Key role for store-operated Ca2+ channels in activating gene expression in human airway bronchial epithelial cells.
Ca2+ entry into airway epithelia is important for activation of the NFAT family of transcription factors and expression of genes including epidermal growth factor that help orchestrate local inflammatory responses. However, the identity of epithelial Ca2+ channel that activates these transcriptional responses is unclear. In many other non-excitable cells, store-operated Ca2+ entry is a major route for Ca2+ influx and is mediated by STIM1 and Orai1 proteins. This study was performed to determine if store-operated Ca2+ channels were expressed in human bronchial epithelial cells and, if so, whether they coupled Ca2+ entry to gene expression. Cytoplasmic Ca2+ measurements, patch clamp recordings, RNAi knockdown and functional assays were used to identify and then investigate the role of these Ca2+ channels in activating the NFAT and c-fos pathways and EGF expression. STIM1 and Orai1 mRNA transcripts as well as proteins were robustly in epithelial cells and formed functional Ca2+ channels. Ca2+ entry through the channels activated expression of c-fos and EGF as well as an NFAT-dependent reporter gene. Store-operated Ca2+ entry was also important for epithelial cell migration in a scrape wound assay. These findings indicate that store-operated Ca2+ channels play an important role in stimulating airway epithelial cell gene expression and therefore comprise a novel potential therapeutic target for the treatment of chronic asthma and related airway disorders.


Inhaled anesthetic responses of recombinant receptors and knockin mice harboring α2(S270H/L277A) GABA(A) receptor subunits that are resistant to isoflurane.
The mechanism by which the inhaled anesthetic isoflurane produces amnesia and immobility is not understood. Isoflurane modulates GABA(A) receptors (GABA(A)-Rs) in a manner that makes them plausible targets. We asked whether GABA(A)-R α2 subunits contribute to a site of anesthetic action in vivo. Previous studies demonstrated that Ser270 in the second transmembrane domain is involved in the modulation of GABA(A)-Rs by volatile anesthetics and alcohol, either as a binding site or a critical allosteric residue. We engineered GABA(A)-Rs with two mutations in the α2 subunit, changing Ser270 to His and Leu277 to Ala. Recombinant receptors with these mutations demonstrated normal affinity for GABA, but substantially reduced responses to isoflurane. We then produced mutant (knockin) mice in which this mutated subunit replaced the wild-type α2 subunit. The adult mutant mice were overtly normal, although there was evidence of enhanced neonatal mortality and fear conditioning. Electrophysiological recordings from dentate granule neurons in brain slices confirmed the decreased actions of isoflurane on mutant receptors contributing to inhibitory synaptic currents. The loss of righting reflex EC(50) for isoflurane did not differ between genotypes, but time to regain the righting reflex was increased in N(2) generation knockins. This effect was not observed at the N(4) generation. Isoflurane produced immobility (as measured by tail clamp) and amnesia (as measured by fear conditioning) in both wild-type and mutant mice, and potencies (EC(50)) did not differ between the strains for these actions of isoflurane. Thus, immobility or amnesia does not require isoflurane potentiation of the α2 subunit.


Northern Hemisphere forcing of climatic cycles in Antarctica over the past 360,000 years.
The Milankovitch theory of climate change proposes that glacial-interglacial cycles are driven by changes in summer insolation at high northern latitudes. The timing of climate change in the Southern Hemisphere at glacial-interglacial transitions (which are known as terminations) relative to variations in summer insolation in the Northern Hemisphere is an important test of this hypothesis. So far, it has only been possible to apply this test to the most recent termination, because the dating uncertainty associated with older terminations is too large to allow phase relationships to be determined. Here we present a new chronology of Antarctic climate change over the past 360,000 years that is based on the ratio of oxygen to nitrogen molecules in air trapped in the Dome Fuji and Vostok ice cores. This ratio is a proxy for local summer insolation, and thus allows the chronology to be constructed by orbital tuning without the need to assume a lag between a climate record and an orbital parameter. The accuracy of the chronology allows us to examine the phase relationships between climate records from the ice cores and changes in insolation. Our results indicate that orbital-scale Antarctic climate change lags Northern Hemisphere insolation by a few millennia, and that the increases in Antarctic temperature and atmospheric carbon dioxide concentration during the last four terminations occurred within the rising phase of Northern Hemisphere summer insolation. These results support the Milankovitch theory that Northern Hemisphere summer insolation triggered the last four deglaciations.


GABA receptors as broadcasters of sexually differentiating signals in the brain.
Epileptic seizures are more common in males than in females. One of the areas that has recently been implicated in the higher susceptibility of males to seizures is the substantia nigra reticulata (SNR). Several studies support the existence of phenotypic differences between male and female infantile SNR neurons, and particularly in several aspects of the GABAergic system, including its ability to control seizures. We have recently found that at postnatal day 14-17 (PN14-17) rats, which are equivalent to infants, activation of GABA(A) receptors has different physiological effects in male and female SNR neurons. This is likely due to the differences in the expression of the neuronal-specific potassium-chloride co-transporter KCC2, which regulates the intracellular chloride concentration. In male PN14-17 SNR neurons, GABA(A)-receptor activation with muscimol causes depolarization and increments in intracellular calcium concentration and the expression of calcium regulated genes, such as KCC2. Blockade of L-type voltage-sensitive calcium channels (L-VSCC) by nifedipine decreases KCC2 mRNA expression. However, in PN14-17 females, muscimol hyperpolarizes the SNR neurons, does not increase intracellular calcium, and decreases KCC2 mRNA expression. In PN15 females, nifedipine has no effect on KCC2 mRNA expression in the SNR. This sexually dimorphic function of GABA(A) receptors also creates divergent patterns of estradiol signaling. In male PN15 rats, estradiol decreases KCC2 mRNA expression in SNR neurons. Pretreatment with the GABA(A)-receptor antagonist bicuculline or with nifedipine, prevents the appearance of estradiol-mediated downregulation of KCC2 mRNA expression. In contrast, in PN15 females, estradiol does not influence KCC2 expression. These findings show that, in infantile rats, drugs or conditions that modulate the activity of GABA(A) receptors or L-VSCCs have different effects on the differentiation of the SNR. As a result, they have the potency of causing long-term changes in the function of the SNR in the control of seizures, movement, and the susceptibility to and course of epilepsy and movement disorders.


Quality indicators for patients with traumatic brain injury in European intensive care units: a CENTER-TBI study.
The aim of this study is to validate a previously published consensus-based quality indicator set for the management of patients with traumatic brain injury (TBI) at intensive care units (ICUs) in Europe and to study its potential for quality measurement and improvement. Our analysis was based on 2006 adult patients admitted to 54 ICUs between 2014 and 2018, enrolled in the CENTER-TBI study. Indicator scores were calculated as percentage adherence for structure and process indicators and as event rates or median scores for outcome indicators. Feasibility was quantified by the completeness of the variables. Discriminability was determined by the between-centre variation, estimated with a random effect regression model adjusted for case-mix severity and quantified by the median odds ratio (MOR). Statistical uncertainty of outcome indicators was determined by the median number of events per centre, using a cut-off of 10. A total of 26/42 indicators could be calculated from the CENTER-TBI database. Most quality indicators proved feasible to obtain with more than 70% completeness. Sub-optimal adherence was found for most quality indicators, ranging from 26 to 93% and 20 to 99% for structure and process indicators. Significant (p < 0.001) between-centre variation was found in seven process and five outcome indicators with MORs ranging from 1.51 to 4.14. Statistical uncertainty of outcome indicators was generally high; five out of seven had less than 10 events per centre. Overall, nine structures, five processes, but none of the outcome indicators showed potential for quality improvement purposes for TBI patients in the ICU. Future research should focus on implementation efforts and continuous reevaluation of quality indicators. The core study was registered with ClinicalTrials.gov, number NCT02210221, registered on August 06, 2014, with Resource Identification Portal (RRID: SCR_015582).


Allograft reconstruction for symptomatic chronic complete proximal hamstring tendon avulsion.
Complete proximal hamstring tendon avulsion is an uncommon injury that can cause significant disability in young, athletic individuals. Surgical reattachment is recommended and can be performed on a delayed basis if the tissue is sufficiently mobile. We report 2-year follow up for two cases where interpositional allograft tissue was used for reconstruction because the tendon was too retracted for primary repair. Two 30-year-old patients with complete proximal hamstring avulsion at least 2 years earlier reported severe hamstring weakness and restrictions with respect to sport and recreational activities. Proximal hamstring tendon reconstruction with Achilles tendon allograft was performed for both patients. They were immobilized for 8 weeks with the hip in extension and the knee in flexion using a custom orthosis, followed by physical therapy and weight bearing as tolerated. The patients were followed for over 2 years after the surgery and were evaluated with physical examination, isokinetic strength testing and detailed questions about their function. Following the procedure, both patients returned to a more active lifestyle that was greatly improved with respect to participation in sport and function. This procedure should be considered as a salvage operation as the patients did not return to completely normal function and demonstrated hamstring weakness on the operated side.


The contribution of predators and scavengers to human well-being.
Predators and scavengers are frequently persecuted for their negative effects on property, livestock and human life. Research has shown that these species play important regulatory roles in intact ecosystems including regulating herbivore and mesopredator populations that in turn affect floral, soil and hydrological systems. Yet predators and scavengers receive surprisingly little recognition for their benefits to humans in the landscapes they share. We review these benefits, highlighting the most recent studies that have documented their positive effects across a range of environments. Indeed, the benefits of predators and scavengers can be far reaching, affecting human health and well-being through disease mitigation, agricultural production and waste-disposal services. As many predators and scavengers are in a state of rapid decline, we argue that researchers must work in concert with the media, managers and policymakers to highlight benefits of these species and the need to ensure their long-term conservation. Furthermore, instead of assessing the costs of predators and scavengers only in economic terms, it is critical to recognize their beneficial contributions to human health and well-being. Given the ever-expanding human footprint, it is essential that we construct conservation solutions that allow a wide variety of species to persist in shared landscapes. Identifying, evaluating and communicating the benefits provided by species that are often considered problem animals is an important step for establishing tolerance in these shared spaces.


Removal of Rhodamine B from aqueous solution using magnetic NiFe nanoparticles.
Surface-modified magnetic nano alloy particles Ni2.33Fe were prepared using a hydrothermal method and they were utilized for removing Rhodamine B (RhB) from aqueous solution. The magnetic nanoparticles were characterized by X-ray diffraction, scanning electron microscopy, thermogravimetric analysis and Fourier transform infrared spectroscopy, which confirmed that the surface of the magnetic product with a face-centered cubic-type structure was successfully modified by sodium citrate. Kinetics studies were conducted. The pseudo-second-order kinetic model was used for fitting the kinetic data successfully. The Freundlich and Langmuir adsorption models were employed for the mathematical description of adsorption equilibrium. It was found that the adsorption isotherm can be very satisfactorily fitted by the Freundlich model.


