Intraarticular corticosteroid injection for first carpometacarpal osteoarthritis.
To evaluate the efficacy of intraarticular corticosteroid injections for osteoarthritis (OA) of the first carpometacarpal (CMC) joint. This was a prospective case series of patients presenting to a community rheumatology clinic with OA of the 1st CMC joint. A total of 0.25 ml of methylprednisolone acetate was injected into the 1st CMC joint in 25 patients, who were followed for one year. A significant improvement in the visual analog scale (VAS) for pain was noted at one month but not at 3, 6, or 12 months postinjection. However, 5 patients were pain-free at 12 months postinjection. Many patients noted improvement in tasks such as lifting a full cup and turning a faucet. Injections were well tolerated, with only 2 patients noting minor side effects. These results suggest that intraarticular corticosteroid injection for 1st CMC OA is a well tolerated procedure. A significant longterm benefit of corticosteroid injection for 1st CMC OA was not observed.


Hypoglycemia Among Patients with Type 2 Diabetes: Epidemiology, Risk Factors, and Prevention Strategies.
Hypoglycemia is the most common and often treatment-limiting serious adverse effect of diabetes therapy. Despite being potentially preventable, hypoglycemia in type 2 diabetes incurs substantial personal and societal burden. We review the epidemiology of hypoglycemia in type 2 diabetes, discuss key risk factors, and introduce potential prevention strategies. Reported rates of hypoglycemia in type 2 diabetes vary widely as there is marked heterogeneity in how hypoglycemia is defined, measured, and reported. In randomized controlled trials, rates of severe hypoglycemia ranged from 0.7 to 12 per 100 person-years. In observational studies, hospitalizations or emergency department visits for hypoglycemia were experienced by 0.2 (patients treated without insulin or sulfonylurea) to 2.0 (insulin or sulfonylurea users) per 100 person-years. Patient-reported hypoglycemia is much more common. Over the course of 6 months, 1-4% non-insulin users reported need for medical attention for hypoglycemia; 1-17%, need for any assistance; and 46-58%, any hypoglycemia symptoms. Similarly, over a 12-month period, 4-17% of insulin-treated patients reported needing assistance and 37-64% experienced any hypoglycemic symptoms. Hypoglycemia is most common among older patients with multiple or advanced comorbidities, patients with long diabetes duration, or patients with a prior history of hypoglycemia. Insulin and sulfonylurea use, food insecurity, and fasting also increase hypoglycemia risk. Clinical decision support tools may help identify at-risk patients. Prospective trials of efforts to reduce hypoglycemia risk are needed, and there is emerging evidence supporting multidisciplinary interventions including treatment de-intensification, use of diabetes technologies, diabetes self-management, and social support. Hypoglycemia among patients with type 2 diabetes is common. Patient-centered multidisciplinary care may help proactively identify at-risk patients and address the multiplicity of factors contributing to hypoglycemia occurrence.


Flutter in flow-limited collapsible tubes: a mechanism for generation of wheezes.
We studied flutter in collapsible tubes as a possible mechanism for the generation of respiratory wheezes. The pressure-flow relationships and the wall oscillations of thick-walled [wall thickness (h)-to-lumen radius (r) ratio 1:1.7 to 1.3] self-supporting latex and Silastic tubes mounted between rigid pipes were measured. A high-impedance vacuum pump was connected to the downstream end. Upstream and downstream valves were used to control corresponding resistances. We found loud honking sounds and tube wall oscillations that occurred only when the tubes were buckled and flow limiting, i.e., when the flow became constant and independent of downstream driving pressure. The overall range of oscillatory frequencies was 260-750 Hz for airflow, presenting as sharp peaks of power on the frequency spectrum. The oscillatory frequencies (f) were higher at higher fluid velocities (u) and with narrower distance between opposing flattened walls (2b), resulting from increasing downstream suction pressure and the transmural pressure becoming more negative. The effect of u and b on f for a latex tube (h-to-r ratio 1:1.7) were found to be f = 228 + 0.021 (u/b). These relationships were valid throughout the range of oscillations in this tube (283-720 Hz) and with flow rates of 12-64 l/min. The experimental data were compared with predictions of the fluid dynamic flutter theory and the vortex-induced wall vibrations mechanism. We conclude that viscid flutter in soft tubes is the more probable mechanism for the generation of oscillations in the soft tube model and is a possible mechanism for the generation of respiratory wheezes.


Direct conversion of fibroblasts to functional neurons by defined factors.
Cellular differentiation and lineage commitment are considered to be robust and irreversible processes during development. Recent work has shown that mouse and human fibroblasts can be reprogrammed to a pluripotent state with a combination of four transcription factors. This raised the question of whether transcription factors could directly induce other defined somatic cell fates, and not only an undifferentiated state. We hypothesized that combinatorial expression of neural-lineage-specific transcription factors could directly convert fibroblasts into neurons. Starting from a pool of nineteen candidate genes, we identified a combination of only three factors, Ascl1, Brn2 (also called Pou3f2) and Myt1l, that suffice to rapidly and efficiently convert mouse embryonic and postnatal fibroblasts into functional neurons in vitro. These induced neuronal (iN) cells express multiple neuron-specific proteins, generate action potentials and form functional synapses. Generation of iN cells from non-neural lineages could have important implications for studies of neural development, neurological disease modelling and regenerative medicine.


Guns and butter: a no cost defense against predation for Chrysomela confluens.
Chrysomela confluens produces a salicylaldehyde-based defensive secretion which is very effective against generalist predators and apparently produced at no cost. If no cost defenses are common, then one of the basic assumptions in the plant-herbivore literature, i.e. tradeoffs among defense, reproduction, and growth, must be reconsidered. We examined the effectiveness of this defense by exposing defended larvae and larvae whose secretion had been removed to a generalist predator. Larvae which had their secretions intact were attacked by only 7% of the ants which encountered them, and none of these larvae suffered serious damage. In contrast, those which had been "milked" of their secretions immediately prior to exposure were attacked in 48% of such encounters, and two-thirds of the larvae were killed. Larvae which had been milked 24 or 72 h before exposure, then allowed to regenerate their defenses, were attacked at rates indistinguishable from larvae that had not been milked. Thus regenerated defenses are just as effective as original defenses. We also tested the hypothesis that the cost of defense production and maintainence would be reflected in reductions in developmental rates and final adult mass and increases in leaf consumption rate. We found that larvae which were milked daily of their secretions manifested no measurable cost of recharging reservoirs. Milked larvae grew and fed at the same rates as their control sibs, and became adults of equal or slightly larger size. The liberation of glucose from salicin, a precursor present in leaves of salicaceous hosts, during the production of salicylaldehyde apparently provides enough of an energetic benefit to offset the cost of maintaining an effective defense. Consistent with this hypothesis, we did not find that milked larvae compensated for increased nutritional or salicin demands by increasing their feeding rates. Although this patterns is familiar to chemical ecologists it is generally unappreciated in the plant-herbivore literature. It is likely that many arthropod herbivore defensive systems come at little or no cost, given the intimacy of association between herbivores and their food plants. Sequestration of host plant defensive chemicals which eliminates the cost of synthesis is common in arthropods. The de novo synthesis of chemical defenses may be less costly than expected if it is integrated into other parts of an insects metabolism. Calculations based on the bond energies or molecular constitution of the compounds will not yield a complete perception of cost. Tests over the life of the herbivore, coupled with an understanding of the herbivore's metabolism, are necessary.


Genetic population structure of the prosobranch snail Potamopyrgus antipodarum (Gray) in Denmark using PCR-RAPD fingerprints.
Parthenogenetic species are often more widely distributed geographically than their sexual relatives. This success in colonizing can be explained either by dispersal of one or a few clones of wide physiological tolerance or by the distribution of many locally adapted clones. Here we test the hypothesis that successfully invading clones of Potamopyrgus antipodarum (Gray) are composed of a few broadly adapted genotypes by using polymerase chain reaction random amplified polymorphic DNA (PCR-RAPD) fingerprinting on six different populations of P. antipodarum from Denmark and three morphotypes of P. antipodarum from Britain. We detected two genotypes of P. antipodarum in six populations examined across Denmark using four decamer primers. The two genotypes were found to be morphologically and genetically indistinguishable from British P. antipodarum. In five of the six Danish populations only one genotype was found; at the remaining site, the two genotypes occurred sympatrically. The present study suggests that P. antipodarum successfully invaded Europe by the proliferation of very few clones.


Preimmune Control of the Variance of TCR CDR-B3: Insights Gained From Germline Replacement of a TCR Dβ Gene Segment With an Ig D H Gene Segment.
We have previously shown that the sequence of the immunoglobulin diversity gene segment (D H ) helps dictate the structure and composition of complementarity determining region 3 of the immunoglobulin heavy chain (CDR-H3). In order to test the role of germline D sequence on the diversity of the preimmune TCRβ repertoire of T cells, we generated a mouse with a mutant TCRβ DJC locus wherein the Dβ2-Jβ2 gene segment cluster was deleted and the remaining diversity gene segment, Dβ1 (IMGT:TRDB1), was replaced with DSP2.3 (IMGT:IGHD2-02), a commonly used B cell immunoglobulin D H gene segment. Crystallographic studies have shown that the length and thus structure of TCR CDR-B3 places amino acids at the tip of CDR-B3 in a position to directly interact with peptide bound to an MHC molecule. The length distribution of complementarity determining region 3 of the T cell receptor beta chain (CDR-B3) has been proposed to be restricted largely by MHC-specific selection, disfavoring CDR-B3 that are too long or too short. Here we show that the mechanism of control of CDR-B3 length depends on the Dβ sequence, which in turn dictates exonucleolytic nibbling. By contrast, the extent of N addition and the variance of created CDR3 lengths are regulated by the cell of origin, the thymocyte. We found that the sequence of the D and control of N addition collaborate to bias the distribution of CDR-B3 lengths in the pre-immune TCR repertoire and to focus the diversity provided by N addition and the sequence of the D on that portion of CDR-B3 that is most likely to interact with the peptide that is bound to the presenting MHC.


