[Prehospital management of acute coronary syndrome in patients on long-term direct oral anticoagulant treatment]. / Prähospitale Behandlung des akuten Koronarsyndroms unter DOAK-Dauertherapie.

BACKGROUND: Despite an increasing incidence of patients suffering from acute coronary syndrome (ACS) under simultaneous treatment with direct oral anticoagulants (DOAC), neither sufficient scientific data nor uniform guidelines for the anticoagulation treatment of these patients are currently available. OBJECTIVE: The aim of this study was to determine the current practice of preclinical treatment of ACS in patients under DOAC treatment. MATERIAL AND METHODS: An internet and paper-based survey of emergency physicians, specialists of internal medicine, anesthesiologists, emergency and intensive care physicians was performed concerning the prehospital treatment of ACS in patients under long-term DOAC treatment. RESULTS: Overall, 284 questionnaires were answered. Substantial differences in the current treatment of ACS under long-term DOAC therapy were identified. While 39% of the respondents stated that they administer a combination treatment of heparin and acetylsalicylic acid (ASA), 36% renounced the administration of heparin. If a dose reduction was performed, 71% answered that they reduce the heparin dosage. Also, in cases of ST-segment elevation myocardial infarction 48% of the physicians renounced the administration of heparin. CONCLUSION: In Germany there is currently a heterogeneous practice of emergency treatment of ACS patients under DOAC therapy with respect to the administration of heparin and ASA. Therefore, guidelines of the specialist medical societies should address the prehospital emergency anticoagulation management of ACS in patients under therapy with DOAC, which correspond to the needs of patients and emergency physicians.

Low-temperature plasma treatment induces DNA damage leading to necrotic cell death in primary prostate epithelial cells.

BACKGROUND: In recent years, the rapidly advancing field of low-temperature atmospheric pressure plasmas has shown considerable promise for future translational biomedical applications, including cancer therapy, through the generation of reactive oxygen and nitrogen species. METHOD: The cytopathic effect of low-temperature plasma was first verified in two commonly used prostate cell lines: BPH-1 and PC-3 cells. The study was then extended to analyse the effects in paired normal and tumour (Gleason grade 7) prostate epithelial cells cultured directly from patient tissue. Hydrogen peroxide (H2O2) and staurosporine were used as controls throughout. RESULTS: Low-temperature plasma (LTP) exposure resulted in high levels of DNA damage, a reduction in cell viability, and colony-forming ability. H2O2 formed in the culture medium was a likely facilitator of these effects. Necrosis and autophagy were recorded in primary cells, whereas cell lines exhibited apoptosis and necrosis. CONCLUSIONS: This study demonstrates that LTP treatment causes cytotoxic insult in primary prostate cells, leading to rapid necrotic cell death. It also highlights the need to study primary cultures in order to gain more realistic insight into patient response.

[Patient with a Fontan circulation undergoing caesarean section: Anesthesiological management]. / Sectio caesarea bei einer Patientin mit Fontan-Zirkulation : Anästhesiologisches Management.

Adults suffering from congenital heart diseases (CHD) represent a challenge to anesthesiologists because of the diverse pathologies, complex pathophysiology and special treatment strategies. Due to improved therapeutic options for CHD, patient quality of life and life expectancy is increasing, leaving them as a growing population including pregnant patients with CHD. This article presents the main principles of the pathophysiology and anesthesiological management of pregnant patients living with a Fontan circulation based on a case report, which was complicated by an aortic coarctation and atonic uterine hemorrhage.

Hypercalcemia in renal transplant patients: prevalence and management in Canadian transplant practice.

