A novel smartphone app for blood pressure measurement: a proof-of-concept study against an arterial catheter.

Smartphones may provide a highly available access to simplified hypertension screening in environments with limited health care resources. Most studies involving smartphone blood pressure (BP) apps have focused on validation in static conditions without taking into account intraindividual BP variations. We report here the first experimental evidence of smartphone-derived BP estimation compared to an arterial catheter in a highly dynamic context such as induction of general anesthesia. We tested a smartphone app (OptiBP) on 121 patients requiring general anesthesia and invasive BP monitoring. For each patient, ten 1-min segments aligned in time with ten smartphone recordings were extracted from the continuous invasive BP. A total of 1152 recordings from 119 patients were analyzed. After exclusion of 2 subjects and rejection of 565 recordings due to BP estimation not generated by the app, we retained 565 recordings from 109 patients (acceptance rate 51.1%). Concordance rate (CR) and angular CR demonstrated values of more than 90% for systolic (SBP), diastolic (DBP) and mean (MBP) BP. Error grid analysis showed that 98% of measurement pairs were in no- or low-risk zones for SBP and MBP, of which more than 89% in the no-risk zone. Evaluation of accuracy and precision [bias ± standard deviation (95% limits of agreement)] between the app and the invasive BP was 0.0 ± 7.5 mmHg [- 14.9, 14.8], 0.1 ± 2.9 mmHg [- 5.5, 5.7], and 0.1 ± 4.2 mmHg [- 8.3, 8.4] for SBP, DBP and MBP respectively. To the best of our knowledge, this is the first time a smartphone app was compared to an invasive BP reference. Its trending ability was investigated in highly dynamic conditions, demonstrating high concordance and accuracy. Our study could lead the way for mobile devices to leverage the measurement of BP and management of hypertension.

[Implementation of regularly performed resuscitation training at a hyperbaric treatment center]. / Implementierung eines regelmäßigen Reanimationstrainings an einem Druckkammerzentrum.

BACKGROUND: Medical emergency situations and even cardiac arrest can occur during treatment of patients in therapeutic hyperbaric chambers just as in other clinical departments; therefore, high quality structured management should be implemented for dealing with emergencies in this special working area. To ensure this the emergency medical treatment should not only be performed according to the current state of medical knowledge but needs to take the special features of the hyperbaric environment including safety aspects into account. METHOD: This article presents a description of the implementation and effects of routine emergency and resuscitation training at a center for hyperbaric medicine. RESULTS: By simulation of emergencies in a hyperbaric chamber it rapidly became clear that the treatment of medical emergencies and cardiac arrest under hyperbaric conditions has some special features and due to safety aspects cannot always be performed according to current medical guidelines. At the time of this simulation in a real life working environment, previously unknown structural and logistic problems became obvious whereby the solutions contributed to a significant improvement of structural and process quality and could potentially also improve the outcome quality. Furthermore, a positive and lasting learning effect in the fields of quality of resuscitation measures, organization of the workplace, communication skills, logistics and safety aspects was detectable by analyzing participant performance over a period of 4 years. On the part of the participating staff a positive feedback and high acceptance of emergency simulator training was confirmed. CONCLUSION: Through annual compulsory emergency training of the complete staff of the hyperbaric unit at the actual workplace, a structural and confident approach to dealing with emergencies and resuscitation situations was observed. By the use of on-site simulator training even in specialized hospital units, deficits and tentativeness regarding logistics, course of action, organization and communication in emergency situations can be minimized to provide optimum patient care in a real life emergency situation by focusing on the medical measures.

Long-term outcome after surgical treatment of cranial cruciate ligament rupture in small breed dogs. Comparison of tibial plateau leveling osteotomy and extra-articular stifle stabilization.

