Evaluation of a continual compliance monitoring program for dapsone in an outpatient Hansen's disease clinic.
Guidelines for the assessment of patient compliance to dapsone were developed and evaluated. The urinary dapsone-to-creatinine (D/C) ratio following standardization by dose, ideal body weight, and time since last dose was used for assessment of compliance. Compliance standards were established in 12 patients of known compliance and confirmed prospectively in nine inpatients on 14 occasions. Compliance increased significantly among outpatients (N = 30) attending the University of Illinois Hansen's Disease Clinic from 47% at base line to 73% at 6 months and 80% at 18 months after establishing the monitoring program. In a subgroup of 18 patients, a similar increase in compliance was observed from 50% to 80%. A good therapeutic response was seen in the subgroup patients who were compliant. A poor therapeutic response was seen in the consistently noncompliant patients. These results demonstrate that use of a continual compliance monitoring program can improve patient drug compliance in an outpatient Hansen's disease clinic.


Role of interleukin-1 in stress responses. A putative neurotransmitter.
Recently, the central roles of interleukin-1 (IL-1) in physical stress responses have been attracting attention. Stress responses have been characterized as central neurohormonal changes, as well as behavioral and physiological changes. Administration of IL-1 has been shown to induce effects comparable to stress-induced changes. IL-1 acts on the brain, especially the hypothalamus, to enhance release of monoamines, such as norepinephrine, dopamine, and serotonin, as well as secretion of corticotropin-releasing hormone (CRH). IL-1-induced activation of the hypothalamo-pituitary-adrenal (HPA) axis in vivo depends on secretion of CRH, an intact pituitary, and the ventral noradrenergic bundle that innervates the CRH-containing neurons in the paraventricular nucleus of the hypothalamus. Recent studies have shown that IL-1 is present within neurons in the brain, suggesting that IL-1 functions in neuronal transmission. We showed that IL-1 in the brain is involved in the stress response, and that stress-induced activation of monoamine release and the HPA axis were inhibited by IL-1 receptor antagonist (IL-1Ra) administration directly into the rat hypothalamus. IL-1Ra has been known to exert a blocking effect on IL-1 by competitively inhibiting the binding of IL-1 to IL-1 receptors. In the latter part of this review, we will attempt to describe the relationship between central nervous system diseases, including psychological disorders, and the functions of IL-1 as a putative neurotransmitter.


Developing a collaborative research system for Aboriginal health.
Investigator-driven research and the use of peer review are contentious in community-based research and are particularly problematic in Indigenous research. In this project, we conducted a qualitative study among stakeholders in an Australian Aboriginal majority-controlled research-funding organisation to examine the research funding process. A steering group guided the project and contributed to the research findings. In-depth interviews (n=18) with stakeholders in the Cooperative Research Centre for Aboriginal Health were conducted to canvass views on the research funding process and options for alternate processes. A discussion document, supported by an extensive literature review, was provided prior to interview. This research was an iterative process where the discussion document and interview schedule were revised as the research findings informed the project. Participants overwhelmingly endorsed a move to a more collaborative research culture, although the form the culture might take varied. Suggested elements included involvement of grant funding bodies as brokers in building collaborative networks and the substitution of named 'critical friends' for blinded peer review. Barriers to changing the research culture to a more collaborative model were described. A collaborative structure with targeted project development would permit redistribution of the time and effort (previously expended on peer review) into research development and would increase community participation in decision-making in the research funding process.


Emergency esophagectomy: Experience of a high volume esophageal cancer center.
The objective of the study was to review the experience of dealing oncological emergency esophagectomies at a dedicated Cancer hospital. We performed a retrospective review of data of eleven esophagectomies at the Department of Surgical Oncology, Shaukat Khanum Memorial Cancer Hospital and Research Centre, Lahore (SKMCH&RC) Pakistan, from 1st January, 2009 to 30th June, 2019. Out of 590 oncological esophagectomies, eleven patients had emergency resection. We collected the data of demographics, primary disease, comorbidities, location of tumor and perforation, cause of perforations, radiological and endoscopic findings, clinical findings and follow-up visits after discharge. Data was analyzed by SPSS version 21 for windows. All 11 patients out of five hunded ninety had esophageal cancer. At the time of initial staging, eight (72%) had locally advanced stage (stage III and IV). Open transhiatal approach was used in six (55%) patients, and the rest had three stage esophagectomies. Primary reconstructions with gastric conduit were performed in all, except in two (18%) patients, Respiratory complications were the most common of the encountered complications, seven (63%) of the patients had palliative resection. Ninety day mortality was observed in 3(27.3%) patients. On long term follow up, six patients had recurrence, with median Disease-Free Survival (DFS) 5.88 months and Median Overall Survival (OS) was 6.37 months. Out of 11, only three patients are alive without disease, while one patient is lost during follow-up. Emergency esophagectomy is a lifesaving procedure; there should be multidisciplinary team approach towards the management. Early diagnosis and management is of paramount importance.


Progress in cardiovascular bioprinting.
Cardiovascular disease has been the leading cause of death globally for the past 15 years. Following a major cardiac disease episode, the ideal treatment would be the replacement of the damaged tissue, due to the limited regenerative capacity of cardiac tissues. However, we suffer from a chronic organ donor shortage which causes approximately 20 people to die each day waiting to receive an organ. Bioprinting of tissues and organs can potentially alleviate this burden by fabricating low cost tissue and organ replacements for cardiac patients. Clinical adoption of bioprinting in cardiovascular medicine is currently limited by the lack of systematic demonstration of its effectiveness, high costs, and the complexity of the workflow. Here, we give a concise review of progress in cardiovascular bioprinting and its components. We further discuss the challenges and future prospects of cardiovascular bioprinting in clinical applications.


The evaluation and cluster analysis of parapneumonic effusion in childhood.
We studied 80 children with parapneumonic effusion (PPE) with respect to the clinical manifestations and treatment alternatives as well as prospective follow-up for 1 year. Out of the 80 patients, 59 per cent were male. The mean age of the patients was 4.0 +/- 3.1 years. Mild effusion was successfully treated by antibiotic alone in 33 per cent of the patients. Tube thoracostomy (TT) was utilized in 63 per cent of the patients. In this group, 11 healed completely, 13 patients required surgical treatment, and 25 required fibrinolytic therapy (FT). FT was successful in 18, and no complication due to FT was observed. Six patients who received FT required surgical therapy later in the course of treatment. Cluster analysis revealed a group of patients with rapid progression and a short history of symptoms (4-6 days) that showed significantly higher rate of complicated prognosis (p<0.05). Successful FT prevented surgical operation in 22 per cent of the patients who were candidates for surgical treatment. The follow-up for one year revealed sequelae on chest X-ray in 28 per cent of the patients most of whom had an operation for necrotizing pneumonia. Pulmonary function tests performed over seven years of age were abnormal in 57 per cent of the patients.


Isothermal Titration Calorimetry for the Screening of Aflatoxin B1 Surface-Enhanced Raman Scattering Sensor Affinity Agents.
In this work, isothermal titration calorimetry (ITC) is employed as an affinity agent screening method for the surface-enhanced Raman scattering (SERS) detection of aflatoxin B1 (AFB1). AFB1, a potent carcinogen produced by a fungus that infects crops, is an important target due to the monitoring required based on its FDA regulation. Polymer affinity agents, like those studied here, have the potential to enable separation and detection of relevant small molecules such as pesticides, drugs, and biological toxins, like AFB1, especially when paired with a vibrational spectroscopy technique such as SERS. Herein, seven homopolymers were synthesized to be evaluated as AFB1 affinity agents based on hypothetical hydrogen bonding interactions. Nitrogen-inclusive poly( N-(2-aminoethyl) methacrylamide) (pAEMA) polymers and their oxygen analogs, poly(2-hydroxyethyl methacrylate) (pHEMA) were evaluated. ITC was demonstrated as an effective method for rapid screening among the polymer affinity agents. Chain lengths between seven and 39 repeat units were synthesized to study length-based variance in affinity agent performance. An ITC method was optimized and used for the rapid screening of polymer affinity agents. The results were compared to those generated by SERS. Good agreement between the ITC results and follow-up SERS sensing experiments showcased ITC's screening potential for analytical applications such as separation and detection.


Nitrogen-doped semiconducting oxides. Implications on photochemical, photocatalytic and electronic properties derived from EPR spectroscopy.
Engineering defects in semiconducting metal oxides is a challenge that remains at the forefront of materials chemistry research. Nitrogen has emerged as one of the most attractive elements able to tune the photochemical and photocatalytic properties of semiconducting oxides, boosting visible-light harvesting and charge separation events, key elements in promoting solar driven chemical reactions. Doping with nitrogen is also a strategy suggested to obtain p-type conduction properties in oxides showing n-type features in their pristine state and to impart collective magnetic properties to the same systems. Here, we review the evolution in the understanding of the role of nitrogen doping in modifying the photochemical and electronic properties of the most common semiconducting oxides used in mentioned applications including: TiO2, ZnO, SnO2 and zirconium titanates. With an emphasis on polycrystalline materials, we highlight the unique role of Electron Paramagnetic Resonance (EPR) spectroscopy in the direct detection of open-shell N-based defects and in the definition of their structural and electronic properties. Synthetic strategies for the insertion of nitrogen defects in the various matrices are also discussed, along with the influence of the corresponding low-lying energy states on the general electronic properties of the doped solids.


Sucrose permeability as a means of detecting diseases of the upper digestive tract.
The healthy gastric epithelium will not allow easy permeation of a disaccharide-sized molecule such as sucrose. However, during gastric damage, intact sucrose can pass the gastric epithelium and ultimately appear in the urine. We examined the relationship between total urinary sucrose excretion and various diseases. We used 149 patients (105 had upper gastrointestinal disease, 12 had gastric cancer and 32 were normal). Subjects were given a solution containing 100 g sucrose in 450 c.c. water. All urine was collected for 7.5 h. The urinary sucrose concentration was determined by anion exchange high-performance liquid chromatography. Total urinary sucrose excretion was significantly higher in patients with gastric ulcer and those with gastric cancer than in endoscopically normal controls. In the 34 patients with gastric ulcer, the total sucrose excretion was closely correlated with ulcer size. Ulcer location did not affect urinary sucrose excretion. A strong correlation was also observed between sucrose excretion and lesion size in the 12 patients with gastric cancer. The sucrose permeability test may be a relatively sensitive method to detect gastric disease.


