Characteristics and Outcomes Reported by Patients With Primary Sclerosing Cholangitis Through an Online Registry.
Primary sclerosing cholangitis (PSC) is a fibrostenosing disease of the bile ducts associated with inflammatory bowel disease (IBD), for which the only treatment is liver transplantation. PSC progression has been defined in cohorts from referral centers or single-nation population databases. However, observations made from these cohorts have limited applicability owing to referral bias and demographic confounders. We analyzed data from the worldwide PSC Partners Patient Registry, an international online database established in 2014 to obtain information from individuals with PSC or their caretakers and compare symptoms, disease progression, and treatments of PSC in the United States and other countries. We analyzed demographic and clinical characteristics, symptoms, and clinical outcomes of patients with PSC using the PSC Partners Patient Registry. Participants completed an online standardized questionnaire and electronic case report, providing information on age, age at symptom onset, age at PSC diagnosis, methods of diagnosis, concurrent diagnoses, family history, and medication use. Of 873 registrants, 811 (92.9%) had completed questionnaires and 528 (65.1%) had their PSC diagnosis confirmed; we found no significant demographic or clinical differences between patients with vs without a confirmed diagnosis. In contrast to other studies, we found a higher proportion of individuals with PSC to be female (52.5%). However, the mean age at PSC diagnosis (32.4 ± 14.7 y) and the proportion of individuals with PSC and IBD (67.1%) were similar to those from prior reports. Most cases in the database were from the United States (74.9%). More than half of the participants reported having pruritus, abdominal pain, fatigue, or sleep disturbances; rates were not significantly different among participants within vs outside the United States. There was no significant difference in treatment with ursodeoxycholic acid between participants within vs outside the United States (50.0% and 57.8%; P = .07). The median time of transplant-free survival was 21 years; transplant-free survival was associated with female sex and Crohn's disease. Our findings from an analysis of data from the PSC Partners Patient Registry confirm those from previous studies, although we found a higher proportion of individuals with PSC to be female. In addition to allowing efficient collection of patient-reported outcomes, the patient-driven registry allows for inclusion of previously under-represented cases of PSC.


Standing at work and varicose veins.
This study attempts to determine whether or not prolonged standing at work involves an excess risk for the occurrence of varicose veins. A cohort of 1.6 million 20-to-59-year-old Danes gainfully employed in 1991 were followed for 3 years according to first hospitalization due to varicose veins of the lower extremities. The exposure data came from a representative sample of the baseline population. Altogether 5940 people were interviewed about occupational exposure and confounding factors. For men working mostly in a standing position, the risk ratio for varicose veins was 1.85 [95% confidence interval (95% CI) 1.33-2.36] in a comparison with all other men. The corresponding risk ratio for women was 2.63 (95% CI 2.25-3.02). The results were adjusted for age, social group, and smoking. Working in a standing position is associated with subsequent hospitalization due to varicose veins for both men and women.


Global, regional, and national burden of low back pain, 1990-2020, its attributable risk factors, and projections to 2050: a systematic analysis of the Global Burden of Disease Study 2021.
Low back pain is highly prevalent and the main cause of years lived with disability (YLDs). We present the most up-to-date global, regional, and national data on prevalence and YLDs for low back pain from the Global Burden of Diseases, Injuries, and Risk Factors Study 2021. Population-based studies from 1980 to 2019 identified in a systematic review, international surveys, US medical claims data, and dataset contributions by collaborators were used to estimate the prevalence and YLDs for low back pain from 1990 to 2020, for 204 countries and territories. Low back pain was defined as pain between the 12th ribs and the gluteal folds that lasted a day or more; input data using alternative definitions were adjusted in a network meta-regression analysis. Nested Bayesian meta-regression models were used to estimate prevalence and YLDs by age, sex, year, and location. Prevalence was projected to 2050 by running a regression on prevalence rates using Socio-demographic Index as a predictor, then multiplying them by projected population estimates. In 2020, low back pain affected 619 million (95% uncertainty interval 554-694) people globally, with a projection of 843 million (759-933) prevalent cases by 2050. In 2020, the global age-standardised rate of YLDs was 832 per 100 000 (578-1070). Between 1990 and 2020, age-standardised rates of prevalence and YLDs decreased by 10·4% (10·9-10·0) and 10·5% (11·1-10·0), respectively. A total of 38·8% (28·7-47·0) of YLDs were attributed to occupational factors, smoking, and high BMI. Low back pain remains the leading cause of YLDs globally, and in 2020, there were more than half a billion prevalent cases of low back pain worldwide. While age-standardised rates have decreased modestly over the past three decades, it is projected that globally in 2050, more than 800 million people will have low back pain. Challenges persist in obtaining primary country-level data on low back pain, and there is an urgent need for more high-quality, primary, country-level data on both prevalence and severity distributions to improve accuracy and monitor change. Bill and Melinda Gates Foundation.


Risk factors, complications and survival after upper abdominal surgery: a prospective cohort study.
Preoperative weight loss and abnormal serum-albumin have traditionally been associated with reduced survival. More recently, a correlation between postoperative complications and reduced long-term survival has been reported and the significance of the relative proportion of skeletal muscle, visceral and subcutaneous adipose tissue has been examined with conflicting results. We investigated how preoperative body composition and major non-fatal complications related to overall survival and compared this to established predictors in a large cohort undergoing upper abdominal surgery. From 2001 to 2006, 447 patients were included in a Norwegian multicenter randomized controlled trial in major upper abdominal surgery. Patients were now, six years later, analyzed as a single prospective cohort and overall survival was retrieved from the National Population Registry. Body composition indices were calculated from CT images taken within three months preoperatively. Preoperative serum-albumin <35 g/l (HR = 1.52, p = 0 .014) and weight loss >5 % (HR = 1.38, p = 0.023) were independently associated with reduced survival. There was no association between any of the preoperative body composition indices and reduced survival. Major postoperative complications were independently associated with reduced survival but only as long as patients who died within 90 days were included in the analysis. Our study has confirmed the robust significance of the traditional indicators, preoperative serum-albumin and weight loss. The body composition indices did not prove beneficial as global indicators of poor prognosis in upper abdominal surgery. We found no association between non-fatal postoperative complications and long-term survival.


Inflammation-associated gene expression in RAW 264.7 macrophages induced by toxins from fungi common on damp building materials.
Most fungi that grow on damp building materials produce low molecular weight compounds, some of which are known to be toxic. In this study, we tested the hypothesis that exposure to some metabolites of fungi common on damp building materials would result in time-, dose-, and compound-specific responses in the production of various chemokines by RAW 264.7 cells. Cell cultures were exposed to a 10-7M or 10-8M metabolite dose for 2, 4, 8 or 24h. Metabolite concentrations used were based on those that might be expected in alveolar macrophages due to inhalation exposure from living or working in a damp building. Compared to controls, exposure provoked significant time-, dose- and compound-specific responses manifest as differentially elevated secretion of three of nine cytokines tested in culture supernatant of treated cells. The greatest number of cytokines produced in response to the metabolites tested were in andrastin A-treated cells (GM-CSF, TGFβ1, Tnf-α) followed by koninginin A (TGFβ1 and Tnf-α) and phomenone (GM-CSF, TGFβ1). Chaetoglobosin A, chaetomugilin D and walleminone exposures each resulted in significant time-specific production of Tnf-α only. This investigation adds to a body of evidence supporting the role of low molecular weight compounds from damp building materials as pathogen associated molecular patterns (PAMPs). Along with fungal glucan and chitin, these compounds contribute to the non-allergy based respiratory outcomes for people living and working in damp buildings.


Effects of Air Pollution on Lung Innate Lymphoid Cells: Review of In Vitro and In Vivo Experimental Studies.
Outdoor air pollution is associated with respiratory infections and allergies, yet the role of innate lymphoid cells (ILCs) in pathogen containment and airway hyperresponsiveness relevant to effects of air pollutants on ILCs is poorly understood. We conducted a systematic review to evaluate the available evidence on the effect of outdoor air pollutants on the lung type 1 (ILC1) and type 2 ILCs (ILC2) subsets. We searched five electronic databases (up to Dec 2018) for studies on the effect of carbon monoxide (CO), sulfur dioxide (SO2), nitrogen dioxide (NO2), diesel exhaust particles (DEP), ozone (O3), and particulate matter (PM) on respiratory ILCs. Of 2209 identified citations, 22 full-text papers were assessed for eligibility, and 12 articles describing experimental studies performed in murine strains (9) and on human blood cells (3) were finally selected. Overall, these studies showed that exposure to PM, DEP, and high doses of O3 resulted in a reduction of interferon gamma (IFN-γ) production and cytotoxicity of ILC1. These pollutants and carbon nanotubes stimulate lung ILC2s, produce high levels of interleukin (IL)-5 and IL-13, and induce airway hyperresponsiveness. These findings highlight potential mechanisms by which human ILCs react to air pollution that increase the susceptibility to infections and allergies.


Resection of pediatric intracerebral tumors with the aid of intraoperative real-time 3-D ultrasound.
Intraoperative ultrasound (IOUS) has become a useful tool employed daily in neurosurgical procedures. In pediatric patients, IOUS offers a radiation-free and safe imaging method. This study aimed to evaluate the use of a new real-time 3-D IOUS technique (RT-3-D IOUS) in our pediatric patient cohort. Over 24 months, RT-3-D IOUS was performed in 22 pediatric patients (8 girls and 14 boys) with various brain tumors. These lesions were localized by a standard navigation system followed by analyses before, intermittently during, and after neurosurgical resection using the iU22 ultrasound system (Philips, Bothell, USA) connected to the RT-3-D probe (X7-2). In all 22 patients, real-time 3-D ultrasound images of the lesions could be obtained during neurosurgical resection. Based on this imaging method, rapid orientation in the surgical field and the approach for the resection could be planned for all patients. In 18 patients (82%), RT-3-D IOUS revealed a gross total resection with a favorable neurological outcome. RT-3-D IOUS provides the surgeon with advanced orientation at the tumor site via immediate live two-plane imaging. However, navigation systems have yet to be combined with RT-3-D IOUS. This combination would further improve intraoperative localization.