OGG1 is a novel prognostic indicator in acute myeloid leukaemia.
OGG1 (8-oxoguanine DNA glycosylase) constitutes a key component of the DNA base excision repair pathway, catalysing the removal of 8-oxoguanine nucleotides from DNA, thereby suppressing mutagenesis and cell death. We found that OGG1 expression was significantly downregulated by the RUNX1-ETO fusion protein product of the t(8;21) chromosome translocation in normal haematopoietic progenitor cells and in patients with acute myeloid leukaemia (AML). Further examination of OGG1 expression in 174 AML trial patients using Affymetrix microarrays showed that the prevalence rate of OGG1 expression was 33% and correlated strongly with adverse cytogenetics. OGG1-expressing patients had a worse relapse-free survival and overall survival and an increased risk of relapse at 5-years of follow-up. There remained a trend towards increased relapse rate among OGG1-expressing patients, even after adjusting for other known risk factors in comprehensive stratified analyses. We also determined a trend for OGG1 expression to have a more adverse impact on disease outcome in the context of the FLT3-ITD mutation. This study highlights OGG1 as a valuable prognostic marker that could be used to sub-stratify AML patients to predict those likely to fail conventional chemotherapies but those likely to benefit from novel therapeutic approaches that modulate DNA repair activity.


Autologous bone-marrow stem-cell transplantation for myocardial regeneration.
Implantation of bone-marrow stem cells in the heart might be a new method to restore tissue viability after myocardial infarction. We injected up to 1.5x10(6) autologous AC133+ bone-marrow cells into the infarct border zone in six patients who had had a myocardial infarction and undergone coronary artery bypass grafting. 3-9 months after surgery, all patients were alive and well, global left-ventricular function was enhanced in four patients, and infarct tissue perfusion had improved strikingly in five patients. We believe that implantation of AC133+ stem cells to the heart is safe and might induce angiogenesis, thus improving perfusion of the infarcted myocardium. See Commentary page 11


[Comparison of three surgical techniques for controlling nasal width after Le Fort I osteotomy].
To identify nasal width changes occurring after Le Fort I osteotomy and to compare prospectively the effect of three surgical techniques for controlling postoperative nasal width. In the study, 79 patients (22 male and 57 female, mean age 23.2 ± 3.4 years), who received Le Fort I osteotomy at Peking University Hospital of Stomatology from 2011 to 2014, were randomly divided into three groups. Group 1 was treated with traditional intraoral alar base cinch suture (ABCS); Group 2 with extraoral ABCS, and Group 3 with traditional ABCS plus an extra intraoral suture at points G.lat. All the patients had taken 3D photos using 3dMD camera before operation, and 3, and 6 months after operation. The nasal widths, which were indicated as distances between Sbal-Sbal, G.lat-G.lat and Al-Al, were measured by two examiners in the 3D photos three times with a time-interval of one week. SPSS 13.0 was used to do the statistic analysis. At the end of the postoperative 6 months, the nasal widths lessened as compared with the postoperative 3 months. No significant differences were found between the three groups 6 months after the operation. The degree of the postoperative nasal width widening had positive correlation with that of the intraoperative nasal width widening, and had negative correlation with the initial nasal width and the amount of post-suture narrowing. There is no difference between three suturing techniques for controlling nasal width widening after Le Fort I osteotomy. The postoperative nasal width-widening can't be totally avoided, and the alteration might last at least 6 months after the operation. For patients with narrow nasal width and need to move maxilla forward, more overcorrection of ABCS is needed to control the postoperative nasal base widening.


Shock Index Predicts Patient-Related Clinical Outcomes in Stroke.
Background The prognostic value of shock index ( SI ), heart rate divided by systolic blood pressure, in stroke for clinical outcomes other than mortality is not well understood. Methods and Results We examined the Get With The Guidelines-Stroke ( GWTG -Stroke) data to explore the usefulness of SI in predicting in-hospital outcomes in 425 808 acute stroke cases (mean age: 71.0±14.5 years; 48.8% male; 89.7% ischemic stroke and 10.3% intracerebral hemorrhage) admitted between October 2012 and March 2015. Compared with patients with SI of 0.5 to 0.7, patients with SI >0.7 (13.6% of the sample) had worse outcomes, with adjusted odds ratios of 2.00 (95% confidence interval [ CI ], 1.92-2.08) for in-hospital mortality, 1.46 (95% CI , 1.43-1.49) for longer length of hospital stay >4 days, 1.50 (95% CI , 1.47-1.54) for discharge destination other than home, 1.41 (95% CI , 1.38-1.45) for inability to ambulate independently at discharge, and 1.52 (95% CI , 1.47-1.57) for modified Rankin Scale score of 3 to 6 at discharge. Results were similar when analyses were confined to those with available National Institutes of Health Stroke Scale (NIHSS) or within individual stroke subtypes or when SI was additionally included in the models with or without blood pressure components. Every 0.1 increase in SI >0.5 was associated with significantly worse outcomes in linear spline models. The addition of SI to existing GWTG -Stroke mortality prediction models without NIHSS demonstrated modest improvement, but little to no improvement was noted in models with NIHSS . Conclusions SI calculated at the point of care may be a useful prognostic indicator to identify those with high risk of poor outcomes in acute stroke, especially in hospitals with limited experience with NIHSS assessment.


An integrated osteopathic treatment approach in acute otitis media.
Ear pain is a common patient complaint in the practice of the primary care physician. Acute otitis media can affect a person of any age, although it is more often seen in children than in adults. The disease is usually caused by Streptococcus pneumoniae (Diplococcus pneumoniae) or Haemophilus influenzae. The differential diagnosis and subsequent treatment of otitis media is approximately the same for children and adults. First-line therapy usually consists of an antibiotic regimen of amoxicillin in combination with autoinflation exercises. In the case presented, a pharmacologic regimen was combined with osteopathic manipulation.


Simulation of therapeutic electron beam tracking through a non-uniform magnetic field using finite element method.
In radiotherapy, megaelectron volt (MeV) electrons are employed for treatment of superficial cancers. Magnetic fields can be used for deflection and deformation of the electron flow. A magnetic field is composed of non-uniform permanent magnets. The primary electrons are not mono-energetic and completely parallel. Calculation of electron beam deflection requires using complex mathematical methods. In this study, a device was made to apply a magnetic field to an electron beam and the path of electrons was simulated in the magnetic field using finite element method. A mini-applicator equipped with two neodymium permanent magnets was designed that enables tuning the distance between magnets. This device was placed in a standard applicator of Varian 2100 CD linear accelerator. The mini-applicator was simulated in CST Studio finite element software. Deflection angle and displacement of the electron beam was calculated after passing through the magnetic field. By determining a 2 to 5cm distance between two poles, various intensities of transverse magnetic field was created. The accelerator head was turned so that the deflected electrons became vertical to the water surface. To measure the displacement of the electron beam, EBT2 GafChromic films were employed. After being exposed, the films were scanned using HP G3010 reflection scanner and their optical density was extracted using programming in MATLAB environment. Displacement of the electron beam was compared with results of simulation after applying the magnetic field. Simulation results of the magnetic field showed good agreement with measured values. Maximum deflection angle for a 12 MeV beam was 32.9° and minimum deflection for 15 MeV was 12.1°. Measurement with the film showed precision of simulation in predicting the amount of displacement in the electron beam. A magnetic mini-applicator was made and simulated using finite element method. Deflection angle and displacement of electron beam were calculated. With the method used in this study, a good prediction of the path of high-energy electrons was made before they entered the body.


Trauma and dissociation: implications for borderline personality disorder.
Psychological trauma can have devastating consequences on emotion regulatory capacities and lead to dissociative processes that provide subjective detachment from overwhelming emotional experience during and in the aftermath of trauma. Dissociation is a complex phenomenon that comprises a host of symptoms and factors, including depersonalization, derealization, time distortion, dissociative flashbacks, and alterations in the perception of the self. Dissociation occurs in up to two thirds of patients with borderline personality disorder (BPD). The neurobiology of traumatic dissociation has demonstrated a heterogeneity in posttraumatic stress symptoms that, over time, can result in different types of dysregulated emotional states. This review links the concepts of trauma and dissociation to BPD by illustrating different forms of emotional dysregulation and their clinical relevance to patients with BPD.


Molecular characterization of antimicrobial resistance in non-typhoidal salmonellae associated with systemic manifestations from India.
Extended-spectrum cephalosporins and fluoroquinolones are essential antimicrobials for treating invasive salmonellosis, although emerging resistance to these antimicrobials is of growing concern, especially in India. Therefore, a study was conducted to characterize the antimicrobial susceptibility phenotypes, types of extended-spectrum β-lactamase (ESBL) gene plasmids and serological relationships of 21 non-typhoidal Salmonella isolates from patients who attended three different hospitals in India from 2006 to 2008. The isolates were cultured from stool, blood and cerebrospinal fluid samples obtained from patients presenting with diarrhoea and accompanying systemic manifestations such as fever, vomiting and meningism. Non-typhoidal Salmonella isolates were investigated using serotyping and antimicrobial susceptibility testing. PCR screening was also performed to detect the β-lactamase, qnr and aac(6')-Ib-cr genes and class 1 integrons. Sequencing for quinolone resistance mutations and plasmid replicon typing were also performed. An antimicrobial resistance microarray was used for preliminary screening and identification of bla(TEM) and bla(SHV) genes, and phenotypic testing for the presence of efflux pumps was also performed. Ten out of 21 isolates (48%) possessed the extended-spectrum cephalosporin resistance phenotype, with PCR amplification and sequencing revealing that isolates possessed TEM-1, SHV-12, DHA-1, OXA-1-like and CTX-M-15 ESBL genes. FII(s) plasmid replicons were detected in seven isolates (33%). The involvement of efflux pumps was detected in four isolates (19%) resistant to ciprofloxacin. It was concluded that SHV-12-carrying Salmonella serotype Agona may play an important role in ESBL-mediated resistance in non-typhoidal salmonellae in India. The very high percentage (48%) of ESBL-producing non-typhoidal salmonellae isolated from these patients represents a real and immediate challenge to the effective antimicrobial therapy of Salmonella infections associated with systemic manifestations. Continued surveillance for the presence of ESBL-producing (non-typhoidal) salmonellae in India is essential.