Variation in compulsory psychiatric inpatient admission in England: a cross-classified, multilevel analysis.
The increasing rate of compulsory admission to psychiatric inpatient beds in England is worrying. Studying variation between places and services could be key to identifying targets for interventions to reverse this trend. We modelled spatial variation in compulsory admissions in England using national patient-level data and quantified the extent to which patient, local-area, and service-setting characteristics accounted for this variation. This study is a cross-sectional, multilevel analysis of the 2010-11 Mental Health Minimum Data Set (MHMDS). Data from eight provider trusts were excluded, including three independent provider trusts that lacked spatial identification codes. We excluded patients detained under sections of the Mental Health Act concerned only with conveyance to, or assessment in, a registered Place of Safety, or for short-term (≤72 h) assessment only, as these do not in themselves necessarily mean that the person will be admitted to an inpatient mental health bed. MHMDS contained reasonably complete data for a limited number of patient characteristics, namely age, sex, and ethnicity; however, several patient-level variables could not be included in our analysis because of high levels of missing data. Multilevel models were applied with MLwiN to estimate variation in compulsory admission, starting with null (unconditional) models that partitioned total variance in compulsory admission between each level in the model. The primary outcome was compulsory admission to a psychiatric inpatient bed, compared with people admitted voluntarily or receiving only community-based care. Data were available for 1 238 188 patients, covering 64 National Health Service provider trusts (93%) and 31 865 census lower super output areas (LSOAs; 98%). 7·5% and 5·6% of the variance in compulsory admission occurred at LSOA level and provider trust levels, respectively, after adjusting for patient characteristics. Black patients were almost three times more likely to be admitted compulsorily than were white patients (odds ratio [OR] 2·94, 95% CI 2·90-2·98). Compulsory admission was greater in more deprived areas (OR 1·22, 1·18-1·27) and in areas with more non-white residents (OR 1·51, 1·43-1·59), after adjusting for confounders. Rates of compulsory admission to inpatient psychiatric beds vary significantly between local areas and services, independent of patient, area, and service characteristics. Compulsory admission rates seem to reflect local factors, especially socioeconomic and ethnic population composition. Understanding how these factors condition access to, and use of, mental health care is likely to be important for developing interventions to reduce compulsion. National Institute for Health Research Health Services and Delivery Research Programme.


Control of Cell-Cell Distance and Cell Densities in Millimeter-Sized 3D Tissues Constructed by Collagen Nanofiber Coating Techniques.
In the field of tissue engineering, fabrication of millimeter- or centimeter-sized three-dimensional (3D) human tissues with controlled 3D cell density, 3D cell components, and 3D cell locations has been a goal of researchers. In this study, we fabricated collagen nanofiber layers of varying thicknesses on cell surfaces by previously reported collagen-coating techniques and then constructed millimeter-sized 3D human tissues, controlling the 3D cell-cell distance. In these 3D tissues, cells in the constructed 3D tissues kept a constant cell-cell distance after 24 h of incubation. Thickness of the obtained 3D tissues was controlled successfully by altering the number of cells seeded and coating times. The maximum thickness was over 2 mm when coating was carried out three times. Cell-cell distance was also controllable from approximately 15-30 μm. When cells coated once and twice with collagen nanofibers were used for the continuous construction of 3D tissues, millimeter-sized 3D tissues with areas of different cell density were obtained. The interfaces between higher and lower cell density areas were slightly mixed, but more than 80% viability was maintained after 4 days of incubation. The results suggested that stable millimeter-sized 3D tissues can be achieved using collagen nanofiber-coated cells. In addition, 3D tissues constructed by collagen-coated iPS-CM and human cardiac fibroblast were also successful.


Skeletal muscle-restricted expression of human SOD1 causes motor neuron degeneration in transgenic mice.
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease of motor neurons (MNs) that causes skeletal muscle paralysis. Familial forms of ALS are linked to mutations in the superoxide dismutase-1 (SOD1) gene. The mechanisms of human SOD1 (hSOD1) toxicity to MNs are unknown. We hypothesized that skeletal muscle is a primary site of pathogenesis in ALS that triggers MN degeneration. We created transgenic (tg) mice expressing wild-type-, G37R- and G93A-hSOD1 gene variants only in skeletal muscle. These tg mice developed age-related neurologic and pathologic phenotypes consistent with ALS. Affected mice showed limb weakness and paresis with motor deficits. Skeletal muscles developed severe pathology involving oxidative damage, protein nitration, myofiber cell death and marked neuromuscular junction (NMJ) abnormalities. Spinal MNs developed distal axonopathy and formed ubiquitinated inclusions and degenerated through an apoptotic-like pathway involving capsase-3. Mice expressing wild-type and mutant forms of hSOD1 developed MN pathology. These results demonstrate that human SOD1 in skeletal muscle has a causal role in ALS and identify a new non-autonomous mechanism for MN degeneration explaining their selective vulnerability. The discovery of instigating molecular toxicities or disease progression determinants within skeletal muscle could be very valuable for the development of new effective therapies for the treatment and cure of ALS.


Accuracy of postcontrast 3D turbo spin-echo MR sequence for the detection of enhanced inflammatory lesions in patients with multiple sclerosis.
Therapeutic strategies for patients with MS partly rely on contrast-enhanced MR imaging. Our aim was to assess the diagnostic performance of 3D turbo spin-echo MR imaging with variable refocusing flip angles at 3T for the detection of enhanced inflammatory lesions in patients with multiple sclerosis. Fifty-six patients with MS were prospectively investigated by using postcontrast T1-weighted axial 2D spin-echo and 3D TSE MR images. The order in which both sequences were performed was randomized. Axial reformats from 3D T1 TSE were generated to match the 2D spin-echo images. The reference standard was defined by using clinical data and all MR images available. Three separate sets of MR images (2D spin-echo images, axial reformats, and multiplanar images from 3D TSE sequences) were examined in a blinded fashion by 2 neuroradiologists separately for the detection of enhanced MS lesions. Image artifacts and contrast were evaluated. No artifacts related to vascular pulsation were observed on 3D TSE images, whereas image artifacts were demonstrated on 2D spin-echo images in 41 patients. One hundred twelve enhanced MS lesions were identified in 19 patients. Sixty-four lesions were correctly diagnosed by using 2D spin-echo images; 90, by using 3D TSE axial reformatted views; and 106, by using multiplanar analysis of the 3D TSE sequence. Multiplanar analysis was 94.7% sensitive and 100% specific for the diagnosis of patients with at least 1 enhanced lesion. Contrast of enhanced MS lesions was significantly improved by using the 3D TSE sequence (P < .011). The 3D TSE sequence with multiplanar analysis is a useful tool for the detection of enhanced MS lesions.


TRIAD IX: Can a Patient Testimonial Safely Help Ensure Prehospital Appropriate Critical Versus End-of-Life Care?
The present study sought to assess the clarity of Physician Orders for Life-Sustaining Treatment (POLST) or Living Will (LW) documents alone or in combination with a video message/testimonial (VM). Emergency medical services (EMS) personnel responded to survey questions about the meaning of stand-alone POLST and LW documents and those used in conjunction with emergent care scenarios. Personnel were randomized to receive documents only or documents with VM. Questions sought a code status for each scenario and a resuscitation decision. Code status responses were analyzed for consensus (95% response rate), resuscitation responses for correct treatment decisions. The survey response rate was 85%. Approximately half of emergency medical technician (EMT) respondents were EMT basic, and half EMT respondents were paramedic, with an average age of 42 years. Less than half had previous POLST/LW training averaging 2 hours. Consensus failed to be reached for stand-alone documents. For clinical scenarios, responses to POLST documents specifying do not resuscitate/comfort measures only or cardiopulmonary resuscitation/full treatment exceeded 80% for code status designation and correct resuscitation decisions. Other POLST resuscitation/treatment combinations showed more disparate responses, and most benefited from VM with changes in responses of 20% or more (P ≤ 0.025). Code status responses to LW-based scenarios evidenced a nonconsensus majority (79%-83%) that was significantly affected with VMs (≥12%, P ≤ 0.004); half evidenced large changes in resuscitation decisions (49%, P < 0.001). Document clarity, judged by consensus response, was rarely evidenced. video message/testimonial seems to be a helpful aid to both POLST and LWs. Standardized education and training reveal opportunities to improve patient safety to ensure patient wishes.


Down-regulation of progesterone receptor membrane component 1 (PGRMC1) in peripheral nucleated blood cells associated with premature ovarian failure (POF) and polycystic ovary syndrome (PCOS).
Progesterone receptor membrane component 1 (PGRMC1) is a member of a progesterone-binding complex implicated in female reproduction. We aimed i) to determine the natural expression of PGRMC1 in peripheral nucleated blood cells throughout the menstrual cycle and ii) to investigate any association between PGRMC1 levels in leukocytes and conditions characterized by reduced fertility. We analyzed PGRMC1 expression in peripheral leukocytes from 15 healthy cycling women over four weeks. Additionally, we determined PGRMC1 levels in samples from patients with premature ovarian failure (POF) and polycystic ovary syndrome (PCOS) as well as in healthy postmenopausal women and male controls. The levels of PGRMC1 protein in nucleated peripheral blood cells were quantified by Western blot analysis. PGRMC1 levels did not vary significantly throughout the menstrual cycle. We observed a significant down-regulation of PGRMC1 in postmenopausal women and in patients with premature ovarian failure (POF) and polycystic ovary syndrome (PCOS) when compared to early follicular phase of healthy women. This study suggests that reduced levels of PGRMC1 in peripheral leukocytes are associated with perturbed ovulatory function.


Depressive symptoms among children attending community based support in South Africa - pathways for disrupting risk factors.
Children in Southern Africa are exposed to high rates of structural and family adversities. This study tests whether services from Community Based Organisations (CBOs) in South Africa can promote children's resilience against depression exposed to such adversities. Two linked longitudinal studies were conducted, comprising n = 1848 children aged 9 to 13 years. One group received CBO services, whilst the other (quasi-control) did not. Analyses used interaction terms in regression models to test for potential moderation effects of CBO attendance, and marginal effects models to interpret significant interactions. Two interaction effects were shown, demonstrating moderation effects of CBO attendance on common structural disadvantages. First, children exposed to community violence showed increased depression (contrast = 0.62 [95%CI 0.43, 0.82], p < .001), but this association was removed by CBO access (contrast = 0.07 [95%CI -0.28, 0.43], p = .682). Second, children living in informal housing showed increased depression (contrast = 0.63 [95%CI 0.42, 0.85], p < .001), however, this association was removed by CBO access (contrast = 0.01 [95%CI -0.55, 0.56], p = .977). CBO attendance is associated with fewer depressive symptoms, and can buffer against important structural adversities of poor housing and violence that are common in high HIV-prevalence areas. However, CBO attendance was not able to remove the increased psychosocial distress associated with some family-level vulnerabilities such as orphanhood and abuse. These findings highlight the centrality of CBO-provided psychosocial support for children in Southern Africa, and suggest areas for bolstering provision.