Clinical perspectives in congenital adrenal hyperplasia due to 3β-hydroxysteroid dehydrogenase type 2 deficiency.
3β-hydroxysteroid dehydrogenase type 2 deficiency (3βHSD2D) is a very rare variant of congenital adrenal hyperplasia (CAH) causing less than 0.5% of all CAH. The aim was to review the literature. PubMed was searched for relevant articles. 3βHSD2D is caused by HSD3B2 gene mutations and characterized by impaired steroid synthesis in the gonads and the adrenal glands and subsequent increased dehydroepiandrosterone (DHEA) concentrations. The main hormonal changes observed in patients with 3βHSD2D are elevated ratios of the Δ5-steroids over Δ4-steroids but molecular genetic testing is recommended to confirm the diagnosis. Several deleterious mutations in the HSD3B2 gene have been associated with salt-wasting (SW) crisis in the neonatal period, while missense mutations have been associated with a non-SW phenotype. Boys may have ambiguous genitalia, whereas girls present with mild or no virilization at birth. The existence of non-classic 3βHSD2D is controversial. In an acute SW crisis, the treatment includes prompt rehydration, correction of hypoglycemia, and parenteral hydrocortisone. Similar to other forms of CAH, glucocorticoid and mineralocorticoid replacement is needed for long-term management. In addition, sex hormone replacement therapy may be required if normal progress through puberty is failing. Little is known regarding possible negative long-term consequences of 3βHSD2D and its treatments, e.g., fertility, final height, osteoporosis and fractures, adrenal and testicular tumor risk, and mortality. Knowledge is mainly based on case reports but many long-term outcomes could be presumed to be similar to other types of CAH, mainly 21-hydroxylase deficiency, although in 3βHSD2D it seems to be more difficult to suppress the androgens.


Neuroinflammatory responses in diabetic retinopathy.
Diabetic retinopathy (DR) is a common complication of diabetes and has been recognized as a vascular dysfunction leading to blindness in working-age adults. It becomes increasingly clear that neural cells in retina play an important role in the pathogenesis of DR. Neural retina located at the back of the eye is part of the brain and a representative of the central nervous system. The neurosensory deficits seen in DR are related to inflammation and occur prior to the clinically identifiable vascular complications. The neural deficits are associated with abnormal reactions of retina glial cells and neurons in response to hyperglycemia. Improper activation of the innate immune system may also be an important contributor to the pathophysiology of DR. Therefore, DR manifests characteristics of both vasculopathy and chronic neuroinflammatory diseases. In this article, we attempt to provide an overview of the current understanding of inflammation in neural retina abnormalities in diabetes. Inhibition of neuroinflammation may represent a novel therapeutic strategy to the prevention of the progression of DR.


Ciprofloxacin inhibition of experimental fracture healing.
Fluoroquinolones, such as ciprofloxacin, have an adverse effect on growing cartilage and endochondral ossification in children. This study was carried out to determine whether ciprofloxacin also has an adverse effect on the healing of experimental fractures. Sixty male 300-gram Wistar rats were divided equally into three groups, which received ciprofloxacin, cefazolin, or no treatment for three weeks, beginning seven days after production of a closed, nondisplaced, bilateral femoral fracture. The serum concentrations of the ciprofloxacin and the cefazolin were 2.4 and 146 micrograms per milliliter, respectively. Radiographic, histological, and biomechanical studies were used to evaluate fracture-healing. Radiographs revealed significantly more advanced healing of the control fractures compared with the fractures in the ciprofloxacin-treated group (average stage, 2.1 compared with 1.5, p = 0.01). The cefazolin-treated group was not different from the controls with respect to radiographic healing (average stage, 1.8 compared with 2.1, p = 0.18). Torsional strength-testing of fracture callus exposed to ciprofloxacin revealed a 16 percent decrease in strength compared with the controls (284 compared with 338 newton-millimeters, p = 0.04) and a 49 percent decrease in stiffness (twenty compared with thirty-nine newton-millimeters per degree, p = 0.001). The biomechanical strength in the cefazolin-treated group was not different from that of the controls. Fracture calluses in the animals treated with ciprofloxacin showed abnormalities in cartilage morphology and endochondral bone formation and a significant decrease in the number of chondrocytes compared with the controls (0.77 x 10(4) compared with 1.3 x 10(4) cells per square millimeter, p = 0.004). These data suggest that experimental fractures exposed to therapeutic concentrations of ciprofloxacin in serum demonstrate diminished healing during the early stages of fracture repair. The administration of ciprofloxacin during early fracture repair may compromise the clinical course of fracture-healing.


Insight in frontotemporal dementia: conceptual analysis and empirical evaluation of the consensus criterion "loss of insight" in frontotemporal dementia.
The objective of this study was to suggest a new formulation of the core research diagnostic consensus criterion "loss of insight" in frontotemporal dementia (FTD). Eight patients with FTD (diagnoses made by interviews, medical and neuropsychological examination, CT scan, and regional cerebral glucose metabolism measured by positron emission tomography (PET) participated in the study). The results indicated that insight was present in three out of eight patients, and that insight appears to be a heterogeneous concept. Two types of insight emerged: Emotional insight associated with frontotemporal functions, and cognitive insight, related to posterior cognitive functions. These results suggest that loss of insight should not serve as a core criterion on FTD, but serves well as a supportive criterion of the disease.


Alterations of INPP4B, PIK3CA and pAkt of the PI3K pathway are associated with squamous cell carcinoma of the lung.
The aim of the study was to investigate how alterations in the PI3K pathway correlate with non-small cell lung cancer subtypes squamous cell carcinoma (SSC) and adenocarcinoma (ADCA). We analyzed copy number variation and protein expression of INPP4B, protein expression of pAkt, PDPK1, and PTEN and mutational status of PIK3CA and PTEN in 180 cases. Nineteen% displayed loss of INPP4B copy, whereas 47% lacked expression, both showing correlation with SCC. Elevated pAkt expression was seen in 63% of all cases, also correlating to SCC. PDPK1 was expressed in 70%, more in male than female patients. Regarding PTEN, 50% displayed loss of expression, of which seven were identified with mutations in the phosphatase domain. We detected nine cases (5%) of PIK3CA mutations, all identified as the E545K hot spot mutation in the helical domain, all except one in SCC. When analyzing all PI3K pathway components together, we show that patients with at least one alteration in the PI3K pathway are twice as likely to have SCC, than ADCA. Interestingly, we also found a strong correlation between high pAkt expression and PTEN expression. As comparison, we also analyzed mitogen-activated protein kinase (MAPK) pathway genes, where we identified fifteen KRAS mutations (8%) and one BRAF mutation (1%), significantly associated to ADCA. No association was found to the Gly972Arg polymorphism of IRS-1, involved in activation of both PI3K and MAPK pathways. In conclusion, we show here that several components of the PI3K pathway, alone and in combination, are correlated to development of SCC of the lung.


Boundary-induced segregation in nanoscale thin films of athermal polymer blends.
The surface segregation of binary athermal polymer blends confined in a nanoscale thin film was investigated by dissipative particle dynamics. The polymer blend included linear/linear, star/linear, bottlebrush/linear, and rod-like/linear polymer systems. The segregation was driven by purely entropic effects and two different mechanisms were found. For the linear/linear and star/linear polymer blends, the smaller sized polymers were preferentially segregated to the boundary because their excluded volumes were smaller than those of the matrix polymers. For the bottlebrush/linear and rod-like/linear polymer blends, the polymers with a larger persistent length were preferentially segregated to the boundary because they favored staying in the depletion zone by alignment with the wall. Our simulation outcome was consistent with experimental results and also agreed with theoretical predictions - that is, a surface excess dictated by the chain ends for the branch/linear system. These consequences are of great importance in controlling the homogeneity and surface properties of polymer blend thin films.


A qualitative study of health professions curricula and management of lateral ankle ligament sprain demonstrates inconsistency.
Health educators aim to graduate students who are safe, effective and practice evidence-based medicine (EBM). Clinical Practice Guidelines (CPGs) are tools for translating evidence into clinical practice for health professionals and educators who lack time to appraise the evidence. There have been CPGs published for lateral ankle ligament sprains (LALS) for physiotherapists, nurses, and doctors. Clinical decision rules have also been developed for LALS to increase the safety of practice. The Ottawa Ankle Rules (OAR) were developed to screen for the need for an x-ray following an ankle or foot injury. Educators from the Australasian College of Sports and Exercise Physicians (ACSEP), St John Ambulance first aiders, pharmacy, nursing, and physiotherapy disciplines were participants in this study. Using purposeful sampling with semi-structured questions and a LALS case study, 19 Australian educators were interviewed. Curricula and textbooks were also collected and analysed. Two researchers independently analysed the data using a deductive method. Analysis found that no educator used a CPG to inform their teaching. There was no common LALS curriculum for the five groups studied. There were two approaches: a triage curriculum (St John Ambulance, pharmacy, nursing) and a reflective curriculum (ASCEP and physiotherapy). Textbooks influenced curriculum for physiotherapy, pharmacy and first aid educators. The triage curricula recommend rest, ice, compression and elevation (RICE) alone, while the reflective curricula uses OAR, RICE, immobilisation if the LALS is severe, functional support (brace), exercises and manual therapy. In addition, ACSEP and physiotherapy do not recommend electrotherapy. All five groups were cautious about the use of non-steroidal anti-inflammatory drugs (NSAIDs). Physiotherapy and ACSEP educators teach OAR. Despite not using the CPGs to inform curriculum, physiotherapy and ACSEP have unintentionally aligned their curriculum with current LALS CPG recommendations. However, nursing, pharmacy and first aid trainers are not teaching OAR or aligned with LALS CPGs. Educators in pharmacy, nursing and first aid should re-examine their curricula and consider possibly teaching OAR and using CPG. Clinical practice guideline developers should consider pharmacists and first aiders as users of their LALS CPGs.


Trigger point dry needling for the treatment of myofascial pain syndrome: current perspectives within a pain neuroscience paradigm.
Myofascial pain syndrome is a pain condition characterized by the presence of trigger points. Current evidence, mostly experimental studies, clearly supports a role of trigger points on peripheral and central sensitization since they are able to contribute to sensitization of peripheral nociceptors, spinal dorsal horn neurons, and the brainstem. Several interventions are proposed for treating trigger points, dry needling being one of the most commonly used by clinicians. There is no consensus on the clinical application of trigger point dry needling: some authors propose that local twitch responses should be elicited during the needling intervention to be effective, whereas others do not. The application of trigger point dry needling is able to reduce the excitability of the central nervous system by reducing peripheral nociception associated to the trigger point, by reducing dorsal horn neuron activity, and by modulating pain-related brainstem areas. However, the effects are mainly observed in the short-term, and effect sizes are moderateto small. Therefore, the current review proposes that the application of trigger point dry needling should be integrated into current pain neuroscience paradigm by combining its application with pain neuroscience education, graded exercise and manual therapy. Additionally, patient's expectations, beliefs, previous experiences and patient-clinician interaction should be considered when integrating trigger point dry needling into a comprehensive treatment approach.


