Direct 16S/18S rRNA Gene PCR Followed by Sanger Sequencing as a Clinical Diagnostic Tool for Detection of Bacterial and Fungal Infections: a Systematic Review and Meta-Analysis.

rRNA gene Sanger sequencing is being used for the identification of cultured pathogens. A new diagnostic approach is sequencing of uncultured samples by using the commercial DNA extraction and sequencing platform SepsiTest (ST). The goal was to analyze the clinical performance of ST with a focus on nongrowing pathogens and the impact on antibiotic therapy. A literature search used PubMed/Medline, Cochrane, Science Direct, and Google Scholar. Eligibility followed PRISMA-P criteria. Quality and risk of bias were assessed drawing on QUADAS-2 (quality assessment of diagnostic accuracy studies, revised) criteria. Meta-analyses were performed regarding accuracy metrics compared to standard references and the added value of ST in terms of extra found pathogens. We identified 25 studies on sepsis, infectious endocarditis, bacterial meningitis, joint infections, pyomyositis, and various diseases from routine diagnosis. Patients with suspected infections of purportedly sterile body sites originated from various hospital wards. The overall sensitivity (79%; 95% confidence interval [CI], 73 to 84%) and specificity (83%; 95% CI, 72 to 90%) were accompanied by large effect sizes. ST-related positivity was 32% (95% CI, 30 to 34%), which was significantly higher than the culture positivity (20%; 95% CI, 18 to 22%). The overall added value of ST was 14% (95% CI, 10 to 20%) for all samples. With 130 relevant taxa, ST uncovered high microbial richness. Four studies demonstrated changes of antibiotic treatment at 12% (95% CI, 9 to 15%) of all patients upon availability of ST results. ST appears to be an approach for the diagnosis of nongrowing pathogens. The potential clinical role of this agnostic molecular diagnostic tool is discussed regarding changes of antibiotic treatment in cases where culture stays negative.

Baseline values but not treatment-induced changes in carotid intima-media thickness predict incident cardiovascular events in treated hypertensive patients: findings in the European Lacidipine Study on Atherosclerosis (ELSA).

BACKGROUND: Baseline carotid intima-media thickness (IMT) and plaques are considered predictors of cardiovascular events, but whether they maintain predictive value in treated hypertensive patients and whether time-related (or treatment-induced) IMT changes are additional predictors are unknown. METHODS AND RESULTS: Analyses were performed of the data from the European Lacidipine Study on Atherosclerosis (ELSA), a large, randomized, intervention trial in which 2334 hypertensive patients from 7 European countries were followed up under effective antihypertensive treatment for 3.75 years. Kaplan-Meier curves indicated progressively lower survival free of any type of outcome except stroke, with increasing baseline IMT quartiles or increasing IMT values, even after adjustment for major baseline risk factors. Incidence of any outcome except stroke also was related to baseline number of carotid plaques. However, when both baseline and on-treatment IMT values were entered in Cox proportional-hazards models, differences in IMT compared with baseline did not predict cardiovascular outcomes. Although on-treatment rather than baseline IMT values significantly entered some of the proportional-hazards models, baseline and on-treatment IMTs were highly correlated, and therefore these results are inconclusive. CONCLUSIONS: ELSA shows that carotid intima-media thickening and plaques are important added risks of cardiovascular outcomes in a treated hypertensive population independently of blood pressure and traditional risk factors. However, the analysis failed to show a predictive role of treatment-dependent IMT changes. These negative conclusions should be tempered by the limitations inherent in the smallness of these changes compared with the large individual differences in baseline IMTs.

The 5.5-year results of MegaOATS--autologous transfer of the posterior femoral condyle: a case-series study.