Hypercalcemia, occurring in up to 25% of patients within 12 months following renal transplantation, and persistent hyperparathyroidism were evaluated following renal transplantation, by retrospective chart review of 1000 adult patients transplanted between January 1, 2003 and January 31, 2008 with at least six months follow-up. Serum calcium, parathyroid hormone, and phosphate levels were recorded at 12, 24, 36, and 48 months. Average follow-up was 766 (535) d (mean (SD); median 668 d). Majority were first transplants (85%); deceased donor 57%. Point prevalence of hypercalcemia (serum Ca(2+) > 2.6 mM) was 16.6% at month 12, 13.6% at month 24, 9.5% at month 36, and 10.1% at month 48. Point prevalence of serum parathyroid hormone (PTH) > 10 pM was 47.6% at month 12, 51.1% at month 24, 43.4% at month 36, and 39.3% at month 48. Estimated glomerular filtration rate (GFR) was maintained throughout and was not different between patients with or without hypercalcemia or elevated PTH. Cinacalcet was prescribed in 12% of patients with hypercalcemia and persistent hyperparathyroidism; parathyroidectomy was performed in 112/1000 patients, 15 post-transplant. Persistent hyperparathyroidism, often accompanied by hypercalcemia, is common following successful renal transplantation, but the lack of clear management suggests the need for further study and development of evidence-based guidelines.

HDAC inhibitor confers radiosensitivity to prostate stem-like cells.

BACKGROUND: Radiotherapy can be an effective treatment for prostate cancer, but radiorecurrent tumours do develop. Considering prostate cancer heterogeneity, we hypothesised that primitive stem-like cells may constitute the radiation-resistant fraction. METHODS: Primary cultures were derived from patients undergoing resection for prostate cancer or benign prostatic hyperplasia. After short-term culture, three populations of cells were sorted, reflecting the prostate epithelial hierarchy, namely stem-like cells (SCs, α2ß1integrin(hi)/CD133(+)), transit-amplifying (TA, α2ß1integrin(hi)/CD133(-)) and committed basal (CB, α2ß1integrin(lo)) cells. Radiosensitivity was measured by colony-forming efficiency (CFE) and DNA damage by comet assay and DNA damage foci quantification. Immunofluorescence and flow cytometry were used to measure heterochromatin. The HDAC (histone deacetylase) inhibitor Trichostatin A was used as a radiosensitiser. RESULTS: Stem-like cells had increased CFE post irradiation compared with the more differentiated cells (TA and CB). The SC population sustained fewer lethal double-strand breaks than either TA or CB cells, which correlated with SCs being less proliferative and having increased levels of heterochromatin. Finally, treatment with an HDAC inhibitor sensitised the SCs to radiation. INTERPRETATION: Prostate SCs are more radioresistant than more differentiated cell populations. We suggest that the primitive cells survive radiation therapy and that pre-treatment with HDAC inhibitors may sensitise this resistant fraction.

[Supraglottic airway devices in emergency medicine : impact of gastric drainage]. / Supraglottische Atemwegshilfen in der Notfallmedizin : Stellenwert der Magendrainage.

This case report describes a life-saving use of a supraglottic airway device (LT-D™-Larynxtubus, VBM Medizintechnik, Sulz, Germany) in an out-of-hospital emergency patient suffering from severe traumatic brain injury. Mechanical ventilation with the laryngeal tube was complicated by repeated airway obstructions and pronounced gastric distension with air as a consequence of oropharyngeal leakage. In this situation pulmonary ventilation of the patient was compromised so that emergency endotracheal intubation became necessary in the resuscitation area with vital indications. In this context the status of supraglottic airway devices in emergency medicine is discussed as well as the reasons for the gastric distension. Besides the immediate drastic consequences of gastric distension with respect to pulmonary ventilation, potential deleterious non-pulmonary consequences of this complication are highlighted. The clinical relevance of the described complications as well as the associated possibility of an optimized position control necessitate the recommendation only to use second generation supraglottic airway devices with integrated gastric access in (out-of-hospital) emergency medicine.

Assessment of a preclinical model for studying the survival and engraftment of human stem cell derived osteogenic cell populations following orthotopic implantation.