Chemokine production by G protein-coupled receptor activation in a human mast cell line: roles of extracellular signal-regulated kinase and NFAT.
Chemoattractants are thought to be the first mediators generated at sites of bacterial infection. We hypothesized that signaling through G protein-coupled chemoattractant receptors may stimulate cytokine production. To test this hypothesis, a human mast cell line (HMC-1) that normally expresses receptors for complement components C3a and C5a at low levels was stably transfected to express physiologic levels of fMLP receptors. We found that fMLP, but not C3a or C5a, induced macrophage inflammatory protein (MIP)-1ss (CCL4) and monocyte chemoattractant protein-1 (CCL2) mRNA and protein. Although fMLP stimulated both sustained Ca(2+) mobilization and phosphorylation of extracellular signal-regulated kinase (ERK), these responses to C3a or C5a were transient. However, transient expression of C3a receptors in HMC-1 cells rendered the cells responsive to C3a for sustained Ca(2+) mobilization and MIP-1ss production. The fMLP-induced chemokine production was blocked by pertussis toxin, PD98059, and cyclosporin A, which respectively inhibit G(i)alpha activation, mitgen-activated protein kinase kinase-mediated ERK phosphorylation, and calcineurin-mediated activation of NFAT. Furthermore, fMLP, but not C5a, stimulated NFAT activation in HMC-1 cells. These data indicate that chemoattractant receptors induce chemokine production in HMC-1 cells with a selectivity that depends on the level of receptor expression, the length of their signaling time, and the synergistic interaction of multiple signaling pathways, including extracellular signal-regulated kinase phosphorylation, sustained Ca(2+) mobilization and NFAT activation.


Improved robustness of target location in nonhomogeneous backgrounds by use of the maximum-likelihood ratio test location algorithm.
We address the problem of localizing small targets with random gray levels that appear in random background clutter. We consider the recently proposed maximum-likelihood ratio test (MLRT) algorithm, which scans the observed scene with an estimation window in which the local statistics are estimated. In the presence of a spatially homogeneous background, we show that if the estimation window is a few times larger than the target itself, the MLRT is quasi-equivalent to the optimal maximum-likelihood (ML) algorithm, which uses the whole scene for estimating the background statistics. The MLRT thus constitutes an efficient alternative to the ML algorithm and is more robust in dealing with spatially nonhomogeneous clutter since it utilizes a small estimation window.


Adaption of the ex vivo mycobacterial growth inhibition assay for use with murine lung cells.
In the absence of a correlate(s) of protection against human tuberculosis and a validated animal model of the disease, tools to facilitate vaccine development must be identified. We present an optimised ex vivo mycobacterial growth inhibition assay (MGIA) to assess the ability of host cells within the lung to inhibit mycobacterial growth, including Bacille Calmette-Guérin (BCG) and Mycobacterium tuberculosis (MTB) Erdman. Growth of BCG was reduced by 0.39, 0.96 and 0.73 log10 CFU following subcutaneous (s.c.) BCG, intranasal (i.n.) BCG, or BCG s.c. + mucosal boost, respectively, versus naïve mice. Comparatively, a 0.49 (s.c.), 0.60 (i.n.) and 0.81 (s.c. + mucosal boost) log10 reduction in MTB CFU was found. A BCG growth inhibitor, 2-thiophenecarboxylic acid hydrazide (TCH), was used to prevent quantification of residual BCG from i.n. immunisation and allow accurate MTB quantification. Using TCH, a further 0.58 log10 reduction in MTB CFU was revealed in the i.n. group. In combination with existing methods, the ex vivo lung MGIA may represent an important tool for analysis of vaccine efficacy and the immune mechanisms associated with vaccination in the organ primarily affected by MTB disease.


Epidemiologic study of cats and dogs affected by the 1991 Oakland fire.
To identify risk factors of pets affected by the 1991 Oakland, Calif fire for being lost, found, adopted, or reunited with owners. Retrospective cohort study. 1,075 cats and 197 dogs affected by the fire and 221 cats and 128 dogs not affected by the fire. Records compiled from 1991 to 1995 by the Oakland Firestorm Pet Hotline were analyzed. Peak activity for the hotline was on days 3 and 4 after the fire, but decreased to a low, steady rate by day 21. Many pets were found that had been abandoned or were part of a large free-roaming population that existed at the time of the fire. Many were missing after the fire and presumed killed. The longer owners delayed looking for their pet, the lower the chance of being reunited. Pets wearing collars with the owners' names and addresses had a more than 10-fold chance of being reunited, compared with pets without collars. Increasing odds for adoption of lost pets was associated with their proximity to the fire. Hotlines set up after sudden impact disasters, such as the Oakland Firestorm Pet Hotline, will probably register primarily abandoned animals. Reunion of pets with their owners appears to be most likely for those that receive an overall better level of care than pets found at other times. Adoption of pets after this disaster was primarily by the person who found that animal and principally resulted when the pet was found close to the disaster area.


Differential requirements for semaphorin 3F and Slit-1 in axonal targeting, fasciculation, and segregation of olfactory sensory neuron projections.
The formation of precise stereotypic connections in sensory systems is critical for defining accurate internal representations of the external world; however, the molecular mechanisms underlying the formation of sensory maps are poorly understood. Here, we examine the roles of two structurally unrelated repulsive guidance cues, semaphorin 3F (Sema3F) and Slit-1, in olfactory sensory axon fasciculation, targeting, and segregation. Using sema3F-/- mice, we show that Sema3F is critical for vomeronasal sensory neuron axonal fasciculation and for segregation of these sensory afferents from the main olfactory system; however, Sema3F plays only a minor role in targeting of apical vomeronasal neuron axons to the anterior accessory olfactory bulb (AOB). In addition, we show that Sema3F is required for lamina-specific targeting of olfactory sensory axons within the main olfactory system. In contrast to Sema3F, Slit-1 is dispensable for fasciculation of basal vomeronasal neuron axons but is critical for targeting these axons to the posterior AOB. These results reveal discrete and complementary roles for secreted semaphorins and slits in axonal targeting, fasciculation, and segregation of olfactory sensory neuron projections.


Randomized double-blind study of the nutritional efficacy and bifidogenicity of a new infant formula containing partially hydrolyzed protein, a high beta-palmitic acid level, and nondigestible oligosaccharides.
The aim of this study was to evaluate the nutritional efficacy and bifidogenic characteristics of a new infant formula containing partially hydrolyzed whey protein, modified vegetable oil with a high beta-palmitic acid content, prebiotic oligosaccharides, and starch. In a double-blind study, healthy formula-fed term infants aged younger than 2 weeks were randomized to receive either the new infant formula (NF) or a standard formula (SF) until the age of 12 weeks. Anthropometric measurements were taken at enrollment, 6 weeks, and 12 weeks. In a subsample of infants, blood samples were taken at 6 weeks and stool samples were taken at enrollment and 6 weeks. Blood samples were analyzed for biochemical measures of protein status and amino acids, and stools were analyzed for total bacteria and bifidobacteria. Mothers completed a feeding diary and questionnaire at 6 and 10 weeks. One hundred fifty-four infants were enrolled in the study; 102 completed the trial. The growth of infants in both formula groups was in line with published growth curves. During the first 6 weeks, NF girls gained more weight and head circumference than the SF girls. These velocity differences were not maintained throughout the 12-week study period. The NF stools had a higher proportion of bifidobacteria at 6 weeks compared with the SF stools, and they were softer. There were no clinically significant differences in the blood biochemical and amino acid values between groups. Both formulas were well tolerated by the infants. When compared with a standard infant formula, the new formula supported satisfactory growth, led to higher counts of bifidobacteria in the feces, produced blood bio-chemical values typical of formula-fed infants, and was well tolerated.


[Detailed evaluation of sexual function after radical prostatectomy: is patient satisfaction correlated with the quality of erections?].
Post radical prostatectomy potency rates, quantified on the basis of physician survey, have ranged up to 80%. Physician derived potency data, however, may not be representative of true post-prostatectomy potency rates or more importantly may not accurately portray patients' post-operative sexual satisfaction. We conducted a pilot study combining physician derived and patient derived subjective data with objective measures of erectile function. Eleven men, mean age of 59 years, who were treated with nerve sparing radical retropubic prostatectomy formed the study group. Initially, the patients responded to a physician directed telephone survey on sexual status. Potency was then objectively assessed utilizing Rigiscan testing on two consecutive evenings. Lastly, the patients completed a validated short questionnaire directed to obtain a patients' subjective perception of sexual function. All the patients responded to the first part of the study by informing the physician that they were sexually active or potent after radical prostatectomy. Of these 11 patients, however, only 2 (18%) were mostly satisfied with their sex life according to the quality of life questionnaire. Rigiscan testing revealed that 8 of the 11 patients had nocturnal erections which were adequate for vaginal penetration. Of the 5 patients who stated that they were mostly dissatisfied with their sexual functioning, 3 had objective evidence of adequate erectile ability as documented by Rigiscan. Three of the four patients who were ambivalent with respect to their sexual function also demonstrated objective evidence of normal erectile activity. Although a patient may inform his care provider that he is sexually active or potent, he may not be satisfied with his present level of sexual functioning. In addition, we observed that some dissatisfied patients do have normal Rigiscan patterns indicating that a percentage of patients who are not happy with their level of sexual function after radical prostatectomy may have a psychogenic component to their problem.


The LAS VEGAS risk score for prediction of postoperative pulmonary complications: An observational study.
Currently used pre-operative prediction scores for postoperative pulmonary complications (PPCs) use patient data and expected surgery characteristics exclusively. However, intra-operative events are also associated with the development of PPCs. We aimed to develop a new prediction score for PPCs that uses both pre-operative and intra-operative data. This is a secondary analysis of the LAS VEGAS study, a large international, multicentre, prospective study. A total of 146 hospitals across 29 countries. Adult patients requiring intra-operative ventilation during general anaesthesia for surgery. The cohort was randomly divided into a development subsample to construct a predictive model, and a subsample for validation. Prediction performance of developed models for PPCs. Of the 6063 patients analysed, 10.9% developed at least one PPC. Regression modelling identified 13 independent risk factors for PPCs: six patient characteristics [higher age, higher American Society of Anesthesiology (ASA) physical score, pre-operative anaemia, pre-operative lower SpO2 and a history of active cancer or obstructive sleep apnoea], two procedure-related features (urgent or emergency surgery and surgery lasting ≥ 1 h), and five intra-operative events [use of an airway other than a supraglottic device, the use of intravenous anaesthetic agents along with volatile agents (balanced anaesthesia), intra-operative desaturation, higher levels of positive end-expiratory pressures > 3 cmH2O and use of vasopressors]. The area under the receiver operating characteristic curve of the LAS VEGAS risk score for prediction of PPCs was 0.78 [95% confidence interval (95% CI), 0.76 to 0.80] for the development subsample and 0.72 (95% CI, 0.69 to 0.76) for the validation subsample. The LAS VEGAS risk score including 13 peri-operative characteristics has a moderate discriminative ability for prediction of PPCs. External validation is needed before use in clinical practice. The study was registered at Clinicaltrials.gov, number NCT01601223.


