The role of antioxidants in ischaemia-reperfusion in a human DIEP flap model.

BACKGROUND: A better understanding of the pathophysiology of ischaemia-reperfusion injury and a possible treatment for it, is of great importance. The deep inferior epigastric perforator (DIEP) flap is an innovative, clinical model of ischaemia-reperfusion. There has been an ongoing interest in the health benefits and medical applications of antioxidants. We hypothesised that during ischaemia-reperfusion, specific antioxidants are depleted. METHODS: Seventeen DIEP flaps were performed in 15 patients undergoing breast reconstruction. In each free flap, 3-mm skin biopsies were taken from the DIEP flap at four different time points during and after surgery. In those tissue biopsies, concentrations of the antioxidants vitamin E, glutathione (GSH) and uric acid and total antioxidant capacity (TEAC) were measured. RESULTS: Unexpectedly, no immediate change was observed in GSH concentrations during ischaemia-reperfusion. Uric acid concentrations were significantly increased at all time points following reperfusion. Vitamin E concentrations also showed a significant incline 30min and 1h after reperfusion. However, 1h after reperfusion, a significant decrease in total hydrophilic antioxidant capacity (TEAC) was seen. In the next hour, this capacity recovered. CONCLUSIONS: During ischaemia-reperfusion, a deficiency in hydrophilic antioxidant capacity develops. This is a potential cause for the development of ischaemia-reperfusion injury by reactive oxygen species. This clinical trial is registered on Clinical Trials: http://www.clinicaltrials.gov/. Trial registry name: The DIEP-flap as a model of ischaemia-reperfusion. Registration identification number (NCT): 00482469.

The Effect of Statins and Other Cardiovascular Medication on Ischemia-Reperfusion Damage in a Human DIEP Flap Model: Theoretical and Epidemiological Considerations.

Background. Statins and other cardiovascular medication possess antioxidant capacity. It was examined whether chronic use of these medications protects from the development of ischemia-reperfusion (I/R) related complications after DIEP (Deep Inferior Epigastric Perforator Free Flap) surgery. This paper contains a literature study on the antioxidant working mechanisms of these drugs. Methods. Medical information of 134 DIEP patients (173 flaps) was studied from their medical files. Patient and operative characteristics were registered, as well as I/R related complications. Results. Of the group that didnot use statins, 16.3% developed complications versus 30.8% amongst patients that did use these drugs (P = 0.29). Amongst patients that chronically use other cardiovascular medication, 26.8% developed I/R related complications versus 14.4% of the patients without medication (P = 0.10). Conclusions. Chronic use of statins or other cardiovascular medication didnot decrease the occurrence of I/R related complications after DIEP surgery. Therefore, research should be aimed at evaluating short-term pre-treatment with statins.

Prevalence of urinary incontinence among community-dwelling adults receiving home care.

We conducted a cross-sectional survey in 2005 to determine the prevalence of and factors associated with urinary incontinence (UI) in adults receiving home care. Of the 2,866 patients surveyed, 46% suffered from UI; 6.5% had stress, 16.6% had urge, 9% had mixed, and 17.6% had functional incontinence. No diagnosis regarding type of UI had been established in 50.2%. Factors associated with UI were advanced age, higher body mass index, and impaired mobility. UI is prevalent in older persons receiving home care, but the lack of diagnosis of type of UI in half of the participants surveyed impedes management of UI.

Mitotic counting in surgical pathology: sampling bias, heterogeneity and statistical uncertainty.

Mitotic counting in surgical pathology: sampling bias, heterogeneity and statistical uncertainty Although several articles on the methodological aspects of mitotic counting have been published, the effects of macroscopic sampling and tumour heterogeneity have not been discussed in any detail. In this review the essential elements for a standardized mitotic counting protocol are described, including microscopic calibration, specific morphological criteria, macroscopic selection, counting procedure, effect of biological variation, threshold, and the setting of an area of uncertainty ('grey area'). We propose that the use of a standard area for mitotic quantification and of a grey area in mitotic counting protocols will facilitate the application of mitotic counting in diagnostic and prognostic pathology.

Does the risk of childhood diabetes mellitus require revision of the guideline values for nitrate in drinking water?