Supercooled liquid-like solvent in trypsin crystals: implications for crystal annealing and temperature-controlled X-ray radiation damage studies.
The study of temperature-dependent physical changes in flash-cooled macromolecular crystals is pertinent to cryocrystallography and related issues such as crystal annealing, X-ray radiation damage and kinetic crystallography. In this context, the unit-cell volume of flash-cooled trigonal and orthorhombic trypsin crystals has been monitored upon warming from 100 to 200 K and subsequent re-cooling to 100 K. Crystals of both forms were obtained under the same crystallization conditions, yet they differ in solvent content and channel size. An abrupt non-reversible unit-cell volume decrease is observed at 185 K in orthorhombic and at 195 K in trigonal crystals as the temperature is increased; this result is consistent with ultra-viscous solvent leaving the crystals. Concomitant appearance of ice rings in the diffraction patterns suggests that the transported solvent forms crystalline ice. These results demonstrate that solvent in flash-cooled protein crystals is liquid-like near its crystallization temperature, as has been proposed, yet controversially discussed, for the case of pure water. The use of mineral oil prevents the unit-cell volume decrease in trigonal but not in orthorhombic crystals. The observation of liquid-like solvent has implications in the development of annealing protocols and points a way to the rational design of temperature-controlled crystallographic studies that aim either at studying specific radiation damage or at trapping enzymatic intermediate states.


Activation of the sympathetic nervous system modulates neutrophil function.
Emerging evidence has revealed that noradrenaline (NA), the main neurotransmitter of the sympathetic nervous system (SNS), regulates a variety of immune functions via binding to adrenergic receptors present on immune cells. In this study, we examined the role of NA in the regulation of neutrophil functions. Neutrophils were isolated from the bone marrow of naïve mice and treated with NA at various concentrations to assess the effect on various neutrophil functions. Additionally, we performed cremaster intravital microscopy to examine neutrophil-endothelial cell interactions following NA superfusion in vivo. In a separate group of animals, mice were subjected to an experimental model of stroke and at 4 and 24 h neutrophils were isolated for assessment on their ability to migrate toward various chemokines. Treatment of neutrophils with NA for 4 h significantly impaired neutrophil chemotaxis and induced an N2 neutrophil phenotype with reduced expression of the genes critical for cytoskeleton remodeling and inflammation. Prolonged NA administration promoted neutrophils to release myeloperoxidase and IL-6, but suppressed the production of interferon-γ and IL-10, reduced neutrophil activation and phagocytosis. Superfusion of NA over the cremaster muscle almost completely inhibited fMLP-induced neutrophil adhesion/arrest and transmigration. Furthermore, using a mouse model of stroke, a pathological condition in which SNS activation is evident, neutrophils isolated from poststroke mice showed markedly reduced chemotaxis toward all of the chemokines tested. The findings from our study indicate that neutrophil chemotaxis, activation, and phagocytosis can all be negatively regulated in an NA-dependent manner. A better understanding of the relationship between sympathetic activation and neutrophil function will be important for the development of effective antibacterial interventions.


Improved multimodal prediction of progression from MCI to Alzheimer's disease combining genetics with quantitative brain MRI and cognitive measures.
There is a pressing need for non-invasive, cost-effective tools for early detection of Alzheimer's disease (AD). Using data from the Alzheimer's Disease Neuroimaging Initiative (ADNI), Cox proportional models were conducted to develop a multimodal hazard score (MHS) combining age, a polygenic hazard score (PHS), brain atrophy, and memory to predict conversion from mild cognitive impairment (MCI) to dementia. Power calculations estimated required clinical trial sample sizes after hypothetical enrichment using the MHS. Cox regression determined predicted age of onset for AD pathology from the PHS. The MHS predicted conversion from MCI to dementia (hazard ratio for 80th versus 20th percentile: 27.03). Models suggest that application of the MHS could reduce clinical trial sample sizes by 67%. The PHS alone predicted age of onset of amyloid and tau. The MHS may improve early detection of AD for use in memory clinics or for clinical trial enrichment. A multimodal hazard score (MHS) combined age, genetics, brain atrophy, and memory. The MHS predicted time to conversion from mild cognitive impairment to dementia. MHS reduced hypothetical Alzheimer's disease (AD) clinical trial sample sizes by 67%. A polygenic hazard score predicted age of onset of AD neuropathology.


Molecular dynamics simulations and conductance studies of the interaction of VP1 N-terminus from Polio virus and gp41 fusion peptide from HIV-1 with lipid membranes.
The icosahedral Polio virus capsid consists of 60 copies of each of the coat proteins VP1, VP2, VP3 and myristolyated VP4 (myrVP4). Catalyzed by the host cell receptor the Polio virus enters the host cell via externalization of myrVP4 and the N terminal part of VP1. There are several assumptions about the individual role of both of the proteins in the mechanism of membrane attachment and genome injection. We use the first 32 N terminal amino acids of VP1 and applied molecular dynamics simulations to assess its mechanism of function when attached and inserted into hydrated lipid membranes (POPC). Helical models are placed in various positions in regard to the lipid membrane to start with. As a comparison, the first 33 amino acids of the fusion peptide of gp41 of HIV-1 are simulated under identical conditions. Computational data support the idea that VP1 is not penetrating into the membrane to form a pore; it rather lays on the membrane surface and only perturbs the membrane. Furthermore, this idea is strengthened by channel recordings of both peptides showing irregular openings.


Mono- and bichromatic electron dynamics: LiH, a test case.
This paper describes an electron dynamics method where the time dependence of an external oscillating electric field is the perturbing part of the Hamiltonian. Application of the electric field induces charge movement inside the molecule and electronic transitions between the molecular orbitals. The test system is the neutral LiH molecule. The method is applied to wave functions calculated using the B3LYP (hybrid) density functional, with the STO-3G and the 6-31+G basis sets. The molecule undergoes full population inversion between the HOMO and the LUMO when the electric field is in resonance with the HOMO-LUMO energy gap. The magnitude of the electric field directly affects the rate at which electronic transitions occur and the rate at which charges move between lithium and hydrogen atoms. The method is used to model both monochromatic and bichromatic multiphoton effects in LiH. Monochromatic one-, two- and three-photon transitions occur between the HOMO, LUMO and two other virtual orbitals. There is evidence of both [1+2] direct and [1+1+1] stepwise multiphoton transitions. Bichromatically, two "laser" pulses are applied at different frequencies. Electronic transitions can be fine-tuned to occur via pre-specified pathways of virtual molecular orbitals.


Are acid secretory tests of diagnostic value in patients with benign-looking gastric ulcers?
In 41 patients in whom endoscopy showed benign gastric ulcer, gastric secretory function was assessed before and after stimulation with pentagastrin (6 micrograms/kg); they were then followed up for a period of 2-4 years. Of these 41 patients, 37 had benign and the remaining 4 malignant ulcers. Of the 37 patients with benign gastric ulcers, 11 had basal achlorhydria but all secreted acid after pentagastrin stimulation. Seven of these 37 patients (19%) secreted less than 5 mmol/h in response to stimulation with pentagastrin. Of the 4 patients with malignant gastric ulcer, 3 had basal achlorhydria. After pentagastrin stimulation 3 of these patients (75%) secreted up to 5 mmol/h and 1 patient secreted 17 mmol/h. Biopsy evidence of malignancy was only obtained after repeat endoscopy and biopsy in 2 of these 4 patients. The present study confirms that a malignant ulcer may simulate a benign ulcer on endoscopy, and shows that the finding of hypochlorhydria in such patients warrants a cautious approach with repeat endoscopy and biopsy to exclude a malignant lesion.


Metabolic and renal outcomes of empagliflozin in patients with type 2 diabetes mellitus attending Armed Forces Hospital in Saudi Arabia: Retrospective cohort study.
To explore the effects of empagliflozin (25 mg) on metabolic and renal parameters in patients with type 2 diabetes mellitus (T2DM). This retrospective observational comparative study was conducted at a military hospital in southern Saudi Arabia. All adults (aged >18 years) with T2DM who attended diabetic clinics between October 2021 to March 2022 (6 months), with or without insulin treatment, were eligible for inclusion in the study. Following the initiation of empagliflozin treatment, statistically significant reductions in patient weight (kg) were observed at 1, 3-5, and 6 months. In addition, low-density lipoprotein levels significantly decreased 3-5 months post-treatment initiation (p=0.011). However, serum creatinine level decreased gradually with time during the treatment with empagliflozin, from 87.45±31.78 (0.105) to 78.39±27.43 (0.033). Furthermore, after empagliflozin treatment, the urinary albumin-to-creatinine ratio significantly decreased at 3-5 and 6 months. Moreover, HbA1c levels exhibited statistically significant decreases at 3-5 months (p<0.001) and at 6 months (p<0.001) following the initiation of empagliflozin treatment. Notably, systolic and diastolic blood pressure significantly reduced 6 months after empagliflozin treatment. In the current study, empagliflozin has demonstrated efficacy in controlling blood pressure and body weight, and improving renal function, short-term dyslipidemia, and glycemic control in patients with T2DM.


[The effect of feeding mangel-wurzels on blood beta-hydroxybutyrate level of milk-producing cows].
The effect of feeding dairy cattle mangel-wurzels on the serum increased concentration of beta-hydroxybutyrate was investigated in serum is normally indicative of an increased catabolism of fats in the liver as a result of an energy shortage. Feeding cattle large amounts of mangel-wurzels can cause the serum beta-hydroxybutyrate concentration to increase above normal reference values, thus seemingly indicating that there is a (sub)clinical acetonaemia. In fact the increased concentration of beta-hydroxybutyrate is probably the result of an increased production of butyrate in the rumen.


Pseudomonas attachment to new hydrogel contact lenses.
The adhesion of two strains of Pseudomonas aeruginosa to unworn hydrogel contact lenses (CLs) was studied by scanning electron microscopy. New daily-wear polymacon lenses (38.6% water content) and extended-wear lidofilcon A lenses (70% water content) were exposed to a suspension of 10(8) colony-forming units per milliliter of P aeruginosa for 2, 5, 15, 30, and 60 minutes. Increasing numbers of bacteria were found to attach to the lenses with time, and no significant differences between CL types were demonstrated. Attachment was considered to be irreversible since washing did not remove the bacteria. We conclude that P aeruginosa can actively attach to new, unworn hydrogel CLs.