INTRODUCTION: Preclinical studies with osteoprogenitor cells derived from human embryonic stem cells (hESC) do not lead to substantial bone regeneration in vivo. The degree of survival following implantation might play a role in their long term efficiency. We investigated the initial engraftment of hESCs-derived cells during two weeks post-implantation and compared it to such response for adult bone marrow stromal cells (hBMSC)-derived osteoprogenitor cells. METHODS: hBMSC and H9-hES cells pre-treated with osteogenic factors were implanted into a calvarial defect in both adult WT and nude rats. At days 7 and 14 post-implantation, samples were analysed for persistence of implanted cells, initiation of regeneration of host bone, angiogenesis and apoptosis. RESULTS: At day 7, hESC and hBMSC were detected within defects in both rat strains. By day 14 human cells were only detected in immune-deficient rats whilst still maintaining an osteoblastic phenotype and engendered a significant increase in bone formation. In WT animals, the participation of implanted cells was very limited due to their poor survival. CONCLUSION: This study demonstrates the ability of hESC and hBMSC derived osteoprogenitor cells to survive transplantation, to engraft and to develop an osteogenic phenotype during the early stage following implantation, validating the appropriate preclinical model.

The effect of a cervical collar on the seal pressure of the LMA Supreme™: a prospective, crossover trial.

For personnel inexperienced in airway management, supraglottic airway devices may be the first choice in an emergency. Changing head position is known to reduce the seal pressure of a laryngeal mask airway. The aim of this study was to investigate whether the use of a cervical collar improves the stability of airways secured with the LMA Supreme™ (The Laryngeal Mask Company Limited, Mahé, Seychelles). In this crossover trial, the primary endpoint was the difference in the seal pressure of the LMA Supreme in anaesthetised patients in maximum passive extension of the neck, with and without a cervical collar. The median (IQR [range]) seal pressure was 18 (13.8-22.1 [0-30]) cmH(2) O in maximum passive extension without a cervical collar. With a cervical collar in place, the seal pressure increased to 28 (22.8-30 [17-30]) cmH(2) O (p<0.001). In the neutral head position, the seal pressure was 22 (17.6-24.5 [12-30]) cmH(2) O without and 27 (22-30 [12-30]) cmH(2) O with a cervical collar in place (p<0.001). We found that a cervical collar stabilises the airway with an LMA Supreme in place and we recommend this combination for (pre-hospital) emergency cases.

A prospective study of the factors affecting outcomes of nonsurgical root canal treatment: part 1: periapical health.

AIM: To investigate the probability of and factors influencing periapical status of teeth following primary (1°RCTx) or secondary (2°RCTx) root canal treatment. METHODOLOGY: This prospective study involved annual clinical and radiographic follow-up of 1°RCTx (1170 roots, 702 teeth and 534 patients) or 2°RCTx (1314 roots, 750 teeth and 559 patients) carried out by Endodontic postgraduate students for 2-4 (50%) years. Pre-, intra- and postoperative data were collected prospectively on customized forms. The proportion of roots with complete periapical healing was estimated, and prognostic factors were investigated using multiple logistic regression models. Clustering effects within patients were adjusted in all models using robust standard error. RESULTS: proportion of roots with complete periapical healing after 1°RCTx (83%; 95% CI: 81%, 85%) or 2°RCTx (80%; 95% CI: 78%, 82%) were similar. Eleven prognostic factors were identified. The conditions that were found to improve periapical healing significantly were: the preoperative absence of a periapical lesion (P = 0.003); in presence of a periapical lesion, the smaller its size (P ≤ 0.001), the better the treatment prognosis; the absence of a preoperative sinus tract (P = 0.001); achievement of patency at the canal terminus (P = 0.001); extension of canal cleaning as close as possible to its apical terminus (P = 0.001); the use of ethylene-diamine-tetra-acetic acid (EDTA) solution as a penultimate wash followed by final rinse with NaOCl solution in 2°RCTx cases (P = 0.002); abstaining from using 2% chlorexidine as an adjunct irrigant to NaOCl solution (P = 0.01); absence of tooth/root perforation (P = 0.06); absence of interappointment flare-up (pain or swelling) (P =0.002); absence of root-filling extrusion (P ≤ 0.001); and presence of a satisfactory coronal restoration (P ≤ 0.001). CONCLUSIONS: Success based on periapical health associated with roots following 1°RCTx (83%) or 2°RCTx (80%) was similar, with 10 factors having a common effect on both, whilst the 11th factor 'EDTA as an additional irrigant' had different effects on the two treatments.

A prospective study of the factors affecting outcomes of non-surgical root canal treatment: part 2: tooth survival.