INTRODUCTION: Large osteochondral defects of the weight-bearing zones of femoral condyles in young and active patients were treated by autologous transfer of the posterior femoral condyle (large osteochondral autogenous transplantation system (MegaOATS)). The technique presented is a sound and feasible salvage procedure to address large osteochondral defects in weight-bearing zones. METHODS: Thirty-six patients between July 1996 and December 2000 were included. Thirty-three patients (10 females, 23 males) were evaluated by the Lysholm score and X-ray scans. A random sample of 16 individuals underwent magnetic resonance imaging analysis. The average age at the date of surgery was 34.3 (15 to 59) years, and the mean follow up was 66.4 (46 to 98) months. The mean defect size was 6.2 (2 to 10.5) cm2, in 27 patients affecting the medial femoral condyle and in six patients affecting the lateral femoral condyle. Trauma or osteochondrosis dissecans were pathogenetic in 82%. RESULTS: The Lysholm score in all 33 individuals showed a highly significant increase from a preoperative median 49.0 points to a median 86.0 points (P < or = 0.001). Twenty-seven patients returned to recreational sports. X-ray scans showed a rounding of the osteotomy edge in 24 patients, interpreted as a partial remodelling of the posterior femoral condyle. Preoperative osteoarthritis in 17 individuals was related to significant lower Lysholm scores (P = 0.014), but progression in 17 patients did not significantly influence the score results (P = 0.143). All 16 magnetic resonance imaging examinations showed vital and congruent grafts. CONCLUSION: Patients significantly improve in the Lysholm score, in daily-life activity levels and in return to recreational sports. Thirty-one out of 33 patients were comfortable with the results and would undergo the procedure again. The MegaOATS technique is therefore recommended as a salvage procedure for young individuals with large osteochondral defects in the weight-bearing zone of the femoral condyle.

The E-cadherin repressor snail plays a role in tumor progression of endometrioid adenocarcinomas.

Endometrial cancer is the most common gynecologic cancer in the developed world. The cell-adhesion protein E-cadherin acts as a tumor-suppressor protein and is down-regulated by the transcription factor Snail, whose expression was shown to be associated with estrogen receptor signaling. This study aimed to investigate the expression of E-cadherin, Snail, and estrogen-receptor alpha in 87 primary tumors and 26 metastases of endometroid endometrial carcinomas. Reduced E-cadherin immunoreactivity was seen in 44.8% of the primary tumors and 65.4% of the metastases with a statistical correlation to higher tumor grade (P=0.003) only in metastatic lesions. About 28.7% of primary tumor specimens showed a positive Snail immunoreactivity that was correlated with reduced estrogen-receptor alpha expression (P=0.047). Positive Snail immunoreactivity was also seen in 53.8% of the metastases where it was correlated with higher tumor grade (P=0.003) and abnormal E-cadherin expression (P=0.003). Interestingly, a Snail expressing endometrial carcinoma-cell line showed a higher migration potential than a variant of this cell line with low levels of Snail. Taken together, our data are in line with a proposed role for Snail in endometrial tumor progression.

Submucosal endoscopy: a novel approach to en bloc endoscopic mucosal resection (with videos).

BACKGROUND: The submucosal layer is of eminent importance for endoscopic mucosal resection (EMR) in the GI tract. OBJECTIVE: Development of submucosal endoscopy, which allows diagnostic and therapeutic endoscopy of the submucosal space (SS) in the esophagus. DESIGN: Acute experiments in a live porcine model. INTERVENTIONS: An area in the esophagus was marked with a diathermic probe to define a mucosal piece for resection. After local infiltration, a 1- to 2-cm transverse incision was performed 1 to 2 cm proximal and distal of these margins. We entered the SS with a flexible small-caliber videoendoscope through the proximal incision and dissected the fibrous submucosal connective tissue in a longitudinal direction with a blunt forceps. For EMR, the lifted mucosa was subsequently separated by use of an insulated-tip hook needle-knife. MAIN OUTCOME MEASUREMENTS: En bloc resection of prespecified mucosal areas. RESULTS: A total of 15 mucosal pieces were resected in 4 pigs. The size of the resected pieces varied from 1.6 cm x 0.9 cm to 7.4 cm x 1.7 cm ex vivo. In a fifth pig, 2 circular mucosectomies (lengths 3.0 cm and 1.6 cm) were done. All mucosal pieces could be completely resected en bloc. The endoscopic view in the SS was excellent. There were no procedure-related complications. LIMITATIONS: The method has not yet been evaluated in humans. CONCLUSIONS: Entering the SS for submucosal endoscopy is a novel, innovative, and practicable method for the dissection of mucosal neoplastic lesions. We demonstrated that mucosal areas of various sizes could be resected en bloc without complications.

Midazolam-pain, but one cannot remember it: a survey among Southern German endoscopists.