OBJECTIVE: Cranial cruciate ligament rupture is the most frequently occurring disease of the canine stifle. After introduction of corrective osteotomies of the proximal tibia as surgical treatment option in medium, large and giant breeds, the tibial plateau leveling osteotomy (TPLO) evolved into one of the favorite procedures. In the past small breed dogs have usually been treated by extra-articular stabilization techniques. TPLO has recently become an accepted treatment method in these breeds as well as, and is nowadays used with increasing frequency. The purpose of this study was to evaluate long-term outcomes of TPLO compared to an extra-articular stabilization technique in small breed dogs. MATERIAL AND METHODS: A total of 40 stifles of 30 small breed dogs weighing ≤ 15 kg were treated for cranial cruciate ligament rupture with either TPLO (n = 23) or a lateral capsular-fascial imbrication technique (CFI; n = 17). Dogs were clinically examined before and at least 6 months after surgery by treadmill analysis and radiography. Moreover a questionnaire was provided to the owners to assess postoperative function and overall satisfaction. RESULTS: Twenty-one of 23 operated limbs (91.3%) treated with TPLO and five of 17 (29.4%) treated with the CFI showed absolute values comparable to healthy dogs when evaluated by peak vertical force, vertical impulse and its symmetry index during objective gait analysis. Both groups showed mild, but continuous progression of osteoarthritis. TPLO led to a significantly faster recovery and a higher degree of owner satisfaction. CONCLUSION: Based on clinical examination and objective gait analysis TPLO yielded excellent long-term results and a high degree of owner satisfaction in small breed dogs ≤ 15 kg treated for cranial cruciate ligament rupture. CLINICAL RELEVANCE: TPLO seems to be superior to CFI treatment of small breed dogs in this study, although CFI should be considered as treatment option under certain circumstances.

[Aetiology, pathogenesis, diagnostics and therapy of cranial cruciate ligament rupture in dogs]. / Ätiologie, Pathogenese, Diagnostik und Therapie der Ruptur des kranialen Kreuzbandes beim Hund.

Cranial cruciate ligament rupture is the most common cause of hindlimb disease in dogs and is often associated with an additional lesion of the medial meniscus. The exact aetiopathogenesis of this condition in dogs is unknown. Normally, the degeneration of the ligament first leads to a partial rupture which progresses to a complete rupture following an unspectacular trauma. The positive cranial drawer test confirms the cranial cruciate ligament rupture, however, in some cases, particularly in partial ligament lesions, the diagnosis is not obvious. Therefore, the reexamination of the sedated patient is recommended to generally increase the sensitivity of clinical tests. Magnetic resonance imaging and stifle joint arthroscopy are useful methods to evaluate the cruciate ligaments in indistinct cases or to assess the joint for secondary changes. The therapy of cranial cruciate ligament ruptures can be divided into extra- and intracapsular as well as tibial osteotomies. The principle of the different tibial osteotomies is the muscular compensation of stifle joint instability. The results are successful but not uncritical. This article presents a short review on the substantial problems associated with cranial cruciate ligament rupture in dogs. A more detailed description of secondary problems and complications would exceed the scope of this article and should be considered in another study.

Determination of genotoxicity by the Comet assay applied to murine precision-cut lung slices.

Precision-cut lung slices (PCLSs) are an organotypic lung model that is widely used in pharmacological, physiological, and toxicological studies. Genotoxicity testing, as a pivotal part of early risk assessment, is currently established in vivo in various organs including lung, brain, or liver, and in vitro in cell lines or primary cells. The aim of the present study was to provide the three-dimensional organ culture PCLS as a new ex vivo model for determination of genotoxicity using the Comet assay. Murine PCLS were exposed to increasing concentrations of ethyl methane sulfonate 'EMS' (0.03-0.4%) and formalin (0.5-5mM). Tissue was subsequently dissociated, and DNA single-strand breaks were quantified using the Comet assay. Number of viable dissociated lung cells was between 4×10(5) and 6.7×10(5)cells/slice. Even treatment with EMS did not induce toxicity compared to untreated tissue control. As expected, DNA single-strand breaks were increased dose-dependently and significantly after exposure to EMS. Here, tail length rose from 24µm to 75µm. In contrast, formalin resulted in a significant induction of DNA cross-links. The effects induced by EMS and formalin demonstrate the usefulness of PCLS as a new ex vivo lung model for genotoxicity testing in the early risk assessment of airborne substances in the future.

Effects of acute in vitro exposure of murine precision-cut lung slices to gaseous nitrogen dioxide and ozone in an air-liquid interface (ALI) culture.

