim of this study was to investigate the frequency of three (epi)genetic alterations (p53 and K-ras mutations and p16(INK4a) promoter hypermethylation) in symptomatic chronic smokers compared with patients with lung cancer and to evaluate the use of exfoliative material for such analyses. PATIENTS AND METHODS: Fifty-one patients with histologically confirmed lung cancer and 25 chronic smokers (> 20 pack-years) were investigated for mutations in the K-ras (codon 12) and p53 (codons 248, 249, and 273) genes and for allelic hypermethylation of the p16(INK4a) gene. DNA was isolated from sputum and bilateral bronchial lavage, and brushings were taken at bronchoscopy. RESULTS: Forty-one genetic lesions were detected within exfoliative material from the group of 51 patients with lung cancer and 10 lesions in the chronic smoker group. K-ras mutations occurred exclusively in the lung cancer group, whereas p53 mutations and p16(INK4a) promoter hypermethylation were also found in chronic smokers. Three of eight chronic smokers who harbored an (epi)genetic alteration were subsequently diagnosed with lung cancer. Analysis of sputum yielded information equivalent to that of samples obtained during bronchoscopy. CONCLUSION: p16(INK4a) promoter hypermethylation and p53 mutations can occur in chronic smokers before any clinical evidence of neoplasia and may be indicative of an increased risk of developing lung cancer or of early disease. K-ras mutations occur exclusively in the presence of clinically detectable neoplastic transformation. Molecular analysis of sputum for such markers may provide an effective means of screening chronic smokers to enable earlier detection and therapeutic intervention of lung cancer.

Facilitated detection of oncogene mutations from exfoliated tissue material by a PNA-mediated 'enriched PCR' protocol.

An 'enriched polymerase chain reaction (PCR)' protocol has been established for the sensitive detection of oncogene mutations in body fluid samples from cancer patients. This two-step protocol combines an allele-specific PCR clamping step followed by a PCR-RFLP (restriction fragment length polymorphism) confirmatory step. The method thus resembles a nested PCR technique starting directly from genomic DNA material and, in no more than 54 PCR cycles, allows the sensitive detection of one mutant allele in 10(3) normal alleles. This protocol was tested on bronchial cytology samples and sputum taken from lung cancer patients and point mutations could be detected both in codon 12 of K-ras and in three codons (248, 249, and 273) of the p53 gene. Comparing this protocol with a different 'enriched PCR' method based on repetitive PCR-RFLP steps, a high concordance was noted between the two methods. Although the present protocol seems to be less sensitive by approximately one order of magnitude, it is much easier to perform and thus could be applied to the rapid but sensitive detection of allelic subfractions in a population of cells derived from exfoliative material.

Expression of alternatively spliced mdm2 transcripts correlates with stabilized wild-type p53 protein in human glioblastoma cells.

A puzzling finding in various human tumors, including glioblastoma multiforme (GBM), is the stabilization of wild-type (wt) p53 protein. The biological significance of this phenomenon and the mechanism by which it occurs are unexplained. Recent reports have revealed that mdm2 exerts its negative regulation on the p53 signal by directly binding p53 protein and thereby instigating its proteasomal degradation. mdm2 has been shown to exist in alternatively spliced forms in human ovarian and bladder carcinomas, and recently in GBM, with loss or disruption of its p53 binding domain. Here we report that alternatively spliced transcripts of mdm2 are present in 7 of 16 human GBM primary cell cultures and in the established GBM cell lines LN 229 and LN 18. Sequencing demonstrated loss of the amino terminal p53 binding domain in these alternatively spliced mdm2 transcripts, and an out-of-frame splicing in the majority of cases. A significant correlation between the presence of mdm2 splice variants and increased expression of wt p53 protein was observed. Furthermore, in the presence of an mdm2 splice variant, wt p53 stabilization occurred despite coincident MDM2 amplification. Our findings suggest that wt p53 protein stabilization may arise as a consequence of alternative splicing of mdm2. Such a mechanism might account for wt p53 protein accumulation in GBM cells, even in the presence of MDM2 gene amplification.

Functional domains in cyclin D1: pRb-kinase activity is not essential for transformation.

Although cyclin D1 plays a major role during cell cycle progression and is involved in human tumourigenesis, its domain structure is still poorly understood. In the present study, we have generated a series of cyclin D1 N- and C-terminal deletion constructs. These mutants were used to define the domains required for transformation of rat embryonal fibroblasts (REF) in cooperation with activated Ha-ras and and to establish correlations with defined biochemical properties of cyclin D1. Protein binding and REF assays showed that the region of the cyclin box required for the interaction with CDK4 as well as C-terminal sequences determining protein stability were crucial for transformation. Surprisingly, however, the N-terminal deletion of 20 amino acids which impaired pRb kinase activity did not affect the transforming ability of cyclin D1. Likewise, no effect on transformation was observed with mutants defective in p21CIP interaction. These observations argue against a crucial role of pRb inactivation or p21CIP squelching in cyclin D1-mediated transformation.