AIM: To investigate the probability of and factors influencing tooth survival following primary (1°RCTx) or secondary (2°RCTx) root canal treatment. METHODOLOGY: This prospective study involved annual follow-up of 2 (100%) to 4 years (50%) of 1°RCTx (759 teeth, 572 patients) and 2°RCTx (858 teeth, 642 patients) carried out by Endodontic postgraduate students. Pre-, intra- and post-operative data were collected prospectively from consented patients. Information about extraction of the root filled tooth was sought from the patient, the referring dentist or derived from the patient's records and included the timing and reasons for extraction. Tooth survival was estimated and prognostic factors were investigated using Cox regression. Clustering effects within patients were adjusted in all models using robust standard error. RESULTS: The 4-year cumulative tooth survival following 1°RCTx [95.4% (93.6%, 96.8%)] or 2°RCTx [95.3% (93.6%, 96.5%)] was similar. Thirteen prognostic factors were identified. Significant patient factors included history of diabetes and systemic steroid therapy. Significant pre-operative factors included narrow but deep periodontal probing depth; pain; discharging sinus; and iatrogenic perforation (for 2°RCTx cases only). Significant intra-operative factors included iatrogenic perforation; patency at apical terminus; and extrusion of root fillings. Significant post-operative restorative factors included presence of cast restoration versus temporary restoration; presence of cast post and core; proximal contacts with both mesial and distal adjacent teeth; and terminal location of the tooth. The presence of pre-operative pain had a profound effect on tooth loss within the first 22 months after treatment [hazard ratio (HR) = 3.1; P = 0.001] with a lesser effect beyond 22 months (HR = 2.4; P = 0.01). Patency at the apical terminus reduced tooth loss (HR = 0.3; P < 0.01) within the first 22 months after treatment but had no significant effect on tooth survival beyond 22 months. Extrusion of gutta-percha root filling did not have any effect on tooth survival (HR = 1.1; P = 0.2) within the first 22 months but significantly increased the hazard of tooth loss beyond 22 months (HR = 3.0; P = 0.003). CONCLUSIONS: The 4-year tooth survival following primary or secondary root canal treatment was 95%, with thirteen prognostic factors common to both.

Tooth survival following non-surgical root canal treatment: a systematic review of the literature.

AIMS: To investigate (i) the effect of study characteristics on reported tooth survival after root canal treatment (RCTx) and (ii) the effect of clinical factors on the proportion of root filled teeth surviving after RCTx. METHODOLOGY: Longitudinal human clinical studies investigating tooth survival after RCTx which were published up to the end of 2007 were identified electronically (MEDLINE and Cochrane database 1966-2007 December, week 4). In addition, four journals (Dental Traumatology, International Endodontic Journal, Journal of Endodontics, Oral Surgery Oral Medicine Oral Pathology Oral Radiology & Endodontics), bibliographies of all relevant articles and review articles were hand searched. Two reviewers (Y-LN, KG) assessed and selected the studies based on specified inclusion criteria and extracted the data onto a pre-designed proforma, independently. The criteria were as follows: (i) clinical study on RCTx; (ii) stratified analysis of primary and secondary RCTx available; (iii) sample size given and larger than 10; (iv) at least 6-month postoperative review; (v) success based on survival of tooth; and (vi) proportion of teeth surviving after treatment given or could be calculated from the raw data. Three strands of evidence or analyses were used to triangulate a consensus view. The reported findings from individual studies, including those excluded for quantitative analysis, were utilized for the intuitive synthesis, which constituted the first strand of evidence. Secondly, the pooled weighted proportion of teeth surviving and thirdly the combined effects of potential prognostic factors were estimated using the fixed and random effects meta-analyses on studies fulfilling all the inclusion criteria. RESULTS: Of the 31 articles identified, 14 studies published between 1993 and 2007 were included. The majority of studies were retrospective (n = 10) and only four prospective. The pooled percentages of reported tooth survival over 2-3, 4-5 and 8-10 years following RCTx were 86% (95% CI: 75%, 98%), 93% (95% CI: 92%, 94%) and 87% (95% CI: 82%, 92%), respectively. Substantial differences in study characteristics were found to hinder effective direct comparison of findings. Evidence for the effect of prognostic factors on tooth survival was weak. Based on the data available for meta-analyses, four conditions were found to significantly improve tooth survival. In descending order of influence, the conditions increasing observed proportion of survival were as follows: (i) a crown restoration after RCTx; (ii) tooth having both mesial and distal proximal contacts; (iii) tooth not functioning as an abutment for removable or fixed prosthesis; and (iv) tooth type or specifically non-molar teeth. Statistical heterogeneity was substantial in some cases but its source could not be investigated because of insufficient available information. CONCLUSIONS: The pooled proportion of teeth surviving over 2-10 years following RCTx ranged between 86% and 93%. Four factors (listed above) were identified as significant prognostic factors with concurrence between all three strands of evidence.