Prognosis of cirrhotic patients admitted to Emergency Departments: A multicenter study.
Life threatening complications can occur at any stage of cirrhosis progression. There are few studies on the prognosis of cirrhotic patients managed in an Emergency Department (ED) although management of patients will occur in the ED. The objective of our study was to determine the risk factors for mortality in cirrhotic patients who visited to the ED. All cirrhotic patients attending ED in three different university hospitals of Assistance Publique - Hôpitaux de Paris between January 2014 and June 2015 were identified by a retrospective analysis of digital records and included in the study. The primary end-point was 30-day mortality in all cirrhotic patients who visited the ED. A total of 609 ED visits were analyzed among 224 patients: 115 (51%) presented a cirrhosis of alcoholic origin, 43 (19%) were caused by Hepatitis C, 28 (13%) of mixed origin (viral and alcoholic), 17 (8%) were caused by Hepatitis B and 21 (9%) of other origins. Fifty-five (25%) of these patients died within 30 days of their initial presentation to the ED. In multivariate analysis, the age (Odds Ratio: 1.04 [1.01-1.07]), cirrhosis associated with hepatocellular carcinoma (OR: 3.07 [1.37-6.91]), serum creatinine at admission (OR: 1.01 [1.01-1.02]), serum bilirubin at admission (OR: 1.01 [1.01-1.02]) and health impairment (OR: 2.57 [1.28-5.16]) were associated with mortality. The mortality rate of cirrhotic patients attending an ED was high. The prognosis of cirrhotic patients admitted to the ED depended on the severity of the liver and other organ dysfunction. The presence of a hepatocellular carcinoma on admission was also a risk factor for death.


Prolonged constant load cycling exercise is associated with reduced gross efficiency and increased muscle oxygen uptake.
This study investigated the effects of prolonged constant load cycling exercise on cycling efficiency and local muscle oxygen uptake responses. Fourteen well-trained cyclists each completed a 2-h steady-state cycling bout at 60% of their maximal minute power output to assess changes in gross cycling efficiency (GE) and muscle oxygen uptake (mVO2 ) at time points 5, 30, 60, 90, and 120 min. Near-infrared spatially resolved spectroscopy (NIRS) was used to continually monitor tissue oxygenation of the Vastus Lateralis muscle, with arterial occlusions (OCC) applied to assess mVO2 . The half-recovery time of oxygenated hemoglobin (HbO2 ) was also assessed pre and post the 2-h cycling exercise by measuring the hyperemic response following a 5-min OCC. GE significantly declined during the 2-h cycling bout (18.4 ± 1.6 to 17.4 ± 1.4%; P < 0.01). Conversely, mVO2 increased, being significantly higher after 90 and 120 min than at min 5 (+0.04 mlO2 /min/100 g; P = 0.03). The half-recovery time for HbO2 was increased comparing pre and post the 2-h cycling exercise (+7.1 ± 19s), albeit not significantly (d: 0.48; P = 0.27). This study demonstrates that GE decreases during prolonged constant load cycling exercise and provides evidence of an increased mVO2 , suggestive of progressive mitochondrial or contractile inefficiency.


Cross-reactions in Legionella antisera with Bordetella pertussis strains.
While preparing slide agglutination test antisera and immunofluorescence conjugates for the identification of Legionella species and serogroups, we found that several of the reagents cross-reacted with Bordetella pertussis strains. To determine the extent of this problem and to estimate the specificity of Legionella reagents, we tested slide agglutination test antisera against 22 species and 35 serogroups with 92 bacterial strains representing 19 genera. The only cross-reactions observed were with Legionella pneumophila serogroup 10, L. maceachernii, L. gormanii, and L. feeleii serogroup 1 antisera and 4 of 10 B. pertussis strains. Nineteen conjugates, previously available from the Centers for Disease Control but no longer distributed as reference reagents, were tested with the four cross-reactive B. pertussis strains. Two conjugates, L. micdadei and L. wadsworthii, stained three of the B. pertussis strains at a fluorescence intensity of greater than or equal to 3+. All cross-reactions were removed from the antisera and conjugates by absorption with the cross-reacting strain without diminishing the homologous reaction. Special emphasis should be placed on the identification and removal of cross-reactions in Legionella reagents with strains that have similar morphologic and growth characteristics.


DNA methylation signatures identify biologically distinct subtypes in acute myeloid leukemia.
We hypothesized that DNA methylation distributes into specific patterns in cancer cells, which reflect critical biological differences. We therefore examined the methylation profiles of 344 patients with acute myeloid leukemia (AML). Clustering of these patients by methylation data segregated patients into 16 groups. Five of these groups defined new AML subtypes that shared no other known feature. In addition, DNA methylation profiles segregated patients with CEBPA aberrations from other subtypes of leukemia, defined four epigenetically distinct forms of AML with NPM1 mutations, and showed that established AML1-ETO, CBFb-MYH11, and PML-RARA leukemia entities are associated with specific methylation profiles. We report a 15 gene methylation classifier predictive of overall survival in an independent patient cohort (p < 0.001, adjusted for known covariates).


A role for Cdk2 kinase in negatively regulating DNA replication during S phase of the cell cycle.
Using cell-free extracts made from Xenopus eggs, we show that cdk2-cyclin E and A kinases play an important role in negatively regulating DNA replication. Specifically, we demonstrate that the cdk2 kinase concentration surrounding chromatin in extracts increases 200-fold once the chromatin is assembled into nuclei. Further, we find that if the cdk2-cyclin E or A concentration in egg cytosol is increased 16-fold before the addition of sperm chromatin, the chromatin fails to initiate DNA replication once assembled into nuclei. This demonstrates that cdk2-cyclin E or A can negatively regulate DNA replication. With respect to how this negative regulation occurs, we show that high levels of cdk2-cyclin E do not block the association of the protein complex ORC with sperm chromatin but do prevent association of MCM3, a protein essential for replication. Importantly, we find that MCM3 that is prebound to chromatin does not dissociate when cdk2-cyclin E levels are increased. Taken together our results strongly suggest that during the embryonic cell cycle, the low concentrations of cdk2-cyclin E present in the cytosol after mitosis and before nuclear formation allow proteins essential for potentiating DNA replication to bind to chromatin, and that the high concentration of cdk2-cyclin E within nuclei prevents MCM from reassociating with chromatin after replication. This situation could serve, in part, to limit DNA replication to a single round per cell cycle.


Cytochemical flow analysis of intracellular G6PD and aggregate analysis of mosaic G6PD expression.
Medicines that exert oxidative pressure on red blood cells (RBC) can cause severe hemolysis in patients with glucose-6-phosphate dehydrogenase (G6PD) deficiency. Due to X-chromosome inactivation, females heterozygous for G6PD with 1 allele encoding a G6PD-deficient protein and the other a normal protein produce 2 RBC populations each expressing exclusively 1 allele. The G6PD mosaic is not captured with routine G6PD tests. An open-source software tool for G6PD cytofluorometric data interpretation is described. The tool interprets data in terms of % bright RBC, or cells with normal G6PD activity in specimens collected from 2 geographically and ethnically distinct populations, an African American cohort (USA) and a Karen and Burman ethnic cohort (Thailand) comprising 242 specimens including 89 heterozygous females. The tool allowed comparison of data across 2 laboratories and both populations. Hemizygous normal or deficient males and homozygous normal or deficient females cluster at narrow % bright cells with mean values of 96%, or 6% (males) and 97%, or 2% (females), respectively. Heterozygous females show a distribution of 10-85% bright cells and a mean of 50%. The distributions are associated with the severity of the G6PD mutation. Consistent cytofluorometric G6PD analysis facilitates interlaboratory comparison of cellular G6PD profiles and contributes to understanding primaquine-associated hemolytic risk.


Manipulating objects and telling words: a study on concrete and abstract words acquisition.
Four experiments (E1-E2-E3-E4) investigated whether different acquisition modalities lead to the emergence of differences typically found between concrete and abstract words, as argued by the words as tools (WAT) proposal. To mimic the acquisition of concrete and abstract concepts, participants either manipulated novel objects or observed groups of objects interacting in novel ways (Training 1). In TEST 1 participants decided whether two elements belonged to the same category. Later they read the category labels (Training 2); labels could be accompanied by an explanation of their meaning. Then participants observed previously seen exemplars and other elements, and were asked which of them could be named with a given label (TEST 2). Across the experiments, it was more difficult to form abstract than concrete categories (TEST 1); even when adding labels, abstract words remained more difficult than concrete words (TEST 2). TEST 3 differed across the experiments. In E1 participants performed a feature production task. Crucially, the associations produced with the novel words reflected the pattern evoked by existing concrete and abstract words, as the first evoked more perceptual properties. In E2-E3-E4, TEST 3 consisted of a color verification task with manual/verbal (keyboard-microphone) responses. Results showed the microphone use to have an advantage over keyboard use for abstract words, especially in the explanation condition. This supports WAT: due to their acquisition modality, concrete words evoke more manual information; abstract words elicit more verbal information. This advantage was not present when linguistic information contrasted with perceptual one. Implications for theories and computational models of language grounding are discussed.


Advances in cell-free protein array methods.
Cell-free protein microarrays represent a special form of protein microarray which display proteins made fresh at the time of the experiment, avoiding storage and denaturation. They have been used increasingly in basic and translational research over the past decade to study protein-protein interactions, the pathogen-host relationship, post-translational modifications, and antibody biomarkers of different human diseases. Their role in the first blood-based diagnostic test for early stage breast cancer highlights their value in managing human health. Cell-free protein microarrays will continue to evolve to become widespread tools for research and clinical management. Areas covered: We review the advantages and disadvantages of different cell-free protein arrays, with an emphasis on the methods that have been studied in the last five years. We also discuss the applications of each microarray method. Expert commentary: Given the growing roles and impact of cell-free protein microarrays in research and medicine, we discuss: 1) the current technical and practical limitations of cell-free protein microarrays; 2) the biomarker discovery and verification pipeline using protein microarrays; and 3) how cell-free protein microarrays will advance over the next five years, both in their technology and applications.