Siah2 regulates stability of prolyl-hydroxylases, controls HIF1alpha abundance, and modulates physiological responses to hypoxia.
Hypoxia-inducible factor-1alpha (HIF1alpha) is a central regulator of the cellular response to hypoxia. Prolyl-hydroxylation of HIF1alpha by PHD enzymes is prerequisite for HIF1alpha degradation. Here, we demonstrate that the abundance of PHD1 and PHD3 are regulated via their targeting for proteasome-dependent degradation by the E3 ubiquitin ligases Siah1a/2, under hypoxia conditions. Siah2 null fibroblasts exhibit prolonged PHD3 half-life, resulting in lower levels of HIF1alpha expression during hypoxia. Significantly, hypoxia-induced HIF1alpha expression was completely inhibited in Siah1a/2 null cells, yet could be rescued upon inhibition of PHD3 by RNAi. Siah2 targeting of PHD3 for degradation increases upon exposure to even mild hypoxic conditions, which coincides with increased Siah2 transcription. Siah2 null mice subjected to hypoxia displayed an impaired hyperpneic respiratory response and reduced levels of hemoglobin. Thus, the control of PHD1/3 by Siah1a/2 constitutes another level of complexity in the regulation of HIF1alpha during hypoxia.


Retrievals of aerosol optical depth and Angström exponent from ground-based Sun-photometer data of Singapore.
The role of aerosols in climate and climate change is one of the factors that is least understood at the present. Aerosols' direct interaction with solar radiation is a well understood mechanism that affects Earth's net radiative forcing. However, quantifying its magnitude is more problematic because of the temporal and spatial variability of aerosol particles. To enhance our understanding of the radiative effects of aerosols on the global climate, Singapore has joined the AERONET (Aerosol Robotic Network) worldwide network by contributing ground-based direct Sun measurements performed by means of a multiwavelength Sun-photometer instrument. Data are collected on an hourly basis, then are uploaded to be fully screened and quality assured by AERONET. We use a one year data record (level 1.5/2.0) of measured columnar atmospheric optical depth, spanning from November 2006 to October 2007, to study the monthly and seasonal variability of the aerosol optical depth and the Angström exponent. We performed independent retrievals of these parameters (aerosol optical depth and Angström exponent) by using the photometer's six available bands covering the near-UV to near-IR (380-1080 nm). As a validation, our independent retrievals were compared with AERONET 1.5/2.0 level direct Sun product.


Knowledge sharing networks related to hospital quality measurement and reporting.
With the growing momentum toward hospital quality reporting by public payers, hospitals face increasing pressures to improve their medical record documentation and administrative data coding performance. The literature on "professional complex systems" has put forth various strategies for improving the performance of professional organizations. In doing so, it has emphasized the importance of creating effective structures for knowledge sharing and organizational learning. This study integrates knowledge networks and professional organizations literatures to develop hypotheses related to knowledge sharing network effectiveness in professional organizations. Correspondingly, this study explores the relationship between the organizational knowledge sharing structure related to quality and hospital coding performance related to quality. Simultaneously, this study seeks to identify other organizational characteristics associated with coding for quality measurement. The purpose is to identify strategies not only for improving hospital coding performance but also for the organization to adapt to the changing environment. An exploratory and comparative research design is used. The sample is composed of four hospitals, two showing "good-coding" performance for quality measurement and two showing "poor-coding" performance. Interviews and surveys are conducted with administrators and staff in the quality, medical staff, and coding subgroups in each facility. Survey data are subjected to social network analysis to examine knowledge sharing structures. This study finds that good-coding performance is systematically associated with a knowledge sharing network structure rich in brokerage and hierarchy (with leaders connecting different professional subgroups to each other and to the external environment) rather than in density (where everyone is directly connected to everyone else). From a health care management perspective, this study suggests that to improve hospital coding performance, senior administrators must undertake proactive and unceasing efforts to coordinate knowledge exchange across physician and coding subgroups and connect these subgroups with the changing external environment.


Randomized phase II designs.
As the use of molecularly targeted agents, which are anticipated to increase overall survival (OS)and progression-free survival (PFS) but not necessarily tumor response, has increased in oncology, there has been a corresponding increase in the recommendation and use of randomized phase II designs. Such designs reduce the potential for bias, existent in comparisons with historical controls, but also substantially increase the sample size requirements. We review the principal statistical designs for historically controlled and randomized phase II trials, along with their advantages, disadvantages, and statistical design considerations. We review the arguments for and against the use of randomization in phase II studies, the situations in which the use of historical controls is preferred, and the situations in which the use of randomized designs is preferred. We review methods used to calculate predicted OS or PFS values from historical controls, adjusted so as to be appropriate for an experimental sample with particular prognostic characteristics. We show how adjustment of the type I and type II error bounds for randomized studies can facilitate the detection of appropriate target increases in median PFS or OS with sample sizes appropriate for phase II studies. Although there continue to be differences among investigators concerning the use of randomization versus historical controls in phase II trials, there is agreement that each approach will continue to be appropriate, and the optimal approach will depend upon the circumstances of the individual trial.


Family medicine obstetrics. Collaborative interdisciplinary program for a declining resource.
A continuing decline in the number of family physicians in Canada providing obstetric, and particularly intrapartum, care. The Maternity Centre of Hamilton in Ontario was a pilot project initiated to help family physicians provide full obstetric care through a collaborative interdisciplinary model and shared call. Eleven family physicians provided care in collaboration with a nurse practitioner and other health professionals. Women came from the Maternity Centre's own practices, community physicians, or agencies, or through self-referral. More than a quarter of the women were considered psychosocially high-risk patients. Key features of the program included interdisciplinary collaboration and information technology that supported prenatal and birth documentation. The program has helped family physicians, and even recruited some, to practise full obstetric care and has provided high-quality, accessible services to pregnant women. Physicians experienced increased jab and personal satisfaction, and patients were highly satisfied with the service.


Insulin delivery rate into plasma in normal and diabetic subjects.
Removal of insulin-(131)I from plasma was studied in normal and diabetic subjects with both single injection and continuous infusion of isotope techniques. Patients were studied either in the fasting state or during steady-state hyperglycemia produced by a continuous intravenous glucose infusion. Steady-state plasma insulin concentration during these studies ranged from 10 to 264 muU/ml. Labeled insulin specific activity time curves consisted of more than one exponential, indicating that a multicompartmental system for insulin metabolism exists. A mathematical technique which is applicable to non-first order processes was used to calculate the rate at which insulin was lost irreversibly from the plasma insulin pool. A direct, linear relationship was found between insulin irreversible loss rate and plasma insulin concentration over the range of concentrations studied. This linearity implies lack of saturability of the insulin removal mechanism. Since the plasma insulin pool was in a steady state during these studies, insulin irreversible loss rate was equal to the rate at which newly secreted insulin was being delivered to the general circulation. Therefore, these results indicate that changes in plasma insulin concentration result from parallel changes in the rate of insulin delivery and not from changes in the opposite direction of the rate of insulin removal. A wide range of insulin delivery rates was found among patients with similar plasma glucose concentrations, suggesting that there exists considerable variability in responsiveness to endogenous insulin among these patients.


Prevalent genotypes of meticillin-resistant Staphylococcus aureus: report from Pakistan.
Meticillin-resistant Staphylococcus aureus (MRSA) is a major nosocomial pathogen in Pakistan and is emerging in the community. This is one of the first reports of the prevalent genotypes of MRSA in both hospital and community settings in Pakistan. Isolates collected in 2006-2007 were characterized by PFGE, staphylococcal cassette chromosome mec (SCCmec) typing and multilocus sequence typing (MLST). PFGE identified nine pulsotypes, the majority of isolates belonging to pulsotypes A (n=70) and B (n=38), which were predominant among hospital-onset MRSA (HO-MRSA) and community-onset MRSA (CO-MRSA) isolates, respectively. Among the HO-MRSA isolates, variants of SCCmec type III were prevalent, whilst SCCmec type IV or variants were predominant in the CO-MRSA isolates. MLST identified two principal sequence types, ST8 and ST239. An association was observed between ST8, PFGE pulsotype B and SCCmec type IV in the CO-MRSA (ST8-MRSA-IV). Similarly, ST239, PFGE pulsotype A and SCCmec type III were associated with HO-MRSA (ST239-MRSA-III). Therefore, the prevalent genotypes circulating in Pakistan at the time of study were ST8-MRSA-IV and ST239-MRSA-III in the community and hospital settings, respectively. A set of HO-MRSA isolates collected in 1997 were characterized by PFGE and SCCmec typing for comparison. The isolates belonged to two PFGE pulsotypes (A, n=28; B, n=11) and contained just two SCCmec types. These results suggest that an increase in genetic diversity occurred over the period 1997-2007 as a result of either microevolution or the importation of strains from surrounding areas.


Identification of retinoyl complexes as the autofluorescent component of the neuronal storage material in Batten disease.
Cytosomes filled with intensely fluorescent material in the form of curvilinear bodies were isolated by density gradient centrifugation followed by pronase digestion from the cerebral cortex of a child who had died at age 7 from the late infantile form of Batten disease. Forty-three percent of the dry weight of the storage material was extracted by a mixture of chloroform and methanol, leaving a waterinsoluble amorphous fluorescent residue. Infrared spectroscopy, proton magnetic resonance spectrscopy, and mass spectrometry of this residue strongly suggested the presence of retinoyl polyenes linked to a small peptide. Base hydrolysis and methanolysis yielded retinoic acid and methyl retinoate, respectively. Ozonolysis yielded a product derived from the substituted cyclohexenyl ring of vitamin A. The results indicate that the fluorescent component of the neuronal storage material is a retinoyl complex and is not derived from peroxidized polyunsatured fatty acids as previously thought.


Tissue regeneration and biomineralization in sea urchins: role of Notch signaling and presence of stem cell markers.
Echinoderms represent a phylum with exceptional regenerative capabilities that can reconstruct both external appendages and internal organs. Mechanistic understanding of the cellular pathways involved in regeneration in these animals has been hampered by the limited genomic tools and limited ability to manipulate regenerative processes. We present a functional assay to investigate mechanisms of tissue regeneration and biomineralization by measuring the regrowth of amputated tube feet (sensory and motor appendages) and spines in the sea urchin, Lytechinus variegatus. The ability to manipulate regeneration was demonstrated by concentration-dependent inhibition of regrowth of spines and tube feet by treatment with the mitotic inhibitor, vincristine. Treatment with the gamma-secretase inhibitor DAPT resulted in a concentration-dependent inhibition of regrowth, indicating that both tube feet and spine regeneration require functional Notch signaling. Stem cell markers (Piwi and Vasa) were expressed in tube feet and spine tissue, and Vasa-positive cells were localized throughout the epidermis of tube feet by immunohistochemistry, suggesting the existence of multipotent progenitor cells in these highly regenerative appendages. The presence of Vasa protein in other somatic tissues (e.g. esophagus, radial nerve, and a sub-population of coelomocytes) suggests that multipotent cells are present throughout adult sea urchins and may contribute to normal homeostasis in addition to regeneration. Mechanistic insight into the cellular pathways governing the tremendous regenerative capacity of echinoderms may reveal processes that can be modulated for regenerative therapies, shed light on the evolution of regeneration, and enable the ability to predict how these processes will respond to changing environmental conditions.