G protein-coupled receptor120 (GPR120) transcription in intestinal epithelial cells is significantly affected by bacteria belonging to the Bacteroides, Proteobacteria, and Firmicutes phyla.
Free fatty acids (FFA) are produced in the intestine by microbial fermentation. Recently, a family of G protein-coupled receptors (GPR) acting as FFA transporters has been reported including GPR120, which is expressed by intestinal epithelial cells. The GPR120 has been reported to affect the expression of glucagon-like peptide (GLP)-1 as well as function as a control point for anti-inflammatory effects. The aim of the present study was to evaluate whether 12 selected intestinal bacteria, representing the 4 major phyla present in the intestine, affect intestinal epithelial cell GPR120 and GLP-1 mRNA abundance. Supernatants of the 12 bacteria were added to differentiated Caco-2 intestinal epithelial cells cultured on filter inserts in concentrations corresponding to a cell:bacteria ratio of 1:200. After 4 h of incubation, changes in cellular mRNA of GLP-1 and GPR120 by bacterial supernatant were examined using real-time reverse transcriptase polymerase chain reaction. The abundance of GLP-1 mRNA decreased when cells were exposed to 4 of the 12 supernatants (P ≤ 0.05) compared with cells without bacteria added. Supernatants from 8 of the 12 bacteria analyzed increased the mRNA level of GPR120 (P ≤ 0.05) compared with cells without bacteria added. The alteration in cellular GPR120 mRNA was observed with bacteria categorized as either probiotics or bacteria capable of inducing an anti-inflammatory effect. The beneficial effect of these bacteria may very well be mediated by regulation of GPR120. The regulation of GPR120 by intestinal microbiota represents a direct signaling pathway for gut bacteria to affect host health and metabolism.


The effectiveness and cost-effectiveness of hospital-based specialist palliative care for adults with advanced illness and their caregivers.
Serious illness is often characterised by physical/psychological problems, family support needs, and high healthcare resource use. Hospital-based specialist palliative care (HSPC) has developed to assist in better meeting the needs of patients and their families and potentially reducing hospital care expenditure. There is a need for clarity on the effectiveness and optimal models of HSPC, given that most people still die in hospital and also to allocate scarce resources judiciously. To assess the effectiveness and cost-effectiveness of HSPC compared to usual care for adults with advanced illness (hereafter patients) and their unpaid caregivers/families. We searched CENTRAL, CDSR, DARE and HTA database via the Cochrane Library; MEDLINE; Embase; CINAHL; PsycINFO; CareSearch; National Health Service Economic Evaluation Database (NHS EED) and two trial registers to August 2019, together with checking of reference lists and relevant systematic reviews, citation searching and contact with experts to identify additional studies. We included randomised controlled trials (RCTs) evaluating the impact of HSPC on outcomes for patients or their unpaid caregivers/families, or both. HSPC was defined as specialist palliative care delivered by a palliative care team that is based in a hospital providing holistic care, co-ordination by a multidisciplinary team, and collaboration between HSPC providers and generalists. HSPC was provided to patients while they were admitted as inpatients to acute care hospitals, outpatients or patients receiving care from hospital outreach teams at home. The comparator was usual care, defined as inpatient or outpatient hospital care without specialist palliative care input at the point of entry into the study, community care or hospice care provided outside of the hospital setting. We used standard methodological procedures expected by Cochrane. We assessed risk of bias and extracted data. To account for use of different scales across studies, we calculated standardised mean differences (SMDs) with 95% confidence intervals (CIs) for continuous data. We used an inverse variance random-effects model. For binary data, we calculated odds ratio (ORs) with 95% CIs. We assessed the evidence using GRADE and created a 'Summary of findings' table. Our primary outcomes were patient health-related quality of life (HRQoL) and symptom burden (a collection of two or more symptoms). Key secondary outcomes were pain, depression, satisfaction with care, achieving preferred place of death, mortality/survival, unpaid caregiver burden, and cost-effectiveness. Qualitative data was analysed where available. We identified 42 RCTs involving 7779 participants (6678 patients and 1101 caregivers/family members). Twenty-one studies were with cancer populations, 14 were with non-cancer populations (of which six were with heart failure patients), and seven with mixed cancer and non-cancer populations (mixed diagnoses). HSPC was offered in different ways and included the following models: ward-based, inpatient consult, outpatient, hospital-at-home or hospital outreach, and service provision across multiple settings which included hospital. For our main analyses, we pooled data from studies reporting adjusted endpoint values. Forty studies had a high risk of bias in at least one domain. Compared with usual care, HSPC improved patient HRQoL with a small effect size of 0.26 SMD over usual care (95% CI 0.15 to 0.37; I2 = 3%, 10 studies, 1344 participants, low-quality evidence, higher scores indicate better patient HRQoL). HSPC also improved other person-centred outcomes. It reduced patient symptom burden with a small effect size of -0.26 SMD over usual care (95% CI -0.41 to -0.12; I2 = 0%, 6 studies, 761 participants, very low-quality evidence, lower scores indicate lower symptom burden). HSPC improved patient satisfaction with care with a small effect size of 0.36 SMD over usual care (95% CI 0.41 to 0.57; I2 = 0%, 2 studies, 337 participants, low-quality evidence, higher scores indicate better patient satisfaction with care). Using home death as a proxy measure for achieving patient's preferred place of death, patients were more likely to die at home with HSPC compared to usual care (OR 1.63, 95% CI 1.23 to 2.16; I2 = 0%, 7 studies, 861 participants, low-quality evidence). Data on pain (4 studies, 525 participants) showed no evidence of a difference between HSPC and usual care (SMD -0.16, 95% CI -0.33 to 0.01; I2 = 0%, very low-quality evidence). Eight studies (N = 1252 participants) reported on adverse events and very low-quality evidence did not demonstrate an effect of HSPC on serious harms. Two studies (170 participants) presented data on caregiver burden and both found no evidence of effect of HSPC (very low-quality evidence). We included 13 economic studies (2103 participants). Overall, the evidence on cost-effectiveness of HSPC compared to usual care was inconsistent among the four full economic studies. Other studies that used only partial economic analysis and those that presented more limited resource use and cost information also had inconsistent results (very low-quality evidence). Quality of the evidence The quality of the evidence assessed using GRADE was very low to low, downgraded due to a high risk of bias, inconsistency and imprecision. Very low- to low-quality evidence suggests that when compared to usual care, HSPC may offer small benefits for several person-centred outcomes including patient HRQoL, symptom burden and patient satisfaction with care, while also increasing the chances of patients dying in their preferred place (measured by home death). While we found no evidence that HSPC causes serious harms, the evidence was insufficient to draw strong conclusions. Although these are only small effect sizes, they may be clinically relevant at an advanced stage of disease with limited prognosis, and are person-centred outcomes important to many patients and families. More well conducted studies are needed to study populations with non-malignant diseases and mixed diagnoses, ward-based models of HSPC, 24 hours access (out-of-hours care) as part of HSPC, pain, achieving patient preferred place of care, patient satisfaction with care, caregiver outcomes (satisfaction with care, burden, depression, anxiety, grief, quality of life), and cost-effectiveness of HSPC. In addition, research is needed to provide validated person-centred outcomes to be used across studies and populations.


Comparing scales of environmental effects from gasoline and ethanol production.
Understanding the environmental effects of alternative fuel production is critical to characterizing the sustainability of energy resources to inform policy and regulatory decisions. The magnitudes of these environmental effects vary according to the intensity and scale of fuel production along each step of the supply chain. We compare the spatial extent and temporal duration of ethanol and gasoline production processes and environmental effects based on a literature review and then synthesize the scale differences on space-time diagrams. Comprehensive assessment of any fuel-production system is a moving target, and our analysis shows that decisions regarding the selection of spatial and temporal boundaries of analysis have tremendous influences on the comparisons. Effects that strongly differentiate gasoline and ethanol-supply chains in terms of scale are associated with when and where energy resources are formed and how they are extracted. Although both gasoline and ethanol production may result in negative environmental effects, this study indicates that ethanol production traced through a supply chain may impact less area and result in more easily reversed effects of a shorter duration than gasoline production.


Active repression of methylated genes by the chromosomal protein MBD1.
MBD1 belongs to a family of mammalian proteins that share a methyl-CpG binding domain. Previous work has shown that MBD1 binds to methylated sites in vivo and in vitro and can repress transcription from methylated templates in transcription extracts and in cultured cells. In the present study we established by several experimental criteria that, contrary to a previous report, MBD1 is not a component of the MeCP1 repressor complex. We identified a powerful transcriptional repression domain (TRD) at the C terminus of MBD1 that can actively repress transcription at a distance. Methylation-dependent repression in vivo depends on the presence of both the TRD and the methyl-CpG binding domain. The mechanism is likely to involve deacetylation, since the deacetylase inhibitor trichostatin A can overcome MBD1-mediated repression. Accordingly, we found that endogenous MBD1 is particularly concentrated at sites of centromeric heterochromatin, where acetylated histone H4 is deficient. Unlike MBD2 and MeCP2, MBD1 is not depleted by antibodies to the histone deacetylase HDAC1. Thus, the deacetylase-dependent pathway by which MBD1 actively silences methylated genes is likely to be different from that utilized by the methylation-dependent repressors MeCP1 and MeCP2.


Experimental in vivo measurements of light emission in plants: a perspective dedicated to David Walker.
This review is dedicated to David Walker (1928-2012), a pioneer in the field of photosynthesis and chlorophyll fluorescence. We begin this review by presenting the history of light emission studies, from the ancient times. Light emission from plants is of several kinds: prompt fluorescence (PF), delayed fluorescence (DF), thermoluminescence, and phosphorescence. In this article, we focus on PF and DF. Chlorophyll a fluorescence measurements have been used for more than 80 years to study photosynthesis, particularly photosystem II (PSII) since 1961. This technique has become a regular trusted probe in agricultural and biological research. Many measured and calculated parameters are good biomarkers or indicators of plant tolerance to different abiotic and biotic stressors. This would never have been possible without the rapid development of new fluorometers. To date, most of these instruments are based mainly on two different operational principles for measuring variable chlorophyll a fluorescence: (1) a PF signal produced following a pulse-amplitude-modulated excitation and (2) a PF signal emitted during a strong continuous actinic excitation. In addition to fluorometers, other instruments have been developed to measure additional signals, such as DF, originating from PSII, and light-induced absorbance changes due to the photooxidation of P700, from PSI, measured as the absorption decrease (photobleaching) at about 705 nm, or increase at 820 nm. In this review, the technical and theoretical basis of newly developed instruments, allowing for simultaneous measurement of the PF and the DF as well as other parameters is discussed. Special emphasis has been given to a description of comparative measurements on PF and DF. However, DF has been discussed in greater details, since it is much less used and less known than PF, but has a great potential to provide useful qualitative new information on the back reactions of PSII electron transfer. A review concerning the history of fluorometers is also presented.


