Reallocation of time between device-measured movement behaviours and risk of incident cardiovascular disease.
To improve classification of movement behaviours in free-living accelerometer data using machine-learning methods, and to investigate the association between machine-learned movement behaviours and risk of incident cardiovascular disease (CVD) in adults. Using free-living data from 152 participants, we developed a machine-learning model to classify movement behaviours (moderate-to-vigorous physical activity behaviours (MVPA), light physical activity behaviours, sedentary behaviour, sleep) in wrist-worn accelerometer data. Participants in UK Biobank, a prospective cohort, were asked to wear an accelerometer for 7 days, and we applied our machine-learning model to classify their movement behaviours. Using compositional data analysis Cox regression, we investigated how reallocating time between movement behaviours was associated with CVD incidence. In leave-one-participant-out analysis, our machine-learning method classified free-living movement behaviours with mean accuracy 88% (95% CI 87% to 89%) and Cohen's kappa 0.80 (95% CI 0.79 to 0.82). Among 87 498 UK Biobank participants, there were 4105 incident CVD events. Reallocating time from any behaviour to MVPA, or reallocating time from sedentary behaviour to any behaviour, was associated with lower CVD risk. For an average individual, reallocating 20 min/day to MVPA from all other behaviours proportionally was associated with 9% (95% CI 7% to 10%) lower risk, while reallocating 1 hour/day to sedentary behaviour from all other behaviours proportionally was associated with 5% (95% CI 3% to 7%) higher risk. Machine-learning methods classified movement behaviours accurately in free-living accelerometer data. Reallocating time from other behaviours to MVPA, and from sedentary behaviour to other behaviours, was associated with lower risk of incident CVD, and should be promoted by interventions and guidelines.


Limb development in a "nonmodel" vertebrate, the direct-developing frog Eleutherodactylus coqui.
Mechanisms that mediate limb development are regarded as highly conserved among vertebrates, especially tetrapods. Yet, this assumption is based on the study of relatively few species, and virtually none of those that display any of a large number of specialized life-history or reproductive modes, which might be expected to affect developmental pattern or process. Direct development is an alternative life history found in many anuran amphibians. Many adult features that form after hatching in metamorphic frogs, such as limbs, appear during embryogenesis in direct-developing species. Limb development in the direct-developing frog Eleutherodactylus coqui presents a mosaic of apparently conserved and novel features. The former include the basic sequence and pattern of limb chondrogenesis, which are typical of anurans generally and appear largely unaffected by the gross shift in developmental timing; expression of Distal-less protein (Dlx) in the distal ectoderm; expression of the gene Sonic hedgehog (Shh) in the zone of polarizing activity (ZPA); and the ability of the ZPA to induce supernumerary digits when transplanted to the anterior region of an early host limb bud. Novel features include the absence of a morphologically distinct apical ectodermal ridge, the ability of the limb to continue distal outgrowth and differentiation following removal of the distal ectoderm, and earlier cessation of the inductive ability of the ZPA. Attempts to represent tetrapod limb development as a developmental "module" must allow for this kind of evolutionary variation among species.


Defining electronic-prescribing and infusion-related medication errors in paediatric intensive care - a Delphi study.
The use of health information technology (HIT) to improve patient safety is widely advocated by governmental and safety agencies. Electronic-prescribing and smart-pump technology are examples of HIT medication error reduction strategies. The introduction of new errors on HIT implementation is, however, also recognised. To determine the impact of HIT interventions, clear medication error definitions are required. This study aims to achieve consensus on defining as medication errors a range of either technology-generated, or previously unaddressed infusion-related scenarios, common in the paediatric intensive care setting. This study was conducted in a 23-bed paediatric intensive care unit (PICU) of an Irish tertiary paediatric hospital. A modified Delphi technique was employed: previously undefined medication-incidents were identified by retrospective review of voluntary incident reports and clinical pharmacist interventions; a multidisciplinary expert panel scored each incident using a 9-point Likert scale over a number of iterative rounds; levels of agreement were assessed to produce a list of medication errors. Differences in scoring between healthcare professionals were assessed. Seventeen potential errors or 'scenarios' requiring consensus were identified, 13 of which related to technology recently implemented into the PICU. These were presented to a panel of 37 participants, comprising of doctors, nurses and pharmacists. Consensus was reached to define as errors all reported smart-pump scenarios (n = 6) and those pertaining to the pre-electronic process of prescribing weight-based paediatric infusions (n = 4). Of 7 electronic-prescribing scenarios, 4 were defined as errors, 2 were deemed not to be and consensus could not be achieved for the last. Some differences in scoring between healthcare professionals were found, but were only significant (p < 0.05) for two and three scenarios in consensus rounds 1 and 2 respectively. The list of medication errors produced using the Delphi technique highlights the diversity of previously undefined medication errors in PICU. The increased complexity of electronic-prescribing processes is evident from the difficulty in achieving consensus on those scenarios. Reducing ambiguity in defining medication errors should assist future research on the impact of HIT medication safety initiatives in critical care. The increasing use of HIT and associated new errors will necessitate further similar studies.


BMP13 prevents the effects of annular injury in an ovine model.
Chronic back pain is a global health problem affecting millions of people worldwide and carries significant economic and social morbidities. Intervertebral disc damage and degeneration is a major cause of back pain, characterised by histological and biochemical changes that have been well documented in animal models. Recently there has been intense interest in early intervention in disc degeneration using growth factors or stem cell transplantation, to replenish the diseased tissues. Bone Morphogenetic Proteins (BMPs) have been approved for clinical use in augmenting spinal fusions, and may represent candidate molecules for intervertebral disc regeneration. BMP13 has an important role in embryonic development and recent genetic evidence shows a role in the development of the human spine. This study explores the effect of BMP13 on a damaged intervertebral disc in an ovine model of discal degeneration. We found that, when injected at the time of injury, BMP13 reversed or arrested histological changes that occurred in the control discs such as loss of extracellular matrix proteins. In addition, BMP13 injected discs retained greater hydration after 4 months, and possessed more cells in the NP. Taken together, BMP13 may be a potent clinical therapeutic agent when used early in the degeneration cascade to promote healthy disc tissue.


Characteristics and outcomes of patients with a haematological malignancy admitted to the intensive care unit for a neurological event.
Patients with haematological malignancies are at risk of concomitant critical neurological events warranting intensive care unit admission. We aimed to examine the characteristics and outcomes of this patient population, as more knowledge could facilitate decision making on ICU admission and treatment. A retrospective cohort study of 68 patients in adult ICUs of six Dutch university hospitals between 2003 and 2011. The median Acute Physiology and Chronic Health Evaluation (APACHE) II score was 23 (IQR, 16-27), and 77% of patients needed mechanical ventilation within the first 24 hours of admission. Forty percent of patients had received an allogeneic stem cell transplantation, and 22% were neutropenic on admission. The most frequent underlying haematological condition was non-Hodgkin lymphoma (27%). Seizures were the most common neurological event for ICU admission (29%). The median ICU length of stay was 5 days (IQR, 1-13 days). ICU mortality (28%), hospital mortality (37%) and 3-month mortality (50%) were comparable with other studies of ICU patients with haematological malignancies. Factors associated with 3-month survival were baseline platelet count (113×10(9)/L in survivors v 39×10(9)/L in non-survivors, P<0.01) and APACHE II score (20 in survivors v 25 in non-survivors, P=0.02). Patients with a history of haematological malignancy presenting with a critical neurological event have comparable survival rates with other patients with a haematologic malignancy admitted to the ICU. Our findings suggest that restrictions in ICU care are not justified for this patient population.


Modulation of transient pressure by an air pocket in a horizontal pipe with an end orifice.
In urban drainage systems, a sudden increase in the flow rate can cause the transition of the flow from open channel to pipe flow, and the entrapment of large air pockets in sewers, which might result in serious geysers and water-hammer like pressure events. This paper presents a numerical analysis of flow processes associated with the pressurization and release of an air pocket in order to study its influence on transient pressure in a horizontal pipe with an end orifice. The influence of the air pocket inside the pipe on the peak pressure can be described in two distinct regimes. In regime I for the pipe with a small orifice, the peak pressure is modulated by the pressurization and expansion of the air pocket and its subsequent damping. In regime II for the pipe with a large orifice, air can be quickly expelled, and the water column directly impinges on the pipe end wall and causes water-hammer like pressure. With the increase of the orifice size, the peak pressure decreases due to the change in the water velocity. In the study cases, the peak pressure in regime I is about two times the inlet pressure, while it can be more than forty times in regime II.


Comparative analysis of quality-of-life metrics after endoscopic surgery for sinonasal neoplasms.
The objective of this study was to evaluate the postoperative quality of life (QOL) after endoscopic resection of sinus and skull base neoplasms using validated outcomes measures and to perform correlation of the various metrics to better discern their efficacy. Prospective QOL data collection and retrospective chart review were performed. QOL data were collected using the 20-item Sino-Nasal Outcome Test (SNOT-20), Anterior Skull Base Questionnaire (ASBQ), European Quality-of-Life-5 Dimension (EQ-5D) questionnaire, and Lund-Kennedy endoscopic (LKE) score in 71 patients with sinonasal and skull base tumors. The mean age was 53 years and mean follow-up was 14.5 months at the time QOL data were collected. Benign and malignant tumors represented 39 (54.9%) and 32 (45.1%) cases, respectively. Twenty malignancies (62.5%) were stage T3 or T4, and 23 required postoperative chemotherapy and radiation (CRT). Factors indicating worsened postoperative QOL included malignant histopathology, T3 or T4 tumors, and the use of postoperative CRT (p < 0.05). There was a strong correlation of ASBQ with EQ-5D and SNOT-20 scores (r < -0.5) and a moderate correlation between the SNOT-20 and EQ-5D (r > 0.3), and the LKE had moderate correlation with SNOT-20 (r > 0.3) and weak correlation to the ASBQ (r > -0.3) and EQ-5D (r < 0.3). Patients who have undergone endoscopic resection of sinonasal tumors have quantifiable QOL changes as measured by various validated metrics. This study shows that concurrent use of these instruments may better discern QOL outcomes after endoscopic tumor surgery.