[Treatment of polytrauma in the intensive care unit]. / Polytraumaversorgung auf der Intensivstation.

The treatment of severely injured trauma patients (polytrauma) is one of the outstanding challenges in medical care. Early in the initial course the patient's diagnostics have to be scrupulously reevaluated by an interdisciplinary team (tertiary trauma survey) to reduce deleterious sequelae of missed injuries after the initial assessment. Severely injured patients stay in intensive care for an average of 11 days. During this time the patient's therapy has to ensure a high quality evidence-based intensive care treatment and simultaneously has to be tailored to the current individual injuries. Because of the fact that the damage control strategy is gaining increasing acceptance, the intensive care unit plays a pivotal role in the critical time between emergency and elective surgery. Therefore a close cooperation between physicians of the intensive care unit and all surgical disciplines involved is essential to reach the aim of therapeutic efforts. After survival of emergency treatment patients with severe trauma should be reintegrated into social and occupational life as soon as possible.

[Skill retention using extraglottic airways in out-of-hospital emergencies: efficacy and long-term results of simulator-based medical education : A prospective follow-up study]. / Routineerhalt beim Umgang mit extraglottischen Atemwegen im Rettungsdienst: Effektivität und Nachhaltigkeit einer simulatorbasierten Ausbildung : Eine prospektive 2­zeitige Simulatorstudie.

OBJECTIVE: For emergency medicine personnel (EMP), there is little evidence concerning the adequate timing for refresher courses to maintain routine in the application of extraglottic airways. The aim of this study was to evaluate the efficacy and long-term results of a simulator-based education concept teaching the basic airway management skills with extraglottic airways for EMP and also to draw conclusions concerning the adequate time interval for refresher courses. METHODS: By use of an explorative, prospective simulator-study with nonphysician EMP, airway management skills using the Larynxmaske Supreme® (LMA­S) after an introduction lecture were examined. The application of an endotracheal tube (ETT) served as control. Time for preparation of the airway devices, insertion success, and resulting apnea time were assessed immediately after the first introduction lecture (t1) and unannounced 9-12 months thereafter (t2). RESULTS: Comparison of the times for preparation of the LMA­S at t1 and t2 demonstrated similar results. After the introduction lecture, all paramedics were able to insert the LMA­S successfully after maximal 2 attempts; 9-12 months later success rates with the LMA­S were unchanged. Apnea time during airway management was shorter with the LMA­S compared to the ETT (p < 0.01). Times needed for preparation of the airway devices were similar. CONCLUSION: The results of this simulator study indicate that a standardized introduction lecture is appropriate to ensure long-lasting procedural skills up to 12 months, so that subsequent refresher courses in basic airway management with the LMA­S once a year may be adequate. A simulator-based education in basic airway management skills with extraglottic airways is recommended for facilitation of further clinical education according to the current guidelines.

Fecal elastase 1 and vitamin D3 in patients with osteoporotic bone fractures.