Cooperative DNA binding mediated by KicGAS/ORF52 oligomerization allows inhibition of DNA-induced phase separation and activation of cGAS.
Cyclic GMP-AMP synthase (cGAS) is a key DNA sensor that detects aberrant cytosolic DNA arising from pathogen invasions or genotoxic stresses. Upon binding to DNA, cGAS is activated and catalyzes the synthesis of cyclic GMP-AMP (cGAMP), which induces potent antimicrobial and antitumor responses. Kaposi sarcoma-associated herpesvirus (KSHV) is a human DNA tumor virus that causes Kaposi sarcoma and several other malignancies. We previously reported that KSHV inhibitor of cGAS (KicGAS) encoded by ORF52, inhibits cGAS enzymatic activity, but the underlying mechanisms remained unclear. To define the inhibitory mechanisms, here we performed in-depth biochemical and functional characterizations of KicGAS, and mapped its functional domains. We found KicGAS self-oligomerizes and binds to double stranded DNA cooperatively. This self-oligomerization is essential for its DNA binding and cGAS inhibition. Interestingly, KicGAS forms liquid droplets upon binding to DNA, which requires collective multivalent interactions with DNA mediated by both structured and disordered domains coordinated through the self-oligomerization of KicGAS. We also observed that KicGAS inhibits the DNA-induced phase separation and activation of cGAS. Our findings reveal a novel mechanism by which DNA viruses target the host protein phase separation for suppression of the host sensing of viral nucleic acids.


Errata.
My attention has been drawn by correspondents to two incorrect statements in my article "Research and Politics" (Science, March 4, 1949). First, it was stated that Vavilov died in a concentration camp. This should have read "died in exile in a remote region of the USSR," since the details are unknown. Second, I said that Kammerer committed suicide after going to Russia. Actually, he did so while still in Vienna, at the moment when he was finally to move to Moscow.


Remote ischemic preconditioning for prevention of contrast-induced acute kidney injury in diabetic patients.
There are some clinical trials showing that short-term ischemia in one organ can protect different organs against higher intensity and longer ischemic insult. We designed a study to assess whether remote ischemic preconditioning (RIPC) on one organ can decrease the rate of contrast-induced acute kidney injury (AKI) in diabetic patients who undergo coronary artery angiography (CAA). This randomized control trial included 96 diabetic patients who were candidates for CAA. Exclusion criteria were congestive heart failure and complications during CAA. All of the patients received 1000 mL of normal saline before CAA. The RIPC group underwent 3 cycles of 5-minute ischemia in their right arm. Serum creatinine was measured before and 24 hours after CAA. Contrast-induced AKI was reported in 5 cases in the control group and 1 case in the RIPC group (P = .13, odds ratio, 5.4). The differences in serum creatinine level before and after the procedure was significantly lower in RIPC group than that in the control group (P = .04, odds ratio, 0.08). Serum creatinine rise significantly correlated with contrast dose (P = .02) and a history of hypertension (P = .02) in both groups. Ischemic preconditioning had a protective effect on contrast-induced AKI in our study. Since this method is harmless and cost effective, further studies on patients with chronic kidney disease is required to evaluate addition of ischemic preconditioning to our clinical practice for prevention of contrast-induced AKI.


An optical clearing technique for plant tissues allowing deep imaging and compatible with fluorescence microscopy.
We report on a nondestructive clearing technique that enhances transmission of light through specimens from diverse plant species, opening unique opportunities for microscope-enabled plant research. After clearing, plant organs and thick tissue sections are amenable to deep imaging. The clearing method is compatible with immunocytochemistry techniques and can be used in concert with common fluorescent probes, including widely adopted protein tags such as GFP, which has fluorescence that is preserved during the clearing process.


Mobile phones to support adherence to antiretroviral therapy: what would it cost the Indian National AIDS Control Programme?
Adherence to antiretroviral treatment (ART) is critical to maintaining health and good clinical outcomes in people living with HIV/AIDS. To address poor treatment adherence, low-cost interventions using mobile communication technology are being studied. While there are some studies that show an effect of mobile phone reminders on adherence to ART, none has reported on the costs of such reminders for national AIDS programmes. This paper aims to study the costs of mobile phone reminder strategies (mHealth interventions) to support adherence in the context of India's National AIDS Control Program (NACP). The study was undertaken at two tertiary level teaching hospitals that implement the NACP in Karnataka state, South India. Costs for a mobile phone reminder application to support adherence, implemented at these sites (i.e. weekly calls, messages or both) were studied. Costs were collected based on the concept of avoidable costs specific to the application. The costs that were assessed were one-time costs and recurrent costs that included fixed and variable costs. A sequential procedure for costing was used. Costs were calculated at national-programme level, individual ART-centre level and individual patient level from the NACP's perspective. The assessed costs were pooled to obtain an annual cost per patient. The type of application, number of ART centres and number of patients on ART were varied in a sensitivity analysis of costs. The Indian NACP would incur a cost of between 79 and 110 INR (USD 1.27-1.77) per patient per year, based on the type of reminder, the number of patients on ART and the number of functioning ART centres. The total programme costs for a scale-up of the mHealth intervention to reach the one million patients expected to be on treatment by 2017 is estimated to be 0.36% of the total five-year national-programme budget. The cost of the mHealth intervention for ART-adherence support in the context of the Indian NACP is low and is facilitated by the low cost of mobile communication in the country. Extending the use of mobile communication applications beyond adherence support under the national programme could be done relatively inexpensively.


Blocking inducible co-stimulator in the absence of CD28 impairs Th1 and CD25+ regulatory T cells in murine colitis.
Several autoimmune disease models depend on an imbalance in the activation of aggressor T(h)1 and CD4(+)CD25(+) regulatory T (T(reg)) cells. Here we compare the requirement for signals through the co-stimulatory molecules CD28 and inducible co-stimulator (ICOS) in chronic murine colitis, a model for inflammatory bowel disease. We used a colitis model in which disease-causing CD45RB(hi) T cells alone or in combination with CD4(+)CD25(+) T cells from either CD28-deficient or wild-type donors were transferred into T cell-deficient animals, half of which were treated with ICOS-blocking reagents. Blocking ICOS on the surface of CD28-deficient T(h)1 cells abrogated development of colitis, whereas blocking CD28 or ICOS alone had little to no effect on disease induction. In contrast to T(h)1 cells, regulatory T cell functioning depended mostly on CD28 signaling with only a minor contribution for ICOS. We conclude that CD28 and ICOS collaborate to development of murine colitis by aggressor T(h)1 cells, and that CD28 is required for T(reg) cells, which should caution against the use of CD28-blocking reagents in inflammatory bowel disease.


[Dandy-Walker malformation associated with extracranial abnormalities in a newborn].
Hydrocephalus is a clinical condition that consists of an accumulation of cerebrospinal fluid around the brain; Dandy- Walker syndrome is a rare cause of it. We present the case of a newborn with prenatal diagnose of hydrocephalus due to a Dandy-Walker malformation, as well as ambiguous genitalia. After birth, diagnosis of Dandy-Walker malformation associated with uncommon extracranial manifestations is confirmed. Specifically, the baby presents interscrotal hypospadias and coronary sinus dilatation due to the persistence of the left superior vena cava. With the exposition of this case, we bring Malformación de Dandy-Walker asociada a malformaciones extracraneales en un neonato Dandy-Walker malformation associated with extracranial abnormalities in a newborn out the complexity of the Dandy-Walker syndrome due to the malformations associated with it; the ones that will determine the prognosis and the need of a multidisciplinary treatment.


Adipose tissue extracellular matrix and vascular abnormalities in obesity and insulin resistance.
Insulin resistance is associated with inflammation, fibrosis, and hypoxia in adipose tissue. This study was intended to better characterize the extracellular matrix (ECM) and vascularity of insulin-resistant adipose tissue. Adipose expression of collagens, elastin, and angiogenic factors was assessed using real-time RT-PCR and immunohistochemistry (IHC) in abdominal sc adipose tissue. Adipocyte-macrophage coculture experiments examined the effects of polarized macrophages on adipose ECM gene expression, and the effects of collagens were measured in an angiogenesis assay. A total of 74 nondiabetic subjects participated at a University Clinical Research Center. Interventions included baseline adipose biopsy and measurement of insulin sensitivity. Outcome measures included characterization of vascularity and ECM in adipose tissue. CD31 (an endothelial marker) mRNA showed no significant correlation with body mass index or insulin sensitivity. In a subgroup of 17 subjects (nine obese, eight lean), CD31-positive capillary number in obese was decreased by 58%, whereas larger vessels were increased by 70%, accounting for the lack of change in CD31 expression with obesity. Using IHC, obese (compared with lean) subjects had decreased elastin and increased collagen V expression, and adipocytes cocultured with M2 macrophages had reduced elastin and increased collagen V expression. In obese subjects, collagen V was colocalized with large blood vessels, and the addition of collagen V to an angiogenesis assay inhibited endothelial budding. The adipose tissue from obese/insulin-resistant subjects has fewer capillaries and more large vessels as compared with lean subjects. The ECM of adipose tissue may play an important role in regulating the expandability as well as angiogenesis of adipose tissue.


Baseline pain as a moderator of hypnotic analgesia for burn injury treatment.
Sixty-one patients hospitalized for severe burns were randomly assigned to conditions in which they received either hypnosis or a control condition in which they received attention, information, and brief relaxation instructions from a psychologist. The posttreatment pain scores of the 2 groups did not differ significantly when all patients were considered. However, when a subset of patients who reported high levels of baseline pain were examined, it was found that patients in the hypnosis group reported less posttreatment pain than did patients in the control group. The findings are used to replicate earlier studies of burn pain hypnoanalgesia, explain discrepancies in the literature, and highlight the potential importance of motivation with this population.


Financial Stress Among Latino Adults in California During COVID-19.
We study the impact of COVID-19 on stress, and especially on financial stress, among Latino adults in California. We take a mixed methods approach and rely on quantitative and qualitative data for our analysis. We recruited 84 low- and moderate-income (LMI) Latino adults in California through the Understanding America Study (UAS) Internet Panel who also participated in the Mobile Financial Diary (MFD) project, which took place during 2018 - 2019. We analyze data about personal experiences during COVID-19 in October 2020 and compare this to data collected during the period from August to October 2018. Our study portrays the experiences of California Latino adults who were predominantly born in the USA and are likely to be working and speak English. We also observe that a large percentage of our participants had health insurance and relatively high levels of educational attainment. We find contradictory results from our quantitative measures, where one of our indicators of financial behavior and well-being showed a significant increase (Financial Health Score), and the other (Financial Well-Being Scale) showed a significant decrease during COVID-19. Anxiety (GAD-7) and depression (PHQ) measures show no significant changes during COVID-19 in comparison to 2018. Nonetheless, our qualitative data analysis shows that many of our participants were experiencing major stressors during the pandemic associated with labor market experiences and family circumstances. In our qualitative data analysis, we also observe that women seemed to have been affected the most by the pandemic. The online version contains supplementary material available at 10.1007/s41996-021-00087-0.