BACKGROUND: Benzodiazepines, especially midazolam, are the most frequently used agents for gastrointestinal endoscopy worldwide. Among other parameters the quality of sedation is determined by patients' satisfaction assessed after endoscopy. This approach is misleading as the potent amnestic effect of midazolam conceals pain actually suffered during the endoscopic procedure involving distraction of the endoscopists from their actual tasks by audible reactions and defense movements. In this study, we eliminated the influence of patients' amnesia on the assessment of the quality of sedation and rather interviewed endoscopists and their assistant personnel about their experience with midazolam sedation. We replaced the mostly vague term 'compliance' by terms which unequivocally describe the reactions of the patient during an unpleasant endoscopy. METHODS: A short survey consisting of 12 questions was developed. The questionnaires were distributed to the participants - 115 endoscopists and their assistants - of a tutorial about sedation for gastrointestinal endoscopy in three major Southern German cities. The questionnaire retrieved the endoscopists' experience regarding patients' discomfort or pain under sedation with midazolam, their wish for better sedative agents, their preferred sedative regimens, their medical specialty and their professional experience. RESULTS: Participants were highly experienced with the majority having more than 10,000 procedures and a median of 18 years of endoscopic experience; 77% of endoscopists utilized midazolam for sedation. Ninety-eight percent of the questioned physicians felt that patients have pain during endoscopy with midazolam+/-opioid, but do not remember later. Ninety-two percent reported that it happens that patients moan aloud because of pain and almost half of the endoscopists (48%) reported of screaming. The majority of the endoscopists (91%) reported fierce defense movements with midazolam or the need to hold the patient down on the examination couch because of fierce movements, respectively (75%). Seventy percent of the endoscopists wished to have the rooms for endoscopy preferably soundproof away from the waiting room and 93% wished for better sedative agents. CONCLUSIONS: Midazolam was rated as insufficient for sedation by both endoscopists and their assistant personnel. A wish for better sedative drugs exists.

Sevoflurane affects neurogenesis after forebrain ischemia in rats.

BACKGROUND: The effect of sevoflurane on the neuroregenerative potential after neuronal injury is unclear. We investigated the effect of low and high concentrations of sevoflurane on endogenous neurogenesis after cerebral ischemia. METHODS: Anesthetized and ventilated rats were randomized to four different treatment groups. Groups 1 and 2: 1.4% sevoflurane; Groups 3 and 4: 2.8% sevoflurane. In Groups 1 and 3, no cerebral ischemia was induced (sham-operated). In Groups 2 and 4, 10 min of forebrain ischemia was induced by bilateral carotid artery occlusion plus hemorrhagic hypotension. Physiological variables were maintained constant. Bromodeoxyuridine was given as a marker of neurogenesis. After 28 days brains were perfused. Histopathological damage of the hippocampus was evaluated in hematoxylin and eosin (HE) stained sections using the HE-index (0 = no damage; 1 = 1%-10% damage; 2 = 11%-50% damage; 3 = 51%-100% damage). Immunohistochemistry was used to detect bromodeoxyuridine-positive neurons. Eight untreated rats were investigated as naive controls (Group 5). RESULTS: In neither sham-operated group was histopathological damage or change in neurogenesis observed compared to naive controls. In rats anesthetized with 1.4% sevoflurane, cerebral ischemia caused mild neuronal damage (HE-index of 0.64 +/- 0.84) and increased neurogenesis by 60% when compared with respective sham-operated animals; with 2.8% sevoflurane, the HE-index was 1.22 +/- 1.14, and the number of newly generated neurons increased by 230% when compared with respective sham-operated animals. CONCLUSION: The present data suggest that high concentrations of sevoflurane stimulate neurogenesis in the dentate gyrus after cerebral ischemia.

Comparison between breast volume measurement using 3D surface imaging and classical techniques.