BACKGROUND AND AIMS: The aim of the present study was to clarify if patients with osteoporotic bone fractures have exocrine pancreatic insufficiency, especially reduced fecal elastase 1, connected with lowered serum levels of vitamin D3 that could be relevant for predominant osteoporosis. METHODS: Between October 1999 and September 2001, we investigated on 167 patients with an average age of approx. 69 years suffering from typical osteoporotic bone fractures, as well as 20 healthy controls with an average age of 53 years. A standardized osteodensitometry via dual energy X-ray absorptiometry (DEXA) was performed in all participants. Levels of PTH, 1,25(OH)(2) Vitamin D(3), 25(OH) Vitamin D(3), calcium and phosphate in serum, elastase 1 in feces as well as the body mass index were determined in all patients and controls. RESULTS: In patients 25(OH)D3 was more than 60% and 1,25(OH)(2)D(3) was more than 53% decreased compared to controls. Fecal elastase 1 was lower than the lowest reference of 200 microg/g feces in more than 34% of the patients and it was more than 65% reduced in comparison to healthy controls (fecal elastase 1 patients: 240.7 +/- 96.3 microg/g; controls 694.9 +/- 138.6 microg/g). Separation of the patients in accordance with the elastase 1 contend in feces into four groups (below 100 microg/g, between 100 and 200 microg/g, between 201 and 300 microg/g and above 300 microg/g) resulted in significant variations for 25(OH)D(3), 1,25(OH)(2)D(3), calcium and PTH between these groups (p < 0.01). Furthermore 25(OH)D(3), 1,25(OH)(2)D(3), calcium and PTH correlated significantly with elastase 1 in feces (p < 0.01) the way, that lower fecal elastase 1 was connected with lower levels of the other parameters. BMI shows no relevant differences within the patients or between patients and controls. CONCLUSION: Exocrine pancreatic insufficiency, especially lowered fecal elastase 1, may be much more frequent in patients with osteoporotic bone fractures than suggested so far. Lowered exocrine pancreatic function with lowered fecal elastase 1 seems to be relevant as a reason for reduced levels of circulating vitamin D3 metabolites being an appropriate additional cause for predominant osteoporosis.

Outcome of secondary root canal treatment: a systematic review of the literature.

UNLABELLED: AIMS (I): To investigate the effects of study characteristics on the reported success rates of secondary root canal treatment (2 degrees RCT or root canal retreatment); and (ii) to investigate the effects of clinical factors on the success of 2 degrees RCT. METHODOLOGY: Longitudinal human clinical studies investigating outcome of 2 degrees RCT which were published upto the end of 2006 were identified electronically (MEDLINE and Cochrane database 1966-2006 Dec, week 4). Four journals (Dental Traumatology, International Endodontic Journal, Journal of Endodontics, Oral Surgery Oral Medicine Oral Pathology Endodontics Radiology), bibliographies of all relevant papers and review articles were hand-searched. Two reviewers (Y-LN, KG) independently assessed and selected the studies based on specified inclusion criteria and extracted the data onto a pre-designed proforma, independently. The criteria were: (i) Clinical studies on 2 degrees RCT; (ii) Stratified analyses available for 2 degrees RCT where 1 degrees RCT data included; (iii) Sample size given and larger than 10; (iv) At least 6-month post-operative review; (v) Success based on clinical and/or radiographic criteria (strict = absence of apical radiolucency; loose = reduction in size of radiolucency); and (vi) Overall success rate given or could be calculated from the raw data. Three strands of evidence or analyses were used to triangulate a consensus view. The reported findings from individual studies, including those excluded for quantitative analysis, were utilized for the intuitive synthesis which constituted the first strand of evidence. Secondly, the pooled weighted success rates by each study characteristic and potential prognostic factor were estimated using the random effect model. Thirdly, the effects of study characteristics and prognostic factors (expressed as odds ratios) on success rates were estimated using fixed and random effects meta-analysis with DerSimonean and Laird's methods. Meta-regression models were used to explore potential sources of statistical heterogeneity. Study characteristics considered in the meta-regression analyses were: decade of publication, study-specific criteria for success (radiographic, combined radiographic & clinical), unit of outcome measure (tooth, root), duration after treatment when assessing success ('at least 4 years' or '<4 years'), geographic location of the study (North American, Scandinavian, other countries), and qualification of the operator (undergraduate students, postgraduate students, general dental practitioners, specialist or mixed group). RESULTS: Of the 40 papers identified, 17 studies published between 1961 and 2005 were included; none were published in 2006. The majority of studies were retrospective (n = 12) and only five prospective. The pooled weighted success rate of 2 degrees RCT judged by complete healing was 76.7% (95% CI 73.6%, 89.6%) and by incomplete healing, 77.2% (95% CI 61.1%, 88.1%). The success rates by 'decade of publication' and 'geographic location of study' were not significantly different at the 5% level. Eighteen clinical factors had been investigated in various combinations in previous studies. The most frequently and thoroughly investigated were 'periapical status' (n = 13), 'size of lesion' (n = 7), and 'apical extent of RF' (n = 5) which were found to be significant prognostic factors. The effect of different aspects of primary treatment history and re-treatment procedures has been poorly tested. CONCLUSIONS: The pooled estimated success rate of secondary root canal treatment was 77%. The presence of pre-operative periapical lesion, apical extent of root filling and quality of coronal restoration proved significant prognostic factors with concurrence between all three strands of evidence whilst the effects of 1 degrees RCT history and 2 degrees RCT protocol have been poorly investigated.