Virulence and vaccine potential of Salmonella typhimurium mutants deficient in the expression of the RpoS (sigma S) regulon.
The alternative sigma factor RpoS (sigma S) is required for Salmonella virulence in mice. We report the immunizing capacity of Salmonella typhimurium rpoS and rpoS aroA mutants to protect susceptible BALB/c mice against subsequent oral challenge with virulent S. typhimurium. When administered orally or intraperitoneally, rpoS derivatives of the mouse-virulent S. typhimurium strains, C52 and SL1344, were highly attenuated and were efficient single-dose live vaccines. rpoS aroA mutants were more attenuated than corresponding single aroA or rpoS mutants, as assessed after oral or intraperitoneal administration, but retained significant ability to protect mice against salmonellosis. Salmonella rpoS and rpoS aroA mutants therefore deserve serious consideration for rational vaccine design. Consistent with this, Salmonella typhi Ty2, a 'wild-type' strain used widely for the development of human live-vaccine candidates against typhoid fever, was shown to be defective for rpoS. In addition, our results demonstrate that rpoS not only controls the growth and persistence of S. typhimurium in deep lymphoid organs, but also plays a role during the initial stages of oral infection.


Dimensional variation tolerant silicon-on-insulator directional couplers.
We design silicon ridge/rib waveguide directional couplers which are simultaneously tolerant to width, height, coupling gap, and etch depth variations. Using wafer-scale measurements of structures fabricated in the IMEC Standard Passives process, we demonstrate the normalized standard deviation in the per-length coupling coefficient (a metric for the splitting ratio variation) of the variation-tolerant directional couplers is up to 4 times smaller than that of strip waveguide designs. The variation-tolerant couplers are also the most broadband and the deviation in the coupling coefficient shows the lowest spectral dependence.


Risk factors associated with struvite urolithiasis in dogs evaluated at general care veterinary hospitals in the United States.
To identify factors associated with development of struvite urolithiasis in dogs evaluated at general care veterinary hospitals in the United States. Retrospective case-control study. 508 dogs with a first-time diagnosis of struvite urolithiasis and 7,135 control dogs. Electronic medical records of all dogs evaluated at 787 general care veterinary hospitals in the United States between October 2007 and December 2010 were reviewed to identify dogs that developed struvite urolithiasis and 2 groups of control dogs with no history of urolithiasis. Information extracted included diet, age, sex, neuter status, breed size category, hospital location, and date of diagnosis. Urinalysis results, urolith composition, and other disease conditions were recorded if applicable. Potential risk factors were assessed with univariable and multivariable regression analysis. Toy- or small-sized breeds had significantly greater odds of struvite urolithiasis, compared with medium- or large-sized breeds. Neutering significantly increased the odds of this outcome in females only; sexually intact females were more likely to develop struvite urolithiasis than were sexually intact males, but only up to 5 years of age. Urinary factors significantly associated with the outcome were basic (vs acidic) pH, presence of RBCs or WBCs, protein concentration > 30 mg/dL, and ketone concentration ≥ 5 mg/dL. Evaluation of demographic characteristics and urinalysis results may be useful in the early identification of struvite urolithiasis in dogs. Periodic urinalysis in dogs is recommended because of the potential health impact of a late diagnosis of urolithiasis.


Impact of temperature decline from the previous day as a trigger of spontaneous subarachnoid hemorrhage: case-crossover study of prefectural stroke database.
Several environmental factors have been reported to correlate with incidence of spontaneous subarachnoid hemorrhage (SAH). However, because of different patient selection and study designs among these studies, meteorological factors that trigger the incidence of SAH in a short hazard period remain unknown. Among meteorological factors, daily temperature changes may disrupt and violate homeostasis and predispose to cerebrovascular circulatory disturbances and strokes. The authors aimed to investigate whether a decline in the temperature from the highest of the previous day to the lowest of the event day (temperature decline from the previous day [TDP]) triggers SAH in the prefecture-wide stroke database. All 28 participating institutions with primary or comprehensive stroke centers located throughout Kochi Prefecture, Japan, were included in the study. Data collected between January 2012 and December 2016 were analyzed, and 715 consecutive SAH patients with a defined date of onset were enrolled. Meteorological data in this period were obtained from the Kochi Local Meteorological Observatory. A case-crossover study was performed to investigate association of TDP and other environmental factors with onset of SAH. The increasing TDP in 1°C on the day of the SAH event was associated with an increased incidence of SAH (OR 1.041, 95% CI 1.007-1.077) after adjustment for other environmental factors. According to the stratified analysis, a significant association between TDP and SAH was observed in women, patients < 65 years old, and patients with weekday onset. Among these factors, increasing TDP had a great impact on SAH onset in patients < 65 years old (p = 0.028, Mann-Whitney U-test). TDP, temperature decline from the highest of the previous day to the lowest of the day, was correlated with the incidence of spontaneous SAH, particularly in younger patients < 65 years old.


Pulsed Doppler echocardiographic assessment of portal venous flow patterns in patients after the Fontan operation.
To assess the effect of the condition of the right heart after the Fontan operation on portal venous flow, and to determine whether the characteristics of portal venous flow were different when there was an atriopulmonary connection with atrial septal closure rather than an atriopulmonary or total cavopulmonary connection with intra-atrial routing. After the Fontan operation six patients with an atriopulmonary connection (group 1), three patients with an atriosubpulmonary connection (group 2), four patients with intra-atrial routing and an atriopulmonary connection (group 3), and five patients with a total cavopulmonary connection (group 4) were studied by pulsed Doppler echocardiography. The flow signals were recorded for the pulmonary artery, hepatic vein, and intrahepatic portal vein in each patient. Postoperative cardiac catheterisation was performed in 16 of the 18 patients. The Doppler findings were compared with those of 14 controls. The portal flow was pulsatile in 13 patients and constant in five patients. Reversed flow was shown at or just after the QRS wave after hepatic venous regurgitation in two group 1 patients. The flow signal was interrupted in two group 1 patients and two group 2 patients. Decrease in velocity of flow was recorded in two group 1 patients, one group 2 patients, three group 3 patients, and one group 4 patient. Portal flow was constant in one group 3 patient and four group 4 patients. The pulsatility ratio ranged from -0.46 to 0.49 (mean (SD), 0.03 (0.32)) in the patients from group 1 and 2, from 0.41 to 0.76 (0.54 (0.15)) in group 3, and from 0.70 to 0.80 (0.75 (0.04)) in group 4. The ratio in the controls ranged from 0.29 to 0.83 (0.61 (0.13)). The ratio was significantly lower in groups 1 and 2 than in group 3 (p < 0.01), group 4 (p < 0.01), or the controls (p < 0.005). There was no significant difference in the ratio between group 3 and group 4 and no correlation between the pulsatility ratio and the cardiac index (r = 0.26), mean right atrial pressure (r = 0.25), or pulmonary vascular resistance (r = 0.17). The larger hepatic venous regurgitation caused by atrial contraction in patients with an atriopulmonary connection correlated with the higher portal pulsatility and a total cavopulmonary connection reduced portal pulsatility.


A Prognostic Model for Development of Profound Shock among Children Presenting with Dengue Shock Syndrome.
To identify risk factors and develop a prediction model for the development of profound and recurrent shock amongst children presenting with dengue shock syndrome (DSS). We analyzed data from a prospective cohort of children with DSS recruited at the Paediatric Intensive Care Unit of the Hospital for Tropical Disease in Ho Chi Minh City, Vietnam. The primary endpoint was "profound DSS", defined as ≥2 recurrent shock episodes (for subjects presenting in compensated shock), or ≥1 recurrent shock episodes (for subjects presenting initially with decompensated/hypotensive shock), and/or requirement for inotropic support. Recurrent shock was evaluated as a secondary endpoint. Risk factors were pre-defined clinical and laboratory variables collected at the time of presentation with shock. Prognostic model development was based on logistic regression and compared to several alternative approaches. The analysis population included 1207 children of whom 222 (18%) progressed to "profound DSS" and 433 (36%) had recurrent shock. Independent risk factors for both endpoints included younger age, earlier presentation, higher pulse rate, higher temperature, higher haematocrit and, for females, worse hemodynamic status at presentation. The final prognostic model for "profound DSS" showed acceptable discrimination (AUC=0.69 for internal validation) and calibration and is presented as a simple score-chart. Several risk factors for development of profound or recurrent shock among children presenting with DSS were identified. The score-chart derived from the prognostic models should improve triage and management of children presenting with DSS in dengue-endemic areas.


Relevance of prostate cancer in patients with synchronous invasive bladder urothelial carcinoma: a monocentric retrospective analysis.
We retrospectively reviewed data of patients with incidental prostate cancer (PCa) who underwent radical cystoprostatectomy (RCP) for invasive bladder cancer and we analyzed their features with regard to incidence, pathologic characteristics, clinical significance, and implications for management. Clinical data and pathological features of 64 patients who underwent standard RCP for bladder cancer were included in this study. Besides the urothelial carcinoma of the urinary bladder, the location and tumor volume of the PCa, prostate apex involvement, Gleason score, pathological staging and surgical margins were evaluated. Clinically significant PCa was defined as a tumor with a Gleason 4 or 5 pattern, stage ≥ pT3, lymph node involvement, positive surgical margin or multifocality of three or more lesions. Postoperative follow-up was scheduled every 3 months in the first year, every 6 months in the second and third year, annually thereafter. 11 out of 64 patients (17.2%) who underwent RCP had incidentally diagnosed PCa. 3 cases (27.3%) were diagnosed as significant PCa, while 8 cases (72.7%) were clinically insignificant. The positive surgical margin of PCa was detected in 1 patient with significant disease. The prostate apex involvement was present in 1 patient of the significant PCa group. Median follow-up period was 47.8 ± 29.2 (range 4-79). During the follow-up, biochemical recurrence occurred in 1 patient (9%). Concerning the cancer specific survival there was no statistical significance (P = 0.326) between the clinically significant and clinical insignificant cancer group. In line with published studies, incidental PCa does not impact on the prognosis of bladder cancer of patients undergoing RCP.