The aim of this study was to establish an air-liquid interface (ALI) culture of precision-cut lung slices (PCLS) for direct exposure of lung cells to gaseous contaminants. Nitrogen dioxide (NO(2)) and ozone (O(3)) were selected as model gas compounds. Acute pro-inflammatory and toxic effects of NO(2) and O(3) on live lung tissue were investigated. Murine PCLS were exposed to different flow rates (3-30mL/min) of synthetic air, O(3) (3.5-8.5ppm), or NO(2) (1-80ppm). Tissue survived ex vivo in ALI culture and resisted exposure to NO(2) (1-10ppm) and O(3) (3.5-8.5ppm) for 1h. Longer exposure to NO(2) resulted in a clear loss of viability, whereas exposure to O(3) was less effective. Exposure to NO(2) dose-dependently induced release of the pro-inflammatory IL-1alpha (40%), whereas RANTES, IL-12, and eotaxin remained unchanged. Early secretion of IL-1alpha (80%), RANTES (>800%), MIP-1beta (44%), and MCP-1 (60%) was already detected after 1h of exposure to O(3). The obtained data showed that direct exposure to O(3) and NO(2) induced cytotoxicity and pro-inflammatory responses in PCLS with ALI culture. This provides a model that more closely resembles in vivo exposure of airborne contaminants, and thus should be appropriate for toxicity testing.

Programmable CSF shunt valve: in vitro assessment of MR imaging safety at 3T.

A programmable CSF shunt valve was assessed for magnetic field interactions, heating (transmit-receive body radio-frequency coil; whole-body averaged specific absorption rate, 2.1 W/kg), functional alterations, and artifacts at 3T. The programmable valve showed minor magnetic field interactions and heating was not excessive (+0.8 degrees C). The function of the programmable valve was not altered by multiple exposures to the 3T scanner or from exposure to various MR imaging conditions. Therefore, this implant is safe for a patient undergoing MR imaging at 3T or less when the radiologist follows specific safety guidelines. Artifacts for the programmable valve were relatively large in relation to the size and shape of the valve; this finding may impact the diagnostic use of MR imaging if the area of interest is in proximity to this implant.

Novel approaches for studying pulmonary toxicity in vitro.

The in vitro study of adverse cellular effects induced by inhaled pollutants poses a special problem due to the difficulties of exposing cultured cells of the respiratory tract directly to test atmospheres that can include complex gaseous and particulate mixtures. In general, there is no widely accepted in vitro exposure system. However, in vitro methods offer the unique possibility for use of human cells, developed and validated cell culture and exposure device (CULTEX(1)) using the principle of the air/liquid exposure technique. Cells of the respiratory tract are grown on porous membranes in transwell inserts. After removal of the medium, the cells can be treated on their superficial surfaces with the test atmosphere, and at the same time they are supplied with nutrients through the membrane below. In comparison with other experimental approaches, the goal of our studies is to analyze the biological effects of test atmospheres under environmental conditions, i.e. without humidifying the atmosphere or adding additional CO(2). The system used is small and flexible enough independent of a cultivation chamber and thus offers the opportunity for onsite study of indoor and outdoor atmospheres in the field. The efficacy of the exposure device has already been demonstrated in the analysis of dose-dependent cytotoxic and genotoxic effects of exposure of epithelial lung cells to complex mixtures such as native diesel exhaust and side-stream smoke.

Exposure of human lung cells to native diesel motor exhaust--development of an optimized in vitro test strategy.

To investigate the effects of native diesel motor exhaust on human lung cells in vitro, a new experimental concept was developed using an exposure device on the base of the cell cultivation system CULTEX (Patent No. DE19801763.PCT/EP99/00295) to handle the cells during a 1-h exposure period independent of an incubator and next to an engine test rig. The final experimental set-up allows the investigation of native (chemically and physically unmodified) diesel exhaust using short distances for the transportation of the gas to the target cells. The analysis of several atmospheric compounds as well as the particle concentration of the exhaust was performed by online monitoring in parallel. To validate the complete system we concentrated on the measurement of two distinct viability parameters after exposure to air and undiluted, diluted and filtered diesel motor exhaust generated under different engine operating conditions. Cell viability was not influenced by the exposure to clean air, whereas dose-dependent cytotoxicity was found contingent on the dosage of exhaust. Additionally, the quality of exhaust, represented by two engine operating conditions (idling, higher load), also showed well-distinguishable cytotoxicity. In summary, the experimental set-up allows research on biological effects of native engine emissions using short exposure times.