Postoperative survival and functional outcomes for patients with metastatic gynecological cancer to the spine: case series and review of the literature.
Spinal metastases from gynecological cancers are rare, with few cases reported in the literature. In this study, the authors examine a series of patients with spinal metastases from gynecological cancer and review the literature. The cases of 6 consecutive patients who underwent spine surgery for metastatic gynecological cancer between 2007 and 2012 at a single institution were retrospectively reviewed. The recorded demographic, operative, and postoperative factors were reviewed, and the functional outcomes were determined by change in Karnofsky Performance Scale and the American Spine Injury Association (ASIA) score during follow-up. A systematic review of the literature was also performed to evaluate outcomes for patients with similar gynecological metastases to the spine. In this series, details regarding metastatic gynecological cancers to the spine are as follows: 2 patients with cervical cancer (both presented at age 46 years, mean postoperative survival of 32 months), 2 patients with endometrial cancer (mean age of 40 years, mean postoperative survival of 26 months), and 2 patients with leiomyosarcoma (mean age of 44 years, mean postoperative survival of 20 months). All patients presented with pain, and no complications were noted following surgery. All patients with known follow-up had stable or improved neurological outcomes, performance status, and improved pain, without local recurrence of tumor. Overall median survival after diagnosis of metastatic spine lesions for all cases in the literature as well as those treated by the authors was 15 months. When categorized by type, median survival of patients with cervical cancer (n = 2), endometrial cancer (n = 26), and leiomyosarcoma (n = 16) was 32, 10, and 22.5 months, respectively. Gynecological cancers metastasizing to the spine are rare. In this series, overall survival following diagnosis of spinal metastasis and surgery was 27 months, with cervical cancer, endometrial cancer, and leiomyosarcoma survival being 32, 26, and 20 months, respectively. Combined with literature cases, survival differs depending on primary histology, with decreasing survival from cervical cancer (32 months) to leiomyosarcoma (22.5 months) to endometrial cancer (10 months). Integrating such information with other patient factors may more accurately guide decision making regarding management of such spinal lesions.


Effect of green tea in the prevention and reversal of fasting-induced intestinal mucosal damage.
Epidemiologic studies have suggested that high consumption of green tea protects against the development of chronic active gastritis and decreases the risk of stomach cancer. The effect of green tea on the intestinal mucosa was not studied previously, so we examined the effects of green tea on the intestinal mucosa of fasting rats in a controlled experimental setting. Two sets of experiments were performed. In the recovery set, rats were fasted for 3 d, after which they were allowed free access to water, black tea, green tea, or vitamin E for 7 d. On day 8, the animals were killed, and small bowels were removed for histologic examination. In the pretreatment set, rats were allowed a normal diet, but the water supply was replaced with green tea, black tea, or vitamin E for 14 d. They were subsequently fasted for 3 d. On day 4, the rats were killed, and small bowels were removed for histologic examination. In the recovery set, fasting for 3 d caused shortening of villi, atrophy, and fragmentation of mucosal villous architecture, with a significant (P < 0.0001) reduction in the length and surface area of the villi. Ingestion of green tea and, to a lesser extent, vitamin E for 7 d helped in the recovery of villi to normal. In the pretreatment set, drinking green tea, black tea, or vitamin E for 14 d before fasting protected intestinal mucosa from damage. The mucosal and villous atrophy induced by fasting was reverted to normal by the ingestion of green tea and, to a lesser extent, vitamin E. Black tea ingestion had no effect. In addition, ingestion of black tea, green tea, and vitamin E before fasting protected the intestinal mucosa against atrophy.


Cell Tracking Accuracy Measurement Based on Comparison of Acyclic Oriented Graphs.
Tracking motile cells in time-lapse series is challenging and is required in many biomedical applications. Cell tracks can be mathematically represented as acyclic oriented graphs. Their vertices describe the spatio-temporal locations of individual cells, whereas the edges represent temporal relationships between them. Such a representation maintains the knowledge of all important cellular events within a captured field of view, such as migration, division, death, and transit through the field of view. The increasing number of cell tracking algorithms calls for comparison of their performance. However, the lack of a standardized cell tracking accuracy measure makes the comparison impracticable. This paper defines and evaluates an accuracy measure for objective and systematic benchmarking of cell tracking algorithms. The measure assumes the existence of a ground-truth reference, and assesses how difficult it is to transform a computed graph into the reference one. The difficulty is measured as a weighted sum of the lowest number of graph operations, such as split, delete, and add a vertex and delete, add, and alter the semantics of an edge, needed to make the graphs identical. The measure behavior is extensively analyzed based on the tracking results provided by the participants of the first Cell Tracking Challenge hosted by the 2013 IEEE International Symposium on Biomedical Imaging. We demonstrate the robustness and stability of the measure against small changes in the choice of weights for diverse cell tracking algorithms and fluorescence microscopy datasets. As the measure penalizes all possible errors in the tracking results and is easy to compute, it may especially help developers and analysts to tune their algorithms according to their needs.


Clinical effectiveness in severe knee osteoarthritis after intra-articular platelet-rich plasma therapy in association with hyaluronic acid injection: three case reports.
Osteoarthritis (OA) is a degenerative disorder resulting from loss of joint cartilage and underlying bone and causes pain and loss of function. The treatment of knee OA is still a challenge because of the poor self-regeneration capacity of cartilage. The nonsurgical interventions include control of the aggravating factor (such as weight control and the use of walking aids), symptomatic treatment (such as acetaminophen or nonsteroidal anti-inflammatory drugs), prolotherapy, and viscosupplementation. However, the combination of platelet-rich plasma (PRP) and hyaluronic acid (HA) has not been widely used because of lack of clinical evidence and several limitations in patients with severe knee OA. Three patients who suffered from knee pain and poor walking endurance were diagnosed with advanced knee OA. They underwent PRP treatment in association with intra-articular HA injection and showed pain relief and functional improvement. The follow-up standard weight-bearing X-ray images of knees also confirmed the improvement and indicated the possibility of regeneration of the articular cartilage. These cases provide clinical and radiographic evidence for a new therapy for advanced knee OA. This treatment strategy of PRP in association with HA injection can offer a chance to treat severe knee OA, rather than immediate surgery, or a chance for those who cannot undergo surgery. It can also postpone the need of arthroplasty and can significantly improve the daily activity function.


GSM base stations: short-term effects on well-being.
The purpose of this study was to examine the effects of short-term GSM (Global System for Mobile Communications) cellular phone base station RF-EMF (radiofrequency electromagnetic fields) exposure on psychological symptoms (good mood, alertness, calmness) as measured by a standardized well-being questionnaire. Fifty-seven participants were selected and randomly assigned to one of three different exposure scenarios. Each of those scenarios subjected participants to five 50-min exposure sessions, with only the first four relevant for the study of psychological symptoms. Three exposure levels were created by shielding devices in a field laboratory, which could be installed or removed during the breaks between sessions such that double-blinded conditions prevailed. The overall median power flux densities were 5.2 microW/m(2) during "low," 153.6 microW/m(2) during "medium," and 2126.8 microW/m(2) during "high" exposure sessions. For scenario HM and MH, the first and third sessions were "low" exposure. The second session was "high" and the fourth was "medium" in scenario HM; and vice versa for scenario MH. Scenario LL had four successive "low" exposure sessions constituting the reference condition. Participants in scenarios HM and MH (high and medium exposure) were significantly calmer during those sessions than participants in scenario LL (low exposure throughout) (P = 0.042). However, no significant differences between exposure scenarios in the "good mood" or "alertness" factors were obtained. We conclude that short-term exposure to GSM base station signals may have an impact on well-being by reducing psychological arousal.


Disruption of Hippocampal Multisynaptic Networks by General Anesthetics.
Previous studies showed that synaptic transmission is affected by general anesthetics, but an anesthetic dose response in freely moving animals has not been done. The hippocampus provides a neural network for the evaluation of isoflurane and pentobarbital on multisynaptic transmission that is relevant to memory function. Male Long-Evans rats were implanted with multichannel and single electrodes in the hippocampus. Spontaneous local field potentials and evoked field potentials were recorded in freely behaving rats before (baseline) and after various doses of isoflurane (0.25 to 1.5%) and sodium pentobarbital (10 mg/kg intraperitoneal). Monosynaptic population excitatory postsynaptic potentials at the basal and apical dendrites of CA1 were significantly decreased at greater than or equal to 0.25% (n = 4) and greater than or equal to 1.0% (n = 6) isoflurane, respectively. The perforant path evoked multisynaptic response at CA1 was decreased by ~50% at greater than or equal to 0.25% isoflurane (n = 5). A decreased population excitatory postsynaptic potential was accompanied by increased paired-pulse facilitation. Population spike amplitude in relation to apical dendritic population excitatory postsynaptic potential was not significantly altered by isoflurane. Spontaneous hippocampal local field potential at 0.8 to 300 Hz was dose-dependently suppressed by isoflurane (n = 6), with local field potential power in the 50- to 150-Hz band showing the highest decrease with isoflurane dose, commensurate with the decrease in trisynaptic CA1 response. Low-dose pentobarbital (n = 7) administration decreased the perforant path evoked trisynaptic CA1 response and hippocampal local field potentials at 78 to 125 Hz. Hippocampal networks are sensitive to low doses of isoflurane and pentobarbital, possibly through both glutamatergic and γ-aminobutyric acid-mediated transmission. Network disruption could help explain the impairment of hippocampal-dependent cognitive functions with low-dose anesthetic.


Interference of some matrix ions in cation-exchange chromatography.
Ion chromatographic analysis of ions in samples containing complex matrix composition strongly depends on the on-column co-processes caused by sample matrix components. In the present publication studies of different separation phenomena in cation-exchange chromatography are described. The studies were performed at 'non-linear' chromatographic conditions, when the concentration of matrix (interfering) ions significantly exceeded the concentration of the eluent ions. During the research work, the processes already identified in anion-exchange chromatography, i.e. self-elution, on-column change of the eluent and sample-induced micro-gradient elution were used to explain the chromatographic behavior of alkaline and earth-alkaline cations when samples with high matrix cation concentration were analyzed. When present at higher concentrations, the ammonium cation was found to cause self-elution (NH4+ fraction) as well as on-column eluent neutralization due to its ability to diffuse into/back from the porous core of the stationary phase (NH3 fraction). Co-elution of a matrix component and analytes of interest caused spectroscopic interferences that additionally influenced the peak shape of each individual analyte.