A large adolescent idiopathic scoliosis curve in a skeletally immature patient: is early surgery the correct approach? Overview of available evidence.
The goal of this study was to determine whether the available studies provide enough evidence that, in a borderline case of adolescent idiopathic scoliosis with a large (35 to 50 degrees) curve in a skeletally immature patient (Risser 0 to 2) with significant growth potential left, a conservative line of management in the form of bracing can be considered, rather than to rush into a potentially unnecessary major spinal surgery. We reviewed the literature spanning the last 20 years for the results of bracing in this specific group of patients. From the 9 studies selected, a group-specific data extraction was carried out. Three hundred and five patients with a 36 to 50 degrees scoliosis curve and Risser stages 0 to 2 were treated by bracing and the treatment was termed successful in 160 patients. Thus, more than half (52.5%) of the patients were successfully managed with a brace and were spared surgery. The current trend for management of these curves is early surgical intervention, the rationale being the ineffectiveness of bracing in preventing the progression of such a large curve and the difficulty in obtaining satisfactory correction by postponing surgery to a later date. On the basis of our results, we propose a conservative line of management for these curves, in contrast with current views, rather than to rush into a major spine surgery, expecting a favorable outcome with a well-supervised bracing program. If the curve progresses, surgery can always be considered later, keeping in mind the excellent correction obtained with the pedicle screw systems even for large curves of 70 to 100 degrees.


European consensus-based interdisciplinary guideline for melanoma. Part 2: Treatment - Update 2019.
A unique collaboration of multidisciplinary experts from the European Dermatology Forum, the European Association of Dermato-Oncology and the European Organization for Research and Treatment of Cancer (EORTC) was formed to make recommendations on cutaneous melanoma diagnosis and treatment, based on systematic literature reviews and the experts' experience. Cutaneous melanomas are excised with 1- to 2-cm safety margins. Sentinel lymph node dissection shall be performed as a staging procedure in patients with tumour thickness ≥1.0 mm or ≥0.8 mm with additional histological risk factors, although there is as yet no clear survival benefit for this approach. Therapeutic decisions in stage III/IV patients should be primarily made by an interdisciplinary oncology team ("Tumor Board"). Adjuvant therapies in stage III/IV patients are primarily anti-PD-1, independent of mutational status, or dabrafenib plus trametinib for BRAF-mutant patients. In distant metastasis, either resected or not, systemic treatment is indicated. For first-line treatment, particularly in BRAF wild-type patients, immunotherapy with PD-1 antibodies alone or in combination with CTLA-4 antibodies shall be considered. In particular scenarios for patients with stage IV melanoma and a BRAF-V600 E/K mutation, first-line therapy with BRAF/MEK inhibitors can be offered as an alternative to immunotherapy. In patients with primary resistance to immunotherapy and harbouring a BRAF-V600 E/K mutation, this therapy shall be offered in second-line. Systemic therapy in stage III/IV melanoma is a rapidly changing landscape, and it is likely that these recommendations may change in the near future.


Regional haemodynamic effects of captopril, enalaprilat and lisinopril in conscious water-replete and water-deprived Brattleboro rats.
1. The regional haemodynamic effects of intravenous bolus doses of captopril, enalaprilat and lisinopril were assessed in conscious Brattleboro (i.e. vasopressin-deficient) rats, chronically instrumented with miniaturized pulsed Doppler probes and intravascular catheters. 2. Responses to incremental doses of each drug (spanning the median effective dose for the inhibition of the pressor response to angiotensin I) were examined in both water-replete and water-deprived states. 3. In the water-replete state, the haemodynamic profiles of captopril, enalaprilat and lisinopril were generally similar, with incremental doses causing rises in mesenteric and renal flow and, to a lesser extent, hindquarters flow. There were small tachycardias and only slight falls in mean blood pressure. 4. In the water-deprived state, the effects of all three drugs were greatly enhanced; tachycardic and hypotensive effects occurred together with increases in mesenteric, renal and hindquarters flows. However, for renal flow and renal vascular conductance the effectiveness of the drugs decreased in the order enalaprilat greater than captopril greater than lisinopril, whereas for mesenteric flow and mesenteric vascular conductance the order was captopril greater than enalaprilat greater than lisinopril. 5. Since marked haemodynamic actions were seen with doses one-tenth of the median effective dose for the inhibition of the pressor effect of angiotensin I, it is likely these effects were due to inhibition of angiotensin-converting enzyme, although not necessarily to inhibition of angiotensin II production.(ABSTRACT TRUNCATED AT 250 WORDS)


Expression of the 1-aminocyclopropane-1-carboxylic acid deaminase gene requires symbiotic nitrogen-fixing regulator gene nifA2 in Mesorhizobium loti MAFF303099.
Many soil bacteria contain 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase, which degrades ACC, a precursor of the phytohormone ethylene. In order to examine the regulation of the acdS gene encoding ACC deaminase in Mesorhizobium loti MAFF303099 during symbiosis with the host legume Lotus japonicus, we introduced the beta-glucuronidase (GUS) gene into acdS so that GUS was expressed under control of the acdS promoter, and we also generated disruption mutants with mutations in a nitrogen fixation regulator gene, nifA. The histochemical GUS assay showed that there was exclusive expression of acdS in mature root nodules. Two homologous nifA genes, mll5857 and mll5837, were found in the symbiosis island of M. loti and were designated nifA1 and nifA2, respectively. Quantitative reverse transcription-PCR demonstrated that nifA2 disruption resulted in considerably diminished expression of acdS, nifH, and nifA1 in bacteroid cells. In contrast, nifA1 disruption slightly enhanced expression of the acdS transcripts and suppressed nifH to some extent. These results indicate that the acdS gene and other symbiotic genes are positively regulated by the NifA2 protein, but not by the NifA1 protein, in M. loti. The mode of gene expression suggests that M. loti acdS participates in the establishment and/or maintenance of mature nodules by interfering with the production of ethylene, which induces negative regulation of nodulation.


The future of cystic fibrosis care: a global perspective.
The past six decades have seen remarkable improvements in health outcomes for people with cystic fibrosis, which was once a fatal disease of infants and young children. However, although life expectancy for people with cystic fibrosis has increased substantially, the disease continues to limit survival and quality of life, and results in a large burden of care for people with cystic fibrosis and their families. Furthermore, epidemiological studies in the past two decades have shown that cystic fibrosis occurs and is more frequent than was previously thought in populations of non-European descent, and the disease is now recognised in many regions of the world. The Lancet Respiratory Medicine Commission on the future of cystic fibrosis care was established at a time of great change in the clinical care of people with the disease, with a growing population of adult patients, widespread genetic testing supporting the diagnosis of cystic fibrosis, and the development of therapies targeting defects in the cystic fibrosis transmembrane conductance regulator (CFTR), which are likely to affect the natural trajectory of the disease. The aim of the Commission was to bring to the attention of patients, health-care professionals, researchers, funders, service providers, and policy makers the various challenges associated with the changing landscape of cystic fibrosis care and the opportunities available for progress, providing a blueprint for the future of cystic fibrosis care. The discovery of the CFTR gene in the late 1980s triggered a surge of basic research that enhanced understanding of the pathophysiology and the genotype-phenotype relationships of this clinically variable disease. Until recently, available treatments could only control symptoms and restrict the complications of cystic fibrosis, but advances in CFTR modulator therapies to address the basic defect of cystic fibrosis have been remarkable and the field is evolving rapidly. However, CFTR modulators approved for use to date are highly expensive, which has prompted questions about the affordability of new treatments and served to emphasise the considerable gap in health outcomes for patients with cystic fibrosis between high-income countries, and low-income and middle-income countries (LMICs). Advances in clinical care have been multifaceted and include earlier diagnosis through the implementation of newborn screening programmes, formalised airway clearance therapy, and reduced malnutrition through the use of effective pancreatic enzyme replacement and a high-energy, high-protein diet. Centre-based care has become the norm in high-income countries, allowing patients to benefit from the skills of expert members of multidisciplinary teams. Pharmacological interventions to address respiratory manifestations now include drugs that target airway mucus and airway surface liquid hydration, and antimicrobial therapies such as antibiotic eradication treatment in early-stage infections and protocols for maintenance therapy of chronic infections. Despite the recent breakthrough with CFTR modulators for cystic fibrosis, the development of novel mucolytic, anti-inflammatory, and anti-infective therapies is likely to remain important, especially for patients with more advanced stages of lung disease. As the median age of patients with cystic fibrosis increases, with a rapid increase in the population of adults living with the disease, complications of cystic fibrosis are becoming increasingly common. Steps need to be taken to ensure that enough highly qualified professionals are present in cystic fibrosis centres to meet the needs of ageing patients, and new technologies need to be adopted to support communication between patients and health-care providers. In considering the future of cystic fibrosis care, the Commission focused on five key areas, which are discussed in this report: the changing epidemiology of cystic fibrosis (section 1); future challenges of clinical care and its delivery (section 2); the building of cystic fibrosis care globally (section 3); novel therapeutics (section 4); and patient engagement (section 5). In panel 1, we summarise key messages of the Commission. The challenges faced by all stakeholders in building and developing cystic fibrosis care globally are substantial, but many opportunities exist for improved care and health outcomes for patients in countries with established cystic fibrosis care programmes, and in LMICs where integrated multidisciplinary care is not available and resources are lacking at present. A concerted effort is needed to ensure that all patients with cystic fibrosis have access to high-quality health care in the future.