Revisiting the Global Epidemiology of Cholera in Conjuction With the Genomics of Vibrio cholerae.
Toxigenic Vibrio cholerae is responsible for 1.4 to 4.3 million cases with about 21,000-143,000 deaths per year. Dominance of O1 and O139 serogroups, classical and El tor biotypes, alterations in CTX phages and the pathogenicity Islands are some of the major features of V. cholerae isolates that are responsible for cholera epidemics. Whole-genome sequencing (WGS) based analyses of single-nucleotide polymorphisms (SNPs) and other infrequent genetic variants provide a robust phylogenetic framework. Recent studies on the global transmission of pandemic V. cholerae O1 strains have shown the existence of eight different phyletic lineages. In these, the classical and El Tor biotype strains were separated as two distinctly evolved lineages. The frequency of SNP accumulation and the temporal and geographical distribution supports the perception that the seventh cholera pandemic (7CP) has spread from the Bay of Bengal region in three independent but overlapping waves. The 2010 Haitian outbreak shared a common ancestor with South-Asian wave-3 strains. In West Africa and East/Southern Africa, cholera epidemics are caused by single expanded lineage, which has been introduced several times since 1970. The Latin American epidemics that occurred in 1991 and 2010 were the result of introductions of two 7CP sublineages. Sublineages representing wave-3 have caused huge outbreaks in Haiti and Yemen. The Ogawa-Inaba serotype switchover in several cholera epidemics are believed to be due to the involvement of certain selection mechanism(s) rather than due to random events. V. cholerae O139 serogroup is phylogenetically related to the 7CP El Tor, and almost all these isolates belonged to the multilocus sequence type-69. Additional phenotypic and genotypic information have been generated to understand the pathogenicity of classical and El Tor vibrios. Presence of integrative conjugative elements (ICE) with antibiotic resistance gene cassettes, clustered regularly interspaced short palindromic repeats-associated protein system and ctxAB promoter based ToxRS expression of cholera toxin (CT) separates classical and El Tor biotypes. With the availability of WGS information, several important applications including, molecular typing, antimicrobial resistance, new diagnostics, and vaccination strategies could be generated.


Treatment of an open medial tibiotalar dislocation with no associated fracture.
Tibiotalar dislocations without associated fractures are very uncommon in adults, and only a few studies have been published regarding this injury. More than 50% of these dislocations will be posteromedial, with a high incidence of open injuries, and 25% are pure posterior dislocations. In the present report, we discuss our experience and management of a medial tibiotalar dislocation with no associated fracture. In the present case, the patient was brought to the operating room on presentation to our facility and underwent irrigation and debridement with primary closure of his wound. He was immobilized postoperatively. The patient tolerated the operation well and did not sustain any postoperative complications. He was able to regain function of the injured extremity until he was lost to follow-up. Regarding treatment, the surgery should be speedy, gentle to the soft tissue, and with as little implanted material as possible. Although we do not advocate that our management and treatment of this injury become the standard, the present case provides a good example of some of the challenges often encountered when treating these injuries.


The Diagnostic Accuracy of Noninvasive Methods to Measure the Intracranial Pressure: A Systematic Review and Meta-analysis.
Although invasive monitoring is the standard method for intracranial pressure (ICP) measurement, it is not without potential for serious complications. Noninvasive methods have been proposed as alternatives to invasive ICP monitoring. The study aimed to investigate the diagnostic accuracy of the currently available noninvasive methods for intracranial hypertension (ICH) monitoring. We searched 5 databases for articles evaluating the diagnostic accuracy of noninvasive methods in diagnosing ICH in PubMed, Institute of Science Index, Scopus, Cochrane Central Register of Controlled Trials (CENTRAL), and Embase. The quantitative analysis was conducted if there were at least 2 studies evaluating a specific method. The accuracy measures included the sensitivity, specificity, likelihood ratios, and diagnostic odds ratio. We included 134 articles. Ultrasonographic optic nerve sheath diameter (US ONSD) had high diagnostic accuracy (estimated sensitivity of 90%; 95% confidence interval [CI], 87-92, estimated specificity of 88%; 95% CI, 84-91) while the magnetic resonance imaging (MRI) ONSD had estimated sensitivity of 77%; 95% CI, 64-87 and estimated specificity of 89%; 95% CI, 84-93, and computed tomography (CT) ONSD had estimated sensitivity of 93%; 95% CI, 90-96 and estimated specificity of 79%; 95% CI, 56-92. All MRI signs had a very high estimated specificity ranging from 90% to 99% but a low estimated sensitivity except for sinus stenosis which had high estimated sensitivity as well as specificity (90%; 95% CI, 75-96 and 96%; 95% CI, 91-99, respectively). Among the physical examination signs, pupillary dilation had a high estimated specificity (86%; 95% CI, 76-93). Other diagnostic tests to be considered included pulsatility index, papilledema, transcranial Doppler, compression or absence of basal cisterns, and ≥10 mm midline shift. Setting the cutoff value of ICH to ≥20 mm Hg instead of values <20 mm Hg was associated with higher sensitivity. Moreover, if the delay between invasive and noninvasive methods was within 1 hour, the MRI ONSD and papilledema had a significantly higher diagnostic accuracy compared to the >1 hour subgroup. Our study showed several promising tools for diagnosing ICH. Moreover, we demonstrated that using multiple, readily available, noninvasive methods is better than depending on a single sign such as physical examination or CT alone.


Invited Commentary: Methods for Estimating Effects of Minimum Wages on Health.
Economists have been researching effects of minimum wages on unemployment, poverty, income inequality, and educational attainment for over 60 years. Epidemiologists have only recently begun researching minimum wages even though unemployment through education are central topics within social epidemiology. Buszkiewicz et al. (Am J Epidemiol. 2021;190(1):21-30) offer a welcome addition to this nascent literature. A commanding advantage of Buszkiewicz et al.'s study over others is its distinction between a "likely affected" group comprised of workers with ≤12 years of schooling versus "not likely affected" groups with ≥13 years of schooling. But there are disadvantages, common to other studies. Buszkiewicz et al. use cross-sectional data; they include the self-employed as well as part-time and part-year workers in their treatment groups. Their definitions of affected groups based on education create samples with 75% or more of workers who earn significantly above minimum wages; definitions are not based on wages. Inclusion of workers not subject to (e.g., self-employed) or affected by minimum wages biases estimates toward the null. Finally, within any minimum wage data set, it is the state-not federal-increases that account for the lion's share of increases and that form the natural experiments; however, state increases can occur annually whereas the development of chronic diseases might take decades.


Bariatric surgery: Asia-Pacific perspective.
There is a world-wide epidemic of overweight, obesity and morbid obesity. Bariatric surgery today, as the only effective therapy for morbid obesity, is expanding exponentially to meet the global epidemic of morbid obesity. Bariatric surgeons in the Asia-Pacific region had founded the Asia-Pacific Bariatric Surgery Group (APBSG) at Seoul, Korea on October 6, 2004. E-mail requests for information were sent to the national bariatric surgery leaders. These requests were followed, if necessary, by second e-mail requests and communications seeking clarification. The summary data was also discussed at the 1st Asia-Pacific Bariatric Consensus Meeting held in Taipei, February 27, 2005. 11 countries or areas in Asia had started bariatric surgery and responded to the general questions. In 2004, 636 bariatric operations were performed by 61 bariatric surgeons. The earliest data for starting bariatric surgery was in 1974 in Taiwan. Following the development of gastric partition, Taiwan performed the first case in 1981, Japan in 1982 and Singapore in 1987. In 2004, 11 countries have started bariatric surgery. The APBSG was founded in 2004. In 2004, 12.1% of operations were open and 87.9% laparoscopic. The 6 most popular operations were: laparoscopic adjustable banding 42.3%; laparoscopic gastric bypass 34.2%; open vertical banded gastroplasty 7.5%; laparoscopic vertical banded gastroplasty 6.3%; laparoscopic sleeve gastrectomy 6.3%; open gastric bypass 4.2%. Pooling open and laparoscopic procedures, relative percentages were gastric banding 42.3%; gastric bypass 38.4%; vertical banded gastroplasty 13.8%. The APBSG consensus meeting recommended bariatric surgery in Asian patients with BMI >37 or >32 with diabetes or two other obesity-related co-morbidities. Bariatric surgery is expanding rapidly in Asia to meet rapidly increasing obesity. The modification of the indications for bariatric surgery in the Asian is proposed.


Cohort Profile: The Barwon Infant Study.
The modern environment is associated with an increasing burden of non-communicable diseases (NCDs). Mounting evidence implicates environmental exposures, experienced early in life (including in utero), in the aetiology of many NCDs, though the cellular/molecular mechanism(s) underlying this elevated risk across the life course remain unclear. Epigenetic variation has emerged as a candidate mediator of such effects. The Barwon Infant Study (BIS) is a population-derived birth cohort study (n = 1074 infants) with antenatal recruitment, conducted in the south-east of Australia (Victoria). BIS has been designed to facilitate a detailed mechanistic investigation of development within an epidemiological framework. The broad objectives are to investigate the role of specific environmental factors, gut microbiota and epigenetic variation in early-life development, and subsequent immune, allergic, cardiovascular, respiratory and neurodevelopmental outcomes. Participants have been reviewed at birth and at 1, 6, 9 and 12 months, with 2- and 4-year reviews under way. Biological samples and measures include: maternal blood, faeces and urine during pregnancy; infant urine, faeces and blood at regular intervals during the first 4 years; lung function at 1 month and 4 years; cardiovascular assessment at 1 month and 4 years; skin-prick allergy testing and food challenge at 1 year; and neurodevelopmental assessment at 9 months, 2 and 4 years. Data access enquiries can be made at [www.barwoninfantstudy.org.au] or via [firstname.lastname@example.com].