Transverse stability of the carpus. An analytical study.
The stabilizing structures of the transverse carpal arch were studied utilizing an analytical model based on a rigid body spring model theory. This method was used to calculate the relative motion between carpal bones, the distribution of compressive forces in the intercarpal joints, and the tensions in the ligaments under simulated dorsopalmar compression. The contribution of each ligament against the effect of compression was determined by removing ligaments from the model. A multivariate analysis of variance disclosed that sectioning the flexor retinaculum alone does not significantly modify the distribution of forces in ligaments and joints (p = 0.33), and decreases dorsopalmar carpal stiffness by only 7.8%. In contrast, isolated removal of the palmar hamate-capitate ligament causes a pronounced weakening of the carpal structure, from an average of 232 to 131 N/mm (p less than 0.01). If the flexor retinaculum is removed in addition to releasing one palmar intercarpal ligament, a gross instability of the carpal arch is created.


Gastrointestinal Handling of Water-Soluble Vitamins.
Nine compounds are classified as water-soluble vitamins, eight B vitamins and one vitamin C. The vitamins are mandatory for the function of numerous enzymes and lack of one or more of the vitamins may lead to severe medical conditions. All the vitamins are supplied by food in microgram to milligram quantities and in addition some of the vitamins are synthesized by the intestinal microbiota. In the gastrointestinal tract, the vitamins are liberated from binding proteins and for some of the vitamins modified prior to absorption. Due to their solubility in water, they all require specific carriers to be absorbed. Our current knowledge concerning each of the vitamins differs in depth and focus and is influenced by the prevalence of conditions and diseases related to lack of the individual vitamin. Because of that we have chosen to cover slightly different aspects for the individual vitamins. For each of the vitamins, we summarize the physiological role, the steps involved in the absorption, and the factors influencing the absorption. In addition, for some of the vitamins, the molecular base for absorption is described in details, while for others new aspects of relevance for human deficiency are included. © 2018 American Physiological Society. Compr Physiol 8:1291-1311, 2018.


Mechanical properties of a biodegradable self-expandable polydioxanone monofilament stent: In vitro force relaxation and its clinical relevance.
Biodegradable stents are promising treatments for many diseases, e.g., coronary artery disease, urethral diseases, tracheal diseases, and esophageal strictures. The mechanical properties of biodegradable stent materials play a key role in the safety and efficacy of treatment. In particular, insufficient creep resistance of the stent material could result in premature stent collapse or narrowing. Commercially available biodegradable self-expandable SX-ELLA stents made of polydioxanone monofilament were tested. A new, simple, and affordable method to measure the shear modulus of tiny viscoelastic wires is presented. The important mechanical parameters of the polydioxanone filament were obtained: the median Young's modulus was [Formula: see text] = 958 (922, 974) MPa and the shear modulus was [Formula: see text] = 357 (185, 387) MPa, resulting in a Poisson's ratio of ν = 0.34. The SX-ELLA stents exhibited significant force relaxation due to the stress relaxation of the polydioxanone monofilament, approximately 19% and 36% 10 min and 48 h after stent application, respectively. However, these results were expected, and the manufacturer and implanting clinician should be aware of the known behavior of these biodegradable materials. If possible, a biodegradable stent should be designed considering therapeutic force rather than initial force. Additionally, new and more advanced biodegradable shape-memory polymers should be considered for future study and use.


Laminitis in Przewalski horses kept in a semireserve.
Semireserves were created by the European Conservation Project for scientific research in preparation for reintroduction in the wilderness. They are defined as enclosures large enough to carry a group of Przewalski horses throughout the year without any additional feeding. The semireserve offers diverse opportunities for significant scientific research. As part of a general screening program, the hoof development in a group of Przewalski horses was investigated in the semireserve Schorfheide near Berlin. Since the foundation of this semireserve in 1992, veterinary treatment was not necessary with the exception of hoof trimming in two animals in 1993. However, major health problems were encountered in the spring of 1999, when three other mares showed signs of laminitis. The initial diagnosis by the authors and the local veterinary surgeon based on observation of behaviour, gait, stance, walk and trot of three mares whose initial weights were higher than those of the healthy mares. The initial diagnosis was confirmed by palpation and the occurrence of very deep horn rings on all hooves and a laminitic horn ring on the right front hoof of one mare. An adequate laminitic therapy was not possible under the conditions of a semireserve. The applied management aimed at two goals: 1. To reduce endotoxin production and acidosis in the horses by reducing the ingestion of carbohydrate rich food. 2. To reduce the mares level of activity and to prevent tearing of the suspensory apparatus of the coffin bone. To achieve these two goals it was decided to remove the three laminitic mares from the rich pasture in the main part of the semireserve and to confine them onto the poorer pasture of the small separately fenced area. All three affected mares had fully recovered from their laminitic condition. Prevention of grass laminitis can be achieved by the following measures: 1. Reduction in grass intake could be achieved by increasing the grazing pressure by an increase in stocking rate of the horses or mixed grazing with another species such as sheep. 2. A longer term solution to the problem may well be to sow specific varieties of grass with lower concentrations of water soluble carbohydrate.


Estimating risk at a Superfund site using passive sampling devices as biological surrogates in human health risk models.
Passive sampling devices (PSDs) sequester the freely dissolved fraction of lipophilic contaminants, mimicking passive chemical uptake and accumulation by biomembranes and lipid tissues. Public Health Assessments that inform the public about health risks from exposure to contaminants through consumption of resident fish are generally based on tissue data, which can be difficult to obtain and requires destructive sampling. The purpose of this study is to apply PSD data in a Public Health Assessment to demonstrate that PSDs can be used as a biological surrogate to evaluate potential human health risks and elucidate spatio-temporal variations in risk. PSDs were used to measure polycyclic aromatic hydrocarbons (PAHs) in the Willamette River; upriver, downriver and within the Portland Harbor Superfund megasite for 3 years during wet and dry seasons. Based on an existing Public Health Assessment for this area, concentrations of PAHs in PSDs were substituted for fish tissue concentrations. PSD measured PAH concentrations captured the magnitude, range and variability of PAH concentrations reported for fish/shellfish from Portland Harbor. Using PSD results in place of fish data revealed an unacceptable risk level for cancer in all seasons but no unacceptable risk for non-cancer endpoints. Estimated cancer risk varied by several orders of magnitude based on season and location. Sites near coal tar contamination demonstrated the highest risk, particularly during the dry season and remediation activities. Incorporating PSD data into Public Health Assessments provides specific spatial and temporal contaminant exposure information that can assist public health professionals in evaluating human health risks.


Repeated fMRI using iron oxide contrast agent in awake, behaving macaques at 3 Tesla.
Iron oxide contrast agents have been employed extensively in anesthetized rodents to enhance fMRI sensitivity and to study the physiology of cerebral blood volume (CBV) in relation to blood oxygen level-dependent (BOLD) signal following neuronal activation. This study quantified the advantages of exogenous agent for repeated neuroimaging in awake, nonhuman primates using a clinical 3 Tesla scanner. A monocrystalline iron oxide nanoparticle (MION) solution was injected at iron doses of 8 to 10 mg/kg in two macaque monkeys. Adverse behavioral effects due to contrast agent were not observed in either monkey using cumulative doses in excess of 60 mg/kg. Relative to BOLD imaging at 3 Tesla, MION increased functional sensitivity by an average factor of 3 across the brain for a stimulus of long duration. Rapid stimulus presentation attenuated MION signal changes more than BOLD signal changes, due to the slower time constant of the blood volume response relative to BOLD signal. Overall, the contrast agent produced a dramatic improvement in functional brain imaging results in the awake, behaving primate at this field strength. (c) 2002 Elsevier Science (USA).


Two receptor forms for ouabain in sarcolemma-enriched preparations from canine ventricle.
Some evidence indicates that the inotropic effect of cardiac glycosides occurs at concentrations too low to affect Na+,K+-ATPase activity. This suggests that some receptor other than Na+,K+-ATPase mediates the inotropic effect. We studied ouabain binding to sarcolemma-enriched preparations from canine ventricle under conditions known to promote binding to Na+,K+-ATPase. Profiles for binding and dissociation were characterized by two kinetic components: (1) fast association and slow dissociation; (2) slow association and fast dissociation. Profiles in the absence of supporting ligands were consistent with a single species of receptors with slow association, fast dissociation and minimal effect on Na+,K+-ATPase activity. Binding supported by magnesium plus inorganic phosphate inhibited Na+,K+-ATPase activity by 86%. The two binding components were affected differentially by heating at 55 degrees C. It was concluded that the preparation possesses two receptors for ouabain: the Na+,K+-ATPase and a "new" receptor. The latter may be different chemically from the Na+,K+-ATPase. The more likely possibility is that the "new" receptor is the Na+,K+-ATPase in a state characterized by low catalytic activity, low affinity for ouabain, and no requirement of specific ligands for ouabain binding. Further, the data suggest an interdependence between the two forms. This leads to a mechanism which allows an inotropic effect to precede loss of Na+,K+-ATPase activity even though both result from glycoside binding to Na+,K+-ATPase. The mechanism involves an equilibrium between inactive and active forms of the Na+,K+-ATPase such that the inactive form buffers loss of the active form upon exposure to a cardiac glycoside.


Monitoring respiratory mechanics during mechanical ventilation: where do the signals come from?
Graphical patient data have become essential to the understanding and management of ventilator-dependent patients. These electronically generated data often reveal clues to subtle complications that, if corrected, could lead to improved patient-ventilator harmony. The apparent precision of the waveforms and the 3- or 4-place display of numeric data imply high accuracy. Laboratory devices and equipment, with their required certification, generally exhibit accuracies of approximately 2% for flow and significantly less than 1% for pressure. But the cost constraints placed on hospital-grade medical equipment dictate the use of commercial sensors-transducers, which means that pressure measurements will range between 3% and 5% of reading, and flow measurements will range between 6% and 10% (+/- 3 standard deviations of the mean). Other direct and indirect influences, such as temperature, humidity, absolute pressure, system pressure, type of gas, contamination, and myriad additional effects further interfere with the transformation of the variable of interest into an electrically equivalent signal. The abundance of viewable information pertinent to the management of the ventilated patient can be traced to the availability of the many types of transducers combined with microprocessor electronics. The process of capturing a variable of interest (sensing and signal transduction), converting it to a digitized electronic signal (analog-to-digital-conversion), operating on that signal (such as for control of the breathing algorithm and checking for violation of alarm thresholds), and finally converting it back to an analog signal that appears on a monitor generally receives scant appreciation. The process, however, lies at the core of data management in modern ICU ventilators.