In recent years, several studies have addressed a possible relationship between nitrate exposure and childhood type 1 insulin-dependent diabetes mellitus. The present ecologic study describes a possible relation between the incidence of type 1 diabetes and nitrate levels in drinking water in The Netherlands, and evaluates whether the World Health Organization and the European Commission standard for nitrate in drinking water (50 mg/L) is adequate to prevent risk of this disease. During 1993-1995 in The Netherlands, 1,104 cases of type 1 diabetes were diagnosed in children 0-14 years of age. We were able to use 1,064 of these cases in a total of 2,829,020 children in this analysis. We classified mean nitrate levels in drinking water in 3,932 postal code areas in The Netherlands in 1991-1995 into two exposure categories. One category was based on equal numbers of children exposed to different nitrate levels (0.25-2.08, 2.10-6.42, and 6.44-41.19 mg/L nitrate); the other was based on cut-off values of 10 and 25 mg/L nitrate. We determined standardized incidence ratios (SIRs) for type 1 diabetes in subgroups of the 2,829,020 children with respect to both nitrate exposure categories, sex, and age and as compared in univariate analysis using the chi-square test for trend. We compared the incidence rate ratios (IRRs) by multivariate analysis in a Poisson regression model. We found an effect of increasing age of the children on incidence of type 1 diabetes, but we did not find an effect of sex or of nitrate concentration in drinking water using the two exposure categories. For nitrate levels > 25 mg/L, an increased SIR and an increased IRR of 1.46 were observed; however, this increase was not statistically significant, probably because of the small number of cases (15 of 1,064). We concluded that there is no convincing evidence that nitrate in drinking water at current exposure levels is a risk factor for childhood type 1 diabetes mellitus in The Netherlands, although a threshold value > 25 mg/L for the occurrence of this disease can not be excluded.

The DD genotype of the angiotensin converting enzyme gene is negatively associated with right ventricular hypertrophy in male patients with chronic obstructive pulmonary disease.

The renin angiotensin system plays an important role in the development of pulmonary artery remodeling and right ventricular hypertrophy in hypoxia-induced pulmonary hypertension as may occur in patients with COPD. Several polymorphisms of genes encoding for components of the renin angiotensin system such as the M235T polymorphism in the angiotensinogen gene, the 287-base-pair insertion (I)/deletion (D) polymorphism at intron 16 of the ACE gene, and the A1166C polymorphism in the angiotensin II type 1 receptor gene have been associated with an increased risk of cardiovascular diseases. With respect to the pulmonary circulation, only limited data exist on possible associations between polymorphisms of these genes and pulmonary hypertension and/or right ventricular hypertrophy. The objective of the present study was to investigate a possible relationship between polymorphisms of the renin angiotensin system and electrocardiographic evidence of right ventricular hypertrophy in patients with COPD. We therefore determined the angiotensinogen (M235T), angiotensin converting enzyme (I/D), and angiotensin II type 1 receptor (A1166C) genotypes in 87 patients with severe COPD and correlated these data with electrocardiographic parameters of right ventricular hypertrophy. Thirty-one patients (36%) of 87 patients with COPD showed electrocardiographic evidence of right ventricular hypertrophy. In the male, but not in the female, subgroup, the angiotensin-converting enzyme DD genotype was negatively associated with electrocardiographic evidence of right ventricular hypertrophy (male: chi2 = 3.8, p = 0.05; female: chi2 = 0.05, p = 0.82). We found no associations between the investigated polymorphisms in the angiotensinogen and angiotensin II type 1 receptor genes and electrocardiographic evidence of right ventricular hypertrophy.

Number and proliferation of clara cells in normal human airway epithelium.

Experimental pathologic studies suggest that Clara cells are one of the types of airway stem cells but the proliferation of Clara cells in human lungs has not yet been examined. The purpose of this study was to assess in conducting airways of normal human lungs: (1) the number of Clara cells; and (2) the contribution of Clara cells to the proliferation compartment. Samples of histologic normal tissue were taken from seven lungs obtained by autopsy. A triple sequential (immuno)histochemical staining was performed, using MIB-1 as a proliferation marker and anti-CC10 for the identification of Clara cells; subsequently, a PAS stain was carried out as a marker for goblet cells, as these cells were reported to be CC10-immunoreactive in an unknown proportion. Clara cells were virtually absent in the proximal airway epithelium. The number of Clara cells in the terminal bronchioles was 11 +/- 3% (mean +/- SD) and in respiratory bronchioles 22 +/- 5%. The overall proliferation compartment of the conducting airway epithelium was 0.83 +/- 0.47%; the contribution of Clara cells was 9%. In the terminal bronchioles 15% of proliferating airway epithelial cells were Clara cells, and in the respiratory bronchioles this number increased to 44%. The contribution of Clara cells to the proliferation compartment of normal human tracheobronchial epithelium is substantial, demonstrating a role of the Clara cell in the maintenance of the normal epithelium of the distal conducting airways in humans.

Validation of the Point-EXACCT method in non-small cell lung carcinomas.