Vasomotion of mice mesenteric arteries during low oxygen levels.

BACKGROUND: Ischemia of intestinal organs is a main cause of complications in surgical intensive care patients. Changes in the tonus of arteries contributing to vascular resistance play an important role in the determination of blood flow and thus oxygen supply of various abdominal organs. It is generally acknowledged that hypoxia itself is able to alter arterial tonus and thus blood flow. METHODS: The present study compared the effects of various degrees of hypoxia on second-order mesenteric arteries from male C57BL/6J mice. After vessel isolation and preparation, we assessed vessel diameter using an arteriograph perfusion chamber. Investigating mechanisms promoting hypoxia-induced vasodilatation, we performed experiments in Ca2+-containing and Ca2+-free solutions, and furthermore, Ca2+-influx was inhibited by NiCl2, eNOS-/--, and TASK1-/--mice were investigated too. RESULTS: Mild hypoxia 14.4% O2 induced, in 50% of mesenteric artery segments from wild-type (wt) mice, a vasodilatation; severe hypoxia recruited further segments responding with vasodilatation reaching 80% under anoxia. However, the extension of dilatation of luminal arterial diameter reduced from 1.96% ± 0.55 at 14.4% O2 to 0.68% ± 0.13 under anoxia. Arteries exposed to hypoxia in Ca2+-free solution responded to lower oxygen levels with increasing degree of vasodilatation (0.85% ± 0.19 at 14.4% O2 vs. 1.53% ± 0.42 at 2.7% O2). Inhibition of voltage-gated Ca2+-influx using NiCl2 completely diminished hypoxia-induced vasodilatation. Instead, all arterial segments investigated constricted. Furthermore, we did not observe altered hypoxia-induced vasomotion in eNOS-/-- or TASK1-/- mice compared to wt animals. CONCLUSIONS: The present study demonstrated that hypoxic vasodilatation in mice mesenteric arteries is mediated by a NO-independent mechanism. In this experimental setting, we found evidence for Ca2+-mediated activation of ion channels causing hypoxic vasodilatation.

A COL1A1 Sp1 binding site polymorphism predisposes to osteoporotic fracture by affecting bone density and quality.

Osteoporosis is a common disease with a strong genetic component. We previously described a polymorphic Sp1 binding site in the COL1A1 gene that has been associated with osteoporosis in several populations. Here we explore the molecular mechanisms underlying this association. A meta-analysis showed significant associations between COL1A1 "s" alleles and bone mineral density (BMD), body mass index (BMI), and osteoporotic fractures. The association with fracture was stronger than expected on the basis of the observed differences in BMD and BMI, suggesting an additional effect on bone strength. Gel shift assays showed increased binding affinity of the "s" allele for Sp1 protein, and primary RNA transcripts derived from the "s" allele were approximately three times more abundant than "S" allele--derived transcripts in "Ss" heterozygotes. Collagen produced from osteoblasts cultured from "Ss" heterozygotes had an increased ratio of alpha 1(I) protein relative to alpha 2(I), and this was accompanied by an increased ratio of COL1A1 mRNA relative to COL1A2. Finally, the yield strength of bone derived from "Ss" individuals was reduced when compared with bone derived from "SS" subjects. We conclude that the COL1A1 Sp1 polymorphism is a functional genetic variant that predisposes to osteoporosis by complex mechanisms involving changes in bone mass and bone quality.

