Single temperature endonuclease‐enhanced single‐strand conformation polymorphism/heteroduplex mutation analysis: evaluation of the method

Circulating neutrophils were investigated in 15 patients with Crohn's disease (CD), 15 with ulcerative colitis (UC), and 15 healthy volunteers. Dose-response curves for Chemotaxis in Boyden chambers were analysed for sensitivity to leukotriene B₄ (LTB₄), its 20-hydroxy-LTB₄ (20-OH-LTB₄) and 20-carboxy-LTB₄ (20-COOH-LTB₄) catabolites, and 5- and 15-hydroxyeicosatetraenoic acids (HETEs). Positive controls included: complement 5a (C5a), formy-L-methionyl-L-leucyl-L-phenylalanine (f-Met-Leu-Phe), and casein. Control Chemotaxis test were performed at concentrations yielding optimal responses in leucocytes of healthy volunteers. Chemotaxis to suboptimal concentrations of LTB₄ 1.0 and 3.2 nmol/l, and 5-HETE 316 nmol/l, was markedly depressed in patients with chronic inflammatory bowel disease (CIBD). Analyses of individual dose-response curves revealed an underlying decreased sensitivity to LTB₄ in 11 out of 30 patients, to 5-HETE in 10 out of 30 patients with a corresponding decrease of median sensitivity to LTB₄ and 5-HETE in both CD and UC. Peak responses to LTB₄, 5-HETE, f-Met-Leu-Phe, and casein were identical in the three groups tested, whereas the C5a values were significantly depressed in both groups of patients (p<0.05). The potency of LTB₄ exceeded that of 5-HETE by a factor of approximately 100 whereas 20-OH-LTB₄ was nearly as potent as LTB₄. 20-COOH-LTB₄ and 15-HETE did not activate Chemotaxis of human neutrophils. These findings are suggestive of a competitive inhibition of receptors with heterogeneity for LTB₄ and 5-HETE. The depressed sensitivity of peripheral neutrophils to the potent lipoxygenase metabolites of arachidonic acid, LTB₄ and 5-HETE, in CIBD may tend to limit their inflammatory recruitment in analogy with previous findings concerning Chemotaxis to C5a.     

PCR–restriction fragment length polymorphism analysis using groEL gene to differentiate pathogenic Vibrio species

Important pathogenic Vibrio species were differentiated by PCR–restriction fragment length polymorphism analysis. A 1117-bp groEL gene product was amplified using universal primers and digested using the restriction enzymes NruI or XbaI, revealing unique digestion patterns for each of the 10 Vibrio species, of which 7 were pathogenic in humans, along with 2 other species pathogenic in fish.     

Report of 2 indigenous cases of leprosy from a European country: use of polymerase chain reaction–restriction fragment length polymorphism analysis of hsp65 gene for identification of Mycobacterium leprae directly from a clinical sample

In this article, we report on 2 indigenous cases of leprosy detected in a European country. We also report on the use of polymerase chain reaction–restriction fragment length polymorphism analysis of hsp65 gene for rapid identification of Mycobacterium leprae directly from the clinical sample.     

Hydrogen breath testing versus LCT genotyping for the diagnosis of lactose intolerance: a matter of age?

BACKGROUND: Two single nucleotide polymorphisms (-13910 C/T and -22018 G/A) upstream of the lactase gene (LCT) have been found to be associated with lactose tolerance in Europeans. METHODS: In one hundred and twenty Austrian outpatients, who visited the physician's office for symptoms of irritable bowel syndrome (IBS), hydrogen breath testing (HBT) and LCT genotyping by polymerase chain reaction and reverse-hybridisation were performed in parallel. RESULTS: The coincidence between a genotype suggesting lactase non-persistence (lactose intolerance) and a positive HBT result was almost perfect (97.4% for LCT-13910 C/T and 100% for LCT-22018 G/A). Between a genotype indicating lactase persistence (lactose tolerance) and a negative HBT result the coincidence was lower (72% and 71.4%, respectively). Among heterozygotes, there was a statistically significant increase in the proportion of positive HBT results with age. Both SNPs were in accordance in 117/120 (97.5%) patients. CONCLUSION: Genetic analysis of LCT-13910 C/T and LCT-22018 G/A is a good indicator for the presence of lactose intolerance. Because age, as well as a number of secondary causes (e.g. celiac disease), can influence HBT results, it is useful to combine HBT and genetic analysis in the diagnostic assessment of IBS.     