Matching Ontologies to HL7 FHIR Towards Their Syntactic and Semantic Similarity.
Current medical systems need to be able to communicate complex and detailed medical data securely and efficiently. However, the quantity of available healthcare data is rising rapidly, far exceeding the capacity to deliver personal or public health benefits from analyzing this data. Thus, a substantial overhaul of methodology is required to address the real complexity of health. This can be achieved by constructing medical domain ontologies for representing medical terminologies, considered to be a difficult task, requiring a profound analysis of the structure and the concepts of medical terminologies. In this paper, a mechanism is presented for constructing healthcare ontologies, while matching them to HL7 FHIR Resources ontologies both in terms of syntactic and semantic similarity, in order to understand their nature and translate them into a common standard to improve the quality of patient care, research, and health service management.


Simultaneous feature selection and clustering using mixture models.
Clustering is a common unsupervised learning technique used to discover group structure in a set of data. While there exist many algorithms for clustering, the important issue of feature selection, that is, what attributes of the data should be used by the clustering algorithms, is rarely touched upon. Feature selection for clustering is difficult because, unlike in supervised learning, there are no class labels for the data and, thus, no obvious criteria to guide the search. Another important problem in clustering is the determination of the number of clusters, which clearly impacts and is influenced by the feature selection issue. In this paper, we propose the concept of feature saliency and introduce an expectation-maximization (EM) algorithm to estimate it, in the context of mixture-based clustering. Due to the introduction of a minimum message length model selection criterion, the saliency of irrelevant features is driven toward zero, which corresponds to performing feature selection. The criterion and algorithm are then extended to simultaneously estimate the feature saliencies and the number of clusters.


Requirement for Pangolin/dTCF in Drosophila Wingless signaling.
The Wingless (Wg) protein is a secreted glycoprotein involved in intercellular signaling. On activation of the Wg signaling pathway, Armadillo is stabilized, causing target genes to be activated by the transcription factor Pangolin (Pan). This study investigated the roles of Pan in the developing wing of Drosophila by clonal analysis. Three different aspects of wing development were examined: cell proliferation, wing margin specification, and wg self-refinement. Our results indicate that Pan function is critically required for all three of these processes. Consequently, lack of pan causes a severe reduction in the activity of the Wg target genes Distalless and vestigial within their normal domain of expression. Loss of pan function does not, however, lead to a derepression of these genes outside this domain. Thus, although Pan is positively required for the induction of Wg targets in the wing imaginal disk, it does not appear to play a default repressor function in the absence of Wg input. In contrast, lack of zygotic pan function causes a milder phenotype than that caused by the lack of wg function in the embryo. We show that this difference cannot be attributed to maternally provided pan product, indicating that a Pan repressor function usually prevents the expression of embryonic Wg targets. Together, our results suggest that for embryonic patterning the activator as well as repressor forms of Pan play important roles, while for wing development Pan operates primarily in the activator mode.


Human immunodeficiency virus type 1 reverse transcriptase genotype and drug susceptibility changes in infected individuals receiving dideoxyinosine monotherapy for 1 to 2 years.
The genetic mechanisms of human immunodeficiency virus type 1 (HIV-1) resistance to dideoxyinosine (ddI) in vivo have been described based on data from primary HIV-1 isolates. To better define the spectrum of HIV-1 reverse transcriptase (RT) changes occurring during ddI therapy, we determined the genotype and ddI susceptibility of the RT gene of HIV RNA isolated from the plasma of 23 patients who had received 1 to 2 years (mean, 87 +/- 16 weeks) of ddI monotherapy. Population-based sequencing of plasma virus showed that 12 of 23 (52%) patients developed known ddI resistance mutations: L74V (7 patients), K65R (2 patients), L74V with M184V (3 patients), and L74V with K65R (1 patient). Five patients developed one or more known zidovudine resistance mutations (at codons 41, 67, 70, 215, and/or 219) during the study. Other amino acid substitutions were found, but only S68G and L210W occurred in more than one patient. Studies of sensitivity to ddI were performed on population-based recombinant-virus stocks generated by homologous recombination between a plasmid containing an HXB2 clone with the RT gene deleted and RT-PCR products of the RT genes from patients' plasma RNA. The sequences of the virus stocks produced by this procedure were typically identical to the sequence of the input PCR product from plasma RNA. Both an MT-2 cell-based culture assay and a cell-free virion-associated RT inhibition assay showed that viruses possessing an L74V and/or M184V mutation or a K65R mutation had reduced sensitivity to ddI. Viruses without these specific mutations had no change in sensitivity to ddI. The results presented here show that the spectrum of RT mutations in a population of patients on ddI monotherapy is more complex than previously described. The development of multiple mutational patterns, including those that confer resistance to other nucleoside analogs, highlights the complexity of using the currently available nucleoside analogs for antiretroviral therapy.


Sequential estimation of optical properties of a two-layered epithelial tissue model from depth-resolved ultraviolet-visible diffuse reflectance spectra.
A method for estimating the optical properties of two-layered media (such as squamous epithelial tissue) over a range of wavelengths in the ultraviolet-visible spectrum is proposed and tested with Monte Carlo modeling. The method first used a fiber-optic probe with angled illumination and the collection fibers placed at a small separation (<or=300 microm) to restrict the transport of detected light to the top layer. A Monte Carlo-based inverse model for a homogeneous medium was employed to estimate the top layer optical properties from the measured diffuse reflectance spectrum. Then a flat-tip probe with a large source-detector separation (>or=1000 microm) was used to detect diffuse reflectance preferentially from the bottom layer. A second Monte Carlo-based inverse model for a two-layered medium was applied to estimate the bottom layer optical properties, as well as the top layer thickness, given that the top layer optical properties have been estimated. The results of Monte Carlo validation show that this method works well for an epithelial tissue model with a top layer thickness ranging from 200 to 500 microm. For most thicknesses within this range, the absorption coefficients were estimated to within 15% of the true values, the reduced scattering coefficients were estimated to within 20% and the top layer thicknesses were estimated to within 20%. The application of a variance reduction technique to the Monte Carlo modeling proved to be effective in improving the accuracy with which the optical properties are estimated.


Multilocus enzyme electrophoresis for identification and typing of Treponema hyodysenteriae and related spirochaetes.
An assessment was made of multilocus enzyme electrophoresis as a means of identifying and typing spirochaetes isolated from pigs. Using five enzyme systems, 36 isolates from Australia, the U.K. and the U.S.A. were divided into 12 electrophoretic types or multilocus genotypes, comprising four major, genetically distinct groups. All 26 isolates of Treponema hyodysenteriae fell into one group, members of which showed relatively little genetic diversity. Ten isolates of non-pathogenic spirochaetes fell into three genetically different groups. Although the technique was capable of typing organisms within the groups, it was not always as discriminatory as DNA-restriction endonuclease analysis. Examination of additional enzyme loci should increase the sensitivity of the method for typing and for overall assessment of genetic relationships between spirochaetes.


Combinatorial regulation of meiotic holliday junction resolution in C. elegans by HIM-6 (BLM) helicase, SLX-4, and the SLX-1, MUS-81 and XPF-1 nucleases.
Holliday junctions (HJs) are cruciform DNA structures that are created during recombination events. It is a matter of considerable importance to determine the resolvase(s) that promote resolution of these structures. We previously reported that C. elegans GEN-1 is a symmetrically cleaving HJ resolving enzyme required for recombinational repair, but we could not find an overt role in meiotic recombination. Here we identify C. elegans proteins involved in resolving meiotic HJs. We found no evidence for a redundant meiotic function of GEN-1. In contrast, we discovered two redundant HJ resolution pathways likely coordinated by the SLX-4 scaffold protein and also involving the HIM-6/BLM helicase. SLX-4 associates with the SLX-1, MUS-81 and XPF-1 nucleases and has been implicated in meiotic recombination in C. elegans. We found that C. elegans [mus-81; xpf-1], [slx-1; xpf-1], [mus-81; him-6] and [slx-1; him-6] double mutants showed a similar reduction in survival rates as slx-4. Analysis of meiotic diakinesis chromosomes revealed a distinct phenotype in these double mutants. Instead of wild-type bivalent chromosomes, pairs of "univalents" linked by chromatin bridges occur. These linkages depend on the conserved meiosis-specific transesterase SPO-11 and can be restored by ionizing radiation, suggesting that they represent unresolved meiotic HJs. This suggests the existence of two major resolvase activities, one provided by XPF-1 and HIM-6, the other by SLX-1 and MUS-81. In all double mutants crossover (CO) recombination is reduced but not abolished, indicative of further redundancy in meiotic HJ resolution. Real time imaging revealed extensive chromatin bridges during the first meiotic division that appear to be eventually resolved in meiosis II, suggesting back-up resolution activities acting at or after anaphase I. We also show that in HJ resolution mutants, the restructuring of chromosome arms distal and proximal to the CO still occurs, suggesting that CO initiation but not resolution is likely to be required for this process.