Quantification of the complex breast region can be helpful in breast surgery, which is shaped by subjective influences. However, there is no generally recognized method for breast volume calculation. Three-dimensional (3D) body surface imaging represents a new alternative for breast volume computation. The aim of this work was to compare breast volume calculation with 3D scanning and three classic methods, focusing on relative advantages, disadvantages, and reproducibility. Repeated breast volume calculations of both breasts in six patients (n=12) were performed using a 3D laser scanner, nuclear magnetic resonance imaging (MRI), thermoplastic castings, and anthropomorphic measurements. Mean volumes (cc) and mean measurement deviations were calculated, and regression analyses were performed. MRI showed the highest measurement precision, with a mean deviation (expressed as a percentage of mean breast volume) of 1.56+/-0.52% compared with 2.27+/-0.99% for the 3D scanner, 7.97+/-3.53% for thermoplastic castings, and 6.26+/-1.56% for the anthropomorphic measurements. Breast volume calculations using MRI showed the best agreement with 3D scanning measurement (r=0.990), followed by anthropomorphic measurement (r=0.947), and thermoplastic castings (r=0.727). Compared with three classical methods of breast volume calculation, 3D scanning provides acceptable accuracy for breast volume measurements, better spatial interpretation of the anatomical area to be operated on (due to lack of chest deformation), non-invasiveness, and good patient tolerance. After this preliminary study and further development, we believe that 3D body surface scanning could provide better preoperative planning and postoperative control in everyday clinical practice.

The effect of electroencephalogram-targeted high- and low-dose propofol infusion on histopathological damage after traumatic brain injury in the rat.

BACKGROUND: Propofol is commonly used to sedate patients after traumatic brain injury. However, the dose-dependent neuroprotective effects of propofol after head trauma are unknown. We compared histopathological damage after 6 h of electroencephalogram-targeted high- and low-dose propofol infusion in rats subjected to controlled cortical impact (CCI). METHODS: Animals were randomly assigned to CCI/propofol with electroencephalogram burst-suppression-ratio 1%-5% (CCI/lowprop), CCI/propofol with burst-suppression-ratio 30%-40% (CCI/highprop), control group CCI/1.0 vol % halothane (CCI/halo), or sham group with halothane anesthesia (SHAM/halo). Brain slices were stained with kresyl violet (KV) and hematoxylin/eosin (HE) to evaluate lesion volume, number of eosinophilic cells, and activation of caspase-3 in the hippocampus. RESULTS: Lesion volume (mm3) and number of eosinophilic cells in the hippocampus did not differ significantly [lesion volumes: CCI/lowprop 31.55 +/- 14.66 (KV) and 53.77 +/- 8.62 (HE); CCI/highprop 33.81 +/- 10.57 (KV) and 52.30 +/- 11.55 (HE); CCI/halo 36.42 +/- 17.06 (KV) and 57.95 +/- 8.49 (HE)]. Activation of caspase-3 occurred in the ipsilateral hippocampus in all CCI-groups. CONCLUSION: Despite different levels of cortical neuronal function, there were no relevant differences in the short-term histopathological damage. These results challenge the view that the neuroprotective effect of propofol relates to the suppression of cerebral metabolic demand.

New aspects of breast volume measurement using 3-dimensional surface imaging.

Precise and objective calculation of breast volume is helpful to evaluate the aesthetic result of breast surgery, but traditional methods are unsatisfactory. Three-dimensional (3D) scanning of the body surface allows reproducible and objective assessment of the complex breast region but requires further investigation before clinical application. The main goal of this study was to investigate the precision and accuracy of breast volume measurement using 3D body scanning. Five independent observers standardized the 3D scanning method using 2 dummy models (n = 200) and examined its applicability with 6 test subjects and 10 clinical patients (n = 2220). Breast volume measurements obtained with the 3D-scanner technology were compared with reference measurements obtained from test subjects through nuclear magnetic resonance imaging. The mean deviation of the breast volume measurements of 1 test subject by all observers, expressed as percentage of volume, was 2.86 +/- 0.98, significantly higher than the deviation for the dummy models, 1.65 +/- 0.42 (P < 0.001). With respect to all clinical patients, the mean measurement precision obtained preoperatively was less precise than that obtained postoperatively (3.31 +/- 1.02 versus 1.66 +/- 0.49, respectively). Interobserver differences in measurement precision were not statistically significant. The mean breast volumes obtained by nuclear magnetic resonance imaging (441.42 +/- 137.05 mL) and 3D scanning (452.51 +/- 141.88 mL) significantly correlated (r = 0.995, P < 0.001). Breast volume measurement with 3D surface imaging represents a sufficiently precise and accurate method to guarantee objective and exact recording.