Nicotine and angiogenesis: a new paradigm for tobacco‐related diseases

The pathophysiology of tobacco‐related diseases is complex and multifactorial. Among the approximately 4,000 compounds in tobacco smoke are carcinogens such as nitro‐samines, irritants such as a variety of phenolic compounds, volatiles such as carbon monoxide, and of course nicotine. Nicotine itself has quite complex actions, mediated in part by nicotinic cholinergic receptors that may have extraneuronal, as well as neuronal distribution. This review discusses the mechanisms by which nicotine contributes to tobacco‐related disease, with a focus on the surprising new finding that nicotine is a potent angiogenic agent. Nicotine hijacks an endogenous nicotinic cholinergic pathway present in endothelial cells that is involved in physiological, as well as pathological angiogenesis.     

The endocrine module: An integrated course for first‐year medical students combining lecture‐based and modified problem‐based curricula

At the University of Miami School of Medicine, a 2‐week endocrine course, designed to combine the desirable aspects of conventional lecture‐based and modified problem‐based curricula, has been offered to first‐year medical students for 7 years. It is team taught and integrates several basic science and clinical disciplines. Independent learning, problem‐solving activities, and small‐group conferences are emphasized. Each conference period is moderated by two or three faculty members, one or two representing the basic sciences and one representing clinical endocrinology. The course has been well received by the students, and the faculty have enjoyed teaching in this fully integrated format.     

Evaluation of 2‐D DIGE for skeletal muscle: Protocol and repeatability

Proteomic analysis has the potential to yield vast amounts of data. The available proteomic methods have been hampered by methodological errors in quantification due to large gel‐to‐gel variations. The inclusion of an internal standard greatly reduces this variation, and therefore the purpose of this investigation was: 1) to develop a sample preparation protocol for human skeletal muscle for two‐dimensional differentiated gel electrophoresis (DIGE) and 2) to investigate the repeatability of one particular system, the Ettan^? DIGE. To test repeatability, nine aliquots from the same homogenate were labelled with three different CyDye^? dyes (Cy2, Cy3, Cy5). Samples were run on 18×24?cm gels, scanned with a Typhoon^? 9410 laser scanner and analysed in the DeCyder^? software. When selecting spots appearing only in triplicate (n = 1314), the mean error was 1.7?% (SD: 10.5?%; 95?% CI: 1.1–2.4?%). When setting the significance level to 99?%, no false‐positive changes in protein volume ratios were detected. In the protocol presented here, only 0.5?mg tissue was used and separation of >2500 distinct protein spots in the pH range 3–11 and MW 10–200?kDa. Changes in protein abundance of <20?% could be detected. The method is especially useful when comparing muscle proteins between different conditions; for example, healthy and diseased tissue, before and after treatment or different exercise protocols.     

The critical path method to analyze and modify OR‐workflow: Integration of an image documentation system

Intraoperative image documentation is becoming more and more important for quality management in medicine, in terms of forensic documentation, research and teaching. Up to now, no software‐based OR‐image‐documentation system fits satisfactorily into an OR‐workflow. The objective of this study is to transparently show system integration in a clinical workflow for evaluating demands on future system developments. An example of the OR‐workflow is presented for the department of obstetrics and gynecology at the University of Tuebingen (Germany). Twelve representative gynecologic laparoscopic surgeries were analyzed by using the critical path method (CPM). CPM network diagrams are shown for an actual laparoscopic workflow and for a workflow including an OR‐image‐documentation system. With the objective not to increase the total time of actual workflow, the maximum system operation time can be calculated for each period of time. Before surgery the maximum system operation time is x_max = 7,3 minutes. After surgery it has to be assumed that system operation will increase total workflow time. Using the CPM to analyze requirements for system integration in a medical workflow has not yet been investigated. It is an appropriate method to transparently show integration possibilities and to define workflow‐based requirements for the development process of new systems.     