Modelling protection from antimicrobial agents in biofilms through the formation of persister cells.
A mathematical model of biofilm dynamics was used to investigate the protection from antimicrobial killing that could be afforded to micro-organisms in biofilms based on a mechanism of 'persister' cell or phenotypic variant formation. The persister state is a hypothetical, highly protected state adopted by a small fraction of the cells in a biofilm. Persisters were assumed to be generated at a fixed rate, independent of the presence of substrate or antimicrobial agent. Cells were assumed to revert from the persister state when exposed to the growth substrate. Persister cells were assumed to be incapable of growth. The model predicted that persister cells increased in numbers in the biofilm, even though they were unable to grow, accumulating in regions of substrate limitation. In these regions, normal cells failed to grow, but did slowly convert to the persister state. Calculations of persister formation in planktonic cultures predicted that persisters would be present in low numbers in growing cultures, but should accumulate under conditions of slow growth, e.g. very low dilution rates in continuous culture or stationary phase in batch culture. When antibiotic treatment was simulated, bacteria near the biofilm surface were killed, but persisters in the depth of the biofilm were poorly killed. After antibiotic treatment ceased, surviving persister cells quickly reverted and allowed the biofilm to regrow. This modelling study provides motivation for further investigation of the hypothetical persister cell state as an explanation for biofilm resistance to antimicrobial agents.


Aortic valve endocarditis complicated by proximal false aneurysm.
Aortic valve endocarditis remains a life-threatening condition, especially in cases of periannular complications. Aorto-ventricular discontinuity associated with proximal false aneurysm represents a severe picture caused by extensive tissue disruption and is usually associated with prosthetic valve infection. Complex surgical repair is required in these cases and continues to be associated with high mortality and morbidity rates. We retrieved information for 32 patients undergoing operation for infective aortic valve/prosthetic valve endocarditis complicated by pseudoaneurysm arising from aorto-ventricular discontinuity. Patients were relatively young, mostly male and most of them had a prior cardiac operation. Aortic root replacement with valve graft conduit was performed in all cases; it was associated with other procedures in seven patients: CABG (n=2), MV surgery (n=3), MV surgery + CABG (n=1) and pulmonary valve replacement (n=1). We reported and analysed patient outcomes at early and mid-term follow-up. Pre-discharge mortality was 22% (n=7). The postoperative course was complicated in 24 (75%) cases. Eighteen patients (56%) sustained low cardiac output resulting in multiple organ failure syndrome and death in five cases. One patient (3%) experienced a major neurologic deficit with a permanent cerebral stroke. Acute kidney injury complicated the course in 12 cases (37%), continuous renal replacement therapy was necessary in four patients (12%). Overall survival and freedom from endocarditis and reoperation at 5-year was 59% and 89%, respectively. Patients with complicated aortic valve endocarditis presented generally in a poor preoperative state. Surgical treatment poses a non-negligible risk of postoperative mortality and morbidity but provides an acceptable survival rate and a satisfactory recovery at mid-term.


Catalytic resonance of ammonia synthesis by simulated dynamic ruthenium crystal strain.
Ammonia affords dense storage for renewable energy as a fungible liquid fuel, provided it can be efficiently synthesized from hydrogen and nitrogen. In this work, the catalysis of ammonia synthesis was computationally explored beyond the Sabatier limit by dynamically straining a ruthenium crystal (±4%) at the resonant frequencies (102 to 105+ Hz) of N2 surface dissociation and hydrogenation. Density functional theory calculations at different strain conditions indicated that the energies of NHx surface intermediates and transition states scale linearly, allowing the description of ammonia synthesis at a continuum of strain conditions. A microkinetic model including multiple sites and surface diffusion between step and Ru(0001) terrace sites of varying ratios for nanoparticles of differing size revealed that dynamic strain yields catalytic ammonia synthesis conversion and turnover frequency comparable to industrial reactors (400°C, 200 atm) but at lower temperature (320°C) and an order of magnitude lower pressure (20 atm).


Increased Long-Term Mortality among Black CABG Patients Receiving Preoperative Inotropic Agents.
The aim of this study was to examine racial differences in long-term mortality after coronary artery bypass grafting (CABG), stratified by preoperative use of inotropic agents. Black and white patients who required preoperative inotropic support prior to undergoing CABG procedures between 1992 and 2011 were compared. Mortality probabilities were computed using the Kaplan-Meier product-limit method. Hazard ratios (HR) and 95% confidence intervals (CI) were computed using a Cox regression model. A total of 15,765 patients underwent CABG, of whom 211 received preoperative inotropic agents within 48 hours of surgery. Long-term mortality differed by race (black versus white) among preoperative inotropic category (inotropes: adjusted HR = 1.6, 95% CI = 1.009-2.4; no inotropes: adjusted HR = 1.15, 95% CI = 1.08-1.2; P(interaction) < 0.0001). Our study identified an independent preoperative risk-factor for long-term mortality among blacks receiving CABG. This outcome provides information that may be useful for surgeons, primary care providers, and their patients.


Natural and Experimental Host Range of Sirococcus clavigignenti-juglandacearum.
The fungus Sirococcus clavigignenti-juglandacearum causes butternut canker in North America and is not known to kill or seriously affect tree species other than butternut (Juglans cinerea). However, this putative exotic pathogen has been found on branches of black walnut (J. nigra) and heartnut (J. ailantifolia var. cordiformis), raising questions regarding the potential host range of the pathogen and its spread to new areas on tree hosts other than butternut. Results of artificial inoculations of several hardwood species indicated that the pathogen, although not causing lethal cankers, has the ability to colonize and survive in the wood of several genera, including Quercus, Carya, Corylus, Prunus, and Castanea. Several commercially important Persian walnut (J. regia) cultivars tested were moderately or highly susceptible, underscoring the importance of preventing the movement of the pathogen into commercial Juglans plantations in California, where its ability to cause disease and damage trees is unknown. Evidence for the presence of variation in aggressiveness among isolates of S. clavigignenti-juglandacearum was detected based on differences in canker lengths resulting from inoculations with two isolates of the pathogen. In a field test of several previously named butternut cultivars, all were found to be highly susceptible to the disease, calling into question the value of these selections for nut production or future restoration uses.


Metabolic rate does not calibrate the molecular clock.
Rates of molecular evolution vary widely among lineages, but the causes of this variation remain poorly understood. It has been suggested that mass-specific metabolic rate may be one of the key factors determining the rate of molecular evolution, and that it can be used to derive "corrected" molecular clocks. However, previous studies have been hampered by a paucity of mass-specific metabolic rate data and have been largely limited to vertebrate taxa. Using mass-specific metabolic rate measurements and DNA sequence data for >300 metazoan species for 12 different genes, we find no evidence that mass-specific metabolic rate drives substitution rates. The mechanistic basis of the metabolic rate hypothesis is discussed in light of these findings.


Activation of coagulation in bullous pemphigoid and other eosinophil-related inflammatory skin diseases.
Bullous pemphigoid (BP) is a skin disease caused by autoantibodies to hemidesmosomal proteins BP180 and BP230, with eosinophils participating in blister formation. Tissue factor (TF), the initiator of coagulation, is embodied within the eosinophil granules and exposed upon activation. We evaluated the coagulation activation in patients with BP (63), chronic urticaria (CU; 20), atopic dermatitis (AD; 14), cutaneous drug reactions (CDRs; six), psoriasis (20), dermatitis herpetiformis (DH; four) and primary cutaneous T cell lymphoma (CTCL; five), and in 40 healthy controls. Prothrombin fragment F1+2 and d-dimer (coagulation markers) were measured by enzyme-linked immunosorbent assay (ELISA) in all plasma samples and BP blister fluid. Skin TF expression was evaluated immunohistochemically in the patients and 20 controls. F1+2 and d-dimer levels were higher in BP plasma than in control plasma (P = 0·0001 for both), and dramatically high in blister fluid; both correlated positively with disease severity, esinophil counts and anti-BP180 antibodies (P = 0·006-0·0001). Plasma F1+2 and d-dimer levels were higher in the CU, AD and CDR patients than in controls (P = 0·0001 for all), but normal in the psoriasis, DH and CTCL patients. Skin TF was expressed in the BP (P = 0·0001), CU (P = 0·0001), AD (P = 0·001) and CDR patients (P = 0·01), but not in the psoriasis, DH or CTCL patients. Co-localization confocal microscopy studies confirmed eosinophils as the source of TF in 10 BP patients. The coagulation cascade is activated in BP and other eosinophil-mediated skin disorders, but not in non-eosinophil driven conditions. This hypercoagulability may contribute to inflammation, tissue damage and, possibly, thrombotic risk.


Thalidomide exerts its inhibitory action on tumor necrosis factor alpha by enhancing mRNA degradation.
We have examined the mechanism of thalidomide inhibition of lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF-alpha) production and found that the drug enhances the degradation of TNF-alpha mRNA. Thus, the half-life of the molecule was reduced from approximately 30 to approximately 17 min in the presence of 50 micrograms/ml of thalidomide. Inhibition of TNF-alpha production was selective, as other LPS-induced monocyte cytokines were unaffected. Pentoxifylline and dexamethasone, two other inhibitors of TNF-alpha production, are known to exert their effects by means of different mechanisms, suggesting that the three agents inhibit TNF-alpha synthesis at distinct points of the cytokine biosynthetic pathway. These observations provide an explanation for the synergistic effects of these drugs. The selective inhibition of TNF-alpha production makes thalidomide an ideal candidate for the treatment of inflammatory conditions where TNF-alpha-induced toxicities are observed and where immunity must remain intact.


Single nucleotide polymorphism (SNP) variation of wolves (Canis lupus) in Southeast Alaska and comparison with wolves, dogs, and coyotes in North America.
There is considerable interest in the genetics of wolves (Canis lupus) because of their close relationship to domestic dogs (C. familiaris) and the need for informed conservation and management. This includes wolf populations in Southeast Alaska for which we determined genotypes of 305 wolves at 173662 single nucleotide polymorphism (SNP) loci. After removal of invariant and linked SNP, 123801 SNP were used to quantify genetic differentiation of wolves in Southeast Alaska and wolves, coyotes (C. latrans), and dogs from other areas in North America. There is differentiation of SNP allele frequencies between the species (wolves, coyotes, and dogs), although differentiation is relatively low between some wolf and coyote populations. There are varying levels of differentiation among populations of wolves, including low differentiation of wolves in interior Alaska, British Columbia, and the northern US Rocky Mountains. There is considerable differentiation of SNP allele frequencies of wolves in Southeast Alaska from wolves in other areas. However, wolves in Southeast Alaska are not a genetically homogeneous group and there are comparable levels of genetic differentiation among areas within Southeast Alaska and between Southeast Alaska and other geographic areas. SNP variation and other genetic data are discussed regarding taxonomy and management.