Characteristics and Outcome of Streptococcus pneumoniae Endocarditis in the XXI Century: A Systematic Review of 111 Cases (2000-2013).
Streptococcus pneumoniae is an infrequent cause of severe infectious endocarditis (IE). The aim of our study was to describe the epidemiology, clinical and microbiological characteristics, and outcome of a series of cases of S. pneumoniae IE diagnosed in Spain and in a series of cases published since 2000 in the medical literature. We prospectively collected all cases of IE diagnosed in a multicenter cohort of patients from 27 Spanish hospitals (n = 2539). We also performed a systematic review of the literature since 2000 and retrieved all cases with complete clinical data using a pre-established protocol. Predictors of mortality were identified using a logistic regression model. We collected 111 cases of pneumococcal IE: 24 patients from the Spanish cohort and 87 cases from the literature review. Median age was 51 years, and 23 patients (20.7%) were under 15 years. Men accounted for 64% of patients, and infection was community-acquired in 96.4% of cases. The most important underlying conditions were liver disease (27.9%) and immunosuppression (10.8%). A predisposing heart condition was present in only 18 patients (16.2%). Pneumococcal IE affected a native valve in 93.7% of patients. Left-sided endocarditis predominated (aortic valve 53.2% and mitral valve 40.5%). The microbiological diagnosis was obtained from blood cultures in 84.7% of cases. In the Spanish cohort, nonsusceptibility to penicillin was detected in 4.2%. The most common clinical manifestations included fever (71.2%), a new heart murmur (55%), pneumonia (45.9%), meningitis (40.5%), and Austrian syndrome (26.1%). Cardiac surgery was performed in 47.7% of patients. The in-hospital mortality rate was 20.7%. The multivariate analysis revealed the independent risk factors for mortality to be meningitis (OR, 4.3; 95% CI, 1.4-12.9; P < 0.01). Valve surgery was protective (OR, 0.1; 95% CI, 0.04-0.4; P < 0.01). Streptococcus pneumoniae IE is a community-acquired disease that mainly affects native aortic valves. Half of the cases in the present study had concomitant pneumonia, and a considerable number developed meningitis. Mortality was high, mainly in patients with central nervous system (CNS) involvement. Surgery was protective.


Fetal hemoglobin reactivation and cell engineering in the treatment of sickle cell anemia.
The natural history of severe hemoglobinopathies like sickle cell disease (SCD) is rather variable, depending on the circumstances, but the main influence on such variability is the level of fetal hemoglobin (HbF) in the patient's red cells. It is well known that a significant HbF level is associated with a milder course of disease and fewer complications. Therefore, attempts have been made to reactivate using various means the HbF production, which is normally switched off perinatally. A pharmacological approach has been attempted since the 1980s, ranging from drugs like 5-azacytidine and its derivative, decitabine, to a series of compounds like hydroxyurea and a number of histone deacetylase inhibitors like butyrate, which seem to act as epigenetic modifiers. Many other disparate agents have been tried with mixed results, but hydroxyurea remains the most effective compound so far available. Combinations of different compounds have also been tried with some success. Established treatments like bone marrow or cord blood transplantation are so far the only real cure for a limited number of patients with severe hemoglobinopathies. Improved chemotherapy regimens of milder toxicity than those employed in the past have made it possible recently to obtain a stable, mixed donor-recipient chimerism, with reversal of the SCD phenotype. However, great effort is directed to cell engineering, searching for an effective gene vector by which a desired gene can be transferred into new classes of vectors for autologous hemopoietic stem cells. Recent studies are also aiming at targeted insertion of the therapeutic gene into hemopoietic cells, which can also be "induced" human stem cells, obtained from somatic dedifferentiated cells. Attention in this area must be paid to the possibility of undesired effects, like the emergence of potentially oncogenic cell populations. Finally, an update is presented on improved HbF determination methods, because common international standards are becoming mandatory.


Immune-related colitis is associated with fecal microbial dysbiosis and can be mitigated by fecal microbiota transplantation.
Colitis induced by treatment with immune checkpoint inhibitors (ICI), termed irColitis, is a substantial cause of morbidity complicating cancer treatment. We hypothesized that abnormal fecal microbiome features would be present at the time of irColitis onset, and that restoring the microbiome with fecal transplant from a healthy donor would mitigate disease severity. Herein, we present fecal microbiota profiles from 18 patients with irColitis from a single center, 5 of whom were treated with healthy donor fecal microbial transplantation (FMT). Although fecal samples collected at onset of irColitis had comparable -diversity to that of comparator groups with gastrointestinal symptoms, irColitis was characterized by fecal microbial dysbiosis. Abundances of Proteobacteria were associated with irColitis in multivariable analyses. Five patients with irColitis refractory to steroids and biologic anti-inflammatory agents received healthy-donor FMT, with initial clinical improvement in irColitis symptoms observed in four of five patients. Two subsequently exhibited recurrence of irColitis symptoms following courses of antibiotics. Both received a second "salvage" FMT that was, again, followed by clinical improvement of irColitis. In summary, we observed distinct microbial community changes that were present at the time of irColitis onset. FMT was followed by clinical improvements in several cases of steroid- and biologic-agent-refractory irColitis. Strategies to restore or prevent microbiome dysbiosis in the context of immunotherapy toxicities should be further explored in prospective clinical trials.


β-Pyridylenolate zinc catalysts for the ring-opening homo- and copolymerization of ε-caprolactone and lactides.
A family of zinc β-pyridylenolates with varying α,β-substituents has been synthesized. A direct reaction of β-pyridyl ketone ligands with ZnEt2 in toluene afforded dimeric [LZnEt]2 (L = (2-C5H4N)-C(R1)[double bond, length as m-dash]C(R2)-O-). X-ray structural data revealed that all the [LZnEt]2 complexes exist as μ-O-bridged dimers in the solid state, although they adopt a different geometry. Each of the [LZnEt]2 complexes is a highly active catalyst for the ring-opening polymerization (ROP) of ε-caprolactone (ε-CL) and racemic lactide (rac-LA), respectively. The donating group on the enolate ligand can accelerate the ROP reaction and control the polymerization process well. All complexes also showed different abilities in the block and random copolymerization of ε-CL and l-LA. Diblock PCL-b-PLLA and PLLA-b-PCL copolymers were easily prepared via sequential addition of ε-CL and l-LA by using complex 10, respectively. Random copolymers of PCL-ran-PLLA were also synthesized from complex 10. The copolymers of PCL-ran-PLLA were shown to possess monomer contents which matched well with their composition in the monomer feed. NMR and DSC characterization confirmed a highly random structure of these copolymers, resulting in closely average lengths of the caproyl and lactidyl sequences (LCL = 2.1; LLA = 1.9). All zinc complexes as efficient single-component initiators in both homo- and copolymerizations showed that polymers were probably susceptible to chain transfer and transesterification due to the close ZnZn proximity in the dimer. The polymers initiated by [LZnEt]2 in the absence of BnOH showed high molecular weight and narrow molecular weight distributions.


Identifying key areas for active interprofessional learning partnerships: A facilitated dialogue.
Student and service user involvement is recognised as an important factor in creating interprofessional education (IPE) opportunities. We used a team-based learning approach to bring together undergraduate health professional students, early career professionals (ECPs), public partners, volunteers, and carers to explore learning partnerships. Influenced by evaluative inquiry, this qualitative study used a free text response to allow participants to give their own opinion. A total of 153 participants (50 public partners and 103 students and professionals representing 11 healthcare professions) took part. Participants were divided into mixed groups of six (n = 25) and asked to identify areas where students, professionals, and public could work together to improve health professional education. Each group documented their discussions by summarising agreed areas and next steps. Responses were collected and transcribed for inductive content analysis. Seven key themes (areas for joint working) were identified: communication, public as partners, standards of conduct, IPE, quality improvement, education, and learning environments. The team-based learning format enabled undergraduate and postgraduate health professionals to achieve consensus with public partners on areas for IPE and collaboration. Some of our results may be context-specific but the approach is generalisable to other areas.


Bioceramic microneedle arrays are able to deliver OVA to dendritic cells in human skin.
Microneedle-based vaccination into skin has several advantages over vaccination using conventional needles for intramuscular or subcutaneous injections. Microneedle (MN) arrays allow the vaccine to be delivered in a minimally invasive manner and directly into the skin, whereby the skin's superficial immune cells are not by-passed. Additionally, a systemic distribution of the vaccine may be avoided, which implies less side effects and less amount of vaccine needed. For a successful delivery, the needles need to penetrate the stratum corneum and reach the potent network of antigen-presenting dendritic cells (DCs). In this study, we evaluated patches covered with biodegradable ceramic (calcium sulphate) MNs with a tip diameter of approximately 3 μm and with two different lengths (300 and 600 μm) for their ability to penetrate and transfer the model allergen ovalbumin (OVA) into epidermis. MNs with a length of 600 μm (MN-600) and a volume average pore size of 12 ± 1 μm were more efficient in crossing the stratum corneum and to deliver OVA into CD1a+ DCs residing in the epidermis of human ex vivo skin, in comparison to MNs with a length of 300 μm. Quantitative in vitro release studies showed that approximately 90% of the loaded OVA could be released from MN-600 within 1 h. These findings support the further development of ceramic MNs for transcutaneous immunization.


The Effect of a Dairy-Based Recovery Beverage on Post-Exercise Appetite and Energy Intake in Active Females.
This study was designed to assess the effect of a dairy-based recovery beverage on post-exercise appetite and energy intake in active females. Thirteen active females completed three trials in a crossover design. Participants completed 60 min of cycling at 65% V̇O2peak, before a 120 min recovery period. On completion of cycling, participants consumed a commercially available dairy-based beverage (DBB), a commercially available carbohydrate beverage (CHO), or a water control (H₂O). Non-esterified fatty acids, glucose, and appetite-related peptides alongside measures of subjective appetite were sampled at baseline and at 30 min intervals during recovery. At 120 min, energy intake was assessed in the laboratory by ad libitum assessment, and in the free-living environment by weighed food record for the remainder of the study day. Energy intake at the ad libitum lunch was lower after DBB compared to H₂O (4.43 ± 0.20, 5.58 ± 0.41 MJ, respectively; p = 0.046; (95% CI: -2.28, -0.20 MJ)), but was not different to CHO (5.21 ± 0.46 MJ), with no difference between trials thereafter. Insulin and GLP-17-36 were higher following DBB compared to H₂O (p = 0.015 and p = 0.001, respectively) but not to CHO (p = 1.00 and p = 0.146, respectively). In addition, glucagon was higher following DBB compared to CHO (p = 0.008) but not to H₂O (p = 0.074). The results demonstrate that where DBB consumption may manifest in accelerated recovery, this may be possible without significantly affecting total energy intake and subsequent appetite-related responses relative to a CHO beverage.