In vitro efficacy of chemotherapeutics as determined by 50% inhibitory concentrations in cell cultures of mammary gland tumors obtained from dogs.

OBJECTIVE: To determine the 50% inhibitory concentration (IC-50) of carboplatin, cisplatin, and doxorubicin in cell cultures of mammary gland tumors obtained from dogs and to assess whether in vitro efficacy was within the range of clinically relevant concentrations, SAMPLE POPULATION: 30 mammary gland tumors excised from dogs. PROCEDURE: Cell cultures were established from the 30 tumors. Cultures then were treated with carboplatin, cisplatin, or doxorubicin. Growth inhibition of cultures was assessed via DNA measurement 24, 48, and 72 hours after treatment. The IC-50 values were calculated by use of linear interpolation. RESULTS: Cultures varied in their pattern of susceptibility. Doxorubicin induced significantly lower IC-50 values than the platinum derivatives. Cisplatin and carboplatin had comparable effects. The IC-50 values for carboplatin and doxorubicin were in the range of clinically relevant concentrations, but only part of the cisplatin cultures had IC-50 values within clinically relevant concentrations. We did not detect differences in the in vitro susceptibility among subtypes of tumors (ie, adenocarcinoma, solid carcinoma, malignant mixed tumor). CONCLUSION AND CLINICAL RELEVANCE: The IC-50 values determined in this study allowed assessment of in vitro drug efficacy of chemotherapeutics in cultures of mammary gland tumors obtained from dogs. Variations in susceptibility were evident and emphasize the importance of assessing susceptibility and resistance patterns for each tumor. Prospective studies to assess direct correlations between in vitro and in vivo efficacy must be performed to determine the clinical predictive value of this in vitro chemosensitivity assay for treatment of dogs with mammary gland tumors.

A method for in vitro analysis of the biological activity of complex mixtures such as sidestream cigarette smoke.

Studies of the cytotoxicity of air contaminants such as gaseous or particulate compounds and complex mixtures have traditionally used in animal experiments because of the difficulties in exposing cell cultures directly to these substances. New cultivation and exposure techniques enhance the efficiency of in vitro methods, as demonstrated by a new system called CULTEX* which uses a transwell membrane technique for direct exposure of complex mixtures like sidestream cigarette smoke at the air/liquid interface. The factors influencing the susceptibility of human bronchial epithelial cells (e.g. gas flow rate or duration of exposure) were studied and the cells were finally exposed for one hour to clean air or different concentrations of sidestream smoke. The biological parameters estimated were number of cells, metabolic activity and glutathione concentration. After exposure of the cells to sidestream cigarette smoke, dose-dependent effects were measured. Thus, the introduction of these cultivation and exposure techniques offers new testing strategies for the toxicological evaluation of a broad range of airborne and inhalable compounds.

In vitro exposure of isolated cells to native gaseous compounds--development and validation of an optimized system for human lung cells.

An exposure system for adherent growing cells to native gaseous compounds was developed using air/liquid culture techniques on the basis of the Cultex system'. In contrast to other exposure systems the reproducible testing of native environmentally relevant gases without changing their physical or chemical properties including heating, CO2- content and humidity is possible. Specially designed systems for medium flow and gas support guarantee the nutrification and humidification as well as the direct gas contact of the exposed cells which are cultivated on microporous membranes (0.4 microm pore size). The system works independently of a cell culture incubator offering the possibility to analyze any relevant gas mixture directly under indoor or outdoor conditions. Several experimental approaches were carried out to characterize the properties of the system. In exploratory experiments without cells, the reproducibility and quality of the gas/membrane contact could be demonstrated. Exposures of human lung fibroblasts (Lk004 cells) and human lung epithelial cells (HFBE-21 cells) to synthetic air, ozone (202 ppb, 510 ppb) and nitrogen dioxide (75 ppb to 1,200 ppb) established that cells could be treated for 120 minutes without significant loss of cellular viability. At the same time, the experiments confirmed that such exposure times are long enough to detect biological effects of environmentally relevant gas mixtures. The analysis of viability (viable cell number, tetrazoliumsalt cleavage) and intracellular end-points (oxidized/reduced glutathione, ATP/ADP) showed that both gases induced relevant cellular changes. In summary, the efficiency and practicability of this newly developed exposure system for adherent human lung cells could be clearly demonstrated.