Brain insulin, energy and glucose homeostasis; genes, environment and metabolic pathologies.
The central nervous system is essential in maintaining energy and glucose homeostasis. In both animals and humans, efficient cerebral insulin signalling is a pivotal control element in these pathophysiological processes. The action of insulin in the brain is under a multilevel control via metabolic, endocrine and neural signals induced by nutrients, integrated mainly by the hypothalamus. Of particular interest is the interaction of insulin with the anabolic and catabolic neuroregulators. The anorexic peptides insulin, leptin and the neurotransmitter serotonin share common signalling pathways involved in food intake, in particular the insulin receptor substrate, phosphatidylinositol-3-kinase (PI3K) pathway. The dialogue of neurotransmitters and peptides via this signalling pathway is potentially of major importance in the pathophysiology of the brain in general and specifically in the regulation of feeding behaviour. At this time, a new concept in the aetiopathology of type 2 diabetes is immerging. This concept proposes that the combination of defective pancreatic beta-cell function and insulin resistance not only in classical insulin target tissues but in every tissue, contributes to the onset of the disease. It highlights the importance of the disruption of cerebral insulin signal transmission and its direct relation to metabolic diseases. Impaired brain insulin signalling, a link coupling obesity to diabetes, may be related to either genetic factors, or environmental factors such as stress, over or under-feeding and unbalanced diets: such factors may work either independently or in concert. Current approaches used for the prevention and treatment of type 2 diabetes are not adequately effective. Most of the anti-diabetic therapies induce many adverse effects, in particular obesity, and thus may initiate a vicious cycle of problems. In order to develop new, more efficient, preventive and therapeutic strategies for metabolic pathologies, there is an urgent need for increased understanding of the complexity of insulin signalling in the brain and on the interactive, central and peripheral effects of insulin.


[Malignant histiocytosis of pulmonary form].
Malignant histiocytosis beginning by a unique pulmonary location is most unusual. The case reported here has seemed worthy because of its strange clinical aspect, the unexpected diagnosis, and because of the way it answered completely and rapidly to the following chemotherapy: Adriamycin 50 mg/m2, Cyclophosphamide 750 mg/m2, Vincristine 2 mg one day every three weeks associated to Prednisone 100 mg/m2 the five first days. A study of literature has allowed us to review all pleuro-pulmonary location initiating malignant histiocytosis and to stress out the errors of diagnosis that can outcome in this case.


Transmissible strains of Pseudomonas aeruginosa in cystic fibrosis lung infections.
Pseudomonas aeruginosa chronic lung infections are the major cause of morbidity and mortality associated with cystic fibrosis. For many years, the consensus was that cystic fibrosis patients acquire P. aeruginosa from the environment, and hence harbour their own individual clones. However, in the past 15 yrs the emergence of transmissible strains, in some cases associated with greater morbidity and increased antimicrobial resistance, has changed the way that many clinics treat their patients. Here we provide a summary of reported transmissible strains in the UK, other parts of Europe, Australia and North America. In particular, we discuss the prevalence, epidemiology, unusual genotypic and phenotypic features, and virulence of the most intensively studied transmissible strain, the Liverpool epidemic strain. We also discuss the clinical impact of transmissible strains, in particular the diagnostic and infection control approaches adopted to counter their spread. Genomic analysis carried out so far has provided little evidence that transmissibility is due to shared genetic characteristics between different strains. Previous experiences with transmissible strains should help us to learn lessons for the future. In particular, there is a clear need for strain surveillance if emerging problem strains are to be detected before they are widely transmitted.


Preliminary evaluation of novel skin closure of Pfannenstiel incisions using cold helium plasma and chitosan films.
To assess the safety and performance of a new energy-based skin closure system (BioWeld1(TM)) for the surgical Pfannenstiel incision in patients scheduled for elective cesarean section. This prospective, single center, non-randomized study included 20 patients who were scheduled for elective cesarean section. The BioWeld1 system was performed after suturing the internal layers of the cesarean section incision. A clinical evaluation of safety and efficacy was performed for 1, 2, 4-7, 21, and 45 d after the procedure. The Vancouver Scar Scale (VSS) was used to evaluate scarring. Up to 21 d after the procedure, no safety device-related adverse events were reported. All patients had full closure of the epidermis, a very low total VSS score, and no evidence of discharge, redness, edema, or thermal damage. None of the patients exhibited more than a mild degree of encrustation. The BioWeld1 System has been shown to be safe and effective for skin closure in cesarean section.


Intravenous iron supplementation with intra-articular administration of tranexamic acid reduces the rate of allogeneic transfusions after simultaneous bilateral total knee arthroplasty.
Peri-operative intravenous administration of iron supplementation seems a good option to reduce allogeneic blood transfusion in major orthopaedic surgery. However, its efficacy in simultaneous bilateral total knee arthroplasty has not been studied. From December 2014 to May 2015, a total of 72 consecutive patients underwent simultaneous bilateral total knee arthroplasty and received peri-operative intravenous iron supplementation (iron isomaltoside 1000: 600 mg pre-operatively and 400 mg 1 week post-operatively) and intra-articular tranexamic acid (2 g in 20 mL saline at the end of surgery), and were managed with a restrictive transfusion trigger (haemoglobin <7 g/dL). Post-operatively, we observed patients closely for symptoms of anaemia and checked their haemoglobin levels on days 1, 6 and 13 after surgery. The mean baseline haemoglobin level was 13.1 g/dL. The levels remained above 7.0 g/dL on post-operative days 1, 6 and 13 (mean, 11.4 g/dL, 9.9 g/dL and 10.4 g/dL, respectively) in all but one patient who experienced melaena and required allogeneic blood transfusion. Intravenous iron supplementation combined with intra-articular administration of tranexamic acid seems to be an effective strategy for reducing the rate of allogeneic blood transfusion in patients undergoing simultaneous bilateral total knee arthroplasty managed with a restrictive transfusion trigger.


Effect of wind farms on wintering ducks at an important wintering ground in China along the East Asian-Australasian Flyway.
Wind farms offer a cleaner alternative to fossil fuels and can mitigate their negative effects on climate change. However, wind farms may have negative impacts on birds. The East China Coast forms a key part of the East Asian-Australasian Flyway, and it is a crucial region for wind energy development in China. However, despite ducks being the dominant animal taxon along the East China Coast in winter and considered as particularly vulnerable to the effects of wind farms, the potential negative impacts of wind farms on duck populations remain unclear. We therefore assessed the effects of wind farms on duck abundance, distribution, and habitat use at Chongming Dongtan, which is a major wintering site for ducks along the East Asian-Australasian Flyway, using field surveys and satellite tracking. We conducted seven paired field surveys of ducks inside wind farm (IWF) and outside wind farm (OWF) sites in artificial brackish marsh, paddy fields, and aquaculture ponds. Duck abundance was significantly higher in OWF compared with IWF sites and significantly higher in artificial brackish marsh than in aquaculture ponds and paddy fields. Based on 1,918 high-resolution satellite tracking records, the main habitat types of ducks during the day and at night were artificial brackish marsh and paddy fields, respectively. Furthermore, grid-based analysis showed overlaps between ducks and wind farms, with greater overlap at night than during the day. According to resource selection functions, habitat use by wintering ducks was impacted by distance to water, land cover, human activity, and wind farm effects, and the variables predicted to have significant impacts on duck habitat use differed between day and night. Our study suggests that wintering ducks tend to avoid wind turbines at Chongming Dongtan, and landscape of paddy fields and artificial wetlands adjoining natural wetlands is crucial for wintering ducks.


Regional ecosystem structure and function: ecological insights from remote sensing of tropical forests.
Ecological studies in tropical forests have long been plagued by difficulties associated with sampling the crowns of large canopy trees and large inaccessible regions, such as the Amazon basin. Recent advances in remote sensing have overcome some of these obstacles, enabling progress towards tackling difficult ecological problems. Breakthroughs have helped transform the dialog between ecology and remote sensing, generating new regional perspectives on key environmental gradients and species assemblages with ecologically relevant measures such as canopy nutrient and moisture content, crown area, leaf-level drought responses, woody tissue and surface litter abundance, phenological patterns, and land-cover transitions. Issues that we address here include forest response to altered precipitation regimes, regional disturbance and land-use patterns, invasive species and landscape carbon balance.


Signaling the trustworthiness of science.
Trust in science increases when scientists and the outlets certifying their work honor science's norms. Scientists often fail to signal to other scientists and, perhaps more importantly, the public that these norms are being upheld. They could do so as they generate, certify, and react to each other's findings: for example, by promoting the use and value of evidence, transparent reporting, self-correction, replication, a culture of critique, and controls for bias. A number of approaches for authors and journals would lead to more effective signals of trustworthiness at the article level. These include article badging, checklists, a more extensive withdrawal ontology, identity verification, better forward linking, and greater transparency.