Long-term outcome of postmenopausal women with proliferative endometrium on endometrial sampling.
Proliferative endometrium has been reported in 15% of endometrial biopsies of women aged 50 years and older. Contrary to endometrial hyperplasia, proliferative endometrium has not been associated with the risk of endometrial cancer. This study aimed to report on the long-term outcome of postmenopausal women who received a diagnosis of proliferative endometrium. This is a retrospective cohort study of 1808 women aged 55 years and older who underwent endometrial sampling between January 1997 and December 2008. Outcome data were available through February 2018. Women with a proliferative endometrium were compared with those with an atrophic endometrium for future development of endometrial hyperplasia or cancer. A subanalysis was performed for those who presented with postmenopausal bleeding. Uni- and multivariable logistic regression analyses were used to assess for confounders. In this study, 297 women (16.4%) received a diagnosis of proliferative endometrium. Furthermore, 962 women met the inclusion criteria. Among those women, 278 had a proliferative endometrium, and 684 had an atrophic endometrium. Women with a proliferative endometrium were younger (61.2 vs 64.5 years; P<.0001) and had a higher body mass index (33.9 vs 30.6 kg/m2; P<.0001). More African American women had a proliferative endometrium. Both groups had a similar length of surveillance (11.9 vs 11.5 years; P=.27). Women with a proliferative endometrium had a higher risk of developing endometrial hyperplasia or cancer (11.9% vs 2.9%; P<.0001), any endometrial cancer (5.8% vs 1.8%; P=.002), atypical endometrial hyperplasia (2.2% vs 0.4%; P=.02), and nonatypical endometrial hyperplasia (2.0% vs 0.7%; P=.001). The risk of developing endometrial cancer and endometrial hyperplasia remained similar after excluding cases on hormonal replacement therapy (12.2% vs 3%; P=.001). On logistic regression analysis, proliferative endometrium histology (odds ratio, 3.89; 95% confidence interval, 2.03-7.49; P<.0001), age >60 years (odds ratio, 1.98; 95% confidence interval, 1.03-3.82; P=.04), and body mass index >35 kg/m2 (odds ratio, 2.3; 95% confidence interval, 1.09-4.83; P<.0001) remained significant risk factors for progression to cancer. One of the 6 postmenopausal women who underwent endometrial sampling had a proliferative endometrium. Furthermore, 11.9% of women developed endometrial hyperplasia or cancer, a 4-fold greater incidence than women with an atrophic endometrium. The findings of this study suggest that long-term monitoring is warranted for women with postmenopausal bleeding and a proliferative endometrium histology. Further studies are needed to examine if a treatment is required to negate the risk of unopposed estrogen.


An observational study of methionine-bound zinc with antioxidants for mild to moderate acne vulgaris.
APC is a novel methionine-based zinc complex with antioxidants that has been used in acne as a nutritional supplement. This is based on the proven role of zinc and antioxidants in improving acne, specially the inflammatory lesions. The objectives of this study are to explore the efficacy, safety, and tolerability of APC in acne patients with mild to moderate facial acne vulgaris. In this exploratory trial, 48 patients were treated with oral APC thrice a day for 3 months followed by a 4-week treatment-free period. At the end of treatment (Week 12), there was a statistically significant improvement in the global acne count (p < 0.05), which began after 8 weeks (p < 0.05). Almost 79% (38/48) of the patients had 80-100% improvement. There was a significant reduction in pustules (8 weeks (p < 0.05) and 12 weeks (p < 0.001)), and papules and closed comedones (8 weeks (p < 0.05) and 12 weeks (p < 0.001)). Only two patients had side effects. The current data indicate that treatment with oral APC thrice daily for 12 weeks in patients with mild to moderate facial acne vulgaris is efficacious and well tolerated. As the onset of action is late, concomitant topical therapy can enhance the results.


Carbidopa pretreatment improves image interpretation and visualisation of carcinoid tumours with 11C-5-hydroxytryptophan positron emission tomography.
Positron emission tomography (PET) with 11C-5-hydroxytryptophan (5-HTP) as tracer is a promising imaging instrument in the management of patients with neuroendocrine tumours (NETs). However, high radioactivity concentrations in the urinary collecting system sometimes produce image reconstruction artefacts that can make detection of small NETs difficult. As a means to decrease urinary excretion of radioactivity and thereby improve image quality, we examined the effect of pretreatment with carbidopa (CD), a peripheral inhibitor of aromatic amino acid decarboxylase (AADC), which converts 5-HTP to serotonin (5-hydroxytryptamine, 5-HT). Six patients with midgut carcinoid metastases were examined with 11C-5-HTP PET before and 1 h after oral administration of 100 or 200 mg of CD. There was a fourfold significant reduction of tracer uptake in the urinary collecting system after CD administration (p=0.0277, n=6), with a mean standard uptake value (SUV) of 155+/-195 before CD and 39+/-14 after CD. In tumour lesions there was a significant increase in SUV after CD administration (p<0. 0001, n=18), with a mean SUV of 11+/-3 before CD and 14+/-3 after CD. There was no difference between the doses (100 and 200 mg) of CD in this respect. In all patients, image interpretation and tumour detection were markedly improved after CD administration. We conclude that CD premedication improves 11C-5-HTP PET image quality and facilitates detection of NET lesions. Because of the similarity of metabolic pathways, this method could probably be applied to improve PET imaging using other tracers like 18F-DOPA and 11C-DOPA.


Growth of Strain SES-3 with Arsenate and Other Diverse Electron Acceptors.
The selenate-respiring bacterial strain SES-3 was able to use a variety of inorganic electron acceptors to sustain growth. SES-3 grew with the reduction of arsenate to arsenite, Fe(III) to Fe(II), or thiosulfate to sulfide. It also grew in medium in which elemental sulfur, Mn(IV), nitrite, trimethylamine N-oxide, or fumarate was provided as an electron acceptor. Growth on oxygen was microaerophilic. There was no growth with arsenite or chromate. Washed suspensions of cells grown on selenate or nitrate had a constitutive ability to reduce arsenate but were unable to reduce arsenite. These results suggest that strain SES-3 may occupy a niche as an environmental opportunist by being able to take advantage of a diversity of electron acceptors.


Mitochondrial Activity and Cyr1 Are Key Regulators of Ras1 Activation of C. albicans Virulence Pathways.
Candida albicans is both a major fungal pathogen and a member of the commensal human microflora. The morphological switch from yeast to hyphal growth is associated with disease and many environmental factors are known to influence the yeast-to-hyphae switch. The Ras1-Cyr1-PKA pathway is a major regulator of C. albicans morphogenesis as well as biofilm formation and white-opaque switching. Previous studies have shown that hyphal growth is strongly repressed by mitochondrial inhibitors. Here, we show that mitochondrial inhibitors strongly decreased Ras1 GTP-binding and activity in C. albicans and similar effects were observed in other Candida species. Consistent with there being a connection between respiratory activity and GTP-Ras1 binding, mutants lacking complex I or complex IV grew as yeast in hypha-inducing conditions, had lower levels of GTP-Ras1, and Ras1 GTP-binding was unaffected by respiratory inhibitors. Mitochondria-perturbing agents decreased intracellular ATP concentrations and metabolomics analyses of cells grown with different respiratory inhibitors found consistent perturbation of pyruvate metabolism and the TCA cycle, changes in redox state, increased catabolism of lipids, and decreased sterol content which suggested increased AMP kinase activity. Biochemical and genetic experiments provide strong evidence for a model in which the activation of Ras1 is controlled by ATP levels in an AMP kinase independent manner. The Ras1 GTPase activating protein, Ira2, but not the Ras1 guanine nucleotide exchange factor, Cdc25, was required for the reduction of Ras1-GTP in response to inhibitor-mediated reduction of ATP levels. Furthermore, Cyr1, a well-characterized Ras1 effector, participated in the control of Ras1-GTP binding in response to decreased mitochondrial activity suggesting a revised model for Ras1 and Cyr1 signaling in which Cyr1 and Ras1 influence each other and, together with Ira2, seem to form a master-regulatory complex necessary to integrate different environmental and intracellular signals, including metabolic status, to decide the fate of cellular morphology.


Disaster case management and individuals with disabilities.
To examine the case management and disaster recovery needs of individuals with disabilities following Hurricane Katrina. The case managers and supervisors in this study provided case management to individuals with disabilities as part of the largest coordinated disaster case management program in U.S. history, the Katrina Aid Today consortium. This study provides an account of the disaster case management needs of individuals with disabilities as well as a picture of their long-term recovery process two years following the disaster. Forty-two case managers and 12 case management supervisors from this program provided services to a collective caseload of 2,047 individuals with disabilities and their families. Interviews and telephone surveys were conducted with these participants 20-24 months after the disaster. The qualitative data were analyzed using grounded theory methodology, and descriptive statistics summarize the demographic data. Findings suggest that the disaster recovery process is typically more complex and lengthy for individuals with disabilities and requires negotiation of a service system sometimes unprepared for disability-related needs. Barriers to disaster recovery for individuals with disabilities included a lack of accessible housing, transportation, and disaster services. Supports to disaster recovery included the individual effort and advocacy of a case manager, connecting with needed resources, collaboration with other agencies, and client motivation and persistence. Results suggest that disaster recovery is facilitated by case managers with disability expertise, including knowledge about the needs of individuals with disabilities and about disability-related services.


Pepper mild mottle virus as an indicator of fecal pollution.
Accurate indicators of fecal pollution are needed in order to minimize public health risks associated with wastewater contamination in recreational waters. However, the bacterial indicators currently used for monitoring water quality do not correlate with the presence of pathogens. Here we demonstrate that the plant pathogen Pepper mild mottle virus (PMMoV) is widespread and abundant in wastewater from the United States, suggesting the utility of this virus as an indicator of human fecal pollution. Quantitative PCR was used to determine the abundance of PMMoV in raw sewage, treated wastewater, seawater exposed to wastewater, and fecal samples and/or intestinal homogenates from a wide variety of animals. PMMoV was present in all wastewater samples at concentrations greater than 1 million copies per milliliter of raw sewage. Despite the ubiquity of PMMoV in human feces, this virus was not detected in the majority of animal fecal samples tested, with the exception of chicken and seagull samples. PMMoV was detected in four out of six seawater samples collected near point sources of secondary treated wastewater off southeastern Florida, where it co-occurred with several other pathogens and indicators of fecal pollution. Since PMMoV was not found in nonpolluted seawater samples and could be detected in surface seawater for approximately 1 week after its initial introduction, the presence of PMMoV in the marine environment reflects a recent contamination event. Together, these data demonstrate that PMMoV is a promising new indicator of fecal pollution in coastal environments.


Severe cases of seasonal influenza in Russia in 2017-2018.
The 2017-2018 influenza epidemic season in Russia was characterized by a relatively low morbidity and mortality. We evaluated herd immunity prior to the 2017-2018 influenza season in hemagglutination inhibition assay, and performed characterization of influenza viruses isolated from severe or fatal influenza cases and from influenza cases in people vaccinated in the fall of 2017. During the 2017-2018 epidemic season, 87 influenza A and B viruses were isolated and viruses of the 75 influenza cases, including selected viral isolates and viruses analyzed directly from the original clinical material, were genetically characterized. The analyzed A(H1N1)pdm09 viruses belonged to clade 6B.1, B/Yamagata-like viruses belonged to clade 3, and B/Victoria-like viruses belonged to clade 1A and they were antigenically similar to the corresponding vaccine strains. A(H3N2) viruses belonged to clade 3C.2a and were difficult to characterize antigenically and the analysis indicated antigenic differences from the corresponding egg-grown vaccine strain. The next generation sequencing revealed the presence of D222/G/N polymorphism in the hemagglutinin gene in 32% of the analyzed A(H1N1)pdm09 lethal cases. This study demonstrated the importance of monitoring D222G/N polymorphism, including detection of minor viral variants with the mutations, in the hemagglutinin gene of A(H1N1)pdm09 for epidemiological surveillance. One strain of influenza virus A(H1N1)pdm09 was resistant to oseltamivir and had the H275Y amino acid substitution in the NA protein. All other isolates were susceptible to NA inhibitors. Prior to the 2017-2018 epidemic season, 67.4 million people were vaccinated, which accounted for 46.6% of the country's population. Just before the epidemic season 33-47% and 24-30% of blood sera samples collected within the territory of Russia showed the presence of protective antibody titers against vaccine strains of influenza A and influenza B/Victoria-like, respectively. Mass vaccination of the population had evidently reduced the severity of the flu epidemic during the 2017-2018 influenza epidemic season in Russia.


