Comparison of different methods for the recovery of DNA from spores of mycotoxin-producing moulds in spiked food samples.

Several food samples were spiked with fungal conidia to test the efficiency of different cell disruption methods and DNA extraction kits for subsequent molecular detection. For disrupting the firm cell walls of the spores, two different pretreatment methods, namely sonication and bead beating, were tested against no pretreatment. The subsequent DNA extraction and purification was performed using three different DNA extraction methods, which are based on a diverse combination of extraction principles, such as precipitation, thermic-enzymatic lysis, pH-enhancement and bonding with a silica membrane. The aim of the study was to find out the suitable pretreatment and DNA extraction method for the recovery of detectable amounts of fungal DNA from different food matrices. Significance and impact of the study: The choice of 'ready-to-use' commercial kits and methods has been of great importance regarding the recovery of extracted DNA. However, these commercially available kits are neither effective nor time-efficient when extracting DNA from fungal spores embedded in complex food matrices. Different extraction principles were compared and their effectiveness tested using real-time PCR. The combination of different principles for the extraction and purification of DNA was found as the most efficient method (quantity and purity) to obtain DNA from moulds and their spores from food samples.

A single gene defect causing claustrophobia.

Claustrophobia, the well-known fear of being trapped in narrow/closed spaces, is often considered a conditioned response to traumatic experience. Surprisingly, we found that mutations affecting a single gene, encoding a stress-regulated neuronal protein, can cause claustrophobia. Gpm6a-deficient mice develop normally and lack obvious behavioral abnormalities. However, when mildly stressed by single-housing, these mice develop a striking claustrophobia-like phenotype, which is not inducible in wild-type controls, even by severe stress. The human GPM6A gene is located on chromosome 4q32-q34, a region linked to panic disorder. Sequence analysis of 115 claustrophobic and non-claustrophobic subjects identified nine variants in the noncoding region of the gene that are more frequent in affected individuals (P=0.028). One variant in the 3'untranslated region was linked to claustrophobia in two small pedigrees. This mutant mRNA is functional but cannot be silenced by neuronal miR124 derived itself from a stress-regulated transcript. We suggest that loosing dynamic regulation of neuronal GPM6A expression poses a genetic risk for claustrophobia.

Biowaiver monographs for immediate release solid oral dosage forms: quinidine sulfate.

Literature data are reviewed relevant to the decision to allow a waiver of in vivo bioequivalence (BE) testing for the approval of new multisource and reformulated immediate release (IR) solid oral dosage forms containing quinidine sulfate. Quinidine sulfate's solubility and permeability, its therapeutic use and index, pharmacokinetics, excipient interactions and reported BE/bioavailability (BA) problems were taken into consideration. The available data are not fully conclusive, but do suggest that quinidine sulfate is highly soluble and moderately to highly permeable and would likely be assigned to BCS Class I (or at worst BCS III). In view of the inconclusiveness of the data and, more important, quinidine's narrow therapeutic window and critical indication, a biowaiver based approval of quinidine containing dosage forms cannot be recommended for either new multisource drug products or for major postapproval changes (variations) to existing drug products.

Determination and evaluation of the phosphorus load of an artificial shallow lake.

Enhanced eutrophication of lakes due to high nutrient loads from anthropogenic sources has become a worldwide problem. Dying ecosystems and limitation of uses are the consequences. In Bochum, Germany, Lake Umminger is an integral part of a recreation area, but also receives high nutrient loads from the local sewer system, as could be shown with the help of water and nutrient balances. Mass algae growth, the dying of fish and production of digestion gas implied a demand to rehabilitate the lake. Primarily, the urgency and sanitation potential as well as the applicability of external and internal enhancement measures had to be evaluated. The trophic classification needed was based upon the German guideline for the classification of the water quality of natural lakes according to trophic criteria, mainly using Vollenweider's eutrophication model. This paper focuses on a description and analysis of the problems that arose during the application of this model to Lake Umminger, stating that shallow, artificial lakes cannot be evaluated correctly with the existing methods. Although some suggestions for further improvement are given, the development of new evaluation criteria was not in the scope of the study presented.

Outcome after laminectomy for lumbar spinal stenosis. Part I: Clinical correlations.

All patients who underwent decompressive lumbar laminectomy in the Washtenaw County, Michigan metropolitan area during a 7-year period were studied for the purpose of defining long-term outcome, clinical correlations, and the need for subsequent fusion. Outcome was determined by questionnaire and physical examination from a cohort of 119 patients with an average follow-up evaluation interval of 4.6 years. Patients graded their outcome as much improved (37%), somewhat improved (29%), unchanged (17%), somewhat worse (5%), and much worse (12%) compared to their condition before surgery. Poor outcome correlated with the need for additional surgery, but there were few additional significant correlations. No patient had a lumbar fusion during the study interval. The outcome after laminectomy was found to be less favorable than previously reported, based on a patient questionnaire administered to an unbiased patient population. Further randomized, controlled trials are therefore necessary to determine the efficacy of lumbar fusion as an adjunct to decompressive lumbar laminectomy.

Outcome after laminectomy for lumbar spinal stenosis. Part II: Radiographic changes and clinical correlations.

The pre- and postoperative lumbar spine radiographs of 119 patients who underwent decompressive lumbar laminectomy were studied to evaluate radiographic changes and to correlate them with clinical outcome. An accurate and reproducible method was used for measuring pre- and postoperative radiographs that were separated by an average interval of 4.6 years. Levels of the spine that underwent laminectomy showed greater change in spondylolisthesis, disc space angle, and disc space height than unoperated levels. Outcome correlated with radiographic changes at operated and unoperated levels. This study demonstrates that radiographic changes are greater at operated than at unoperated levels and that some postoperative symptoms do correlate with these changes. Lumbar fusion should be considered in some patients who undergo decompressive laminectomy. The efficacy of and unequivocal indications for lumbar fusion can only be determined from randomized, prospective, controlled trials, however, and these studies have not yet been undertaken.

Clinical characteristics and surgical management of vascular complications in patients undergoing cardiac catheterization: interventional versus diagnostic procedures.

The purpose of this report is to define the clinical characteristics and outcome of surgical management of vascular complications after interventional cardiac catheterization and to contrast them to those after diagnostic cardiac catheterization. From October 1985 to December 1989, 101 patients were treated for 106 vascular complications after 1866 interventional and 5046 diagnostic cardiac catheterizations at the University of Michigan Medical Center. Interventional catheterizations resulted in 69 vascular complications in 64 patients (frequency 3.4%). The most common interventions included coronary angioplasty (34), of which 10 required percutaneous partial cardiopulmonary bypass, intraaortic balloon pump placement (14), and aortic valvuloplasty (11). Interventional catheter-related complications included hemorrhage (33), arterial thrombosis (18), pseudoaneurysm formation (12), catheter embolization (2), thromboembolism (2), as well as arteriovenous fistula, pseudoaneurysm, and arterial dissection (1 each). Fifteen of these 69 patients (24%) had suffered acute myocardial infarction just before their catheterization. Surgical repair was performed under local anesthesia in 70% of patients. Major vascular reconstructions were required in 9% of patients. Three percent of the involved lower extremities had to be amputated because of complications occurring after arterial puncture. Eight percent of the patients incurring vascular complications after interventional procedures died after operation. Diagnostic catheterizations resulted in 37 vascular complications in 37 patients (frequency 0.7%). In contrast to diagnostic cardiac catheterization, vascular complications after interventional cardiac catheterization occurred more frequently, were most often due to hemorrhage at the vascular access site, and occurred in high-risk, critically ill patients.(ABSTRACT TRUNCATED AT 250 WORDS)