K-ras point mutations are often detected in part of the lung carcinomas. For the validation of a highly sensitive and rapid assay for known point mutations, Point-EXACCT (Biochim Biophys Acta 1998; 1379:42-52), we analyzed 89 non-small cell lung carcinomas and compared the results with two sequencing methods. No point mutations were found with double-stranded sequencing. Single-stranded sequencing detected six patients positive for K-ras codon 12. When Point-EXACCT was used, K-ras codon 12 mutations were detected in 8 of 52 patients with squamous cell carcinomas, 10 of 29 patients with adenocarcinomas, and 3 of 8 patients with large cell carcinomas. The finding of K-ras mutations in squamous cell carcinomas is explained by the high sensitivity of the method. Therefore, Point-EXACCT may be applicable to detection of those alterations occurring at a low frequency among an excess of cells with wild-type DNA.

Number and proliferation of basal and parabasal cells in normal human airway epithelium.

Two roles have been suggested for basal cells on the basis of studies performed with laboratory animals: (1) anchoring of the tracheobronchial epithelium; and (2) being the epithelial stem cell. Parabasal cells located just above the basal cells have also been shown to contribute to cell renewal. However, a systematic study of the composition and proliferation of basal and parabasal cells in normal human lungs is lacking. The aims of this study were to determine in normal human conducting-airway epithelium: (1) the number of basal and parabasal cells; and (2) the contribution of basal and parabasal cells to the proliferation fraction. Samples of histologically normal tissue, free of pulmonary disease, were taken from seven lungs obtained by autopsy. Immunohistochemical staining was performed with the primary antibody MIB-1 as a proliferation marker and the antikeratin antibody 34betaE12 as a marker for basal and parabasal cells. In the largest conducting airways (diameter >= 4 mm), the percentages of basal and parabasal cells were 31% and 7%, respectively; the contribution to the proliferation compartment was 51% for basal and 33% for parabasal cells. In the smallest airways (diameter < 0.5 mm), 6% of epithelial cells were basal cells, with a 30% contribution to the proliferation compartment, whereas parabasal cells were absent. The high fraction of basal and parabasal cells contributing to the proliferation compartment of normal human conducting-airway epithelium supports the theory that cells at or near the basement membrane are likely to be progenitor cells.

The role of "colonization pressure" in the spread of vancomycin-resistant enterococci: an important infection control variable.

OBJECTIVE: The spread of nosocomial multiresistant microorganisms is affected by compliance with infection control measures and antibiotic use. We hypothesized that "colonization pressure" (ie, the proportion of other patients colonized) also is an important variable. We studied the effect of colonization pressure, compliance with infection control measures, antibiotic use, and other previously identified risk factors on acquisition of colonization with vancomycin-resistant enterococci (VRE). METHODS: Rectal colonization was studied daily for 19 weeks in 181 consecutive patients who were admitted to a single medical intensive care unit. A statistical model was created using a Cox proportional hazards regression model including length of stay in the medical intensive care unit until acquisition of VRE, colonization pressure, personnel compliance with infection control measures (hand washing and glove use), APACHE (Acute Physiology and Chronic Health Evaluation) 11 scores, and the proportion of days that a patient received vancomycin or third-generation cephalosporins, sucralfate, and enteral feeding. RESULTS: With survival until colonization with VRE as the end point, colonization pressure was the most important variable affecting acquisition of VRE (hazard ratio [HR], 1.032; 95% confidence interval [C1], 1.012-1.052; P=.002). In addition, enteral feeding was associated with acquisition of VRE (HR, 1.009; 95% CI, 1.000-1.017; P=.05), and there was a trend toward association of third-generation cephalosporin use with acquisition (HR, 1.007; 95% CI, 0.999-1.015; P=.11). The effects of enteral feeding and third-generation cephalosporin use were more important when colonization pressure was less than 50%. Once colonization pressure was 50% or higher, these other variables hardly affected acquisition of VRE. CONCLUSIONS: Acquisition of VRE was affected by colonization pressure, the use of antibiotics, and the use of enteral feeding. However, once colonization pressure was high, it became the major variable affecting acquisition of VRE.

Which cytological criteria are the most discriminative to distinguish carcinoma, lymphoma, and soft-tissue sarcoma? A probabilistic approach.