A combinated approach to investigate the biochemistry and hydrography of a shallow bay in the south Adriatic Sea: the Gulf of Manfredonia (Italy).
The main goal of this study is to understand the hydrological and biochemical set up and processes of a marine coastal area located in the western side of the south Adriatic sea (the gulf of Manfredonia) by the use of both satellite images and in situ investigations A water sampling in the gulf of Manfredonia was performed at 18 fixed stations in June 2003; physico-chemical and biological parameters (temperature, salinity, dissolved oxygen and fluorescence) were measured along the water column and water samples were collected to analyse dissolved nutrients (ammonia, nitrite, nitrate, phosphate and silicate), total nutrients (nitrogen and phosphorus), chlorophyll a and total suspended solids. Surface spatial distributions of field collected data were examined to characterize biochemical and hydrographic conditions of the Gulf of Manfredonia and these results were related with the remote sensing analysis data. Remote sensed data (obtained by Landsat 7 TM and Modis Terra) were processed to obtain maps of chlorophyll a, temperature and optical characteristics of the gulf; these maps were compared to in situ data. From physico-chemical measurements no stratification was observed in the water column except for the south-eastern area. High concentrations of silicate and ammonia were observed in the northern zone of the gulf, while nitrate, phosphate, chlorophyll a and total suspended solids distributions showed higher values in the central coastal zone. These results were confirmed by remote sensing analysis; Modis elaboration showed a distribution with higher concentrations of chlorophyll a near the coast and Landsat images highlighted the extension of the surrounding agricultural areas crossed by the two main rivers which discharge into the gulf. The integration between field data with the remote sensing analysis showed to be a valid support in coastal zone management.


miR-493-5p attenuates the invasiveness and tumorigenicity in human breast cancer by targeting FUT4.
Breast cancer is a leading cause of cancer-related mortality among women. Altered fucosylation was found to be closely associated with tumorigenesis and metastasis of breast cancer. MicroRNAs (miRNAs) are important regulators of cell proliferation and metastasis, and aberrant miRNA expression has been observed in breast cancer. The present study aimed to evaluate the level of fucosyltransferase IV (FUT4) and miR-493-5p in breast cancer and investigate their relationship. In the present study, we demonstrated the differential expressional profiles of FUT4 and miR‑493-5p in 29 clinical breast cancer tissues, matched adjacent tissue samples and two breast carcinoma cell lines (MCF-7 and MDA-MB-231). Briefly, altered expression levels of FUT4 modified the invasive activities and tumorigenicity of the MCF-7 and MDA-MB-231 cells. Further study demonstrated that miR-493-5p plays a role as a suppressor in breast cancer cell invasion and tumorigenicity. Moreover, the expression levels of miR-493-5p were inversely proportional to those of FUT4 both at the mRNA and protein levels. Luciferase reporter assays confirmed that miR‑493-5p bound to the 3'-untranslated (3'-UTR) region of FUT4, and inhibited the expression of FUT4 in breast cancer cells. Taken together, our data suggest that FUT4 may have a potential role in the treatment of breast cancer, as well as miR-493-5p is a novel regulator of invasiveness and tumorigenicity of breast cancer cells through targeting FUT4. The miR-493-5p/FUT4 pathway has therapeutic potential in breast cancer.


"Can you send an ambulance please?": a comparison of callers' requests for emergency medical dispatch in non-stroke and stroke calls.
Identifying 'true stroke' from an emergency medical services (EMS) call is challenging, with over 50% of strokes being misclassified. In a previous study, we examined the relationship between callers' descriptions of stroke symptoms to the emergency medical dispatcher and the subsequent classification and prioritisation of EMS response. The aim of this subsequent study was to explore further the use of keywords by callers when making emergency calls, comparing stroke and non-stroke calls. All non-stroke calls to one EMS dispatch centre between 8 March 2010 and 14 March 2010 were analysed. These were compared with the stroke calls made to one EMS dispatch centre between 1 October 2006 and 30 September 2007. Content analysis was used to explore the problems described by the caller, and findings were compared between non-stroke and stroke calls. 277 non-stroke calls were identified. Only eight (3%) callers mentioned stroke, 12 (4%) and 11 (4%) mentioned limb weakness and speech problems, respectively, while no caller mentioned more than one classic stroke symptom. This contrasted with 473 stroke calls, where 188 (40%) callers mentioned stroke, 70 (15%) limb weakness and 72 (15%) speech problems, and 14 (3%) mentioned more than one classic stroke symptom. People who contact the EMS about non-stroke conditions rarely say stroke, limb weakness, speech problems or facial weakness. These words are more frequently used when people contact the EMS about stroke, although many calls relating to stroke patients do not mention any of these keywords.


Prevalence and Management of Urinary Incontinence in a Brazilian Hospital: A Prospective, Descriptive Study.
Although urinary incontinence (UI) in hospitalized patients is a frequent health problem, research on the subject is limited. A prospective, descriptive study was conducted to evaluate the prevalence of UI, care implemented among hospitalized adult patients, and impact of UI on quality of life (QoL) in a large private institution in Brazil. All patients admitted during November 2018 who were at least 18 years of age, sufficiently conscious and oriented to answer questions, able to undergo skin inspection, and had the ability to urinate (ie, did not use an indwelling bladder catheter or have a nephrostomy, cystostomy, urostomy, or vesicostomy) were eligible to participate. Data were collected through interviews, physical examinations, chart review, and completion of the International Consultation on Incontinence Questionnaire-Short Form (ICQI-SF), a self-diagnostic, 6-item survey of UI frequency, amount of leakage, and overall impact of UI; answers for items 3 (6 answer options), 4 (4 answer options), and 5 (Likert scale ranging from 0 to 10) were considered individually and summed for a total score ranging from 0 to 21. Data were collected using paper forms and then double-entered and validated in a spreadsheet for statistical analysis. Continuous variables were described as mean/median and standard deviation. Categorical variables were described in absolute numbers and percentages. Pearson's chi-squared test and Fischer's exact test were used to investigate the variable statistical differences. The level of significance was 5%, and the intervals were obtained with 95% confidence. Prevalence was defined as the percentage of people with UI over the study period in all admitted patients. Of the 858 eligible patients, 114 were incontinent (13.3%), but 61 were unable to answer the research questions, 2 used catheterization, and 9 refused to participate; therefore 42 patients (age 80 [range 77.6-82.3] years, 30 [71.4%] women) completed the study. All patients (100%) were managed using diapers. Twenty-one (21, 50%) were married, 24 (57.1%) were white, and 25 (59.5%) were retired. Sixteen (16, 38.1%) had urine loss several times a day, and 17 (40.5%) had urine loss in large amounts; 10 (23.8%) had incontinence-associated dermatitis. The ICQI-SF item regarding interference of urine loss in daily life had a reported mean of 4.1 ± 4.0, inferring urine loss interfered with life; mean ICQI-SF score was 12.1 ± 5.86, implying UI had a moderate impact on QoL. The 13.3% prevalence of UI was similar to previous studies. The absence of a protocol for incontinence management was identified. The results suggest additional, larger sample UI prevalence studies need to be conducted.


Comparison of a phytotherapeutic agent (Permixon) with an alpha-blocker (Tamsulosin) in the treatment of benign prostatic hyperplasia: a 1-year randomized international study.
While the lipido-sterolic extract of Serenoa repens (LSESr)-Permixon((R))-has been shown to have an equivalent efficacy to finasteride in patients with benign prostatic hyperplasia (BPH), to date, there has been no valid comparison of phytotherapy with alpha-blockers. The aim of this study was to assess the equivalent efficacy of Permixon and tamsulosin. Eight hundred and eleven men with symptomatic BPH (I-PSS> or =10) were recruited in 11 European countries for a 12-month, double-blind randomized trial. After a 4-week run-in period, 704 patients were randomly assigned to either tamsulosin 0.4mg/day (N=354) or Permixon 320mg/day (N=350). I-PSS, QoL and Q(max) were evaluated at baseline and periodically for 1 year. Prostate volume and serum prostate-specific antigen (PSA) were measured at selection and at endpoint. The endpoint analysis was performed on the per-protocol population of 542 patients (tamsulosin: N=273; Permixon: N=269). At 12 months, I-PSS decreased by 4.4in each group and no differences were observed in either irritative or obstructive symptom improvements. The increase in Q(max) was similar in both treatment groups (1.8ml/s Permixon, 1.9ml/s tamsulosin). PSA remained stable while prostate volume decreased slightly in the Permixon-treated patients. The two compounds were well tolerated, however, ejaculation disorders occurred more frequently in the tamsulosin group. This study demonstrates that Permixon and tamsulosin are equivalent in the medical treatment of lower urinary tract symptoms in men with BPH, during and up to 12 months of therapy.


The Florentine Archives in Transition: Government, Warfare and Communication (1289-1530 ca.).
A turning point in European administrative and documentary practices was traditionally associated, most famously by Robert-Henri Bautier, with the monarchies of the late sixteenth and early seventeenth centuries. By summarizing previous research in this field, as well as by using both published and unpublished sources, this article intends to underline an earlier process of transition connected to the development of significant new techniques for the production and preservation of documents in Renaissance Italian city-states. Focusing on the important case of Florence, the administrative uses of records connected to government, diplomacy and military needs will be discussed, and evidence will be provided that such documentary practices accelerated significantly during the so-called Italian Wars (from 1494 onwards). A particular reason of interest for Florence at this time is that a major role in the production and storage of a large quantity of state papers was played by Niccolò Machiavelli, one of the outstanding political thinkers of the age. This was especially true in connection to the new militia which he himself created in 1506. By stressing the role of information management and the importance of correspondence networks at a time of war and crisis, this article also contributes to recent scholarship which has focused on the growth of public records relating to diplomacy in Italy during the second half of the fifteenth century, as well as to a recent field of historiography which has lately gained importance: namely the 'documentary history of institutions'.


Platform accelerations of three different whole-body vibration devices and the transmission of vertical vibrations to the lower limbs.
Physical whole-body vibration (WBV) exercises become available at various levels of intensity. In a first series of measurements, we investigated 3-dimensional platform accelerations of three different WBV devices without and with three volunteers of different weight (62, 81 and 100 kg) in squat position (150 degrees knee flexion). The devices tested were two professional devices, the PowerPlate and the Galileo-Fitness, and one home-use device, the PowerMaxx. In a second series of measurements, the transmission of vertical platform accelerations of each device to the lower limbs was tested in eight healthy volunteers in squat position (100 degrees knee flexion). The first series showed that the platforms of two professional devices vibrated in an almost perfect vertical sine wave at frequencies between 25-50 and 5-40 Hz, respectively. The platform accelerations were slightly influenced by body weight. The PowerMaxx platform mainly vibrated in the horizontal plane at frequencies between 22 and 32 Hz, with minimal accelerations in the vertical direction. The weight of the volunteers reduced the platform accelerations in the horizontal plane but amplified those in the vertical direction about eight times. The vertical accelerations were highest in the Galileo (approximately 15 units of g) and the PowerPlate (approximately 8 units of g) and lowest in the PowerMaxx (approximately 2 units of g). The second series showed that the transmission of vertical accelerations at a common preset vibration frequency of 25 Hz were largest in the ankle and that transmission of acceleration reduced approximately 10 times at the knee and hip. We conclude that large variation in 3-dimensional accelerations exist in commercially available devices. The results suggest that these differences in mechanical behaviour induce variations in transmissibility of vertical vibrations to the (lower) body.