Caries infiltrant combined with conventional adhesives for sealing sound enamel in vitro.
To test the null hypothesis that combining low-viscosity caries infiltrant with conventional adhesive resins would not improve sealing of sound enamel against demineralization in vitro. Bovine enamel discs (N = 60) with diameter of 3 mm were randomly assigned to six groups (n = 10). The discs were etched with 37% phosphoric acid for 30 seconds and treated with resins of different monomer content forming the following groups: (1) Icon (DMG), (2) Transbond XT Primer (3M ESPE), (3) Heliobond (Ivoclar Vivadent), (4) Icon + Transbond XT Primer, and (5) Icon + Heliobond. Untreated etched samples served as the negative control. Specimens were subjected to demineralization by immersion in hydrochloric acid (pH 2.6) for 80 hours. Calcium dissolution into the acid was assessed by colorimetric analysis using Arsenazo III method at 16-hour intervals. Groups presenting high protection against demineralization were subjected to further acidic challenge for 15 days with calcium measurements repeated at 24-hour intervals. Data were analyzed by Kruskal-Wallis test and Mann-Whitney U-test. Untreated specimens showed the highest amount of demineralization. Icon and Transbond XT primer decreased the mineral loss significantly compared to the control. Heliobond performed significantly better than both Icon and Transbond XT primer. Combination of Icon both with Transbond XT primer or Heliobond served as the best protective measures and maintained the protective effect for the additional 15-day acidic challenge. Within the limitations of this in vitro study, it could be concluded that the use of low-viscosity caries infiltrant prior to application of the tested conventional adhesives increases their protective effect against demineralization.


Attitudes, skills and use of evidence-based practice among UK osteopaths: a national cross-sectional survey.
Evidence-based practice (EBP) is a clinical decision-making framework that supports quality improvement in healthcare. While osteopaths are key providers of musculoskeletal healthcare, the extent to which osteopaths engage in EBP is unclear. Thus, the aim of this cross-sectional study was to investigate UK osteopaths' attitudes, skills and use of EBP, and perceived barriers and facilitators of EBP uptake. UK-registered osteopaths were invited to complete the Evidence-Based Practice Attitude and Utilisation Survey (EBASE) online. Of the 5200 registered osteopaths in the UK, 9.9% (517/5200) responded to the invitation, and 7.2% (375/5200) completed the EBASE (< 20% incomplete answers). The demographic characteristics of the survey sample were largely similar to those of the UK osteopathy workforce. The osteopaths reported overall positive attitudes towards EBP, with most agreeing that EBP improves the quality of patient care (69.3%) and is necessary for osteopathy practice (76.5%). The majority reported moderate-level skills in EBP, and most (80.8%) were interested in improving these skills. Participating osteopaths typically engaged in EBP activities 1-5 times over the last month. Barriers to EBP uptake included a lack of time and clinical evidence in osteopathy. Main facilitators of EBP included having access to online databases, internet at work, full-text articles, and EBP education materials. UK osteopaths were generally supportive of evidence-based practice, had moderate-level skills in EBP and engaged in EBP activities infrequently. The development of effective interventions that improve osteopaths' skills and the incorporation of EBP into clinical practice should be the focus of future research.


Acupuncture to initiate labor (Acumoms 2): a randomized, sham-controlled clinical trial.
To evaluate the efficacy of acupuncture for labor stimulation. Nulliparous women at 38 weeks or greater were randomized to traditional Chinese medicine (TCM) acupuncture, sham acupuncture, or usual care only groups. Acupuncture points LI4, SP6, BL32, and BL54 were needled bilaterally. The primary outcome was time from enrollment to delivery. Secondary outcomes included rates of spontaneous labor and cesarean delivery. Medical records were abstracted for maternal demographic, medical, and delivery outcome data. ANOVA, Student's t-test, Chi-square, and Kaplan-Meier statistics were used to compare groups. Eighty-nine women were enrolled and randomized. Maternal age, gestational age, prior acupuncture experience, tobacco, alcohol and drug use, gravida, and history of gynecological surgery were similar among the groups. There were no statistically significant differences among groups for time from enrollment to delivery (p=0.20), rates of spontaneous labor (p=0.66), or rates of cesarean delivery (p=0.37). Rates of maternal and neonatal outcomes were not significantly different. TCM acupuncture was not effective in initiating spontaneous labor or reducing the rate of cesarean delivery compared with sham acupuncture or usual medical care.


The role of psychomotor activation in discriminating unipolar from bipolar disorders: a classification-tree analysis.
Multiple studies indicate that bipolar disorders are often underrecognized, misdiagnosed, and incorrectly treated. The aim of the present report is to determine which combination of clinical, demographic, and psychopathological factors and corresponding cutoff scores best discriminate patients with unipolar disorder from those with bipolar disorders. The study sample includes outpatients and inpatients (N = 1,158) participating in 5 studies carried out in the United States and Italy between October 2001 and March 2008, one of which was a randomized clinical trial. Diagnostic assessment was carried out with the SCID, which allows diagnoses to be made according to DSM-IV-TR criteria. Using an exploratory statistical approach based on a classification tree, we employed 5 mania spectrum factors and 6 depression spectrum factors derived from the Mood Spectrum Self-Report Instrument (MOODS-SR) in combination with demographic and clinical characteristics to discriminate participants with unipolar versus bipolar disorders. The psychomotor activation factor, assessing the presence of thought acceleration, distractibility, hyperactivity, and restlessness for 1 or more periods of at least 3 to 5 days in the lifetime, identified subgroups with an increasing likelihood of bipolar disorder diagnosis. Mixed instability and suicidality contributed to further subtyping the sample into mutually exclusive groups, characterized by a different likelihood of receiving a diagnosis of bipolar disorder. Of the demographic and clinical characteristics included in the analysis, only sex proved to be useful to improve the discrimination. The psychomotor activation factor proved to be the most potent discriminator of those with unipolar versus bipolar diagnoses. The items that constitute this factor, together with those that constitute the mixed instability, suicidality, and euphoria factors, might be useful in making the differential diagnosis.


Cyclic Adenosine Monophosphate-Mediated Enhancement of Vascular Endothelial Growth Factor Released by Differentiated Human Monocytic Cells: The Role of Protein Kinase A.
Our investigation was designed to examine the signaling pathway involved in the enhancement of vascular endothelial growth factor (VEGF) release by β-adrenoceptor agonists. Human U937 cells differentiated into macrophages were primed with lipopolysaccharide (LPS) in the absence or presence of β-adrenoceptor agonists and antagonists. The VEGF released and the intracellular cyclic adenosine monophosphate (cAMP) generated were assayed by ELISA. Where necessary, differences between mean values were tested for significance using Student's t test. Isoprenaline, procaterol and salbutamol concentration-dependently enhanced the release of VEGF induced by LPS in U937 cells. R*,R*-(±)-4-[2-[(2-(3-chlorophenyl)-2-hydroxyethyl)amino]propyl]phenoxyacetic acid (BRL 37344), a selective β3-adrenoceptor agonist, did not enhance VEGF release. Using isoprenaline as an agonist, propranolol, ICI 118551 and atenolol produced a parallel rightward shift of the concentration-response curve with no reduction in the maximum response. The -logKB values were 8.12 ± 0.17, 8.03 ± 0.05 and 7.23 ± 0.05 for propranolol, ICI 118551 and atenolol, respectively, indicating the possible involvement of both β1- and β2-adrenoceptor subtypes. Isoprenaline and prostaglandin E2 concentration-dependently increased cAMP generation in U937 cells. Isoprenaline, db-cAMP and 6-Bnz-cAMP, a protein kinase A (PKA) activator, all enhanced VEGF release induced by LPS, and this effect was abolished by KT 5720 and Rp-cAMPS, which are both selective PKA inhibitors, suggesting that PKA is the downstream effector of cAMP activity. 8-CPT-cAMP, a selective activator of the Epac system, had no effect on VEGF release induced by LPS, indicating that the Epac pathway played no role in the release process. In this study, we established that β1- and β2- but not β3-adrenoceptors mediated cAMP-dependent enhancement of VEGF release induced by LPS in differentiated U937 cells, and that PKA was the downstream effector of cAMP activity.


Cytokine mRNA levels in rat ocular tissues after systemic endotoxin treatment.
An intertwined cascading network of cytokines is believed to direct the development of endotoxin-induced uveitis (EIU). This study investigated mRNA levels of interleukin (IL) 1 alpha, IL-1 beta, IL-6, tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, transforming growth factor (TGF)-beta 1, and the helper T lymphocyte marker, CD4, during the course of EIU in rats. Reverse transcription followed by polymerase chain reaction amplification was used to determine relative mRNA levels in four ocular tissues (iris/ciliary body, cornea, lens, and neuroretina) at 0, 1, 3, 6, 24, and 48 hours after subcutaneous injection of 200 micrograms of Escherichia coli endotoxin. Four general patterns of mRNA expression were observed: (1) constitutively expressed and unaffected by endotoxin; (2) constitutively expressed but further induced by endotoxin, reaching peak levels at 3 hours postinjection; (3) initially undetectable or marginally detectable and induced by endotoxin, with peak levels occurring 3 hours postinjection; and (4) never present at appreciable levels. The most dramatic responses were seen in the mRNA levels of IL-1 alpha, IL-1 beta, IL-6, TNF-alpha, and IFN-gamma in iris/ciliary body. Lesser mRNA level responses were found for IL-1 beta and IL-6 in cornea and for IL-1 alpha, IL-1 beta, IL-6, TNF-alpha, and IFN-gamma in neuroretina. Little or no changes in mRNA levels were observed for CD4 or TGF-beta 1 in any tissue or for any mRNA examined in lens. These data show that subcutaneous endotoxin induces cytokine mRNA expression differentially in ocular tissues. These data support the hypothesis that induction of cytokine expression in iris/ciliary body plays a major role in the development of EIU.