Achieving proper exposure in surgical simulation.
One important technique common throughout surgery is achieving proper exposure of critical anatomic structures so that their shapes, which may vary somewhat among patients, can be confidently established and avoided. In this paper, we present an algorithm for determining which regions of selected structures are properly exposed in the context of a mastoidectomy simulation. Furthermore, our algorithm then finds and displays all other points along the surface of the structure that lie along a sufficiently short and straight path from an exposed portion such that their locations can be safely inferred. Finally, we present an algorithm for providing realistic visual cues about underlying structures with view-dependent shading of the bone.


Exploring the interaction of SV2A with racetams using homology modelling, molecular dynamics and site-directed mutagenesis.
The putative Major Facilitator Superfamily (MFS) transporter, SV2A, is the target for levetiracetam (LEV), which is a successful anti-epileptic drug. Furthermore, SV2A knock out mice display a severe seizure phenotype and die after a few weeks. Despite this, the mode of action of LEV is not known at the molecular level. It would be extremely desirable to understand this more fully in order to aid the design of improved anti-epileptic compounds. Since there is no structure for SV2A, homology modelling can provide insight into the ligand-binding site. However, it is not a trivial process to build such models, since SV2A has low sequence identity to those MFS transporters whose structures are known. A further level of complexity is added by the fact that it is not known which conformational state of the receptor LEV binds to, as multiple conformational states have been inferred by tomography and ligand binding assays or indeed, if binding is exclusive to a single state. Here, we explore models of both the inward and outward facing conformational states of SV2A (according to the alternating access mechanism for MFS transporters). We use a sequence conservation analysis to help guide the homology modelling process and generate the models, which we assess further with Molecular Dynamics (MD). By comparing the MD results in conjunction with docking and simulation of a LEV-analogue used in radioligand binding assays, we were able to suggest further residues that line the binding pocket. These were confirmed experimentally. In particular, mutation of D670 leads to a complete loss of binding. The results shed light on the way LEV analogues may interact with SV2A and may help with the on-going design of improved anti-epileptic compounds.


2020 American College of Rheumatology Guideline for the Management of Reproductive Health in Rheumatic and Musculoskeletal Diseases.
To develop an evidence-based guideline on contraception, assisted reproductive technologies (ART), fertility preservation with gonadotoxic therapy, use of menopausal hormone replacement therapy (HRT), pregnancy assessment and management, and medication use in patients with rheumatic and musculoskeletal disease (RMD). We conducted a systematic review of evidence relating to contraception, ART, fertility preservation, HRT, pregnancy and lactation, and medication use in RMD populations, using Grading of Recommendations Assessment, Development and Evaluation methodology to rate the quality of evidence and a group consensus process to determine final recommendations and grade their strength (conditional or strong). Good practice statements were agreed upon when indirect evidence was sufficiently compelling that a formal vote was unnecessary. This American College of Rheumatology guideline provides 12 ungraded good practice statements and 131 graded recommendations for reproductive health care in RMD patients. These recommendations are intended to guide care for all patients with RMD, except where indicated as being specific for patients with systemic lupus erythematosus, those positive for antiphospholipid antibody, and/or those positive for anti-Ro/SSA and/or anti-La/SSB antibodies. Recommendations and good practice statements support several guiding principles: use of safe and effective contraception to prevent unplanned pregnancy, pre-pregnancy counseling to encourage conception during periods of disease quiescence and while receiving pregnancy-compatible medications, and ongoing physician-patient discussion with obstetrics/gynecology collaboration for all reproductive health issues, given the overall low level of available evidence that relates specifically to RMD. This guideline provides evidence-based recommendations developed and reviewed by panels of experts and RMD patients. Many recommendations are conditional, reflecting a lack of data or low-level data. We intend that this guideline be used to inform a shared decision-making process between patients and their physicians on issues related to reproductive health that incorporates patients' values, preferences, and comorbidities.


Standards for the uniform reporting of hypertension in adults using population survey data: recommendations from the World Hypertension League Expert Committee.
Surveillance and monitoring of cardiovascular risk factors including raised blood pressure are critical to informing efforts to prevent and control cardiovascular disease. Yet, many countries lack the capacity for adequate national surveillance. Furthermore, hypertension indicators are often reported in different ways, which hampers the ability to compare and assess progress. In order to encourage standardized hypertension surveillance reporting, the World Hypertension League assembled an Expert Committee to develop a standard set of core indicators, definitions, and recommended analyses. The recommended core indicators are: (1) blood pressure distribution, (2) prevalence of hypertension, (3) awareness of the condition, (4) antihypertensive drug treatment, and (5) control of hypertension based on drug therapy. Each of these can be reported overall and by age group and sex, with crude and age-standardized changes tracked over time in order to assess the impact of instituted policies and programs for hypertension prevention and control. An expanded list of indicators can also facilitate tracking of hypertension prevention and control efforts. Widespread adoption of these indicators and analyses could benefit all those conducting and analyzing hypertension surveys and will facilitate hypertension surveillance efforts.


Altering the trajectory of early postnatal cortical development can lead to structural and behavioural features of autism.
Autism is a behaviourally defined neurodevelopmental disorder with unknown etiology. Recent studies in autistic children consistently point to neuropathological and functional abnormalities in the temporal association cortex (TeA) and its associated structures. It has been proposed that the trajectory of postnatal development in these regions may undergo accelerated maturational alterations that predominantly affect sensory recognition and social interaction. Indeed, the temporal association regions that are important for sensory recognition and social interaction are one of the last regions to mature suggesting a potential vulnerability to early maturation. However, direct evaluation of the emerging hypothesis that an altered time course of early postnatal development can lead to an ASD phenotype remains lacking. We used electrophysiological, histological, and behavioural techniques to investigate if the known neuronal maturational promoter valproate, similar to that in culture systems, can influence the normal developmental trajectory of TeA in vivo. Brain sections obtained from postnatal rat pups treated with VPA in vivo revealed that almost 40% of cortical cells in TeA prematurely exhibited adult-like intrinsic electrophysiological properties and that this was often associated with gross cortical hypertrophy and a reduced predisposition for social play behaviour. The co-manifestation of these functional, structural and behavioural features suggests that alteration of the developmental time course in certain high-order cortical networks may play an important role in the neurophysiological basis of autism.


The Impact of Care Specialty on Survival-Adjusted Medical Costs of COPD Patients After a Hospitalization: a longitudinal analysis.
Little is known about the impact of care provider's specialty on the medical costs of COPD patients over time. To compare the long-term medical costs between newly hospitalized COPD patients whose post-discharge care was initiated by a pulmonary specialist versus by a general practitioner. Retrospective matched cohort study. We identified patients with an incident COPD-related hospitalization from the administrative health database (January 1, 1996, to December 31, 2012) of British Columbia, Canada. Patients were categorized as receiving specialist care or primary care within the first 90 days after discharge from an incident COPD-related hospitalization. Using propensity scores, we matched each patient who initially received specialist care to a patient who received primary care based on demographics, COPD severity, comorbidity, and admission time. A survival-adjusted, multi-part generalized linear model was used to estimate direct medical costs (in 2015 Canadian dollars, [$], including inpatient, outpatient, pharmacy, and community care costs) as overall and as COPD-specific and comorbidity-related costs over the following 5 years. The sample included 7710 patients under each group. The initial specialist-care recipients had a modestly higher 5-year survival than the generalist-care recipients (0.564 [95% CI 0.535, 0.634] vs 0.555 [95% CI 0.525, 0.625]; P < .001). Meanwhile, the former incurred $2809 higher all-cause medical costs over 5 years compared to the latter (95% CI $2343, $2913; P < .001), mainly driven by higher medication costs (difference $1782 [95% CI $1658, $1830]; P < .001) particularly related to COPD medications ($1170 [95% CI $1043, $1225]; P < .001). Specialist care recipients also incurred higher costs of COPD-related hospitalization ($1144 [95% CI $650, $1221]; P < .001). Compared to generalist care, specialist care following COPD hospitalization is slightly more expensive, mainly driven by medication costs especially COPD-specific medications. Future studies should compare differences in health-related quality of life and COPD functional status.


The Texas implementation of medication algorithms: update to the algorithms for treatment of bipolar I disorder.
A panel consisting of academic psychiatrists and pharmacist administrators of the Texas Department of State Health Services (formerly Texas Department of Mental Health and Mental Retardation), community mental health physicians, advocates, and consumers met in May 2004 to review new evidence in the pharmacologic treatment of bipolar I disorder (BDI). The goal of the consensus conference was to update and revise the current treatment algorithm for BDI as part of the Texas Implementation of Medication Algorithms, a statewide quality assurance program for the treatment of major psychiatric illness. The guidelines for evaluating possible medications, the criteria for selection and ranking, and the updated algorithms are described. Principles from previous consensus conferences were reviewed and amended. Medication algorithms for the acute treatment of hypomanic/manic or mixed and depressive episodes in BDI were developed after examining recent efficacy and safety and tolerability data. Recommendations for maintenance treatments were developed. The panel updated the 2 primary algorithms (hypomanic/manic/mixed and depressive) based on clinical evidence for efficacy, tolerability, and safety developed since 2000. Expert consensus was utilized where clinical evidence was limited. Prevention of new episodes or prophylaxis treatment recommendations were developed based on recent data from longer-term trials. Maintenance recommendations are provided as levels versus a specified staged algorithm, as for acute treatment, due to the relatively limited database to inform treatment. These algorithms for the treatment of BDI represent the recommendations based on the most recent evidence available. These recommendations are meant to provide a framework for clinical decision making, not to replace clinical judgment. As with any algorithm, treatment practices will evolve beyond the recommendations of this consensus conference as new evidence and additional medications become available.