Unfavorable effects of passive smoking on aortic function in men.
The aorta acts as both a conduit and an elastic buffering chamber that modulates left ventricular function and coronary blood flow. Previous studies have shown that active smoking has unfavorable effects on aortic elasticity. To study the association between passive smoking and the elastic properties of the human aorta. Comparison of nonsmokers during passive smoking studies and smokers during active smoking or sham smoking studies. Academic medical center. 16 male nonsmokers were assigned to passive smoking studies, and 32 current, long-term, male smokers were randomly assigned to either active smoking (16 patients) or sham smoking (16 patients) studies. All participants underwent diagnostic catheterization. In the passive smoking group, environmental tobacco smoke was vented into an exposure chamber for 5 minutes (mean carbon monoxide level, 30 parts per million). Each participant in the active smoking group smoked one filtered cigarette (1.0 mg of nicotine) under standardized conditions within 5 minutes; each participant in the sham smoking group performed a similar pattern of inhalation with one unlit cigarette. Aortic elastic properties were studied by measuring the aortic pressure-diameter relation before and for 20 minutes after passive, active, or sham smoking. Instantaneous diameter of the thoracic aorta was measured with a high-fidelity ultrasonic dimension catheter. Instantaneous aortic pressure and diameter were measured at the same site. Both passive and active smoking were associated with changes in the aortic pressure-diameter relation (change in mean distensibility in the passive smoking group, from 2.02 to 1.59 x 10(-6) cm2 x dyne(-1) [for comparisons of time course between passive and sham smoking groups, P < 0.001]; change in mean distensibility in the active smoking group, from 2.08 to 1.51 x 10(-6) cm2 x dyne(-1) [for comparisons of time course between active and sham smoking groups, P < 0.001]). These changes represent decreases of 21% and 27%, respectively. No changes in aortic elasticity were seen in the sham smoking group. Both passive and active smoking are associated with an acute deterioration in the elastic properties of the aorta. This association between exposure to tobacco smoke and aortic elasticity indicates that aortic function deteriorates during passive or active smoking.


Challenges and opportunities in genetic improvement of local livestock breeds.
Sufficient genetic variation in livestock populations is necessary both for adaptation to future changes in climate and consumer demand, and for continual genetic improvement of economically important traits. Unfortunately, the current trend is for reduced genetic variation, both within and across breeds. The latter occurs primarily through the loss of small, local breeds. Inferior production is a key driver for loss of small breeds, as they are replaced by high-output international transboundary breeds. Selection to improve productivity of small local breeds is therefore critical for their long term survival. The objective of this paper is to review the technology options available for the genetic improvement of small local breeds and discuss their feasibility. Most technologies have been developed for the high-input breeds and consequently are more favorably applied in that context. Nevertheless, their application in local breeds is not precluded and can yield significant benefits, especially when multiple technologies are applied in close collaboration with farmers and breeders. Breeding strategies that require cooperation and centralized decision-making, such as optimal contribution selection, may in fact be more easily implemented in small breeds.


Isolation and Proteomic Analysis of Microvesicles and Exosomes from HT22 Cells and Primary Neurons.
Exosomes and microvesicles are extracellular vesicles (EVs) released by most cell types. The role of EVs as a method of intercellular communication has led to these vesicles becoming a major area of interest in a variety of scientific fields including neuroscience. Emerging evidence is now demonstrating that the biomolecular composition of EVs, especially exosomes, can play a role in the progression of disease including various neurodegenerative diseases and cancer. In addition to the miRNA profiles of EVs, these vesicles also show interesting changes in protein expression profiles under different physiological and pathological conditions. Characterization of these profiles could prove valuable for both understanding disease pathogenesis and for the discovery of new biomarkers of disease. In this chapter, we describe a protocol for isolation of exosomes and microvesicles from immortalized HT22 cells and primary cortical neurons with sufficient yield and low serum contamination required for downstream analysis and label-free relative quantitation by mass spectrometry.


Evaluation of Vasectomy Trends in the United States.
To use the Truven Health MarketScan database to better approximate the annual rate of vasectomies performed in the US population, to determine changes over time, regional differences, providers performing this, and to know if there is any monthly variation in vasectomy rates. Claims data were evaluated from 2007 to 2015 to determine the annual prevalence of vasectomy by patient age and region in the United States. The cohort included men aged 18-64 years with at least 1 claim in any given year in Truven Health MarketScan. Provider type and place of service were evaluated in 2014 and 2015. Monthly evaluation of vasectomy prevalence compared with total claims was performed. The prevalence of vasectomies decreased from 2007 to 2015, across all age groups and in all locations of the country (P <.001). Using these data and the most recent US census data, an estimated 527,476 vasectomies were performed in the United States in 2015. The North Central and West regions (0.64% and 0.60%, respectively) had the highest annual prevalence of vasectomies. The month of March and the end of the year had the highest proportion of vasectomies performed. In both 2014 and 2015, a urologist in the office setting performed 82% of vasectomies. An estimated 527,476 vasectomies were performed in the United States in 2015. From 2007 to 2015 there was a decrease in the proportion of vasectomies performed in all age groups and in all locations of the country. The end of the year and the month of March are when the most vasectomies are performed.


Carcinogenic Air Toxics Exposure and Their Cancer-Related Health Impacts in the United States.
Public health protection from air pollution can be achieved more effectively by shifting from a single-pollutant approach to a multi-pollutant approach. To develop such multi-pollutant approaches, identifying which air pollutants are present most frequently is essential. This study aims to determine the frequently found carcinogenic air toxics or hazardous air pollutants (HAPs) combinations across the United States as well as to analyze the health impacts of developing cancer due to exposure to these HAPs. To identify the most commonly found carcinogenic air toxics combinations, we first identified HAPs with cancer risk greater than one in a million in more than 5% of the census tracts across the United States, based on the National-Scale Air Toxics Assessment (NATA) by the U.S. EPA for year 2005. We then calculated the frequencies of their two-component (binary), and three-component (ternary) combinations. To quantify the cancer-related health impacts, we focused on the 10 most frequently found HAPs with national average cancer risk greater than one in a million. Their cancer-related health impacts were calculated by converting lifetime cancer risk reported in NATA 2005 to years of healthy life lost or Disability-Adjusted Life Years (DALYs). We found that the most frequently found air toxics with cancer risk greater than one in a million are formaldehyde, carbon tetrachloride, acetaldehyde, and benzene. The most frequently occurring binary pairs and ternary mixtures are the various combinations of these four air toxics. Analysis of urban and rural HAPs did not reveal significant differences in the top combinations of these chemicals. The cumulative annual cancer-related health impacts of inhaling the top 10 carcinogenic air toxics included was about 1,600 DALYs in the United States or 0.6 DALYs per 100,000 people. Formaldehyde and benzene together contribute nearly 60 percent of the total cancer-related health impacts. Our study shows that although there are many carcinogenic air toxics, only a few of them affect public health significantly at the national level in the United States, based on the frequency of occurrence of air toxics mixtures and cancer-related public health impacts. Future research is needed on their joint toxicity and cumulative health impacts.


The number of striatal cholinergic interneurons expressing calretinin is increased in parkinsonian monkeys.
The most abundant interneurons in the primate striatum are those expressing the calcium-binding protein calretinin (CR). The present immunohistochemical study provides detailed assessments of their morphological traits, number, and topographical distribution in normal monkeys (Macaca fascicularis) and in monkeys rendered parkinsonian (PD) by MPTP intoxication. In primates, the CR+ striatal interneurons comprise small (8-12μm), medium (12-20μm) and large-sized (20-45μm) neurons, each with distinctive morphologies. The small CR+ neurons were 2-3 times more abundant than the medium-sized CR+ neurons, which were 20-40 times more numerous than the large CR+ neurons. In normal and PD monkeys, the density of small and medium-sized CR+ neurons was twice as high in the caudate nucleus than in the putamen, whereas the inverse occurred for the large CR+ neurons. Double immunostaining experiments revealed that only the large-sized CR+ neurons expressed choline acetyltransferase (ChAT). The number of large CR+ neurons was found to increase markedly (4-12 times) along the entire anteroposterior extent of both the caudate nucleus and putamen of PD monkeys compared to controls. Comparison of the number of large CR-/ChAT+ and CR+/ChAT+ neurons together with experiments involving the use of bromo-deoxyuridine (BrdU) as a marker of newly generated cells showed that it is the expression of CR by the large ChAT+ striatal interneurons, and not their absolute number, that is increased in the dopamine-depleted striatum. These findings reveal the modulatory role of dopamine in the phenotypic expression of the large cholinergic striatal neurons, which are known to play a crucial role in PD pathophysiology.


Topical medication instillation techniques for glaucoma.
Glaucoma is a leading cause of irreversible blindness worldwide and the second most common cause of blindness after cataracts. The primary treatment for glaucoma aims to lower intraocular pressure (IOP) with the use of topical medicines. Topical medication instillation techniques, such as eyelid closure and nasolacrimal occlusion when instilling drops, have been proposed as potential methods to increase ocular absorption and decrease systemic absorption of the drops. To investigate the effectiveness of topical medication instillation techniques compared with usual care or another method of instillation of topical medication in the management of glaucoma or ocular hypertension. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (which contains the Cochrane Eyes and Vision Trials Register) (2016, Issue 12), MEDLINE Ovid (1946 to 8 December 2016), Embase Ovid (1947 to 8 December 2016), PubMed (1948 to 8 December 2016), LILACS (Latin American and Caribbean Health Sciences Literature Database) (1982 to 8 December 2016), International Pharmaceutical Abstracts Database (1970 to 8 December 2016), the metaRegister of Controlled Trials (mRCT) (www.controlled-trials.com) (last searched 13 May 2013), ClinicalTrials.gov (www.clinicaltrials.gov) (searched 8 December 2016) and the WHO International Clinical Trials Registry Platform (ICTRP) (www.who.int/ictrp/search/en) (searched 8 December 2016). We did not use any date or language restrictions in the electronic searches for trials. We included randomized controlled trials which had compared any topical medication instillation technique with usual care or a different method of instillation of topical medication. Two review authors independently screened records from the searches for eligibility, assessed the risk of bias, and extracted data. We followed methods recommended by Cochrane. We identified two trials (122 eyes of 61 participants) that had evaluated a topical medication instillation technique. We also identified two ongoing trials. Both included trials used a within-person design and administered prostaglandin monotherapy for glaucoma or ocular hypertension. Because the trials evaluated different instillation techniques and assessed different outcomes, we performed no meta-analysis.One trial, conducted in the US, evaluated the effect of eyelid closure (one and three minutes) versus no eyelid closure on lowering IOP. At one to two weeks' follow-up, reduction in IOP was similar in the eyelid closure group and the no eyelid closure group (mean difference (MD) -0.33 mmHg, 95% confidence interval (CI) -0.8 to 1.5; 51 participants; moderate-certainty evidence).The second trial, conducted in Italy, evaluated the effect of using an absorbent cloth to wipe excess fluid after instillation (fluid removal) versus not using an absorbent cloth (no removal) on reducing dermatologic adverse events. At four months' follow-up, eyelashes were shorter among eyes in the fluid removal group compared with the no fluid removal group (MD -1.70 mm, 95% CI -3.46 to 0.06; 10 participants; low-certainty evidence). Fewer eyes showed skin hyperpigmentation in the eyelid region towards the nose in the fluid removal group compared with the no removal group (RR 0.07, 95% CI 0.01 to 0.84; 10 participants; low-certainty evidence); however, the difference was uncertain in the eyelid region towards the temples (RR 0.44, 95% CI 0.07 to 2.66; 10 participants; low-certainty evidence). The effect hypertrichosis (excessive hair growth) was uncertain between groups (RR 1.00, 95% CI 0.17 to 5.98; 10 participants; low-certainty evidence).Neither trial reported other outcomes specified for this review, including the proportion of participants with IOP less than 21 mmHg; participant-reported outcomes related to the ease, convenience, and comfort of instillation techniques; physiologic measurements of systemic absorption; escalation of therapy; mean change in visual fields; optic nerve progression; mean change in best-corrected visual acuity; proportion in whom glaucoma developed; quality of life outcomes; or cost-effectiveness outcomes. Neither trial reported data at follow-up times of more than four months. Evidence to evaluate the effectiveness of topical medication instillation techniques for treatment of glaucoma is lacking. It is unclear what, if any, effects instillation techniques have on topical medical therapy for glaucoma.