The reliability of fine-needle aspiration cytology (FNA) for distinguishing between carcinoma, lymphoma, and sarcoma was established in a previous study (Thunnissen et al., Cytopathology 1993; 4:107-114). The purpose of this study was to investigate which criteria were useful for a probabilistic diagnosis. A total of 78 randomly chosen FNA smears (31 carcinomas, 24 lymphomas, and 23 sarcomas) was sent around and read "blindly" by six cytopathologists. Each pathologist completed a list of 16 criteria for every case. Histology was used as a reference standard. A statistical analysis led to the selection of three criteria: "lymphoglandular bodies," "well-defined clusters," and "spindle-cell nuclei," associated with lymphoma, carcinoma, and soft-tissue sarcoma, respectively. Given the scores on these criteria, the probabilities to be assigned to the three diagnostic categories can be read from a table. It turns out, as one might expect, that the classification of the most probable disease is pretty reliable if one cytologic criterion scores much higher than the other two criteria. On other cases, fuzziness appears and misclassifications are far from improbable. This study offers a general cytologic approach. The cytologic criteria "lymphoglandular bodies," "well-defined clusters," and "spindle-cell nuclei" can be used both in daily practice and in education to assign posterior probabilities to carcinoma, lymphoma, and soft-tissue sarcoma.

The risk factor profile of recto-sigmoid adenomas: a prospective screening study of 665 patients in a clinical rehabilitation centre.

During a prospective screening study for recto-sigmoid adenomatous polyps, the influence of the following risk factors was evaluated: age; gender; body mass index; heredity for colorectal malignancy; diabetes; hypertension; constipation; previous gastric surgery; previous gastric acid inhibition; alcohol and cigarette consumption; serum cholesterol; serum triglycerides; and serum gastrin. Screening fibre-sigmoidoscopy of 665 patients (aged between 50 and 60 years) at a clinical rehabilitation centre for gastrointestinal and metabolic diseases showed that 146 had one or several adenomas. The study population was overweight by a mean of about 15%. Comparison of those with and those without adenoma using univariate analysis, showed that the group with adenomas had higher serum triglyceride values, drank more alcohol on a regular or excessive basis, were more frequent smokers, and had a tendency to raised fasting serum glucose. In a multivariate analysis, age, high serum triglycerides and high alcohol consumption were risk factors for recto-sigmoid adenomas. The risk factor profile identified in this study may help in the selection of individuals for screening sigmoidoscopy from a similar background population. It also identifies target conditions for primary prevention of colorectal neoplasia.

Detrusor activity index: quantification of detrusor overactivity by ambulatory monitoring.

PURPOSE: Ambulatory urodynamic monitoring has been developed and evaluated during the last decade, and its appreciation is growing rapidly. We investigated the usefulness of this technique to quantify detrusor activity, and developed a parameter to quantify the grade of detrusor overactivity. MATERIALS AND METHODS: We investigated 167 female patients with lower urinary tract symptoms and 61 asymptomatic volunteers. Detrusor behavior was classified according to the medical history findings and independently based on urodynamic investigation. The possible categories were termed overactive or not overactive. Subjects who were classified consistently formed the 2 reference groups, which were used in the statistical analysis of logistic regression. RESULTS: By logistic regression a scoring rule was developed. The individual score (detrusor activity index 0 to 1) is a measure of detrusor activity during the filling phase. At a cutoff point of 0.41 the sensitivity equaled specificity of 85% in our patient and volunteer groups. CONCLUSIONS: The scoring rule can be applied clinically as a test to detect and quantify detrusor overactivity in patients with urinary incontinence.

Risk factors for pneumonia, and colonization of respiratory tract and stomach in mechanically ventilated ICU patients.

Risk factors for the development of ventilator-associated pneumonia (VAP) and colonization of the respiratory tract and stomach with enteric gram-negative bacteria (EGB) and Pseudomonadaceae were determined in 141 ventilated patients using univariate analysis and the Cox proportional hazards model. VAP was caused by EGB in 14 patients (10%), and by Pseudomonadaceae in 19 patients (13%). The duration of ventilation was a significant risk factor for VAP caused by EGB and Pseudomonadaceae, and for acquired colonization in oropharynx, stomach, and trachea with these species. Of 20 other variables, oropharyngeal colonization with EGB on admission (hazard ratio [HR] = 4.5) and an infection on admission (HR = 2.7) were selected as risk factors for VAP caused by EGB. Acquired colonization with Pseudomonadaceae in oropharynx (HR = 5.0) was the most important risk factor for VAP caused by these species. Gastric colonization with EGB or Pseudomonadaceae were no risk factors for VAP. For acquired oropharyngeal colonization with EGB only the duration of ventilation was a risk factor, whereas preceding colonization of the trachea with Pseudomonodaceae and duration of ventilation were risk factors for acquired oropharyngeal colonization with these species. In the Cox model, only the duration of ventilation was significantly related to acquired gastric colonization with EGB. Preceding colonization of the orophayrnx and of the trachea with Pseudomonadaceae were risk factors for acquired colonization with these species in the stomach. Twelve patients with VAP (46%) and 38 without VAP (33%) died (p = 0.21). In conclusion, duration of ventilation and colonization of the upper respiratory tract are the most important risk factors for VAP caused by EGB or Pseudomonadaceae.