Group level validation of protein intakes estimated by 24-hour diet recall and dietary questionnaires against 24-hour urinary nitrogen in the European Prospective Investigation into Cancer and Nutrition (EPIC) calibration study.
A calibration approach was developed to correct for systematic between-cohort dietary measurement errors in the European Prospective Investigation into Cancer and Nutrition (EPIC), a large multicenter cohort study. To validate the 24-h diet recalls (24-HDRs) as reference measurements for between-cohort calibration, we estimated the agreement between center mean nitrogen (N) and total energy intakes and mean 24-h urinary N. Similar analyses using N and energy intake data from different dietary questionnaires (DQs) used at study baseline were conducted to estimate the effect of the calibration approach. This study was conducted between 1995 and 1999, and involved 1103 volunteers of both genders from 12 centers participating in European Prospective Investigation into Cancer and Nutrition. Pearson's correlation coefficients were weighted for study center sample size. When both genders were considered together (n = 22), the correlation coefficients between the center mean log-transformed urinary estimates and the center mean log-transformed dietary N estimates from the 24-HDRs were 0.86 and 0.94 after exclusion of outliers. The corresponding correlation with the DQs was 0.53. When center mean total energy intakes were regressed on center mean urinary N, the correlation remained slightly higher with 24-HDRs (0.91; 0.95 after exclusion of outliers) than DQs (0.86). When stratified by gender, these correlations were systematically higher in men than women with both dietary methods. The beta regression coefficients were not significantly different from 1 when mean N (or total energy intakes) from 24-HDR or DQ were regressed on urinary estimates, except with N from 24-HDRs in men and, in most cases, after adjustment for age, body mass index, and sex with both genders together. This suggests that overall the systematic bias across centers is of uniform magnitude. Although relatively high correlations were observed between urinary N and both dietary methods in men, the errors in DQs tend to vary in both directions (under- and over-reporting) in contrast with 24-HDRs in women. This observation may have implications on the dietary measurement error characteristics and support the potential benefit of between-cohort calibration.


Diastolic coronary resistance and capacitance are independent of the duration of diastole.
Systolic myocardial contraction may produce changes in coronary resistance and capacitance that persist throughout a normal diastole. In addition, coronary resistance and capacitance as determined in the arrested heart may not accurately describe normal diastolic behavior. To evaluate these possibilities, an identification method capable of characterizing the input impedance of the coronary circulation in as little as 150 ms was developed. Using this method, coronary dynamics were measured during early and late diastoles in the beating heart with tone intact as well as during adenosine-induced maximal vasodilation. Coronary dynamics were also measured in the arrested heart during maximal vasodilation. With vasomotor tone intact, the parameters of a lumped model of the coronary circulation showed no variation from early to late diastole. During maximal vasodilation, model parameters also showed no variation from early to late diastole. Parameters in the arrested heart were not statistically different from those of the beating heart during maximal vasodilation. However, model parameters determined during maximal vasodilation were significantly different from those determined with tone intact. These results suggest that although coronary resistance and capacitance are dependent on vasomotor tone, they remain constant throughout diastole and remain similar in the arrested heart.


Translational neurophysiology in sheep: measuring sleep and neurological dysfunction in CLN5 Batten disease affected sheep.
Creating valid mouse models of slowly progressing human neurological diseases is challenging, not least because the short lifespan of rodents confounds realistic modelling of disease time course. With their large brains and long lives, sheep offer significant advantages for translational studies of human disease. Here we used normal and CLN5 Batten disease affected sheep to demonstrate the use of the species for studying neurological function in a model of human disease. We show that electroencephalography can be used in sheep, and that longitudinal recordings spanning many months are possible. This is the first time such an electroencephalography study has been performed in sheep. We characterized sleep in sheep, quantifying characteristic vigilance states and neurophysiological hallmarks such as sleep spindles. Mild sleep abnormalities and abnormal epileptiform waveforms were found in the electroencephalographies of Batten disease affected sheep. These abnormalities resemble the epileptiform activity seen in children with Batten disease and demonstrate the translational relevance of both the technique and the model. Given that both spontaneous and engineered sheep models of human neurodegenerative diseases already exist, sheep constitute a powerful species in which longitudinal in vivo studies can be conducted. This will advance our understanding of normal brain function and improve our capacity for translational research into neurological disorders.


The effect of school-based kindergarten transition policies and practices on child academic outcomes.
This study examined the effect of school-based kindergarten transition policies and practices on child outcomes. The authors followed 17,212 children from 992 schools in the Early Childhood Longitudinal Study-Kindergarten sample (ECLS-K) across the kindergarten school year. Hierarchical linear modeling revealed that the number of school-based transition practices in the fall of kindergarten was associated with more positive academic achievement scores at the end of kindergarten, even controlling for family socioeconomic status (SES) and other demographic factors. This effect was stronger for low- and middle-SES children than high-SES children. For low-SES children, 7 transition practices were associated with a .21 standard deviation increase in predicted achievement scores beyond 0 practices. The effect of transition practices was partially mediated by an intervening effect on parent-initiated involvement in school during the kindergarten year. The findings support education policies to target kindergarten transition efforts to increase parent involvement in low-SES families.


Linoleate appears to protect against palmitate-induced inflammation in Huh7 cells.
Polyunsaturated fatty acids (PUFAs) may protect against metabolic diseases. Although the benefits of the n-3 family of PUFA have been well investigated in nonalcoholic steatohepatitis (NASH), little is known about the effect of the n-6 family. This study examined the effect of linoleate, a member of the n-6 family, on regulation of the palmitate-induced inflammatory cytokine interleukin-8 (IL8) in hepatocytes. Huh7 cells and HepG2 cells were cultured with and without free fatty acid treatment (palmitate and linoleate, alone or in combination, 100-1000 μM). Inflammatory pathways, lipid accumulation, apoptosis and cell viability were monitored. Dose- and time-related changes of IL8 mRNA expression were examined and 9 h treatment with 500 μM palmitate showed the greatest elevation of IL8. Co-treatment with 500 μM palmitate and 400 μM linoleate significantly suppressed IL8 production below that with palmitate alone in both cells (both mRNA and protein). A quantitative measurement for lipid accumulation showed no significant difference between palmitate-treated cells (1.69 ± 0.21), linoleate-treated cells (1.61 ± 0.16) and palmitate and linoleate-treated cells (1.73 ± 0.22, NS, n = 7). The co-treatment with 400 μM linoleate inhibited phospho-c-Jun N-terminal kinase (pJNK) activation and IkBα reduction caused by 500 μM palmitate treatment. Treatment with 400 μM linoleate alone led to IL8 production (5.48 fold change), similar to co-treatment, with no influence on the expression of pJNK/IkBα. The cell viability was similar between treatment with 500 μM palmitate and with both 500 μM palmitate and 400 μM linoleate, showing no significant changes in the expression of cleaved caspase-3. Linoleate is a potent regulator of the proinflammatory cytokine IL8 via the JNK and nuclear factor kappa B pathways that are involved in the pathophysiology of NASH, suggesting a future recommendation of dietary management.


Role Preferences of People with Multiple Sclerosis: Image-Revised, Computerized Self-Administered Version of the Control Preference Scale.
The Control Preference Scale (CPS) is the most frequently used measure of patients' preferred roles in treatment decisions. We revised the original CPS and developed a new computerized patient self-administered version (eCPS). We used the eCPS to assess role preferences, and their determinants, in Italian and German people with multiple sclerosis (MS). New cartoons were produced, based on MS health professional and patient input/feedback and previous findings, and pilot tested on 26 Italian and German MS patients. eCPS acceptability and reliability (weighted kappa statistic, wK) in comparison to the original tool, was determined in 92 MS patients who received both CPS versions in random order. The new cartoons were well accepted and easily interpreted by patients, who reported they based their choices mainly on the text and considered the images of secondary importance. eCPS reliability was moderate (wK 0.53, 95% confidence interval [CI] 0.40-0.65) and similar to the test-retest reliability of face-to-face administration assessed in a previous publication (wK 0.65, 95% CI 0.45-0.81). Higher education (odds ratio [OR] 3.74, 95% CI 1.00-14.05) and German nationality (OR 10.30, 95% CI 3.10-34.15) were associated with preference for an active role in the logistic model. The newly devised eCPS was well received and considered easy to use by MS patients. Reliability was in line with that of the original version. Role preference appears affected by cultural characteristics and (borderline statistical significance) education.


Effect of Creosote Bush-Derived NDGA on Expression of Genes Involved in Lipid Metabolism in Liver of High-Fructose Fed Rats: Relevance to NDGA Amelioration of Hypertriglyceridemia and Hepatic Steatosis.
Nordihydroguaiaretic acid (NDGA), the main metabolite of Creosote bush, has been shown to have profound effects on the core components of the metabolic syndrome (MetS), lowering blood glucose, free fatty acids (FFA) and triglyceride (TG) levels in several models of dyslipidemia, as well as improving body weight (obesity), insulin resistance, diabetes and hypertension, and ameliorating hepatic steatosis. In the present study, a high-fructose diet (HFrD) fed rat model of hypertriglyceridemia was employed to further delineate the underlying mechanism by which NDGA exerts its anti-hypertriglyceridemic action. In the HFrD treatment group, NDGA administration by oral gavage decreased plasma levels of TG, glucose, FFA, and insulin, increased hepatic mitochondrial fatty acid oxidation and attenuated hepatic TG accumulation. qRT-PCR measurements indicated that NDGA treatment increased the mRNA expression of key fatty acid transport (L-FABP, CD36), and fatty acid oxidation (ACOX1, CPT-2, and PPARα transcription factor) genes and decreased the gene expression of enzymes involved in lipogenesis (FASN, ACC1, SCD1, L-PK and ChREBP and SREBP-1c transcription factors). Western blot analysis indicated that NDGA administration upregulated hepatic insulin signaling (P-Akt), AMPK activity (P-AMPK), MLYCD, and PPARα protein levels, but decreased SCD1, ACC1 and ACC2 protein content and also inactivated ACC1 activity (increased P-ACC1). These findings suggest that NDGA ameliorates hypertriglyceridemia and hepatic steatosis primarily by interfering with lipogenesis and promoting increased channeling of fatty acids towards their oxidation.