Evone® Flow-Controlled Ventilation During Upper Airway Surgery: A Clinical Feasibility Study and Safety Assessment.
Introduction: During upper airway surgery in a narrowed airway due to tumor or stenosis, safe ventilation, good laryngotracheal exposure, and preservation of an adequate surgical working space are of paramount importance. This can be achieved by small-lumen ventilation such as High Frequency Jet Ventilation (HFJV). However, this technique has major drawbacks, such as air-trapping and desaturation in patients with poor pulmonary reserve. Recently, an innovative ventilating system with flow-controlled ventilation (FCV) and a small-lumen endotracheal tube, the Evone® (Ventinova, Eindhoven, The Netherlands), was introduced, claiming to counter the drawbacks of HFJV. Objectives: To evaluate feasibility and safety of the Evone® FCV system in difficult upper airway surgery and to critically appraise this novel ventilation method. Patients and methods: Evone® is a FCV-device using a small-bore cuffed tube (Tritube®). This ventilator actively sucks air out of the lungs, rather than relying on the passive backflow of air like in HFJV. Data related to the medical history, surgery, and anesthesia of all consecutive patients undergoing upper airway surgery with Evone® FCV ventilation were included in a tertiary center retrospective observational study. Results: Fifteen Patients, with a median age of 54 years, were included. Surgical procedures and indications included laser-assisted endoscopic treatment of idiopathic subglottic stenosis (n = 3), tracheal stenosis (n = 1), and posterior glottic stenosis (n = 2), biopsy and/or Transoral Laser Microsurgery for laryngeal (pre)malignancy (n = 7) and resection of benign lesions with posterior (supra)glottic location (n = 2). Mean ventilation duration was 52.0 min (range 30-115 min, SD 19.6 min), mean surgery duration was 31.7 min (range 15-65 min, SD 13.2 min), mean minimal SaO2 was 96.3% (range 89-100%, SD 4.0%) and mean peak pCO2 was 41.4 mmHg (range 31-50 mmHg, SD = 5.5 mmHg). No anesthesia- or surgery-related complications, adverse events or intra-operative difficulties were reported during or after any of the 15 procedures. In all cases, compared to HFJV, Evone® FCV ventilation allowed a superior visualization and working space during the surgical procedure. Conclusion: The Evone® FCV ventilation system provides excellent conditions in patients undergoing upper airway surgery, as it combines excellent accessibility and visibility of the operation site with safe and stable ventilation.


Computerized Neurocognitive Test (CNT) in mild cognitive impairment and Alzheimer's disease.
Currently, computerized batteries are of great value in detecting cognitive impairment. This aim of this review was to compare the computerized neurocognitive batteries used in most studies with cognitive decline over the last 10 years. Using the search words computerized cognitive assessment with: dementia, mild cognitive impairment, and Alzheimer's disease, the CogState, CNS Vital Sings, COGDRAS and Mindstreams batteries were retrieved.


Provision of Private Care by Doctors Employed in Public Health Institutions: Ethical Considerations and Implications for Clinical Training.
This paper summarizes the difficulties that may emerge when the same care-provider attends to private and public patients within the same or different clinical settings. First, I argue that blurring the boundaries between public and private care may start a slippery slope leading to "black" under-the-table payments for preferential patient care. Second, I question whether public hospitals that allow their doctors to attend to private patients provide an appropriate learning environment for medical students and residents. Finally, I propose a way to both maintain the advantages of private care and avoid its negative consequences: complete separation between the public and the private health care systems.


Characterisation and partial sequence analysis of two novel cypoviruses isolated from the winter moth Operophtera brumata (Lepidoptera: Geometridae).
The complete nucleotide sequences of genomic segments S5 to S10 from Operophtera brumata cypovirus 18 (OpbuCPV18), and the complete nucleotide sequences of genomic segments S2, S5, S9 and S10 from Operophtera brumata cypovirus 19 (OpbuCPV19) have been determined. Each genome segment contained a single open reading frame (ORF). Conserved motifs 5' (AGUAAA....GUUAGCU) 3' were found at the ends of each segment of OpbuCPV18, whilst conserved motifs 5' (AACAAA....UUUGC) 3' were found at each segment terminus of OpbuCPV19. The putative proteins were compared with those of other members of the Reoviridae family. Phylogenetic analysis using the polyhedrin gene (S10) indicated that OpbuCPV18 was most closely related to Dendrolimus punctatus cypovirus 1, whilst OpbuCPV19 was most closely related to Trichoplusia ni cypovirus 15. In addition, analysis of S2, which encoded a putative RNA-dependant RNA polymerase gene, confirmed OpbuCPV19 belonged to the genus Cypovirus. Following the expression of the ORF from OpbuCPV19 S10, using a baculovirus expression vector, occlusion bodies were observed in insect cell culture. This demonstrated that segment 10 coded for the polyhedrin gene, capable of forming a polyhedral crystalline matrix.


Up-to-date on the evidence linking miRNA-related epitranscriptomic modifications and disease settings. Can these modifications affect cross-kingdom regulation?
The field of epitranscriptomics is rapidly developing. Several modifications (e.g. methylations) have been identified for different RNA types. Current evidence shows that chemical RNA modifications can influence the whole molecule's secondary structure, translatability, functionality, stability, and degradation, and some are dynamically and reversibly modulated. miRNAs, in particular, are not only post-transcriptional modulators of gene expression but are themselves submitted to regulatory mechanisms. Understanding how these modifications are regulated and the resulting pathological consequences when dysregulation occurs is essential for the development of new therapeutic targets. In humans and other mammals, dietary components have been shown to affect miRNA expression and may also induce chemical modifications in miRNAs. The identification of chemical modifications in miRNAs (endogenous and exogenous) that can impact host gene expression opens up an alternative way to select new specific therapeutic targets.Hence, the aim of this review is to briefly address how RNA epitranscriptomic modifications can affect miRNA biogenesis and to summarize the existing evidence showing the connection between the (de)regulation of these processes and disease settings. In addition, we hypothesize on the potential effect certain chemical modifications could have on the potential cross-kingdom journey of dietary plant miRNAs.


Molecular detection of fungal pathogens in clinical specimens by 18S rDNA high-throughput screening in comparison to ITS PCR and culture.
The rising incidence of invasive fungal infections and the expanding spectrum of fungal pathogens makes early and accurate identification of the causative pathogen a daunting task. Diagnostics using molecular markers enable rapid identification of fungi, offer new insights into infectious disease dynamics, and open new possibilities for infectious disease control and prevention. We performed a retrospective study using clinical specimens (N = 233) from patients with suspected fungal infection previously subjected to culture and/or internal transcribed spacer (ITS) PCR. We used these specimens to evaluate a high-throughput screening method for fungal detection using automated DNA extraction (QIASymphony), fungal ribosomal small subunit (18S) rDNA RT-PCR and amplicon sequencing. Fungal sequences were compared with sequences from the curated, commercially available SmartGene IDNS database for pathogen identification. Concordance between 18S rDNA RT-PCR and culture results was 91%, and congruence between 18S rDNA RT-PCR and ITS PCR results was 94%. In addition, 18S rDNA RT-PCR and Sanger sequencing detected fungal pathogens in culture negative (N = 13) and ITS PCR negative specimens (N = 12) from patients with a clinically confirmed fungal infection. Our results support the use of the 18S rDNA RT-PCR diagnostic workflow for rapid and accurate identification of fungal pathogens in clinical specimens.


Insulin supersensitivity and normoinsulinaemic hypoglycaemia in uncontrolled type 2 diabetes mellitus: clinical usefulness of 3 h assessment in the 75 g oral glucose tolerance test.
A 60-year-old man with uncontrolled type 2 diabetes mellitus (DM) (glycated haemoglobin 11%) had the unusual symptoms of palpitations and sweating after drinking an excessive amount of soft drinks. Three-hour data in the 75-g oral glucose tolerance test (75g-OGTT) repeatedly showed normoinsulinaemic hypoglycaemia. His diabetic disorder was based on a delayed insulin secretory response to hyperglycaemia and daily excessive intake of glucose from a high caloric diet and soft drinks. However, we paradoxically observed increased insulin sensitivity evaluated by a hyperinsulinaemic-euglycaemic clamp (glucose infusion rate: 64.83 μmol/kg/min). We considered that insulin supersensitivity might be involved in the pathogenic mechanisms of his clinical normoinsulinaemic hypoglycaemia. He was successfully treated by diet and exercise therapy without any hypoglycaemic medications or insulin. Assessment after the 75g-OGTT is useful for investigating the pathogenesis of DM. Insulin supersensitivity and normoinsulinaemic hypoglycaemia might play a role in clinical manifestation and pathogenesis of type 2 DM.


Quantum gate operations using midinfrared binary shaped pulses on the rovibrational states of carbon monoxide.
Frequency domain shaped binary laser pulses were optimized to perform 2 qubit quantum gate operations in (12)C(16)O. The qubit rovibrational state representation was chosen so that all gate operations consisted of one-photon transitions. The amplitude and phase varied binary pulses were determined using a genetic algorithm optimization routine. Binary pulses have two possible amplitudes, 0 or 1, and two phases, 0 or pi, for each frequency component of the pulse. Binary pulses are the simplest to shape experimentally and provide a minimum fidelity limit for amplitude and phase shaped pulses. With the current choice of qubit representation and using optimized binary pulses, fidelities of 0.80 and as high as 0.97 were achieved for the controlled-NOT and alternative controlled-NOT quantum gates. This indicates that with a judicious choice of qubits, most of the required control can be obtained with a binary pulse. Limited control was observed for 2 qubit NOT and Hadamard gates due to the need to control multiple excitations. The current choice of qubit representation produces pulses with decreased energies and superior fidelities when compared with rovibrational qubit representations consisting of two-photon transitions. The choice of input pulse energy is important and applying pulses of increased energy does not necessarily lead to a better fidelity.