Use of cultivated osteoprogenitor cells to increase bone formation in segmental mandibular defects: an experimental pilot study in sheep.

The hypothesis of the present experimental pilot study was that autogeneous cultivated osteoprogenitor cells in porous calcium phosphate scaffolds can increase bone formation in segmental defects of the mandible. The autogenous osteoprogenitor cells of eight sheep were cultivated from bone biopsies from the iliac crest and seeded into cylindrical scaffolds of pyrolized bovine bone of an overall length of 35 mm and 13 mm in diameter. Segmental defects of 35 mm length were created unilaterally in the mandibles of the animals. Reconstruction was performed using cylinders with cultivated osteoprogenitor cells in four animals and empty scaffolds in the remaining four sheep, which served as controls. After 5 months, the mandibles were retrieved and the reconstructed areas were analyzed by qualitative and quantitative histology in serial undecalcified thick-section specimens. There was significantly more bone formation in the group that had received scaffolds with cultivated bone cells (P=0.028). Bone formation was present in 34.4% of the evaluated cross-sectional units in the seeded scaffolds, while it was found in 10.4% in the control group. Although the spatial distribution of bone formation was significantly different across the scaffold in both groups, osteoprogenitor cells appeared to have increased bone formation, particularly in the centre of the defect when compared to the control group. It is concluded that the repair of segmental defects of the mandible can be enhanced by the transplantation of autogenous osteoprogenitor cells in a porous calcium phosphate scaffold.

Development of a cell culture model system for routine testing of substances inducing oxidative stress.

The toxicological model of oxidative stress is a mechanism which is currently thought to be involved in the formation and development of many serious human diseases. Little is known about cellular responses to oxidative damage and the related specific toxicological properties of substances such as chemicals or components of the polluted urban atmosphere. On the basis of biological pathways involved in cellular antioxidative mechanisms, we developed a biological model for studying the oxidative properties of substances. This includes human lung cells and methods for biochemical analysis of cellular endpoints. Antioxidative and glycolysis-related enzyme activities (glutathione peroxidase, glutathione reductase, superoxide dismutase, catalase, phosphofructokinase, enolase, glucose-6-phosphate dehydrogenase), ATP/ADP/AMP and glutathione (oxidized and reduced) are determined. Routine testing of substances with high reproducibility and fast screening is provided by adapting methods for biochemical analysis to determinations using cells grown on microtitre plates. First experiments with standard model substances inducing oxidative stress such as H(2)O(2) and tert-butylhydroperoxide show that enzymatic activities can be measured with good reproducibility and their changes can be followed kinetically. The results indicate the relevance of the determined parameters for such toxicological events and the usefulness of the biological indicator system for routine testing.

Development of an in vitro system for studying effects of native and photochemically transformed gaseous compounds using an air/liquid culture technique.

An experimental in vitro model was established to study the effects of environmentally relevant gaseous compounds on lung cells. The technical unit consists of a gas reaction chamber (2400 l) with a sun-simulator to produce and photochemically transform gaseous mixtures and compounds at the upper limit of environmentally relevant concentrations. Rat lung cells were exposed on transwells in a perspex chamber inside an incubator, into which the gaseous mixtures were conducted. Analysis of the gas phase was performed inside the reaction chamber and at the outlet of the exposure box to assess the effective exposure concentrations. The growth of the cells on PET-membranes allowed direct cell exposure with a minimal barrier for contact between gas and cells. To assess the cytotoxicity, the following biochemical markers for the cellular status after exposure were determined: amount of dsDNA, WST, BrdU-incorporation after exposure, LDH release into the culture medium, activity of glutathione S-transferases and esterases. Using this system, dose-dependent cytotoxicity was found for NO2 in the concentration range from 80 to 360 ppb and strong cytotoxic effects for ozone in the concentration range from 225 to 500 ppb. Exposure to purified air did not show significant effects. In addition, some irradiated gas mixtures (photo smog) showed cytotoxicity whereas non-irradiated mixtures did not.

Micronucleus induction in V79 cells after direct exposure to whole cigarette smoke.