Inflammatory biomarkers in heart failure.
Multiple lines of evidence support the "cytokine hypothesis," which suggests that inflammation plays an important role in the development and progression of heart failure. Circulating markers of inflammation, such as tumor necrosis factor alpha, interleukin 6, and C-reactive protein, may be useful in establishing the diagnosis, gauging prognosis, and evaluating the response to therapy in patients with heart failure. In addition to their potential as heart failure biomarkers, inflammatory cytokines have been investigated as targets of heart failure therapy. Although results for therapies directed against specific cytokines (such as tumor necrosis factor alpha) have thus far been disappointing, multiple studies continue to address the therapeutic potential of modulating the immune response in heart failure. In this review, the authors analyze available data supporting the use of inflammatory markers both as biomarkers and as potential therapeutic targets.


Spiroplasma and host immunity: activation of humoral immune responses increases endosymbiont load and susceptibility to certain Gram-negative bacterial pathogens in Drosophila melanogaster.
Spiroplasma poulsonii and its relatives are facultative, vertically transmitted endosymbionts harboured by several Drosophila species. Their long-term survival requires not only evasion of host immunity, but also that Spiroplasma does not have a net detrimental effect on host fitness. These requirements provide the central framework for interactions between host and endosymbiont. We use Drosophila melaogaster as a model to unravel aspects of the mechanistic basis of endosymbiont-host immune interactions. Here we show that Spiroplasma does not activate an immune response in Drosophila and is not susceptible to either the cellular or humoral arms of the Drosophila immune system. We gain unexpected insight into host factors that can promote Spiroplasma growth by showing that activation of Toll and Imd immune pathways actually increases Sprioplasma titre. Spiroplasma-mediated protection is not observed for variety of fungal and bacterial pathogens and Spiroplasma actually increases susceptibility of Drosophila to certain Gram-negative pathogens. Finally, we show that the growth of endosymbiotic Spiroplasma is apparently self-regulated, as suggested by the unhindered proliferation of non-endosymbiotic Spiroplasma citri in fly haemolymph.


LED PEDD Discharge Photometry: Effects of Software Driven Measurements for Sensing Applications.
This work explores the effects of embedded software-driven measurements on a sensory target when using a LED as a photodetector. Water turbidity is used as the sensory target in this study to explore these effects using a practical and important water quality parameter. Impacts on turbidity measurements are examined by adopting the Paired Emitter Detector Diode (PEDD) capacitive discharge technique and comparing common embedded software/firmware implementations. The findings show that the chosen software method can (a) affect the detection performance by up to 67%, (b) result in a variable sampling frequency/period, and (c) lead to an disagreement of the photo capacitance by up to 23%. Optimized code is offered to correct for these issues and its effectiveness is shown through comparative analyses, with the disagreement reduced significantly from 23% to 0.18%. Overall, this work demonstrates that the embedded software is a key and critical factor for PEDD capacitive discharge measurements and must be considered carefully for future measurements in sensor related studies.


Nonrandom X-inactivation patterns in normal females: lyonization ratios vary with age.
The utility of X-inactivation based clonality assays for evaluation of human neoplasia is well-documented. However, excessive Lyonization is a potential limitation of these assays, because it mimics clonal derivation of cells. The incidence of excessive Lyonization in healthy females is controversial, with reported incidence varying from 4% to 33%. Several explanations have been offered for the observed variation, including different criteria for excessive Lyonization, diversity of X-linked clonality assays, small population sizes and more recently, tissue specificity of X-inactivation patterns. However, it is also possible that stem cell depletion, clonal hematopoiesis, or selection pressures on blood cells results in an increased incidence of excessive skewing. If any of the latter is true, then the incidence of excessive skewing should increase with age in blood cells. To test this hypothesis, we determined X-inactivation ratios at the human androgen receptor locus of 295 normal females from three age groups: (1) neonates, (2) females 28 to 32 years old and, (3) females aged > or = 60 years. The incidence of skewing (allele ratios > or = 3:1) was 8.6% (14 of 162) in neonates (P = .104 v 28 to 32); 16.4% (11/67) in 28 to 32 y.o. (P = .0064 v > or = 60), and 37.9% (25 of 66) in women > or = 60 y.o. (P < .0001 v neonates). When a more stringent criterion for skewing was used (allele ratios > or = 10:1), the incidence was 1.9% (3 of 162) in neonates (P = .362 v 28 to 32), 4.5% (3 of 67) in 28 to 32 y.o. (P = .0022 v > or = 60), and 22.7% (15 of 66) in > or = 60 y.o. group (P < .0001 v neonates). Results have been confirmed at the phosphoglycerate kinase locus for 48 heterozygous females. The incidence of excessive skewing increases with age. In neonates, the incidence is low and may correspond to true excessive Lyonization. Acquired skewing occurs with aging in normal females and is present in 38% of females over the age of 60. Further studies are needed to determine whether acquired skewing is a consequence of stem cell depletion, true clonal hematopoiesis, growth advantage conferred by parental-specific X-chromosomes, or other causes. These data provide an explanation for variation in reported incidence of excessive skewing in normal females. Furthermore, these findings suggest that any study of clonality using X-inactivation based assays should incorporate age-matched controls for Lyonization.


Antibiotic resistance: turning evolutionary principles into clinical reality.
Antibiotic resistance is one of the major challenges facing modern medicine worldwide. The past few decades have witnessed rapid progress in our understanding of the multiple factors that affect the emergence and spread of antibiotic resistance at the population level and the level of the individual patient. However, the process of translating this progress into health policy and clinical practice has been slow. Here, we attempt to consolidate current knowledge about the evolution and ecology of antibiotic resistance into a roadmap for future research as well as clinical and environmental control of antibiotic resistance. At the population level, we examine emergence, transmission and dissemination of antibiotic resistance, and at the patient level, we examine adaptation involving bacterial physiology and host resilience. Finally, we describe new approaches and technologies for improving diagnosis and treatment and minimizing the spread of resistance.


Evaluation of clonal origin of malignant mesothelioma.
The hypothesis that most cancers are of monoclonal origin is often accepted as a fact in the scientific community. This dogma arose decades ago, primarily from the study of hematopoietic malignancies and sarcomas, which originate as monoclonal tumors. The possible clonal origin of malignant mesothelioma (MM) has not been investigated. Asbestos inhalation induces a chronic inflammatory response at sites of fiber deposition that may lead to malignant transformation after 30-50 years latency. As many mesothelial cells are simultaneously exposed to asbestos fibers and to asbestos-induced inflammation, it may be possible that more than one cell undergoes malignant transformation during the process that gives rise to MM, and result in a polyclonal malignancy. To investigate the clonality patterns of MM, we used the HUMARA (Human Androgen Receptor) assay to examine 16 biopsies from 14 women MM patients. Out of 16 samples, one was non-informative due to skewed Lyonization in its normal adjacent tissue. Fourteen out of the 15 informative samples revealed two electrophoretically distinct methylated HUMARA alleles, the Corrected Allele Ratio (CR) calculated on the allele peak areas indicating polyclonal origin MM. Our results show that MM originate as polyclonal tumors and suggest that the carcinogenic "field effect" of mineral fibers leads to several premalignant clones that give rise to these polyclonal malignancies.


Interplay between the cell envelope and mobile genetic elements shapes gene flow in populations of the nosocomial pathogen Klebsiella pneumoniae.
Mobile genetic elements (MGEs) drive genetic transfers between bacteria using mechanisms that require a physical interaction with the cellular envelope. In the high-priority multidrug-resistant nosocomial pathogens (ESKAPE), the first point of contact between the cell and virions or conjugative pili is the capsule. While the capsule can be a barrier to MGEs, it also evolves rapidly by horizontal gene transfer (HGT). Here, we aim at understanding this apparent contradiction by studying the covariation between the repertoire of capsule genes and MGEs in approximately 4,000 genomes of Klebsiella pneumoniae (Kpn). We show that capsules drive phage-mediated gene flow between closely related serotypes. Such serotype-specific phage predation also explains the frequent inactivation of capsule genes, observed in more than 3% of the genomes. Inactivation is strongly epistatic, recapitulating the capsule biosynthetic pathway. We show that conjugative plasmids are acquired at higher rates in natural isolates lacking a functional capsular locus and confirmed experimentally this result in capsule mutants. This suggests that capsule inactivation by phage pressure facilitates its subsequent reacquisition by conjugation. Accordingly, capsule reacquisition leaves long recombination tracts around the capsular locus. The loss and regain process rewires gene flow toward other lineages whenever it leads to serotype swaps. Such changes happen preferentially between chemically related serotypes, hinting that the fitness of serotype-swapped strains depends on the host genetic background. These results enlighten the bases of trade-offs between the evolution of virulence and multidrug resistance and caution that some alternatives to antibiotics by selecting for capsule inactivation may facilitate the acquisition of antibiotic resistance genes (ARGs).