Thermodynamically Guided Synthesis of Mixed-Linker Zr-MOFs with Enhanced Tunability.
Guided by thermodynamics, we have synthesized two mixed-linker zirconium-based metal-organic frameworks (Zr-MOFs), namely, PCN-133 and PCN-134. Both of them possess a layer-pillar structure, in which the connection between Zr6 clusters and primary BTB linkers form a (3,6)-connected kdg layer that is further extended into 3D frameworks by auxiliary DCDPS/TCPP linkers (BTB = benzene tribenzoate, DCDPS = 4,4'-dicarboxydiphenyl sulfone, TCPP = tetrakis(4-carboxyphenyl)porphyrin). PCN-134 demonstrates high porosity (N2 uptake of 717 cm(3)·g(-1) and BET surface area of 1946 cm(2)·g(-1)) and excellent chemical stability in aqueous solutions with pH values ranging from 0 to 13. More importantly, PCN-134 tolerates the partial absence of auxiliary linkers leading to structural defects during the assembly process while preserving its framework integrity. Furthermore, the defect density can be systematically controlled by tuning the occupancy of the auxiliary linker, which in turn affects the MOF properties. For instance, the dichromate uptake of PCN-134 is tuned by adjusting the BTB/TCPP ratios, which gives rise to an efficient dichromate absorbent when the TCPP molar ratio in linkers is set as 22%. In addition, the photocatalytic reduction of Cr(VI) in aqueous solution was also performed by PCN-134-22%TCPP which exhibits excellent catalytic activity. This work not only opens up a new synthetic route toward mixed-linker MOFs, but also provides tunable control of MOF defects and, in turn, the properties.


CD40-mediated signal transduction in human airway smooth muscle.
CD40 is a member of the TNF receptor family that was initially described on the surface of B cells. Recently, CD40 has also been described on mesenchymal cells, such as endothelial cells and fibroblasts, where engagement by its ligand CD40 ligand can lead to up-regulation of costimulatory and cell adhesion molecules, as well as secretion of proinflammatory cytokines. Since airway inflammation potentially involves cell-cell interactions of T cells and eosinophils (which express CD40 ligand) with airway smooth muscle (ASM) cells, we postulated that ASM may express CD40 and that engagement of ASM CD40 may modulate smooth muscle cell function. We demonstrate that CD40 is expressed on cultured human ASM and that expression can be increased by treatment with TNF-alpha or IFN-gamma. Cross-linking CD40 on ASM resulted in enhanced IL-6 secretion and an increase in intracellular calcium concentrations, which were dependent on calcium influx. We show that CD40-mediated signaling events include protein tyrosine phosphorylation and activation of NF-kappaB. Pretreatment of ASM with the tyrosine kinase inhibitors genistein or herbimycin inhibited the rapid mobilization of calcium induced via CD40, suggesting that calcium mobilization was coupled to activation of protein tyrosine kinases. In addition, inhibition of calcium influx inhibited both CD40-mediated NF-kappaB activation and enhancement of IL-6 secretion. These results delineate a potentially important CD40-mediated signal-transduction pathway in ASM, involving protein tyrosine kinase-dependent calcium mobilization, NF-kappaB activation, and IL-6 production. Together, these results suggest a mechanism whereby T cell/smooth muscle cell interactions may potentiate airway inflammation.


Upregulation of miR-137 Expression Suppresses Tumor Growth and Progression via Interacting with DNMT3a Through Inhibiting the PTEN/Akt Signaling in HCC.
Downregulation of miR-137 regulates tumor growth in hepatocellular carcinoma (HCC). Yet, the underlying molecular mechanisms stay unclear. miR-137 and DNA methyltransferase 3a (DNMT3a) expression levels were detected by Western blot, immunohistochemistry and qRT-PCR assays. Luciferase reporter and Western blot assays were also carried out to explore the correlation of miR-137 and DNMT3a. Flow cytometry assay, MTT analysis, transwell and wound healing assay were used to evaluate cell apoptosis, proliferation, as well as invasive and migratory abilities. Western blot was used to examine the caspase-3, cleaved caspase-3, PCNA, MMP-2, and MMP-7 protein levels, as well as PTEN/Akt signaling alternations. Methylation-specific PCR was applied to detect the PTEN promoter methylation status. Xenograft tumor assay, Western blot and immunohistochemistry analyses were taken to confirm the miR-137 regulation in vivo. Downregulation of miR-137, upregulation of DNMT3a, as well as an inverse correlation between them were observed in HCC clinical samples and cells. Moreover, miR-137 targeted directly and inhibited DNMT3a in HCC cells, which further retarded cell proliferative, migratory and invasive capabilities, while promoted apoptotic ones. Additionally, miR-137 overexpression inactivated the PTEN/Akt pathway in HCC cell by decreasing DNMT3a expression. Furthermore, miR-137 overexpression inhibited tumor growth in vivo in HCC via interacting with DNMT3a through inhibiting the PTEN/Akt cascades. Our findings suggested that miR-137 inhibited HCC tumor growth and progression via interacting with DNMT3a and suppressing the PTEN/Akt signaling in vitro and in vivo.


Sediment delivery estimates in water quality models altered by resolution and source of topographic data.
Moderate-resolution (30-m) digital elevation models (DEMs) are normally used to estimate slope for the parameterization of non-point source, process-based water quality models. These models, such as the Soil and Water Assessment Tool (SWAT), use the Universal Soil Loss Equation (USLE) and Modified USLE to estimate sediment loss. The slope length and steepness factor, a critical parameter in USLE, significantly affects sediment loss estimates. Depending on slope range, a twofold difference in slope estimation potentially results in as little as 50% change or as much as 250% change in the LS factor and subsequent sediment estimation. Recently, the availability of much finer-resolution (∼3 m) DEMs derived from Light Detection and Ranging (LiDAR) data has increased. However, the use of these data may not always be appropriate because slope values derived from fine spatial resolution DEMs are usually significantly higher than slopes derived from coarser DEMs. This increased slope results in considerable variability in modeled sediment output. This paper addresses the implications of parameterizing models using slope values calculated from DEMs with different spatial resolutions (90, 30, 10, and 3 m) and sources. Overall, we observed over a 2.5-fold increase in slope when using a 3-m instead of a 90-m DEM, which increased modeled soil loss using the USLE calculation by 130%. Care should be taken when using LiDAR-derived DEMs to parameterize water quality models because doing so can result in significantly higher slopes, which considerably alter modeled sediment loss.


Transfer representation learning for medical image analysis.
There are two major challenges to overcome when developing a classifier to perform automatic disease diagnosis. First, the amount of labeled medical data is typically very limited, and a classifier cannot be effectively trained to attain high disease-detection accuracy. Second, medical domain knowledge is required to identify representative features in data for detecting a target disease. Most computer scientists and statisticians do not have such domain knowledge. In this work, we show that employing transfer learning can remedy both problems. We use Otitis Media (OM) to conduct our case study. Instead of using domain knowledge to extract features from labeled OM images, we construct features based on a dataset entirely OM-irrelevant. More specifically, we first learn a codebook in an unsupervised way from 15 million images collected from ImageNet. The codebook gives us what the encoders consider being the fundamental elements of those 15 million images. We then encode OM images using the codebook and obtain a weighting vector for each OM image. Using the resulting weighting vectors as the feature vectors of the OM images, we employ a traditional supervised learning algorithm to train an OM classifier. The achieved detection accuracy is 88.5% (89.63% in sensitivity and 86.9% in specificity), markedly higher than all previous attempts, which relied on domain experts to help extract features.


Large dynamic range autorefraction with a low-cost diffuser wavefront sensor.
Wavefront sensing with a thin diffuser has emerged as a potential low-cost alternative to a lenslet array for aberrometry. Here we show that displacement of caustic patterns can be tracked for estimating wavefront gradient in a diffuser wavefront sensor (DWFS), enabling large dynamic-range wavefront measurements with sufficient accuracy for eyeglass prescription measurements. We compare the dynamic range, repeatability, precision, and number of resolvable prescriptions of a DWFS to a Shack-Hartmann wavefront sensor (SHWFS) for autorefraction measurement. We induce spherical and cylindrical errors in a model eye and use a multi-level Demon's non-rigid registration algorithm to estimate caustic displacements relative to an emmetropic model eye. When compared to spherical error measurements with the SHWFS using a laser diode with a laser speckle reducer, the DWFS demonstrates a ∼5-fold improvement in dynamic range (-4.0 to +4.5 D vs. -22.0 to +19.5 D) with less than half the reduction in resolution (0.072 vs. 0.116 D), enabling a ∼3-fold increase in the number of resolvable prescriptions (118 vs. 358). In addition to being lower-cost, the unique, non-periodic nature of the caustic pattern formed by a diffuser enables a larger dynamic range of aberration measurements compared to a lenslet array.


Case report: Two sisters with light-chain cardiac amyloidosis, a mere coincidence?
Light-chain amyloidosis has always been described as a sporadic disease caused by plasma cell dyscrasia. Cardiac amyloidosis refers to cardiac involvement with infiltration of amyloid fibrils in the myocardium. The degree of cardiac involvement is the greatest predictor of prognosis. To our knowledge, AL cardiac amyloidosis has only been reported once before in first-degree relatives. In this report, we describe the unusual cases of two sisters with light-chain cardiac amyloidosis. The first patient underwent autologous stem cell transplantation and remained in remission for 10 years until the disease relapsed and she died of end-stage heart failure. The second patient was promptly started on a chemotherapy regimen but died shortly after her initial diagnosis due to rapid progression of cardiac dysfunction. Cardiac amyloidosis is a severe life-threatening condition which requires a multidisciplinary diagnostic and therapeutic approach. Based on this case report, a genetic cause for AL amyloidosis might be suspected or is this a purely coincidental finding? Counselling, screening, and follow-up of other family members are very challenging. As is often the case with rare diseases, many unsolved questions remain, representing important challenges for clinicians.


Contemporary Factors Associated with the Use of Laparoscopy for Inguinal Hernia Repair Among Department of Defense Beneficiaries.
The factors that contribute to variation in utilization of laparoscopic inguinal hernia repair are unknown. We sought to determine the current usage patterns of laparoscopic and open surgery in the elective repair of uncomplicated unilateral inguinal hernia in a large population with universal health care coverage comprised of Department of Defense (DoD) beneficiaries. The DoD Military Health System Data Repository (MDR) tracks health care delivered to a universally insured population of active/reserve/retired members of the U.S. Armed Services and their dependents. The MDR was queried for elective unilateral inguinal hernia repair among adult patients between 2008 and 2014. The primary outcome was laparoscopic (vs. open) approach to hernia repair. We conducted univariable and multivariable analyses of patient- and systems-level factors associated with approach to inguinal hernia repair. This research was approved by our institutional review board prior to commencement of the study and need for informed consent was waived given the design of this study. Among 37,742 elective uncomplicated unilateral inguinal hernia repairs, 35% (n = 13,114) were performed laparoscopically. In 2014, 40% of inguinal hernia repairs were performed laparoscopically, compared with 27% of repairs in 2008 (P < 0.01). In multivariable analysis, laparoscopic hernia repair was more likely for male patients (OR = 1.38, 95% CI = 1.23-1.54, P < 0.01), military (vs. civilian) institutions (OR = 1.34, 95% CI = 1.28-1.41, P < 0.01), active-duty officers (vs. active-duty enlisted; OR = 1.21, 95% CI = 1.12-1.30, P < 0.01), and more recent year of surgery (P < 0.01). Laparoscopic repair was significantly less likely among patients with greater than one comorbidity (vs. none; OR = 0.68, 95% CI = 0.61-0.76, P < 0.01). In a large, universally insured population of military service members and their dependents, laparoscopic inguinal repair is increasingly used and was preferred over open repair for younger, healthier, active-duty patients and those treated within the military (vs. non-military) care system.