Metabolic engineering of astaxanthin production in tobacco flowers.

Using metabolic engineering, we have modified the carotenoid biosynthesis pathway in tobacco (Nicotiana tabacum) to produce astaxanthin, a red pigment of considerable economic value. To alter the carotenoid pathway in chromoplasts of higher plants, the cDNA of the gene CrtO from the alga Haematococcus pluvialis, encoding beta-carotene ketolase, was transferred to tobacco under the regulation of the tomato Pds (phytoene desaturase) promoter. The transit peptide of PDS from tomato was used to target the CRTO polypeptide to the plastids. Chromoplasts in the nectary tissue of transgenic plants accumulated (3S,3'S) astaxanthin and other ketocarotenoids, changing the color of the nectary from yellow to red. This accomplishment demonstrates that plants can be used as a source of novel carotenoid pigments such as astaxanthin. The procedures described in this work can serve as a platform technology for future genetic manipulations of pigmentation of fruits and flowers of horticultural and floricultural importance.

Validation of a 7-point Global Overall Symptom scale to measure the severity of dyspepsia symptoms in clinical trials.

BACKGROUND: Currently there is no consensus on the optimal method to measure the severity of dyspepsia symptoms in clinical trials. AIM: To validate the 7-point Global Overall Symptom scale. METHODS: The Global Overall Symptom scale uses a 7-point Likert scale ranging from 1 = no problem to 7 = a very severe problem. Validation was performed in two randomized-controlled trials (n = 1121 and 512). Construct validity: Global Overall Symptom was compared with the Quality of Life in Reflux And Dyspepsia, Gastrointestinal Symptom Rating Scale, Reflux Disease Questionnaire and 10 specific symptoms using Spearman correlation coefficients. Test-retest reliability: The Intraclass Correlation Coefficient was calculated for patients with stable dyspepsia defined by no change in Overall Treatment Effect score over two visits. Responsiveness: effect size and standardized response mean were also calculated. RESULTS: Construct validity: Change in Global Overall Symptom score correlated significantly with Quality of Life for Reflux And Dyspepsia, Gastrointestinal Symptom Rating Scale, Reflux Disease Questionnaire and specific symptoms (all P < 0.0002). Reliability: The Intraclass Correlation Coefficient was 0.62 (n = 205) and 0.42 (n = 270). Responsiveness: There was a positive correlation between change in Global Overall Symptom and change in symptom severity. The effect size and standardized response mean were 1.1 and 2.1, respectively. CONCLUSION: The Global Overall Symptom scale is a simple, valid outcome measure for dyspepsia treatment trials.

The influence of mechanical stimulation on osteocyte apoptosis and bone viability in human trabecular bone.

It has been shown previously using in vivo and ex vivo animal models, that cyclical mechanical stimulation is capable of maintaining osteocyte viability through the control of apoptotic cell death. Here we have studied the effect of mechanical stimulation on osteocyte viability in human trabecular bone maintained in a 3-D bioreactor system. Bone samples, maintained in the bioreactor system for periods of 3, 7 and 27 days, were subjected to either cyclical mechanical stimulation which engendered a maximum of 3,000 microstrain in a waveform corresponding to physiological jumping exercise for 5 minutes daily or control unloading. Unloading resulted in a decrease in osteocyte viability within 3 days that was accompanied by increased levels of cellular apoptosis. Mechanical stimulation significantly reduced apoptosis (p< or =0.032) and improved the maintenance of osteocyte viability in bone from all patient samples. The percentage Alkaline Phosphatase (ALP) labelled bone surface was significantly increased (p< or =0.05) in response to mechanical stimulation in all samples as was the Bone Formation Rate (BFR/BS) (p=0.005) as determined by calcein label incorporation in the 27-day experiment. These data indicate that in this model system, mechanical stimulation is capable of maintaining osteocyte viability in human bone.