Reassessment of a classical single injection 51Cr‐EDTA clearance method for determination of renal function in children and adults. Part I: Analytically correct relationship between total and one‐pool clearance

Background. Total plasma clearance of ⁵¹Cr‐EDTA, Cl, is widely used as a measure of GFR. Commonly, only the final part of the plasma concentration curve is measured, and a one‐pool clearance (slope‐intercept clearance), Cl₁, is computed. Empirically determined second‐order polynomials of the general form Cl = b?Cl₁+c?Cl₁² are usually used to estimate Cl from a measured Cl₁. However, theoretical considerations indicate that such corrections underestimate Cl at high values. Aims. To derive an analytically correct relationship between Cl and Cl₁ and determine the parameters involved for children and adults. Material and methods. Cl was determined in 149 subjects (M/F/children: 71/46/32) from a complete plasma concentration curve followed for 4–5?h after injection of ⁵¹Cr‐EDTA (range of clearance: 8–183?mL/min/1.73?m²). Plasma volume, PV and the “missing” area under the plasma fraction curve, a (minutes), not used for determination of Cl₁, were measured. Results. The true relationship between Cl and Cl₁ is given by Cl = Cl₁/(1+f?Cl₁), where f = a/PV. For men, women and children alike, the equation f = 0.0032?BSA^?1.3 was applicable (BSA = body surface area in m²). Estimation errors on clearance were within ±8?% for adults and ±13?% for children (95?% limits of agreement). Conclusions. The true relationship between Cl and Cl₁ of ⁵¹Cr‐EDTA is given, resulting in a common correction equation applicable for children and adults. The new equation has better mathematical behaviour than quadratic equations on very high values of clearance and takes into account dependence on body size.     

Reassessment of a classical single injection 51Cr‐EDTA clearance method for determination of renal function in children and adults. Part II: Empirically determined relationships between total and one‐pool clearance

Background. The one‐pool or slope‐intercept technique is widely used when determining total ⁵¹Cr‐EDTA plasma clearance (Cl). The one‐pool clearance (Cl₁), which always exceeds Cl, has mostly been corrected to Cl by multiplication by a constant factor = 0.80, suggested by Chantler (CH_0.80), or by using a second‐order polynomial originally proposed by Brøchner‐Mortensen (BM) and later recommended by the British Nuclear Medicine Society (BM_BNMS). Theoretical considerations indicate that the CH correction gives a systematic overestimate of Cl, whereas the BM correction may underestimate Cl at high values. Objective. To assess the accuracy of Cl as estimated from Cl₁ corrected either by CH_0.80 or by second‐order polynomials. Material and methods. Cl_ref was determined in 149 subjects (M/F/children: 71/46/32) from a complete plasma curve followed for 4–5?h after injection of ⁵¹Cr‐EDTA (range of Cl_ref: 8–183?mL/min/1.73?m²). Cl_est was determined from Cl₁ subsequently corrected by CH_0.80 and four second‐order polynomials. Results. Using CH_0.80 correction, Cl_est underestimated Cl_ref (by a maximum of 20?%) at Cl_ref values less than about 100?mL/min/1.73?m² in children and 130?mL/min/1.73?m² in adults. At higher clearance levels, Cl_ref was increasingly overestimated. Taking the BM_BNMS correction as representative of second‐order polynomials, Cl_est increasingly underestimated Cl_ref at high levels, the error being 10?% at a Cl_ref value of about 175?mL/min/1.73?m². Conclusions. We suggest that the tested correction equations are replaced by the given common correction equation based on the “true” relationship between Cl₁ and Cl thoroughly described in part I of this study.     

Gut microbial ecology in critical illness: is there a role for prebiotics,probiotics, and synbiotics?