Simple and reliable factor V genotyping by PNA-mediated PCR clamping.

Resistance to activated protein C (APC resistance) is the most common cause of thrombophilia and linked to a single point mutation in the factor V gene (G-->A transition at nucleotide 1691). In the past, several PCR based methods have been proposed to determine the allelostatus of individual patients from small amounts of blood DNA including PCR followed by restriction fragment length polymorphism detection (PCR-RFLP), PCR using sequence-specific primers (PCR-SSP) and oligonucleotide ligation assay (OLA). Here, we present a novel approach based on the method of peptide nucleic acid(PNA)-mediated PCR clamping which is extremely sensitive to base pair mismatches. If PNAs specific for the two allelic variants are applied separately in each case a clear discrimination between a heterozygous or homozygous normal or homozygous Factor V Leiden status is possible and no further confirmation step is required. In a prospective study, 60 patients with suspected venous thrombosis events were tested and compared to the conventional PCR-RFLP technique. The concordance between both methods was 100%. PNA-based factor V genotyping, therefore, should be considered for large scale screening of those patients considered to be at risk for deep venous thrombosis.

Frequent detection of ras and p53 mutations in brush cytology samples from lung cancer patients by a restriction fragment length polymorphism-based "enriched PCR" technique.

RFLP-mediated PCR has been successfully applied as a reliable tool in the detection of ras mutations in many cancers and provides a basis for "mutant-enriched PCR" protocols. We have, therefore, modified this technique to the sensitive detection of K-ras codon 12 and also p53 "hot spot" mutations, which, frequently in lung cancer, affect codons at the positions 157, 175, 245, 248, 249, and 273. With a high sensitivity of 1 mutant allele in 10(4) normal alleles, our enrichment assay allows the detection of oncogene alleles when only a few tumor cells are present within a normal cell population. Brush cytology material obtained from the tumor site of 20 patients with endoscopically apparent bronchial carcinoma was compared to macroscopically normal mucosa taken from the contralateral bronchus ("control" cytology). We found K-ras codon 12 mutations in 5 cases (25%) and p53 mutations in 13 cases (65%) in the tumor-derived cell material but, with the exception of two cases, not in cell material taken from the control cytology. Seventy-five % of the samples analyzed showed that at least one of the two oncogenes was affected. In several cases, two p53 lesions were detected concomitantly. The majority of the mutations could be reconfirmed by an alternative approach exploiting changes in the genomic RFLP pattern induced by these mutations and were also demonstrated in separate diagnostic biopsies taken. Thus, we conclude that the established enriched PCR protocol ensures a high sensitivity and preserved specificity for the diagnosis of oncogene lesions associated with lung cancer. Because conventional techniques normally yield a lower incidence of corresponding ras and p53 mutations, we think that both the high rate and the heterogeneity of p53 mutations found in some cases are, indeed, related to the increased sensitivity of this new enriched PCR technique.

Sensitive detection of p53 gene mutations by a 'mutant enriched' PCR-SSCP technique.

For the rapid and sensitive detection of p53 'hot spot' mutations, we combined polymerase chain reaction based single-strand conformational polymorphism (PCR-SSCP) analysis with sequence specific-clamping by peptide nucleic acids (PNAs) in a one-step reaction tube protocol. For this purpose, we designed two PNA molecules comprising aa 246-250 of exon 7 and aa 270-275 of exon 8, respectively, to suppress the amplification of wild-type p53 allelic variants during PCR amplification. Using this method in a survey of 20 brush cytology samples from lung cancer patients, we were able to detect five p53 point mutations occurring in codons 248, 249 and 273 which could not be retrieved by conventional PCR-SSCP. Thus, allelic suppression by PNA molecules opens a way to largely improve the sensitivity of existing PCR-SSCP protocols (approximately 10-50-fold) and could be useful in the detection of 'hot spot' oncogene lesions in histological samples containing only a small number of cancer cells.

A novel small-cell lung carcinoma-specific DNA-binding activity related to the transcription factor ap-1.

A novel TRE-binding protein complex was detected specifically in 12 out of 13 small cell lung carcinoma (SCLC) cell lines. This complex was characterised by a lower electrophoretic mobility than the 'ubiquitous' complex present in all other carcinoma cell lines analysed. As shown by UV-crosslinking and South-Western blotting, the SCLC-specific complex contains a protein with an apparent M(r) > 100 kD, which is far bigger than all Fos and Jun proteins described to date. In addition, the DNA-binding specificity of this complex is different from the specificity of the 'ubiquitous' complex or a Fos/Jun heterodimer.