Previous investigations on the effects of cigarette smoke on cultured cells have used mainly smoke condensate dissolved in culture medium. A system has been designed which allows direct exposure of cells to fresh cigarette smoke, without an intervening layer of growth medium between the cells and the smoke. Preliminary results have been obtained which demonstrate the viability of the system. V79 cells were cultured on porous membranes (Transwell; Costar). During smoke exposure only the lower surface of each Transwell is supplied with culture medium from the bottom of the culture chambers. In this way the cells had direct contact with the atmosphere at the upper surface and could be exposed directly to the test compound. The constructed exposure system consists of a smoke generator and an exposure unit containing six Transwells, the latter contained in an incubator. Cigarette smoke was generated using a standard 2 s, 35 ml puff once per min. The puff is diluted with conditioned air from the incubator and injected into the exposure unit. Following exposure of the cells to air only for 3 h there was no effect upon V79 cell viability. However, after exposure to smoke containing between 88 and 224 mg/m3 particulate matter, an inhibition of cell proliferation and induction of micronuclei was measured. When a Cambridge filter pad was placed between the cigarette and the cell exposure system to remove particulate matter cell proliferation was also reduced and an increased frequency of micronuclei above the control value was measured.

Experimental reconstruction of the mandible using polylactic acid tubes and basic fibroblast growth factor in alloplastic scaffolds.

PURPOSE: This study evaluated the use of a mitogenic growth factor in combination with barrier membranes and porous alloplastic scaffolds for the repair of segmental defects of the mandible. MATERIALS AND METHODS: Fifteen Göttingen minipigs were used for the study. In five animals, mandibular defects of 2 cm were created on both sides of the mandible and bridged by a system of polylactic acid (PLA) tubes and pyrolized bovine bone. On one side of the mandible, the alloplastic scaffolds were loaded with 115 microg recombinant human basic fibroblast growth factor (rhbFGF). In five other animals, defects 4 cm in length were created bilaterally, similarly bridged, and loaded with 230 microg of rhFGF. In five control animals, bilateral 2-cm defects were created that were left empty on one side and bridged with an empty PLA tube on the other. Mitogenic efficacy of the growth factor was assessed on fibroblast cultures by di-methyl-thiazol-2-tetrazolium-bromide assay before implantation. RESULTS: After 5 months, there was negligible bone regeneration in the control defects, regardless of whether they had been left completely empty or bridged by empty PLA tubes. The 2-cm defects showed bridging in 8 of 10 tubes, with complete consolidation by bone ingrowth in four defects. The 4-cm defects showed bony union in six cases, with complete bone fill in two tubes, and four defects incompletely filled. The bFGF had no appreciable effect with regard to velocity, quantity, and three-dimensional structure of bone formation, neither in the short nor in the long defects despite clear in vitro efficacy. CONCLUSION: Repair of segmental defects using bioresorbable membranes appears to be possible. However, a single-dose application of bFGF is apparently ineffective, possibly because of rapid dilution.

Sensitivity of a hamster lung cell line to direct and indirect acting carcinogens.

Cytotoxicity of benzo(a)pyrene (B(a)P), 7,12-dimethylbenz(a)anthracene (DMBA), aflatoxin B1 (AB1), and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) was estimated in vitro by using a hamster lung cell line. The studies were conducted to assess the usefulness of an organ-specific cell culture system for demonstrating differences in the cytotoxic potency of diverse chemical carcinogens. Cytotoxicity was determined by using the succinate dehydrogenase assay (MTT assay) after different incubation times and concentrations with the corresponding carcinogens. The effective concentration EC50 as well as the slope of the regression line were used as parameters for the biological effects. The results from these studies indicate a clear dose-dependent reaction after incubation of the cells with aflatoxin B1 (EC50: 2.3 microM) and MNNG (EC50: 4.0 microM). For the polycyclic hydrocarbons benzo(a)pyrene and DMBA, a dose-independent reaction was found. These results indicate that consideration of the EC50 values only might not be sufficient to characterize differences in the cytotoxic activity of different substances. Chemicals can lead to equal values in the EC50, but cells can differ significantly in their biological sensitivity, meaning that the extent of reduction in cell proliferation depends on the chemical used. By considering the two above-mentioned parameters, a ranking for the analyzed substances will be possible in the following way: AB1, MNNG, DMBA and B(a)P. Taken together, our experiments show that it is possible to reveal differences in the cytotoxic potency of chemicals by using in vitro methods.