Curcumin use in ulcerative colitis: is it ready for prime time? A systematic review and meta-analysis of clinical trials.
Curcumin, an active ingredient of the Indian herb turmeric (Curcuma longa), has shown promising anti-inflammatory properties. Studies of its potential benefits in treating patients with ulcerative colitis (UC) are limited. We performed a systematic review and meta-analysis of human randomized placebo controlled trials to evaluate the efficacy of adjunctive therapy with curcumin in treating patients with UC. We conducted a search of several databases (from January 2000 to September 2018). A random-effects model was used for analysis. We assessed heterogeneity between study-specific estimates using the Cochran Q statistical test, 95% prediction interval (PI) and I2 statistics. The outcomes assessed were the pooled odds of clinical response and remission as well as the endoscopic response. A total of 7 studies with 380 patients (curcumin n=188; placebo n=190) were included in the final analysis. The pooled odds ratio for clinical remission with curcumin use was 2.9 (95%CI 1.5-5.5, I2=45, P=0.002), clinical response was 2.6 (95%CI 1.5-4.5, I2=74%, P=0.001), and endoscopic response/remission was 2.3 (95%CI 1.2-4.6, I2=35.5%, P=0.01). Based on our study, combined mesalamine and curcumin therapy was associated with roughly threefold better odds of a clinical response compared to placebo, with minimal side effects. This response was statistically significant, albeit with heterogeneity, probably due to the different severity scoring indices, curcumin dosages and routes of drug delivery used.


Bystander-initiated rescue breathing for out-of-hospital cardiac arrests of noncardiac origin.
Although chest compression-only cardiopulmonary resuscitation (CPR) is effective for adult out-of-hospital cardiac arrest (OHCA) of cardiac origin, it remains uncertain whether bystander-initiated rescue breathing has an incremental benefit for OHCA of noncardiac origin. A nationwide, prospective, population-based, observational study covering the whole population of Japan and involving consecutive OHCA patients with emergency responder resuscitation attempts was conducted from January 2005 through December 2007. The primary outcome was neurologically intact 1-month survival. Multiple logistic regression analysis was used to assess the contribution of bystander-initiated CPR to better neurological outcomes. Among a total of 43 246 bystander-witnessed OHCAs of noncardiac origin, 8878 (20.5%) received chest compression-only CPR, and 7474 (17.3%) received conventional CPR with rescue breathing. The conventional CPR group (1.8%) had a higher rate of better neurological outcome than both the no CPR group (1.4%; odds ratio, 1.58; 95% confidence interval, 1.28 to 1.96) and the compression-only CPR group (1.5%; odds ratio, 1.32; 95% confidence interval, 1.03 to 1.69). However, the compression-only CPR group did not produce better neurological outcome than the no CPR group (odds ratio, 1.19; 95% confidence interval, 0.96 to 1.47). The number of OHCAs needed to treat with conventional CPR versus compression-only CPR to save a life with favorable neurological outcome after OHCA was 290. This nationwide observational study indicates that rescue breathing has an incremental benefit for OHCAs of noncardiac origin, but the impact on the overall survival after OHCA was small.


Modification of breast cancer risk according to age and menopausal status: a combined analysis of five population-based case-control studies.
While several risk factors for breast cancer have been identified, studies have not consistently shown whether these factors operate more strongly at certain ages or for just pre- or postmenopausal women. We evaluated whether risk factors for breast cancer differ according to age or menopausal status. Data from five population-based case-control studies conducted during 1988-2008 were combined and analyzed. Cases (N = 23,959) and population controls (N = 28,304) completed telephone interviews. Logistic regression was used to estimate adjusted odds ratios and 95 % confidence intervals and tests for interaction by age and menopausal status. Odds ratios for first-degree family history of breast cancer were strongest for younger women-reaching twofold elevations-but were still statistically significantly elevated by 58-69 % among older women. Obesity was inversely associated with breast cancer among younger women and positively associated with risk for older women (interaction P < 0.0001). Recent alcohol intake was more strongly related to breast cancer risk among older women, although consumption of 3 or more drinks/day among younger women also was associated with elevated odd ratios (P < 0.0001). Associations with benign breast disease and most reproductive/menstrual factors did not vary by age. Repeating analysis stratifying by menopausal status produced similar results. With few exceptions, menstrual and lifestyle factors are associated with breast cancer risk regardless of age or menopausal status. Variation in the association of family history, obesity, and alcohol use with breast cancer risk by age and menopausal status may need to be considered when determining individual risk for breast cancer.


Resource utilization in children with paracorporeal continuous-flow ventricular assist devices.
Paracorporeal continuous-flow ventricular assist devices (PCF VAD) are increasingly used in pediatrics, yet PCF VAD resource utilization has not been reported to date. Pediatric Interagency Registry for Mechanically Assisted Circulatory Support (PediMACS), a national registry of VADs in children, and Pediatric Health Information System (PHIS), an administrative database of children's hospitals, were merged to assess VAD implants from 19 centers between 2012 and 2016. Resource utilization, including hospital and intensive care unit length of stay (LOS), and costs are analyzed for PCF VAD, durable VAD (DVAD), and combined PCF-DVAD support. Of 177 children (20% PCF VAD, 14% PCF-DVAD, 66% DVAD), those with PCF VAD or PCF-DVAD are younger (median age 4 [IQR 0-10] years and 3 [IQR 0-9] years, respectively) and more often have congenital heart disease (44%; 28%, respectively) compared to DVAD (11 [IQR 3-17] years; 14% CHD); p < 0.01 for both. Median post-VAD LOS is prolonged ranging from 43 (IQR 15-82) days in PCF VAD to 72 (IQR 55-107) days in PCF-DVAD, with significant hospitalization costs (PCF VAD $450,000 [IQR $210,000-$780,000]; PCF-DVAD $770,000 [IQR $510,000-$1,000,000]). After adjusting for patient-level factors, greater post-VAD hospital costs are associated with LOS, ECMO pre-VAD, greater chronic complex conditions, and major adverse events (p < 0.05 for all). VAD strategy and underlying cardiac disease are not associated with LOS or overall costs, although PCF VAD is associated with higher daily-level costs driven by increased pharmacy, laboratory, imaging, and clinical services costs. Pediatric PCF VAD resource utilization is staggeringly high with costs primarily driven by pre-implantation patient illness, hospital LOS, and clinical care costs.


Molecular contribution function in RESOLFT nanoscopy.
The ultimate objective of a microscope of the highest resolution is to map the molecules of interest in the sample. Traditionally, linear imaging systems are characterized by their spatial frequency transfer function, which is given, in real space, by the point spread function (PSF). By extending the concept of the PSF towards the molecular contribution function (MCF), that quantifies the average contribution of a single fluorophore to the image, a straightforward concept for counting fluorophores is obtained. Using reversible saturable optical fluorescence transitions (RESOLFT), fluorophores are effectively activated only in a small, subdiffraction-sized volume before they are read out. During readout the signal exhibits an increased variance due to the stochastic nature of prior activation, which scales quadratically with the brightness of the active fluorophores while the mean of the signal scales only linearly with it. Using a two-state Markov model for the activation, showing comparable behavior to the switching kinetics of the switchable fluorescent protein rsEGFP2, we can approximate quantitatively the MCF of RESOLFT nanoscopy allowing to count the number of fluorophores within a subdiffraction-sized region of the sample. The method is validated on measurements of tubulin structures in Drosophila melagonaster larvae. Modeling and estimation of the MCF is a promising approach to quantitative microscopy.


Personalized prediction of smartphone-based psychotherapeutic micro-intervention success using machine learning.
Tailoring healthcare to patients' individual needs is a central goal of precision medicine. Combining smartphone-based interventions with machine learning approaches may help attaining this goal. The aim of our study was to explore the predictability of the success of smartphone-based psychotherapeutic micro-interventions in eliciting mood changes using machine learning. Participants conducted daily smartphone-based psychotherapeutic micro-interventions, guided by short video clips, for 13 consecutive days. Participants chose one of four intervention techniques used in psychotherapeutic approaches. Mood changes were assessed using the Multidimensional Mood State Questionnaire. Micro-intervention success was predicted using random forest (RF) tree-based mixed-effects logistic regression models. Data from 27 participants were used, totaling 324 micro-interventions, randomly split 100 times into training and test samples, using within-subject and between-subject sampling. Mood improved from pre- to post-intervention in 137 sessions (initial success-rate: 42.3%). The RF approach resulted in predictions of micro-intervention success significantly better than the initial success-rate within and between subjects (positive predictive value: 0.732 (95%-CI: 0.607; 0.820) and 0.698 (95%-CI: 0.564; 0.805), respectively). Prediction quality was highest using the RF approach within subjects (rand accuracy: 0.75 (95%-CI: 0.641; 0.840), Matthew's correlation coefficient: 0.483 (95%-CI: 0.323; 0.723)). The RF approach does not allow firm conclusions about the exact contribution of each factor to the algorithm's predictions. We included a limited number of predictors and did not compare whether predictability differed between psychotherapeutic techniques. Our findings may pave the way for translation and encourage scrutinizing personalized prediction in the psychotherapeutic context to improve treatment efficacy.


A model-based approach to dose selection in early pediatric development.
The establishment of a rationale for determining dosing regimens in pediatric patients remains a challenge in drug development. In this investigation, we explored several methodologies to support bridging studies and evaluated the best descriptor of developmental changes that can be used as a covariate for dose adjustment in children. The proposed approach is illustrated for the antiviral drug abacavir. Using data from six pharmacokinetic studies in adults and one study in children, a model-based analysis was applied in order to characterize differences in parameter distributions and their implications for systemic exposure to abacavir. Simulations were subsequently performed to define the appropriate dosing regimen in children. Although body weight was identified as a covariate for clearance and volume, dosing recommendations calculated on the basis of mg/kg cannot be linearly applied across all weight ranges. Our analysis shows the consequences of empirical dose adjustment and the importance of priors from historical data to support dose selection in children.