Determination of tetracycline antibiotics in cow's milk by liquid chromatography/tandem mass spectrometry.
A liquid chromatographic/tandem mass spectrometric (LC/MS/MS) multiresidue method for the simultaneous quantitative determination of oxytetracycline, 4-epi-oxytetracycline, tetracycline, 4-epi-tetracycline, chlortetracycline, 4-epi-chlortetracycline and doxycycline in milk has been developed. An extraction procedure consisting of a liquid extraction of the milk samples with trichloroacetic acid was performed. The extract was centrifuged and the supernatant was filtered. Solid-phase extraction (SPE) with an OASIS HLB SPE column was used to clean up the sample extracts. The samples were analysed by LC/MS/MS. The LC separation was performed on a reversed-phase C18 column using gradient elution with a mobile phase consisting of water and a mixture of methanol/acetonitrile. The tetracycline analytes were detected with a quadrupole mass spectrometer using positive ion electrospray ionisation. The confirmatory method has acceptable detection limits and the different tetracyclines can be detected at a residue concentration between 5 and 20 microg/L. The method is validated according to the European requirements for veterinary drug residues and all determined parameters were found to conform to the criteria. The recovery values ranged from 90.4 to 101.2% with relative standard deviations (RSDs) no larger than 9.7%. The overall or between-day precision of the analytical assay determined as repeatability at several residue concentrations and expressed as RSD ranged from 3.3 to 10%. This analytical assay is a useful tool within the Belgian monitoring programme for confirmation of samples which have been positively screened for residues of tetracyclines in raw farm cow's milk.


Enterolosaponins A and B, novel triterpene bisdesmosides from Enterolobium contortisiliquum, and evaluation for their macrophage-oriented cytotoxic activity.
Two novel triterpene bisdesmosides, designated as enterolosaponin A (1) and B (2), were isolated from Enterolobium contortisiliquum. The chemical structures of 1 and 2 were determined by analysis of their extensive spectroscopic data, as well as hydrolysis followed by chromatographic study. Enterolosaponins have a 2-amino-2-deoxy-D-glucosyl unit (D-glucosamine) as one of the monosaccharides constituting their oligosaccharide moieties, which have been rarely found in natural product research. Enterolosaponin A (1) exhibited a highly selective cytotoxicity against BAC1.2F5 mouse macrophages, and it should be notable that the macrophage death caused by 1 was shown to be neither necrotic nor due to induction of apoptosis from morphology of the died cells, whose cytosol occurred in vacuolation.


Postprandial lipoprotein, glucose and insulin responses after two consecutive meals containing rapeseed oil, sunflower oil or palm oil with or without glucose at the first meal.
There is increasing evidence that the degree of postprandial lipaemia may be of importance in the development of atherosclerosis and IHD. Postprandial lipid, lipoprotein, glucose, insulin and non-esterified fatty acid (NEFA) concentrations were investigated in eleven healthy young males after randomized ingestion of meals containing rapeseed oil, sunflower oil or palm oil with or without a glucose drink. On six occasions each subject consumed consecutive meals (separated by 1.75 h) containing 70 g (15 g and 55 g respectively) of each oil. On one occasion with each oil 50 g glucose was taken with the first meal. One fasting and fifteen postprandial blood samples were taken over 9 h. There were no statistically significant differences in lipoprotein and apolipoprotein responses after rapeseed, sunflower and palm oils, whereas insulin responses were lower after sunflower oil than after rapeseed oil (ANOVA, P = 0.04). The NEFA and triacylglycerol concentrations at 1.5 h were reduced when 50 g glucose was taken with the first meal (ANOVA, P < 0.0001 and P < 0.05 respectively), regardless of meal fatty acid composition. In conclusion, the consumption of glucose with a mixed meal containing either rapeseed, sunflower or palm oil influenced the immediate triacylglycerol and NEFA responses compared with the same meal without glucose, whereas no significant effect on postprandial lipaemia after a subsequent meal was observed. The fatty acid composition of the meal did not significantly affect the lipid and lipoprotein responses, whereas an effect on insulin responses was observed.


Medication characteristics beyond cost alone influence decisions to underuse pharmacotherapy in response to financial pressures.
Little is known about how patients facing medication cost pressures make choices about whether to underuse one or more of their prescription drugs. We calculated the probability that older adults would underuse prescription medications for common chronic illnesses because of cost concerns. We also identified differences in cost-related underuse between symptom-relief medications (e.g., analgesics) and primarily "preventive" medications (e.g., antihypertensives). Older chronically ill patients using both symptom-relief and preventive medications (N = 2,008) were identified as part of a nationwide survey in the United States and reported information about their cost-related underuse of 16 medication types. We used regression models to estimate the probability of underuse for each medication type, assuming average out-of-pocket costs, no prescription coverage, and the sociodemographic characteristics of a typical American aged 50+. 23% of respondents reported forgoing medication in the prior year due to cost. The likelihood of cost-related underuse was higher for symptom-relief medications (27%) than for primarily preventive medications (20%, P < .001). Among the subset of patients who cut back on adherence due to cost, the likelihood of forgoing symptom-relief medication (69%) was higher than that for preventive drugs (54%, P < .001). Medication characteristics beyond cost alone influence decisions to underuse treatment in response to financial pressures.


Untargeted plasma metabolite profiling reveals the broad systemic consequences of xanthine oxidoreductase inactivation in mice.
A major challenge in systems biology is integration of molecular findings for individual enzyme activities into a cohesive high-level understanding of cellular metabolism and physiology/pathophysiology. However, meaningful prediction for how a perturbed enzyme activity will globally impact metabolism in a cell, tissue or intact organisms is precluded by multiple unknowns, including in vivo enzymatic rates, subcellular distribution and pathway interactions. To address this challenge, metabolomics offers the potential to simultaneously survey changes in thousands of structurally diverse metabolites within complex biological matrices. The present study assessed the capability of untargeted plasma metabolite profiling to discover systemic changes arising from inactivation of xanthine oxidoreductase (XOR), an enzyme that catalyzes the final steps in purine degradation. Using LC-MS coupled with a multivariate statistical data analysis platform, we confidently surveyed >3,700 plasma metabolites (50-1,000 Da) for differential expression in XOR wildtype vs. mice with inactivated XOR, arising from gene deletion or pharmacological inhibition. Results confirmed the predicted derangements in purine metabolism, but also revealed unanticipated perturbations in metabolism of pyrimidines, nicotinamides, tryptophan, phospholipids, Krebs and urea cycles, and revealed kidney dysfunction biomarkers. Histochemical studies confirmed and characterized kidney failure in xor-nullizygous mice. These findings provide new insight into XOR functions and demonstrate the power of untargeted metabolite profiling for systemic discovery of direct and indirect consequences of gene mutations and drug treatments.


Strategic choice handicaps when females seek high male net viability.
We examine a strategic-choice handicap model in which males send costly signals to advertise their quality to females. Females are concerned with the net viability of the male with whom they mate, where net viability is a function of the male's quality and signal. We identify circumstances in which a signaling equilibrium would require high-quality males to send signals so much larger than those of lower-quality males (to deter mimicry by the latter) as to yield lower net viabilities for the former. This causes females to shun males who send large signals, ensuring that there is no signaling equilibrium.


5-Hydroxymethyl-Furfural and Structurally Related Compounds Block the Ion Conductance in Human Aquaporin-1 Channels and Slow Cancer Cell Migration and Invasion.
Aquaporin-1 (AQP1) dual water and ion channels enhance migration and invasion when upregulated in leading edges of certain classes of cancer cells. Work here identifies structurally related furan compounds as novel inhibitors of AQP1 ion channels. 5-Hydroxymethyl-2-furfural (5HMF), a component of natural medicinal honeys, and three structurally related compounds, 5-nitro-2-furoic acid (5NFA), 5-acetoxymethyl-2-furaldehyde (5AMF), and methyl-5-nitro-2-furoate (M5NF), were analyzed for effects on water and ion channel activities of human AQP1 channels expressed in Xenopus oocytes. Two-electrode voltage clamp showed dose-dependent block of the AQP1 ion current by 5HMF (IC50 0.43 mM), 5NFA (IC50 1.2 mM), and 5AMF (IC50 ∼3 mM) but no inhibition by M5NF. In silico docking predicted the active ligands interacted with glycine 165, located in loop D gating domains surrounding the intracellular vestibule of the tetrameric central pore. Water fluxes through separate intrasubunit pores were unaltered by the furan compounds (at concentrations up to 5 mM). Effects on cell migration, invasion, and cytoskeletal organization in vitro were tested in high-AQP1-expressing cancer lines, colon cancer (HT29) and AQP1-expressing breast cancer (MDA), and low-AQP1-expressing SW480. 5HMF, 5NFA, and 5AMF selectively impaired cell motility in the AQP1-enriched cell lines. In contrast, M5NF immobilized all the cancer lines by disrupting actin cytoskeleton. No reduction in cell viability was observed at doses that were effective in blocking motility. These results define furans as a new class of AQP1 ion channel inhibitors for basic research and potential lead compounds for development of therapeutic agents targeting aquaporin channel activity. SIGNIFICANCE STATEMENT: 5-Hydroxymethyl-2-furfural (5HMF), a component of natural medicinal honeys, blocks the ion conductance but not the water flux through human Aquaporin-1 (AQP1) channels and impairs AQP1-dependent cell migration and invasiveness in cancer cell lines. Analyses of 5HMT and structural analogs demonstrate a structure-activity relationship for furan compounds, supported by in silico docking modeling. This work identifies new low-cost pharmacological antagonists for AQP1 available to researchers internationally. Furans merit consideration as a new class of therapeutic agents for controlling cancer metastasis.