Responses of preterm infants to unimodal and multimodal sensory intervention.
To examine the immediate responses of preterm infants to two forms of unimodal [auditory only (A) and tactile only (T)] and two forms of multimodal sensory stimulation [auditory, tactile and visual (ATV); auditory, tactile, visual and vestibular (ATVV)]. A convenience sample of 54 clinically stable preterm infants (33-34 postconceptional weeks) was randomly assigned to 1 of 5 experimental groups [Control (C); (A); (T); (ATV); and (ATVV)]. Stimulation was applied for 15 minutes once daily for 4 consecutive days. Outcome measures included pulse (PR) and respiratory rate (RR), oxygen saturation, behavioral state (BS), and body temperature. Repeated measures ANOVA identified significant differences among the groups during intervention for PR (p < .001), RR (p = .01), and BS (p < .02). Infants receiving any intervention with a tactile component showed increasing arousal (change in BS), and increased PR and RR during stimulation. Group T infants had higher proportions of PR > 180 while Group ATVV had higher proportions of PR < 140 (p = .0001). Group ATVV showed increased alertness following stimulation (24%) in contrast to having the least alertness during stimulation (11%). Tactile stimulation alone may be too arousing for these infants while the addition of vestibular stimulation may modulate arousal and facilitate optimal arousal prior to feeding.


Integrin α1-null mice exhibit improved fatty liver when fed a high fat diet despite severe hepatic insulin resistance.
Hepatic insulin resistance is associated with increased collagen. Integrin α1β1 is a collagen-binding receptor expressed on hepatocytes. Here, we show that expression of the α1 subunit is increased in hepatocytes isolated from high fat (HF)-fed mice. To determine whether the integrin α1 subunit protects against impairments in hepatic glucose metabolism, we analyzed glucose tolerance and insulin sensitivity in HF-fed integrin α1-null (itga1(-/-)) and wild-type (itga1(+/+)) littermates. Using the insulin clamp, we found that insulin-stimulated hepatic glucose production was suppressed by ∼50% in HF-fed itga1(+/+) mice. In contrast, it was not suppressed in HF-fed itga1(-/-) mice, indicating severe hepatic insulin resistance. This was associated with decreased hepatic insulin signaling in HF-fed itga1(-/-) mice. Interestingly, hepatic triglyceride and diglyceride contents were normalized to chow-fed levels in HF-fed itga1(-/-) mice. This indicates that hepatic steatosis is dissociated from insulin resistance in HF-fed itga1(-/-) mice. The decrease in hepatic lipid accumulation in HF-fed itga1(-/-) mice was associated with altered free fatty acid metabolism. These studies establish a role for integrin signaling in facilitating hepatic insulin action while promoting lipid accumulation in mice challenged with a HF diet.


Adenosine receptor agonists differentially regulate IL-10, TNF-alpha, and nitric oxide production in RAW 264.7 macrophages and in endotoxemic mice.
Adenosine released into the extracellular space by immunologic and nonimmunologic stimuli has been shown to regulate various immune functions. In this study we report that i.p. pretreatment of mice with CGS-21680 HCl (CGS), a selective agonist of A2 adenosine receptors, at 0.2 to 2 mg/kg caused an augmentation of plasma IL-10 levels induced by i.p. injection of LPS, but decreased plasma levels of LPS-induced TNF-alpha. 2-Chloro-N6-cyclopentyladenosine (CCPA), an agonist of A1 adenosine receptors, at 0.5 mg/kg diminished LPS-induced plasma TNF-alpha concentrations, but enhanced LPS-induced IL-10 levels only at the highest dose used (2 mg/kg). The specific A3 adenosine receptor agonist 1-deoxy-1-[6-[[(3-iodophenyl)methyl]amino]-9H-purin-9-yl]-N-methyl-beta- D-ribofuranuronamide, at 0.2 and 0.5 mg/kg potentiated LPS-stimulated IL-10 production and inhibited LPS-induced TNF-alpha production. LPS-induced plasma nitrite and nitrate levels (the breakdown products of nitric oxide (NO)) were suppressed by CGS and CCPA. In the RAW 264.7 macrophage cell line, pretreatment of the cells with both CGS and CCPA inhibited LPS-induced IL-10, TNF-alpha, and NO production, each in a concentration-dependent manner. The inhibitory effect of these drugs on cytokine and NO production was associated with improved mitochondrial respiration. Neither CGS nor CCPA affected the LPS-induced nuclear translocation of transcription factor nuclear factor-kappaB in these cells. These results demonstrate that adenosine receptor stimulation differentially modulates the LPS-induced production of IL-10, TNF-alpha, and NO in vitro and in vivo. The increase in LPS-induced IL-10 production and suppression of LPS-induced TNF-alpha and NO production caused by adenosine receptor activation may explain some of the immunomodulatory actions of adenosine released in excess during inflammatory and/or ischemic insult.


Why use indicators to measure and monitor the inclusion of climate change and environmental sustainability in health professions' education?
Currently, health professionals are inadequately prepared to meet the challenges that climate change and environmental degradation pose to health systems. Health professions' education (HPE) has an ethical responsibility to address this and must include the health effects of climate change and environmental sustainability across all curricula. As there is a narrow, closing window in which to take action to avoid the worst health outcomes from climate change, urgent, systematic, system-level change is required by the education sector. Measuring, monitoring, and reporting activity using indicators have been demonstrated to support change by providing a focus for action. A review of the literature on the use of indicators in medical education for climate change and health, however, yielded no publications. The framework of targets and indicators developed for implementation of the Sustainable Development Goals (SDGs) by 2030 and the UNESCO initiative of the Education for Sustainable Development provide a guide for the development of indicators for HPE. Engaging stakeholders and achieving consensus on an approach to indicator development is essential and, where they exist, accreditation standards may have a supporting role. Creating capacity for environmentally sustainable health care at scale and pace should be our collective goal as health professions' educators.


Enteropathogenic Escherichia coli infection inhibits intestinal serotonin transporter function and expression.
Serotonin transporter (SERT) plays a critical role in regulating serotonin (5-hydroxytryptamine [5-HT]) availability in the gut. Elevated 5-HT levels are associated with diarrheal conditions such as irritable bowel syndrome and enteric infections. Whether alteration in SERT activity contributes to the pathophysiology of diarrhea induced by the food-borne pathogen enteropathogenic Escherichia coli (EPEC) is not known. The present studies examined the effects of EPEC infection on SERT activity and expression in intestinal epithelial cells and elucidated the underlying mechanisms. Caco-2 cells as a model of human intestinal epithelia and EPEC-infected C57BL/6J mouse model of infection were utilized. SERT activity was measured as Na(+) and Cl(-) dependent (3)[H] 5-HT uptake. SERT expression was measured by real-time quantitative reverse-transcription polymerase chain reaction, Western blotting, and immunofluorescence studies. Infection of Caco-2 cells with EPEC for 30-120 minutes decreased apical SERT activity (P < .001) in a type 3 secretion system dependent manner and via involvement of protein tyrosine phosphatases. EPEC infection decreased V(max) of the transporter; whereas cell surface biotinylation studies revealed no alteration in the cellular or plasma membrane content of SERT in Caco-2 cells. EPEC infection of mice (24 hours) reduced SERT immunostaining with a corresponding decrease in SERT messenger RNA levels, 5-HT uptake, and mucosal 5-HT content in the small intestine. Our results demonstrate inhibition of SERT by EPEC and define the mechanisms underlying these effects. These data may aid in the development of a novel pharmacotherapy to modulate the serotonergic system in treatment of infectious diarrheal diseases.


Interspecific variation of zona pellucida glycoconjugates in several species of marsupial.
The zona pellucida glycoconjugate content of several marsupial species was investigated using differential lectin histochemistry. Ovaries from fat-tailed dunnarts, a southern brown bandicoot, grey short-tailed opossums, brushtail possums, ringtail possums, koalas and eastern grey kangaroos were fixed, embedded in paraffin wax, sectioned and stained with ten fluorescein isothiocyanate-conjugated lectins. Sections were also incubated with either neuraminidase or saponified, respectively, before incubation with the lectins to identify saccharide residues masked by sialic acids or O-acetyl groups on sialic acids. The zonae pellucidae surrounding the oocytes of the marsupials demonstrated interspecific variation in glycoconjugate content, with mannose-containing glycoconjugates exhibiting the greatest variation. Some of the zona pellucida glycoconjugates of all species, except those of the opossums, were masked by sialic acid with an increase in fluorescence with lectins from Arachis hypogea (PNA), and Glycine max (SBA), after desialylation. The disaccharide beta-galactose(1-4)N-acetyl-D-glucosamine appeared to be conformationally masked by O-acetyl groups of sialic acids in the zonae pellucidae of all species, with an increase in fluorescence with the lectin from Erythrina cristagalli (ECA), after saponification. Similar intensity and localization of beta-(1-4)-N-acetyl-D-glucosamine, as shown by staining of the lectin from Triticum vulgaris (WGA), to the inner and outer regions of the zona pellucida, were found to those reported in eutherian species. WGA fluorescence became uniform throughout the zonae pellucidae after saponification, indicating differential O-acetylation of sialic acids on the internal compartment of the zonae pellucidae.


[Effect of lox-sites of the Cre lox recombination system on promoterless bar gene expression in transgenic plants].
We demonstrate that localization of lox site between the right border of T-DNA and promoterless bar gene (RB-lox-bar-) led to its highly efficient expression in transgenic plants of Nicotiana tabacum and N. africana. Plasmid vectors used in gene integration experiments contained neomycin phosphotransferase II (npt II) gene under nos promoter as well. Transgenic plants were selected according to their capacity to grow on the medium with kanamycin and then they were tested on the selective medium containing phosphinothricin. 80% of transgenic plants expressed bar gene at the level similar to that in plants transformed with the bar gene under widely used constitutive promoter. Transformation of plants with the plasmid vector containing only promoterless bar gene near T-DNA right border (RB-bar-) and with the vector containing lox site and promoterless bar gene in the middle of the construction (-lox-bar-) led to obtaining no more than 4.5% of transgenic plants resistant to phosphinothricin. PCR analyses confirmed both the absence of tandem repeats and of plasmid recombination resulting in transference of bar gene under promoter in plasmid vector. Nos-terminator situated between the lox site and the right border of T-DNA did not decrease bar gene expression.