Fonts Designed for Macular Degeneration: Impact on Reading.
People with macular degeneration (MD) experience difficulties in reading due to central-field loss. Two new fonts, Eido and Maxular Rx, have been designed specifically for individuals with MD. We have compared reading performance of these new fonts with three mainstream fonts (Times-Roman, Courier, and Helvetica). Subjects with MD (n = 19) and normally sighted subjects (n = 40) were tested with digital versions of the MNREAD test using the five fonts. Maximum reading speed (MRS), critical print size (CPS), and reading acuity (RA) were estimated to characterize reading performance. Physical properties of the fonts were quantified by interletter spacing and perimetric complexity. Reading with MD showed font differences in MRS, CPS, and RA. Compared with Helvetica and Times, Maxular Rx permitted both smaller CPS and RA, and Eido permitted smaller RA. However, the two new fonts presented no advantage over Courier. Spacing, but not Complexity, was a significant predictor of reading performance for subjects with MD. The two fonts, designed specifically for MD, permit smaller print to be read, but provide no advantage over Courier.


Elimination of neglected tropical diseases in the South-East Asia Region of the World Health Organization.
The neglected tropical diseases (NTDs), which affect the very poor, pose a major public health problem in the South-East Asia Region of the World Health Organization (WHO). Although more than a dozen NTDs affect the region, over the past five years four of them in particular - leprosy, lymphatic filariasis, visceral leishmaniasis (kala-azar) and yaws - have been targeted for elimination. These four were selected for a number of reasons. First, they affect the WHO South-East Asia Region disproportionately. For example, every year around 67% of all new leprosy cases and 60% of all new cases of visceral leishmaniasis worldwide occur in countries of the region, where as many as 850 million inhabitants are at risk of contracting lymphatic filariasis. In addition, several epidemiological, technological and historical factors that are unique to the region make each of these four diseases amenable to elimination. Safe and effective tools and interventions to achieve these targets are available and concerted efforts to scale them up, singly or in an integrated manner, are likely to lead to success. The World Health Assembly and the WHO Regional Committee, through a series of resolutions, have already expressed regional and global commitments for the elimination of these diseases as public health problems. Such action is expected to have a quick and dramatic impact on poverty reduction and to contribute to the achievement of the Millennium Development Goals. This paper reviews the policy rationale for disease control in the WHO South-East Asia Region, the progress made so far, the lessons learnt along the way, and the remaining challenges and opportunities.


Biological properties of phospholipase C purified from a fleecerot isolate of Pseudomonas aeruginosa.
A role for one of many exocellular enzymes produced by Pseudomonas aeruginosa--phospholipase C (PLC)--as a prime candidate virulence factor in fleecerot dermatitis has been examined. The addition of Tween 80 in tryptose minimal medium effectively perturbed the membrane system of a field isolate of P. aeruginosa, resulting in increased production and release of a periplasmic enzyme marker, alkaline phosphatase (AP), and also of PLC. PLC activity levels in the culture supernatant were 10- to 15-fold higher in the presence of Tween than in its absence. Apart from AP, the culture medium contained little or no detectable proteolytic enzyme activity, thereby facilitating the partial purification of a haemolytic form of PLC by anion-exchange chromatography. This enzyme, when injected intradermally into the skin of sheep, elicited histopathological lesions virtually identical to those seen in naturally occurring fleecerot. In addition, serum from each of eight sheep afflicted with fleecerot contained high levels of circulating anti-PLC antibody activity when assayed by ELISA. Since these antibodies did not affect the enzymic function of PLC, it is likely that they do not bind to, or are incapable of conformational modification of, the active site.


The Anglo-Scandinavian Cardiac Outcomes Trial: blood pressure-lowering limb: effects in patients with type II diabetes.
To compare the effects of two antihypertensive treatment strategies for the prevention of coronary heart disease and other cardiovascular events in the large subpopulation (n=5137) with diabetes mellitus in the blood pressure-lowering arm of the Anglo-Scandinavian Cardiac Outcomes Trial. Patients had either untreated hypertension or treated hypertension. For those with type II diabetes mellitus, inclusion criteria required at least two additional risk factors. Patients were randomized to amlodipine with addition of perindopril as required (amlodipine-based) or atenolol with addition of thiazide as required (atenolol-based). Therapy was titrated to achieve a target blood pressure of less than 130/80 mmHg. The trial was terminated early due to significant benefits on mortality and stroke associated with the amlodipine-based regimen. In patients with diabetes mellitus, the amlodipine-based treatment reduced the incidence of the composite endpoint--total cardiovascular events and procedures--compared with the atenolol-based regimen (hazard ratio 0.86, confidence interval 0.76-0.98, P=0.026). Fatal and nonfatal strokes were reduced by 25% (P=0.017), peripheral arterial disease by 48% (P=0.004) and noncoronary revascularization procedures by 57% (P<0.001). For the other endpoints included in the composite, the endpoint differences were less clear including coronary heart disease deaths and nonfatal myocardial infarctions (the primary endpoint), which were reduced nonsignificantly by 8% (hazard ratio 0.92, confidence interval 0.74-1.15). In the large diabetic subgroup in the blood pressure-lowering arm of the Anglo-Scandinavian Cardiac Outcomes Trial, the benefits of amlodipine-based treatment, compared with atenolol-based treatment, on the incidence of total cardiovascular events and procedures was significant (14% reduction) and similar to that observed in the total trial population (16% reduction).


Atomic-scale dynamics of triangular hole growth in monolayer hexagonal boron nitride under electron irradiation.
The production of holes by electron beam irradiation in hexagonal boron nitride (hBN), which has a lattice similar to that of graphene, is monitored over time using atomic resolution transmission electron microscopy. The holes appear to be initiated by the formation of a vacancy of boron and grow in a manner that retains an overall triangular shape. The hole growth process involves the formation of single chains of B and N atoms and is accompanied by the ejection of atoms and bundles of atoms along the hole edges, as well as atom migration. These observations are compared to density functional theory calculations and molecular dynamics simulations.


Reduction of the Morphine Maintenance by Blockade of the NMDA Receptors during Extinction Period in Conditioned Place Preference Paradigm of Rats.
Activation of N-methyl-d-aspartate (NMDA) glutamate receptors in the nucleus accumbens is a component of drug-induced reward mechanism. In addition, NMDA receptors play a major role in brain reward system and activation of these receptors can change firing pattern of dopamine neurons. Blockade of glutamatergic neurotransmission reduces the expression of conditioned place preference (CPP) induced by morphine. Therefore, in this study, by using an NMDA receptor antagonist, DL-2-Amino-5-phosphonopentanoic acid sodium salt (AP5), the role of NMDA receptors on the maintenance and reinstatement of morphine-CPP was investigated. Forty-three adult male albino Wistar rats were used in this study. After subcutaneous administration of effective dose of morphine (5 mg/kg) during CPP paradigm, the animals received intracerebroventricular doses of AP5(1, 5, and 25 mM/5μL saline) during extinction period (free morphine stage). Conditioning score was recorded during extinction period and reinstatement phase. Besides, another group of the animals received a single dose administration of AP5(5 mM) just before the administration of ineffective dose of morphine (1 mg/kg) in reinstatement phase. The results revealed that two doses of this antagonist (5 and 25 mM) significantly shortened the extinction period of morphine-CPP but did not reduce reinstatement induced by priming dose of morphine. Moreover, the single dose administration of AP5(5 mM) just before prime-morphine injection decreased reinstatement of morphine-CPP. These findings indicate that blockade of NMDA receptors during extinction period reduces maintenance but not reinstatement of morphine. In addition, blocking these receptors in reinstatement phase decreases reinstatement to extinguished morphine.


Clinical and biological features associated with epidermal growth factor receptor gene mutations in lung cancers.
Mutations in the tyrosine kinase (TK) domain of the epidermal growth factor receptor (EGFR) gene in lung cancers are associated with increased sensitivity of these cancers to drugs that inhibit EGFR kinase activity. However, the role of such mutations in the pathogenesis of lung cancers is unclear. We sequenced exons 18-21 of the EGFR TK domain from genomic DNA isolated from 617 non-small-cell lung cancers (NSCLCs) and 524 normal lung tissue samples from the same patients and 36 neuroendocrine lung tumors collected from patients in Japan, Taiwan, the United States, and Australia and from 243 other epithelial cancers. Mutation status was compared with clinicopathologic features and with the presence of mutations in KRAS, a gene in the EGFR signaling pathway that is also frequently mutated in lung cancers. All statistical tests were two sided. We detected a total of 134 EGFR TK domain mutations in 130 (21%) of the 617 NSCLCs but not in any of the other carcinomas, nor in nonmalignant lung tissue from the same patients. In NSCLC patients, EGFR TK domain mutations were statistically significantly more frequent in never smokers than ever smokers (51% versus 10%), in adenocarcinomas versus cancer of other histologies (40% versus 3%), in patients of East Asian ethnicity versus other ethnicities (30% versus 8%), and in females versus males (42% versus 14%; all P < .001). EGFR TK domain mutation status was not associated with patient age at diagnosis, clinical stage, the presence of bronchioloalveolar histologic features, or overall survival. The EGFR TK domain mutations we detected were of three common types: in-frame deletions in exon 19, single missense mutations in exon 21, and in-frame duplications/insertions in exon 20. Rare missense mutations were also detected in exons 18, 20, and 21. KRAS gene mutations were present in 50 (8%) of the 617 NSCLCs but not in any tumors with an EGFR TK domain mutation. Mutations in either the EGFR TK domain or the KRAS gene can lead to lung cancer pathogenesis. EGFR TK domain mutations are the first molecular change known to occur specifically in never smokers.


Widespread Historical Contingency in Influenza Viruses.
In systems biology and genomics, epistasis characterizes the impact that a substitution at a particular location in a genome can have on a substitution at another location. This phenomenon is often implicated in the evolution of drug resistance or to explain why particular "disease-causing" mutations do not have the same outcome in all individuals. Hence, uncovering these mutations and their locations in a genome is a central question in biology. However, epistasis is notoriously difficult to uncover, especially in fast-evolving organisms. Here, we present a novel statistical approach that replies on a model developed in ecology and that we adapt to analyze genetic data in fast-evolving systems such as the influenza A virus. We validate the approach using a two-pronged strategy: extensive simulations demonstrate a low-to-moderate sensitivity with excellent specificity and precision, while analyses of experimentally validated data recover known interactions, including in a eukaryotic system. We further evaluate the ability of our approach to detect correlated evolution during antigenic shifts or at the emergence of drug resistance. We show that in all cases, correlated evolution is prevalent in influenza A viruses, involving many pairs of sites linked together in chains; a hallmark of historical contingency. Strikingly, interacting sites are separated by large physical distances, which entails either long-range conformational changes or functional tradeoffs, for which we find support with the emergence of drug resistance. Our work paves a new way for the unbiased detection of epistasis in a wide range of organisms by performing whole-genome scans.