PI3K/AKT signaling is essential for communication between tissue-infiltrating mast cells, macrophages, and epithelial cells in colitis-induced cancer.
To understand signaling pathways that shape inflamed tissue and predispose to cancer is critical for effective prevention and therapy for chronic inflammatory diseases. We have explored phosphoinositide 3-kinase (PI3K) activity in human inflammatory bowel diseases and mouse colitis models. We conducted immunostaining of phosphorylated AKT (pAKT) and unbiased high-throughput image acquisition and quantitative analysis of samples of noninflamed normal colon, colitis, dysplasia, and colorectal cancer. Mechanistic insights were gained from ex vivo studies of cell interactions, the piroxicam/IL-10(-/-) mouse model of progressive colitis, and use of the PI3K inhibitor LY294002. Progressive increase in densities of pAKT-positive tumor-associated macrophages (TAM) and increase in densities of mast cells in the colonic submucosa were noted with colitis and progression to dysplasia and cancer. Mast cells recruited macrophages in ex vivo migration assays, and both mast cells and TAMs promoted invasion of cancer cells. Pretreatment of mast cells with LY294002 blocked recruitment of TAMs. LY294002 inhibited mast cell and TAM-mediated tumor invasion, and in mice, blocked stromal PI3K, colitis, and cancer. The PI3K/AKT pathway is active in cells infiltrating inflamed human colon tissue. This pathway sustains the recruitment of inflammatory cells through a positive feedback loop. The PI3K/AKT pathway is essential for tumor invasion and the malignant features of the piroxicam/IL-10(-/-) mouse model. LY294002 targets the PI3K pathway and hinders progressive colitis. These findings indicate that colitis and progression to cancer are dependent on stromal PI3K and sensitive to treatment with LY294002.


Widespread sensorimotor and frontal cortical atrophy in Amyotrophic Lateral Sclerosis.
Widespread cortical atrophy in Amyotrophic Lateral Sclerosis (ALS) has been described in neuropathological studies. The presence of cortical atrophy in conventional and scientific neuroimaging has been a matter of debate. In studies using computertomography, positron emission tomography, proton magnetic resonance spectroscopy and conventional T2-weighted and proton-weighted images, results have been variable. Recent morphometric studies by magnetic resonance imaging have produced conflicting results regarding the extent of grey and white matter involvement in ALS patients. The authors used optimized voxel-based morphometry as an unbiased whole brain approach to detect differences between regional grey and white matter volumes. Seventeen patients with a diagnosis of ALS according to El-Escorial criteria and seventeen age-matched controls received a high resolution anatomical T1 scan. In ALS patients regional grey matter volume (GMV) reductions were found in the pre- and postcentral gyrus bilaterally which extended to premotor, parietal and frontal regions bilaterally compared with controls (p < 0.05, corrected for the entire volume). The revised ALS functional rating scale showed a positive correlation with GMV reduction of the right medial frontal gyrus corresponding to the dorsolateral prefrontal cortex. No significant differences were found for white matter volumes or when grey and white matter density images were investigated. There were no further correlations with clinical variables found. In ALS patients, primary sensorimotor cortex atrophy can be regarded as a prominent feature of the disease. Supporting the concept of ALS being a multisystem disorder, our study provides further evidence for extramotor involvement which is widespread. The lack of correlation with common clinical variables probably reflects the fact that heterogeneous disease processes underlie ALS. The discrepancy within all published morphometric studies in ALS so far may be related to differences in patient cohorts and several methodological factors of the data analysis process. Longitudinal studies are required to further clarify the time course and distribution of grey and white matter pathology during the course of ALS.


Moving Objects in the Barn Owl's Auditory World.
Barn owls are keen hunters of moving prey. They have evolved an auditory system with impressive anatomical and physiological specializations for localizing their prey. Here we present behavioural data on the owl's sensitivity for discriminating acoustic motion direction in azimuth that, for the first time, allow a direct comparison of neuronal and perceptual sensitivity for acoustic motion in the same model species. We trained two birds to report a change in motion direction within a series of repeating wideband noise stimuli. For any trial the starting point, motion direction, velocity (53-2400°/s), duration (30-225 ms) and angular range (12-72°) of the noise sweeps were randomized. Each test stimulus had a motion direction being opposite to that of the reference stimuli. Stimuli were presented in the frontal or the lateral auditory space. The angular extent of the motion had a large effect on the owl's discrimination sensitivity allowing a better discrimination for a larger angular range of the motion. In contrast, stimulus velocity or stimulus duration had a smaller, although significant effect. Overall there was no difference in the owls' behavioural performance between "inward" noise sweeps (moving from lateral to frontal) compared to "outward" noise sweeps (moving from frontal to lateral). The owls did, however, respond more often to stimuli with changing motion direction in the frontal compared to the lateral space. The results of the behavioural experiments are discussed in relation to the neuronal representation of motion cues in the barn owl auditory midbrain.


Arbuscular mycorrhizal fungi reduce the differences in competitiveness between dominant and subordinate plant species.
In grassland communities, plants can be classified as dominants or subordinates according to their relative abundances, but the factors controlling such distributions remain unclear. Here, we test whether the presence of the arbuscular mycorrhizal (AM) fungus Glomus intraradices affects the competitiveness of two dominant (Taraxacum officinale and Agrostis capillaris) and two subordinate species (Prunella vulgaris and Achillea millefolium). Plants were grown in pots in the presence or absence of the fungus, in monoculture and in mixtures of both species groups with two and four species. In the absence of G. intraradices, dominants were clearly more competitive than subordinates. In inoculated pots, the fungus acted towards the parasitic end of the mutualism-parasitism continuum and had an overall negative effect on the growth of the plant species. However, the negative effects of the AM fungus were more pronounced on dominant species reducing the differences in competitiveness between dominant and subordinate species. The effects of G. intraradices varied with species composition highlighting the importance of plant community to mediate the effects of AM fungi. Dominant species were negatively affected from the AM fungus in mixtures, while subordinates grew identically with and without the fungus. Therefore, our findings predict that the plant dominance hierarchy may flatten out when dominant species are more reduced than subordinate species in an unfavourable AM fungal relationship (parasitism).


Telephone screening for amnestic mild cognitive impairment.
To evaluate the utility of telephone screening for identifying subjects with amnestic mild cognitive impairment (aMCI) for enrollment in a clinical trial and to identify which elements of the modified Telephone Interview for Cognitive Status (TICS-m) best predicted the in-clinic determination of aMCI. Subjects aged >/=65 years with memory complaints responded to an advertisement for a clinical trial by calling a central telephone recruiting agency. To determine eligibility, subjects went through a stepwise selection procedure involving a review of major protocol inclusion and exclusion criteria, followed by administration of the Category Fluency Test (CFT) and then the TICS-m. Subjects meeting entry criteria, who obtained a score of </=13 on the CFT for "animals" and </=24 on the CFT for "animals" and "fruits" and who scored between 19 and 38 on the TICS-m, were referred for a clinic appointment to determine whether they met clinical criteria for aMCI. Clinical criteria for aMCI required a score of >/=24 on the Mini-Mental State Examination and a score of </=37 on the Rey Auditory Verbal Learning Test. A post hoc analysis was performed using factor analysis and logistic regression models to investigate which elements of the TICS-m best predicted the in-clinic determination of aMCI. Of 16,988 subjects who called the telephone agency, 8,742 passed the review of inclusion/exclusion criteria; 6,090 met the CFT cut scores and received the TICS-m; 5,223 met cut scores on the TICS-m and were referred for an in-clinic appointment; 747 were seen in the clinic; and 324 met clinical criteria for aMCI. Factor analysis indicated three factors on the TICS-m: language/attention, orientation, and memory. The memory factor, comprising immediate and delayed recall of a word list, was the most important contributor for identifying subjects who met clinical criteria for aMCI. Only 2% of subjects who underwent telephone screening were recruited into the study, but 43% of those who passed telephone screening and were seen in the clinic met clinical criteria for aMCI. The word recall tests of the TICS-m were the most important items for identifying which subjects met clinical criteria for aMCI.


Learning from traumatic experiences with brief eclectic psychotherapy for PTSD.
Brief eclectic psychotherapy for PTSD (BEPP) is an evidence-based therapeutic approach that combines and integrates elements from psychodynamic, cognitive-behavioral, and directive psychotherapy. Psychoeducation is done jointly with the patient and his or her partner. Exposure, a structured writing task, and memorabilia are used to help patients accessing, feeling and expressing their suppressed emotions related to the traumatic experience. In the domain of meaning stage, patients will learn how they and their view of the world have changed, and that they have become "sadder but wiser". Much emphasis is put on the vulnerability of human beings. Finally, an individually tailored farewell ritual is done to end treatment, to reunite with loved ones, and to go on with life.


Reduction of reactive oxygen species prevents hypoxia-induced CREB depletion in pulmonary artery smooth muscle cells.
Hypoxia-induced pulmonary arterial hypertension (PAH) is a deadly disease characterized by progressive remodeling and persistent vasoconstriction of the pulmonary arterial system. Remodeling of the pulmonary artery (PA) involves smooth muscle cell (SMC) proliferation, hypertrophy, migration, and elevated extracellular matrix (ECM) production elicited by mitogens and oxidants produced in response to hypoxic insult. We previously reported that the transcription factor cAMP response element binding protein (CREB) is depleted in medial PA SMCs in remodeled, hypertensive vessels in rats or calves exposed to chronic hypoxia. In culture, CREB loss can be induced in PA SMCs by exogenous oxidants or platelet-derived growth factor. Forced depletion of CREB with small interfering RNA (siRNA) in PA SMCs is sufficient to induce their proliferation, hypertrophy, migration, dedifferentiation, and ECM production. This suggests that oxidant and/or mitogen-induced loss of CREB in medial SMCs is, in part, responsible for PA thickening. Here, we tested whether oxidant scavengers could prevent the loss of CREB in PA SMCs and inhibit SMC proliferation, migration, and ECM production using in vitro and in vivo models. Exposure of PA SMCs to hypoxia induced hydrogen peroxide (H2O2) production and loss of CREB. Treatment of SMCs with exogenous H2O2 or a second oxidant, Sin-1, elicited CREB depletion under normoxic conditions. Exogenous H2O2 also induced SMC proliferation, migration, and increased elastin levels as did forced depletion of CREB. In vivo, hypoxia-induced thickening of the PA wall was suppressed by the superoxide dismutase mimetic, Tempol, which also prevented the loss of CREB in medial SMCs. Tempol also reduced hypoxia-induced SMC proliferation and elastin deposition in the PA. The data indicate that CREB levels in the arterial wall are regulated in part by oxidants produced in response to hypoxia and that CREB plays a crucial role in regulating SMC phenotype and PA remodeling.