Clinical biology and surgical therapy of intramucosal adenocarcinoma of the esophagus.
Mucosal ablation and endoscopic mucosal resection have been proposed as alternatives to surgical resection as therapy for intramucosal adenocarcinoma (IMC) of the esophagus. Acceptance of these alternative therapies requires an understanding of the clinical biology of IMC and the results of surgical resection modified for treatment of early disease. Retrospective review of 78 patients (65 men, 13 women; median age 66 years) with IMC who were treated with progressively less-extensive surgical resections (ie, en bloc, transhiatal, and vagal-sparing esophagectomy) from 1987 to 2005. The tumor was located in a visible segment of Barrett's esophagus in 65 (83%) and in cardia intestinal metaplasia in 13 (17%). A visible lesion was present in 53 (68%) and in all but 4 the lesion was cancer. In those patients with visible Barrett's, the tumor was within 3 cm of the gastroesophageal junction in 66% and within 1 cm in 37%. Esophagectomy was en bloc in 23, transhiatal in 31, vagal-sparing in 20, and transthoracic in 4. Operative mortality was 2.6%. Vagal-sparing esophagectomy had less morbidity, a shorter hospital stay, and no mortality. Of the patients who had en bloc resection, a median of 41 nodes were removed. One patient had one lymph node metastasis on hematoxylin and eosin staining and two others, normal on hematoxylin and eosin staining, had micrometastases on immunohistochemistry. Actuarial survival at 5 years was 88% and was similar for all types of resections. Two patients died from systemic metastases and seven from noncancer causes. IMC occurred in cardia intestinal metaplasia and in Barrett's esophagus. Two-thirds of patients with IMC had a visible lesion. Most tumors occurred near the gastroesophageal junction. Node metastases were uncommon, questioning the need for lymphadenectomy. A vagal-sparing technique had less morbidity than other forms of resection and no mortality. Survival after all types of resection was similar. Outcomes of endoscopic techniques should be compared with this benchmark.


Erythropoietic dynamic equilibrium in rats maintained after microwave irradiation.
The aim of study was to define influence of radiofrequency microwave (RF/MW) radiation on erythropoiesis in rats. The kinetics of polychromatic erythrocytes (PCEs) and micronucleated (MN) PCEs in the bone marrow (BM) and peripheral blood (PB) of rats during the intermittent subchronic experiment was followed. Rats were exposed 2h/day, 7 days/week to RF/MW of 2.45 GHz and whole-body specific absorption rate (SAR) of 1.25+/-0.36 W/kg. Control animals were included in the study. Each exposed and control group was killed on the final day of irradiation. Acridine-orange stained BM and blood smears were examined by fluorescence microscope. PCEs were obtained by inspection of 2000 BM and 1000 PB erythrocytes/slides. BMMNs and PBMNs frequency was obtained by observation of 1000 PCEs/slides. BMPCEs were increased on day 8 and 15, and PBPCEs were elevated on days 2 and 8 (p<0.05). The BMMN frequency was increased on experimental day 15, and MNPCEs in the PB was increased on day 8 (p<0.05). Findings of BM and PBPCEs or MNPCEs declined nearly to the control values until the end of the experiment. Such findings are considered to be indicators of radiation effects on BM erythropoiesis consequently reflected in the PB. Rehabilitated dynamic haemopoietc equilibrium in rats by the end of experiment indicates possibility of activation adaptation process in rats to the selected experimental conditions of subchronic RF/MW exposure.


A systematic review of the Trypanosoma cruzi genetic heterogeneity, host immune response and genetic factors as plausible drivers of chronic chagasic cardiomyopathy.
Chagas disease is a complex tropical pathology caused by the kinetoplastid Trypanosoma cruzi. This parasite displays massive genetic diversity and has been classified by international consensus in at least six Discrete Typing Units (DTUs) that are broadly distributed in the American continent. The main clinical manifestation of the disease is the chronic chagasic cardiomyopathy (CCC) that is lethal in the infected individuals. However, one intriguing feature is that only 30-40% of the infected individuals will develop CCC. Some authors have suggested that the immune response, host genetic factors, virulence factors and even the massive genetic heterogeneity of T. cruzi are responsible of this clinical pattern. To date, no conclusive data support the reason why a few percentages of the infected individuals will develop CCC. Therefore, we decided to conduct a systematic review analysing the host genetic factors, immune response, cytokine production, virulence factors and the plausible association of the parasite DTUs and CCC. The epidemiological and clinical implications are herein discussed.


Structure of native lens connexin 46/50 intercellular channels by cryo-EM.
Gap junctions establish direct pathways for cell-to-cell communication through the assembly of twelve connexin subunits that form intercellular channels connecting neighbouring cells. Co-assembly of different connexin isoforms produces channels with unique properties and enables communication across cell types. Here we used single-particle cryo-electron microscopy to investigate the structural basis of connexin co-assembly in native lens gap junction channels composed of connexin 46 and connexin 50 (Cx46/50). We provide the first comparative analysis to connexin 26 (Cx26), which-together with computational studies-elucidates key energetic features governing gap junction permselectivity. Cx46/50 adopts an open-state conformation that is distinct from the Cx26 crystal structure, yet it appears to be stabilized by a conserved set of hydrophobic anchoring residues. 'Hot spots' of genetic mutations linked to hereditary cataract formation map to the core structural-functional elements identified in Cx46/50, suggesting explanations for many of the disease-causing effects.


Combining Targeted Metabolomic Data with a Model of Glucose Metabolism: Toward Progress in Chondrocyte Mechanotransduction.
Osteoarthritis is a debilitating disease likely involving altered metabolism of the chondrocytes in articular cartilage. Chondrocytes can respond metabolically to mechanical loads via cellular mechanotransduction, and metabolic changes are significant because they produce the precursors to the tissue matrix necessary for cartilage health. However, a comprehensive understanding of how energy metabolism changes with loading remains elusive. To improve our understanding of chondrocyte mechanotransduction, we developed a computational model to calculate the rate of reactions (i.e. flux) across multiple components of central energy metabolism based on experimental data. We calculated average reaction flux profiles of central metabolism for SW1353 human chondrocytes subjected to dynamic compression for 30 minutes. The profiles were obtained solving a bounded variable linear least squares problem, representing the stoichiometry of human central energy metabolism. Compression synchronized chondrocyte energy metabolism. These data are consistent with dynamic compression inducing early time changes in central energy metabolism geared towards more active protein synthesis. Furthermore, this analysis demonstrates the utility of combining targeted metabolomic data with a computational model to enable rapid analysis of cellular energy utilization.


Semantic impairment in stroke aphasia versus semantic dementia: a case-series comparison.
Different neuropsychological populations implicate diverse cortical regions in semantic memory: semantic dementia (SD) is characterized by atrophy of the anterior temporal lobes whilst poor comprehension in stroke aphasia is associated with prefrontal or temporal-parietal infarcts. This study employed a case-series design to compare SD and comprehension-impaired stroke aphasic patients directly on the same battery of semantic tests. Although the two groups obtained broadly equivalent scores, they showed qualitatively different semantic deficits. The SD group showed strong correlations between different semantic tasks--regardless of input/output modality--and substantial consistency when a set of items was assessed several times. They were also highly sensitive to frequency/familiarity and made coordinate and superordinate semantic errors in picture naming. These findings support the notion that amodal semantic representations degrade in SD. The stroke aphasia group also showed multimodal deficits and consistency across different input modalities, but inconsistent performance on tasks requiring different types of semantic processing. They were insensitive to familiarity/frequency--instead, tests of semantic association were influenced by the ease with which relevant semantic relationships could be identified and distractors rejected. In addition, the aphasic patients made associative semantic errors in picture naming that SD patients did not make. The aphasic patients' picture naming performance improved considerably with phonemic cues suggesting that these patients retained knowledge that could not be accessed without contextual support. We propose that semantic cognition is supported by two interacting principal components: (i) a set of amodal representations (which progressively degrade in SD) and (ii) executive processes that help to direct and control semantic activation in a task-appropriate fashion (which are dysfunctional in comprehension-impaired stroke aphasic patients).


Synthesis of activated pyrimidine ribonucleotides in prebiotically plausible conditions.
At some stage in the origin of life, an informational polymer must have arisen by purely chemical means. According to one version of the 'RNA world' hypothesis this polymer was RNA, but attempts to provide experimental support for this have failed. In particular, although there has been some success demonstrating that 'activated' ribonucleotides can polymerize to form RNA, it is far from obvious how such ribonucleotides could have formed from their constituent parts (ribose and nucleobases). Ribose is difficult to form selectively, and the addition of nucleobases to ribose is inefficient in the case of purines and does not occur at all in the case of the canonical pyrimidines. Here we show that activated pyrimidine ribonucleotides can be formed in a short sequence that bypasses free ribose and the nucleobases, and instead proceeds through arabinose amino-oxazoline and anhydronucleoside intermediates. The starting materials for the synthesis-cyanamide, cyanoacetylene, glycolaldehyde, glyceraldehyde and inorganic phosphate-are plausible prebiotic feedstock molecules, and the conditions of the synthesis are consistent with potential early-Earth geochemical models. Although inorganic phosphate is only incorporated into the nucleotides at a late stage of the sequence, its presence from the start is essential as it controls three reactions in the earlier stages by acting as a general acid/base catalyst, a nucleophilic catalyst, a pH buffer and a chemical buffer. For prebiotic reaction sequences, our results highlight the importance of working with mixed chemical systems in which reactants for a particular reaction step can also control other steps.


Enhanced Ordering in Monolayers Containing Glycosphingolipids: Impact of Carbohydrate Structure.
The influence of carbohydrate structure on the ordering of glycosphingolipids (GSLs) and surrounding phospholipids was investigated in monolayers at the air-water interface. Binary mixtures composed of GSLs, chosen to span a range of carbohydrate complexity, and zwitterionic dipalmitoylphosphatidylcholine phospholipid, were studied. X-ray reflectivity was used to measure the out-of-plane structure of the monolayers and characterize the extension and conformation of the GSL carbohydrates. Using synchrotron grazing incidence x-ray diffraction, the in-plane packing of the lipid acyl chains and the area per molecule within ordered domains were characterized at different mole ratios of the two components. Our findings indicate that GSL-containing mixtures, regardless of the carbohydrate size, enhance the ordering of the surrounding lipids, resulting in a larger fraction of ordered phase of the monolayer and greater dimensions of the ordered domains. Reduction of the averaged area per molecule within the ordered domains was also observed but only in the cases where there was a size mismatch between the phospholipid headgroups and GSL components, suggesting that the condensation mechanism involves the relief of steric interactions between headgroups in mixtures.