Ambulatory urodynamics: extramural testing of the lower and upper urinary tract by Holter monitoring of cystometrogram, uroflowmetry, and renal pelvic pressures.

This article elucidates the clinical applicability and state of the art of ambulatory urodynamics. Ambulatory urodynamics have evolved into practical investigations like EAC, HFM, and EAC combined with renal pelvimetry. EAC has been shown to be the method of preference if detrusor overactivity is involved. Conventional filling cystometry has proved to be an unreliable way to exclude detrusor instability. De novo instability after suspension surgery often indicates that an existing detrusor overactivity was not identified preoperatively. EAC including flowmetry has shown considerable variance in obstructive and contractility parameters in males with LUTS indicative for BPH. This raises doubt whether the clinical flow analysis is the suitable "gold standard" as advocated by the ICS. For a real break through of EAC, less complex automatic analysis is necessary. HFM is a newer method within the range of ambulatory urodynamic tests. It has not yet been completely evaluated. But, because the technique is analogous to the office flowmetry, noninvasive and very well accepted by the patients, it is expected to be widely used. This expectation is strengthened by the fact that HFM seems to show individual therapeutic efficacy of drugs, such as alpha-blockers. As a research tool to evaluate efficacy, it is far more powerful than conventional methods because of the reduction of within-patient standard deviation to about 10%. Finally, EAC combined with pelvimetry offers a promising method for the clinical evaluation of a combined dysfunction of upper and lower urinary tract.

The discriminative value of patient characteristics and dyspeptic symptoms for upper gastrointestinal endoscopic findings: a study on the clinical presentation of 1,147 patients.

The discriminative value of patient characteristics and dyspeptic symptoms for upper gastrointestinal endoscopic findings was prospectively assessed in 1,147 patients attending for their first diagnostic endoscopy and who answered paper (n = 431) or computerized (n = 716) questionnaires. The questionnaires provided detailed information concerning present dyspeptic symptoms, with special attention to provoking and/or relieving factors, and smoking and/or drinking habits. In logistic regression models each of a number of 'specific endoscopic diagnoses' was contrasted with normal endoscopy (n = 390), and 'relevant endoscopic disease' (oesophagitis, peptic ulcers, cancers; n = 269) was contrasted with 'irrelevant' and normal endoscopic findings (n = 878). From the regression model a receiver operating characteristic (ROC) curve could be constructed, and the area under the ROC curve (AUC) was calculated to summarize the discriminative power of the regression model. The best discrimination from patients with a normal endoscopy was achieved for patients with gastric (AUC = 0.86) or duodenal (AUC = 0.85) ulcers, followed by patients with hiatus hernia (AUC = 0.78 or oesophagitis (AUC = 0.77). The discriminative performance of the regression models was somewhat less for duodenitis/bulbitis (AUC = 0.75) and endoscopic gastritis (AUC = 0.73). In an open-access endoscopy unit setting, the value of preinvestigation history-taking for the prediction of clinically relevant endoscopic disease was very limited (AUC = 0.63).

Appropriateness of indications for diagnostic upper gastrointestinal endoscopy: association with relevant endoscopic disease.

BACKGROUND: Since the institution of open access endoscopy units there has been a considerable increase of referrals for UGI examinations. Therefore, guidelines for the appropriate use of UGI endoscopy are needed. METHODS: The outcome of first diagnostic UGI endoscopy was prospectively assessed for several referral indications in a consecutive series of 2900 patients. Indications were judged "appropriate" when significantly (p < 0.01) associated with clinically "relevant" endoscopic findings. RESULTS: The proportion of relevant disease for various indications was as follows: signs of UGI bleeding (42.2%); history of peptic ulcer (40.5%); dysphagia (31.9%), short-term (24.4%), and without therapy (20.9%). Relevant endoscopic findings were observed in 21.0% of dyspeptic patients aged 45 years or less, and in 25.3% of those older than 45 years of age. CONCLUSIONS: The generally approved alarm symptoms should be a reason to perform endoscopy without hesitation. Dyspeptic symptoms, despite adequate empiric treatment, as well as first dyspeptic symptoms in patients older than 45 years should also be a reason for endoscopic investigation. Our results support the strategy to treat patients younger than 45 years who have isolated dyspepsia by a limited course of antipeptic agents, provided that they are seen for re-evaluation within 4 to 6 weeks.