Approximately 70% of the immune system is localized in the gastrointestinal tract: its glands, mucosa, and mucosa-associated lymphoid system. The system influences health conditions because it produces large amounts of important gastrointestinal secretions as rich as breast milk in health-supporting and disease-preventing factors, and because of its rich gastrointestinal flora. The intestine normally contains 10 times more microbes than there are eukaryotic cells in the entire body. The optimal function of these microbes depends on the supply of food destined for the colonic bacteria (fermentable fibers, complex proteins, gastrointestinal secretions). The consideration of these functions influences outcome. Unfortunately, the conditions (supply of drugs-especially antibiotics, and reduced supply of food-especially fruits and vegetables) in the modern ICU are extremely poor both for optimal gastrointestinal secretion and for flora and need more attention. To improve treatment, a supply of new and effective flora (probiotics) and food for the flora (prebiotics) is needed, from which numerous health-supporting products (synbiotics) will be produced and absorbed at the level of the mucosa, mainly in the lower gastrointestinal tract.     

Raman difference spectroscopy: a non-invasive method for identification of oral squamous cell carcinoma: publisher’s note

The author list appeared incorrectly in [Biomed. Opt. Express 5(9), 3252–3265 (2014)]. The author names were corrected online as of January 17, 2015: https://www.osapublishing.org/boe/abstract.cfm?uri=boe-5-9-3252.OCIS codes: (170.0170) Medical optics and biotechnology, (170.5660) Raman spectroscopy, (170.6935) Tissue characterization, (170.3890) Medical optics instrumentationThe author list appeared incorrectly in [1]. The author names were corrected online as of January 17, 2015: https://www.osapublishing.org/boe/abstract.cfm?uri=boe-5-9-3252.     

Screening for hearing and visual loss among elderly with questionnaires and tests: which method is the most convincing for action?

OBJECTIVE: To examine the extent to which self-report and performance-based measures identify the same people, and also whether one of these methods any more than the other induces the general practitioner (GP) to offer a patient help? DESIGN: Comparison of tests and questionnaire in a screening programme for hearing and visual loss. SETTING: Twelve general practices in the western part of The Netherlands. PATIENTS: 1121 people 75 years of age and older. MAIN OUTCOME MEASURES: Hearing loss according to the whispered voice test and two questions (Q1, Q2) and visual loss according to the Snellen and low vision charts and two questions (Q3, Q4). The extent to which test and questions influence the GP's judgment regarding intervention is calculated using logistic regression. RESULTS: For hearing loss, the concordance of the whispered voice test with Q1 was 67%, and with Q2, 71%. For visual loss, the concordance of the Snellen chart with Q3 was 79% and of the Low Vision Chart with Q4, 69%. For hearing loss, the results of the test were noticeably more significant for the GP in offering help than the answers to the questions, and for visual loss the results of the Snellen chart slightly more convincing than the answers to the questions. CONCLUSION: When in a screening programme with simple instruments for visual and hearing loss the choice has to be made between questionnaire and tests, the tests will be the best choice.     

Pilates method for low back pain in individuals with Parkinson’s disease: A feasibility study

BackgroundPain is one of the most common and troublesome non-motor symptoms in Parkinson's disease (PD), including low back pain (LBP).ObjectiveThis study aims to evaluate the feasibility, acceptability, and safety of using Pilates for individuals with PD and LBP, as well as the effect on pain intensity, disability, motor and nonmotor PD symptoms, and quality of life. METHODS: Participants received 24 sessions of supervised Pilates twice a week for 12 weeks, each session consisting of 60 min. The isometric contractions of the core muscles were emphasized.ResultsFifteen (13%) of the screened patients were enrolled in the study and continued for the entire 2 months (100%). Participants completed the full training in 352 of 360 sessions (98%). Adverse effects were not reported. Pain and disability were significantly improved after intervention (Visual Analog Scale, p = 0.007; McGill Pain Questionnaire, p = 0.034; Roland-Morris Disability Questionnaire, p = 0.035). There were also significant improvements in depressive symptoms (Beck Depression Inventory, p = 0.028) and PD symptoms (Unified Parkinson's Disease Rating Scale – UPDRSI, UPDRSIII, UPDRS Total). However, there were no statistically significant changes in fatigue and quality of life.ConclusionsThis study suggests that the 12-week Pilates program is feasible and well tolerated by people with PD in mild to moderate stages of the disease. It also appears to be a promising strategy to reduce pain intensity and LBP-related disability, as well as PD motor and non-motor symptoms, which can be associated with this painful symptom. This study provides a basis for future investigations, especially randomized clinical trials.