Recovery Sleep versus Emotion Regulation in Predicting Fire Service Shift Workers Stress, Fatigue and Irritability.
Objective/Background: Fire service shift workers are at risk of developing mental health difficulties related to sleep loss and emotion dysregulation. We aimed to clarify the relationship between off-shift recovery sleep and emotion regulation on stress, fatigue and irritability. Participants: A total of 61 fire service shift workers (e.g. firefighter, captain, engineer, paramedic) on a "5/6" shift. Methods: Following five 24-hour shifts, participants reported on emotion regulation as well as daily sleep, stress, fatigue and irritability during six consecutive off-shift recovery days. Mediation analyses examined (1) emotion regulation as a predictor and sleep as a mediator of stress, fatigue and irritability outcomes; and (2) sleep as a predictor and emotion regulation as a mediator of stress, fatigue and irritability outcomes. Results: Greater self-reported total sleep time predicted lower recovery stress, fatigue, and irritability. Greater subjective sleep efficiency predicted lower recovery stress and fatigue, but not irritability. No significant relationships emerged for objective sleep or emotion regulation variables predicting stress, fatigue or irritability. There were no significant findings with either emotion regulation or sleep variables included as mediators. Conclusions: These findings suggest that stress management programs for fire service shift workers may be most effective when targeting sleep efficiency and quantity rather than emotion regulation strategies in the off-shift recovery period.


Lipoic acid stimulates cAMP production in T lymphocytes and NK cells.
The anti-oxidant lipoic acid (LA) potently suppresses clinical and pathologic disease in the animal model of multiple sclerosis, experimental autoimmune encephalomyelitis, by inhibiting the migration of pathogenic T cells to the spinal cord. The mechanism by which this occurs is largely unknown. In this report we demonstrate that LA induces increases in cyclic AMP, a known immunosuppressant, in human T cells. The increase in cAMP is associated with increased adenylyl cyclase activity and is partially blocked by prostanoid receptor antagonists. We present evidence that LA also stimulates cAMP production in natural killer (NK) cells. This novel mechanism of action is highly relevant to the immunomodulatory effects of LA and provides further support for the study of LA as a therapeutic agent for multiple sclerosis and other autoimmune diseases.


Myoadenylate deaminase deficiency. Functional and metabolic abnormalities associated with disruption of the purine nucleotide cycle.
To assess the role of the purine nucleotide cycle in human skeletal muscle function, we evaluated 10 patients with AMP deaminase deficiency (myoadenylate deaminase deficiency; MDD). 4 MDD and 19 non-MDD controls participated in an exercise protocol. The latter group was composed of a patient cohort (n = 8) exhibiting a constellation of symptoms similar to those of the MDD patients, i.e., postexertional aches, cramps, and pains; as well as a cohort of normal, unconditioned volunteers (n = 11). The individuals with MDD fatigued after performing only 28% as much work as their non-MDD counterparts. Muscle biopsies were obtained from the four MDD patients and the eight non-MDD patients at rest and following exercise to the point of fatigue. Creatine phosphate content fell to a comparable extent in the MDD (69%) and non-MDD (52%) patients at the onset of fatigue. Following exercise the 34% decrease in ATP content of muscle from the non-MDD subjects was significantly greater than the 6% decrease in ATP noted in muscle from the MDD patients (P = 0.048). Only one of four MDD patients had a measurable drop in ATP compared with seven of eight non-MDD patients. At end-exercise the muscle content of inosine 5'-monophosphate (IMP), a product of AMP deaminase, was 13-fold greater in the non-MDD patients than that observed in the MDD group (P = 0.008). Adenosine content of muscle from the MDD patients increased 16-fold following exercise, while there was only a twofold increase in adenosine content of muscle from the non-MDD patients (P = 0.028). Those non-MDD patients in whom the decrease in ATP content following exercise was measurable exhibited a stoichiometric increase in IMP, and total purine content of the muscle did not change significantly. The one MDD patient in whom the decrease in ATP was measurable, did not exhibit a stoichiometric increase in IMP. Although the adenosine content increased 13-fold in this patient, only 48% of the ATP catabolized could be accounted for by the combined increases of adenosine, inosine, hypoxanthine, and IMP. Studies performed in vitro with muscle samples from seven MDD and seven non-MDD subjects demonstrated that ATP catabolism was associated with a fivefold greater increase in IMP in non-MDD muscle. There were significant increases in AMP and ADP content of the muscle from MDD patients following ATP catabolism in vitro, while there was no detectable increase in AMP or ADP in non-MDD muscle. Adenosine content of MDD muscle increased following ATP catabolism, but there was no detectable increase in adenosine content of non-MDD muscle following ATP catabolism in vitro. These studies demonstrate that AMP deaminase deficiency leads to reduced entry of adenine nucleotides into the purine nucleotide cycle during exercise. We postulate that the resultant disruption of the purine nucleotide cycle accounts for the muscle dysfunction observed in these patients.


Reference gene selection for qPCR is dependent on cell type rather than treatment in colonic and vaginal human epithelial cell lines.
The ability of commensal bacteria to influence gene expression in host cells under the influence of pathogenic bacteria has previously been demonstrated, however the extent of this interaction is important for understanding how bacteria can be used as probiotics. Real-time quantitative polymerase chain reaction is the most sensitive tool for evaluating relative changes to gene expression levels. However as a result of its sensitivity an appropriate method of normalisation should be used to account for any variation incurred in preparatory experimental procedures. These variations may result from differences in the amount of starting material, quality of extracted RNA, or in the efficiency of the reverse transcriptase or polymerase enzymes. Selection of an endogenous control gene is the preferred method of normalisation, and ideally a proper validation of the gene's appropriateness for the study in question should be performed. In this study we used quantitative polymerase chain reaction data and applied four different algorithms (geNorm, BestKeeper, NormFinder, and comparative ΔCq) to evaluate eleven different genes as to their suitability as endogenous controls for use in studies involving colonic (HT-29) and vaginal (VK2/E6E7) human mucosal epithelial cells treated with probiotic and pathogenic bacteria. We found phosphoglycerate kinase 1 to be most appropriate for HT-29 cells, and ribosomal protein large P0 to be the best choice for VK2/E6E7 cells. We also showed that use of less stable reference genes can lead to less accurate quantification of expression levels of gene of interest (GOI) and also can result in decreased statistical significance for GOI expression levels when compared to control. Additionally, we found the cell type being analysed had greater influence on reference gene selection than the treatment performed. This study provides recommendations for stable endogenous control genes for use in further studies involving colonic and vaginal cell lines after bacterial challenge.


A Real-World Observational Study of Time to Treatment Intensification Among Elderly Patients with Inadequately Controlled Type 2 Diabetes Mellitus.
Among elderly patients, the management of type 2 diabetes mellitus (T2DM) is complicated by population heterogeneity and elderly-specific complexities. Few studies have been done to understand treatment intensification among elderly patients failing multiple oral antidiabetic drugs (OADs). To examine the association between time to treatment intensification of T2DM and elderly-specific patient complexities. In this observational, retrospective cohort study, elderly (aged ≥ 65 years) Medicare beneficiaries (n = 16,653) with inadequately controlled T2DM (hemoglobin A1c ≥ 8.0% despite 2 OADs) were included. Based on the consensus statement for diabetes care in elderly patients published by the American Diabetes Association and the American Geriatric Society, elderly-specific patient complexities were defined as the presence or absence of 5 geriatric syndromes: cognitive impairment; depression; falls and fall risk; polypharmacy; and urinary incontinence. Overall, 48.7% of patients received intensified treatment during follow-up, with median time to intensification 18.5 months (95% CI = 17.7-19.3). Median time to treatment intensification was shorter for elderly patients with T2DM with polypharmacy (16.5 months) and falls and fall risk (12.7 months) versus those without polypharmacy (20.4 months) and no fall risk (18.6 months). Elderly patients with urinary incontinence had a longer median time to treatment intensification (18.6 months) versus those without urinary incontinence (14.6 months). The median time to treatment intensification did not significantly differ by the elderly-specific patient complexities that included cognitive impairment and depression. However, after adjusting for demographic, insurance, clinical characteristics, and health care utilization, we found that only polypharmacy was associated with time to treatment intensification (adjusted hazard ratio, 1.10; 95% CI = 1.04-1.15; P = 0.001). Less than half of elderly patients with inadequately controlled T2DM received treatment intensification. Elderly-specific patient complexities were not associated with time to treatment intensification, emphasizing a positive effect of the integrated health care delivery model. Emerging health care delivery models that target integrated care may be crucial in providing appropriate treatment for elderly T2DM patients with complex conditions.


Lanthanide-based tools for the investigation of cellular environments.
Biological probes constructed from lanthanides can provide a variety of readout signals, such as the luminescence of Eu(iii), Tb(iii), Yb(iii), Sm(iii) and Dy(iii), and the proton relaxation enhancement of Gd(iii) and Eu(ii). For numerous applications the intracellular delivery of the lanthanide probe is essential. Here, we review the methods for the intracellular delivery of non-targeted complexes (i.e. where the overall complex structure enhances cellular uptake), as well as complexes attached to a targeting unit (i.e. to a peptide or a small molecule) that facilitates delivery. The cellular applications of lanthanide-based supramolecules (dendrimers, metal organic frameworks) are covered briefly. Throughout, we emphasize the techniques that can confirm the intracellular localization of the lanthanides and those that enable the determination of the fate of the probes once inside the cell. Finally, we highlight methods that have not yet been applied in the context of lanthanide-based probes, but have been successful in the intracellular delivery of other metal-based probes.