Substrate and mispairing properties of 5-formyl-2'-deoxyuridine 5'-triphosphate assessed by in vitro DNA polymerase reactions.
5-Formyluracil (fU) is one of the thymine lesions produced by reactive oxygen radicals in DNA and its constituents. In this work, 5-formyl-2'-deoxyuridine 5'-triphosphate (fdUTP) was chemically synthesized and extensively purified by HPLC. The electron withdrawing 5-formyl group facilitated ionization of fU. Thus, p K a of the base unit of fdUTP was 8.6, significantly lower than that of parent thymine (p K a = 10.0 as dTMP). fdUTP efficiently replaced dTTP during DNA replication catalyzed by Escherichia coli DNA polymerase I (Klenow fragment), T7 DNA polymerase (3'-5'exonuclease free) and Taq DNA polymerase. fU-specific cleavage of the replication products by piperidine revealed that when incorporated as T, incorporation of fU was virtually uniform, suggesting minor sequence context effects on the incorporation frequency of fdUTP. fdUTP also replaced dCTP, but with much lower efficiency than that for dTTP. The substitution efficiency for dCTP increased with increasing pH from 7.2 to 9.0. The parallel correlation between ionization of the base unit of fdUTP (p K a = 8.6) and the substitution efficiency for dCTP strongly suggests that the base-ionized form of fdUTP is involved in mispairing with template G. These data indicate that fU can be specifically introduced into DNA as unique lesions by in vitro DNA polymerase reactions. In addition, fU is potentially mutagenic since this lesion is much more prone to form mispairing with G than parent thymine.


Network dynamical stability analysis reveals key "mallostatic" natural variables that erode homeostasis and drive age-related decline of health.
Using longitudinal study data, we dynamically model how aging affects homeostasis in both mice and humans. We operationalize homeostasis as a multivariate mean-reverting stochastic process. We hypothesize that biomarkers have stable equilibrium values, but that deviations from equilibrium of each biomarker affects other biomarkers through an interaction network - this precludes univariate analysis. We therefore looked for age-related changes to homeostasis using dynamic network stability analysis, which transforms observed biomarker data into independent "natural" variables and determines their associated recovery rates. Most natural variables remained near equilibrium and were essentially constant in time. A small number of natural variables were unable to equilibrate due to a gradual drift with age in their homeostatic equilibrium, i.e. allostasis. This drift caused them to accumulate over the lifespan course and makes them natural aging variables. Their rate of accumulation was correlated with risk of adverse outcomes: death or dementia onset. We call this tendency for aging organisms to drift towards an equilibrium position of ever-worsening health "mallostasis". We demonstrate that the effects of mallostasis on observed biomarkers are spread out through the interaction network. This could provide a redundancy mechanism to preserve functioning until multi-system dysfunction emerges at advanced ages.


Phase 2 Open-Label Trial Investigating Percutaneous Laser Ablation for Treatment of Early-Stage Breast Cancer: MRI, Pathology, and Outcome Correlations.
An institutional review board-approved, multicenter clinical trial was designed to determine the efficacy and outcome of percutaneous laser ablation (PLA) in the treatment of invasive ductal breast carcinoma (IDC). Post-ablation magnetic resonance imaging (MRI) was compared with surgical pathology in evaluation of residual post-ablation IDC and ductal carcinoma in situ. Patients with a single focus of IDC 20 mm or smaller by pre-ablation MRI were treated with PLA. The patients underwent a 28-day post-ablation MRI, followed by surgical resection. Cell viability criteria were applied to pre- and post-ablation pathology specimens, which evaluated hematoxylin-eosin (H&E), cytokeratin (CK) 8/18, estrogen receptor, and Ki67 staining patterns. In this study, 61 patients were reported as the intention-to-treat cohort for determination of PLA efficacy. Of these 61 patients, 51 (84%) had complete tumor ablation confirmed by pathology analysis. One subject's MRI imaging was not performed per protocol, which left 60 subjects evaluable for MRI pathology correlation. Five patients (8.3%) had residual IDC shown by both MRI and pathology. Post-ablation discordance was noted between MRI and pathology, with four patients (6.7%) false-positive and four patients (6.7%) false-negative. The negative predictive value (NPV) of MRI for all the patients was 92.2% (95% confidence interval [CI], 71.9-91.9%). Of the 47 patients (97.9%) with tumors 15 mm or smaller, 46 were completely ablated, with an MRI NPV of 97.7% (95% CI, 86.2-99.9%). Percutaneous laser ablation is a potential alternative to surgery for treatment of early-stage IDC. Strong correlations exist between post-ablation MRI and pathologic alterations in CK8/18, ER, and Ki67 staining.


Haptoglobin binding to apolipoprotein A-I prevents damage from hydroxyl radicals on its stimulatory activity of the enzyme lecithin-cholesterol acyl-transferase.
Apolipoprotein A-I (ApoA-I), a major component of HDL, binds haptoglobin, a plasma protein transporting to liver or macrophages free Hb for preventing hydroxyl radical production. This work aimed to assess whether haptoglobin protects ApoA-I against this radical. Human ApoA-I structure, as analyzed by electrophoresis and MS, was found severely altered by hydroxyl radicals in vitro. Lower alteration of ApoA-I was found when HDL was oxidized in the presence of haptoglobin. ApoA-I oxidation was limited also when the complex of haptoglobin with both high-density lipoprotein and Hb, immobilized on resin beads, was exposed to hydroxyl radicals. ApoA-I function to stimulate cholesterol esterification was assayed in vitro by using ApoA-I-containing liposomes. Decreased stimulation was observed when liposomes oxidized without haptoglobin were used. Conversely, after oxidative stress in the presence of haptoglobin (0.5 microM monomer), the liposome activity did not change. Plasma of carrageenan-treated mice was analyzed by ELISA for the levels of haptoglobin and ApoA-I, and used to isolate HDL for MS analysis. Hydroxyproline-containing fragments of ApoA-I were found associated with low levels of haptoglobin (18 microM monomer), whereas they were not detected when the haptoglobin level increased (34-70 microM monomer). Therefore haptoglobin, when circulating at enhanced levels with free Hb during the acute phase of inflammation, might protect ApoA-I structure and function against hydroxyl radicals.


Loss of ARNT/HIF1beta mediates altered gene expression and pancreatic-islet dysfunction in human type 2 diabetes.
beta cell dysfunction is a central component of the pathogenesis of type 2 diabetes. Using oligonucleotide microarrays and real-time PCR of pancreatic islets isolated from humans with type 2 diabetes versus normal glucose-tolerant controls, we identified multiple changes in expression of genes known to be important in beta cell function, including major decreases in expression of HNF4alpha, insulin receptor, IRS2, Akt2, and several glucose-metabolic-pathway genes. There was also a 90% decrease in expression of the transcription factor ARNT. Reducing ARNT levels in Min6 cells with small interfering RNA (siRNA) resulted in markedly impaired glucose-stimulated insulin release and changes in gene expression similar to those in human type 2 islets. Likewise, beta cell-specific ARNT knockout mice exhibited abnormal glucose tolerance, impaired insulin secretion, and changes in islet gene expression that mimicked those in human diabetic islets. Together, these data suggest an important role for decreased ARNT and altered gene expression in the impaired islet function of human type 2 diabetes.


Oestrogen plus progestin and lung cancer in postmenopausal women (Women's Health Initiative trial): a post-hoc analysis of a randomised controlled trial.
In the post-intervention period of the Women's Health Initiative (WHI) trial, women assigned to treatment with oestrogen plus progestin had a higher risk of cancer than did those assigned to placebo. Results also suggested that the combined hormone therapy might increase mortality from lung cancer. To assess whether such an association exists, we undertook a post-hoc analysis of lung cancers diagnosed in the trial over the entire follow-up period. The WHI study was a randomised, double-blind, placebo-controlled trial undertaken in 40 centres in the USA. 16 608 postmenopausal women aged 50-79 years with an intact uterus were randomly assigned by a computerised, stratified, permuted block algorithm to receive a once-daily tablet of 0.625 mg conjugated equine oestrogen plus 2.5 mg medroxyprogesterone acetate (n=8506) or matching placebo (n=8102). We assessed incidence and mortality rates for all lung cancer, small-cell lung cancer, and non-small-cell lung cancer by use of data from treatment and post-intervention follow-up periods. Analysis was by intention to treat. This study is registered with ClinicalTrials.gov, number NCT00000611. After a mean of 5.6 years (SD 1.3) of treatment and 2.4 years (0.4) of additional follow-up, 109 women in the combined hormone therapy group had been diagnosed with lung cancer compared with 85 in the placebo group (incidence per year 0.16%vs 0.13%; hazard ratio [HR] 1.23, 95% CI 0.92-1.63, p=0.16). 96 women assigned to combined therapy had non-small-cell lung cancer compared with 72 assigned to placebo (0.14%vs 0.11%; HR 1.28, 0.94-1.73, p=0.12). More women died from lung cancer in the combined hormone therapy group than in the placebo group (73 vs 40 deaths; 0.11%vs 0.06%; HR 1.71, 1.16-2.52, p=0.01), mainly as a result of a higher number of deaths from non-small-cell lung cancer in the combined therapy group (62 vs 31 deaths; 0.09%vs 0.05%; HR 1.87, 1.22-2.88, p=0.004). Incidence and mortality rates of small-cell lung cancer were similar between groups. Although treatment with oestrogen plus progestin in postmenopausal women did not increase incidence of lung cancer, it increased the number of deaths from lung cancer, in particular deaths from non-small-cell lung cancer. These findings should be incorporated into risk-benefit discussions with women considering combined hormone therapy, especially those with a high risk of lung cancer. National Heart, Lung and Blood Institute, National Institutes of Health.