Gene conversion and linkage: effects on genome evolution and speciation.
Crossing over is well known to have profound effects on patterns of genetic diversity and genome evolution. Far less direct attention has been paid to another distinct outcome of meiotic recombination: noncrossover gene conversion (NCGC). Crossing over and NCGC both shuffle combinations of alleles, and this degradation of linkage disequilibrium (LD) has major evolutionary consequences, ranging from immediate effects on nucleotide diversity to long-term consequences that shape genome evolution, species formation and species persistence. Unlike simple crossing over, NCGC has the potential to alter allele frequencies. Gene conversion can also occur in genomic regions where crossing over does not, and it purportedly exhibits more uniform rates across genomes. Considerable progress has been made towards understanding the mechanisms of gene conversion, and this progress enables us to begin exploring how gene conversion affects processes such as molecular evolution and interspecies gene flow. These topics are timely with the recent shift in focus from a primarily neutral null model of molecular evolution and speciation to one incorporating base levels of selection, making it all the more crucial to understand the basis and evolutionary implications of linkage. Here, we discuss the impact of gene conversion on genome structure and evolution and the current methods for detecting these events. We provide a comprehensive review of how gene conversion breaks down LD and affects both short- and long-term evolutionary processes, and we contrast its impact to that expected from crossing over alone.


Prolonged calcitonin receptor signaling by salmon, but not human calcitonin, reveals ligand bias.
Salmon calcitonin (sCT) and human calcitonin (hCT) are pharmacologically distinct. However, the reason for the differences is unclear. Here we analyze the differences between sCT and hCT on the human calcitonin receptor (CT(a)R) with respect to activation of cAMP signaling, β-arrestin recruitment, ligand binding kinetics and internalization. The study was conducted using mammalian cell lines heterologously expressing the human CT(a) receptor. CT(a)R downstream signaling was investigated with dose response profiles for cAMP production and β-arrestin recruitment for sCT and hCT during short term (<2 hours) and prolonged (up to 72 hours) stimulation. CT(a)R kinetics and internalization was investigated with radio-labeled sCT and hCT ligands on cultured cells and isolated membrane preparations from the same cell line. We found that sCT and hCT are equipotent during short-term stimulations with differences manifesting themselves only during long-term stimulation with sCT inducing a prolonged activation up to 72 hours, while hCT loses activity markedly earlier. The prolonged sCT stimulation of both cAMP accumulation and β-arrestin recruitment was attenuated, but not abrogated by acid wash, suggesting a role for sCT activated internalized receptors. We have demonstrated a novel phenomenon, namely that two distinct CT(a)R downstream signaling activation patterns are activated by two related ligands, thereby highlighting qualitatively different signaling responses in vitro that could have implications for sCT use in vivo.


Physician follow-up and long-term use of evidence-based medication for patients with hypertension who were discharged from an emergency department: a prospective cohort study.
More than 25% of the population has hypertension. The number of patients seeking care for hypertension in emergency departments has increased by more than 60% in the last decade, with less than 10% of these patients subsequently admitted to hospital. Managing physicians recommend early follow-up to patients who are discharged from the emergency department, but there is a paucity of literature assessing the impact or timing of follow-up on patient outcomes. Using a population-based cohort design, we included patients more than 65 years of age who were discharged from an Ontario emergency department with a primary diagnosis of hypertension between 2007 and 2014. We identified 2 cohorts: an incident cohort, and a cohort in which patients were on no more than 1 class of evidence-based antihypertensive medication at the time of presentation. Using logistic regression, we assessed the association of early follow-up care (within 7 d) and basic care (8-30 d), compared with no care within 30 days, on patient use of a new evidence-based antihypertensive medication 1 year later. Our study included 2088 patients with a new diagnosis of hypertension (the first cohort), and 6420 patients in the second cohort. Of patients with new diagnoses, 48.2% and 30.2% obtained early and basic follow-up care, respectively, compared with 50.0% and 30.9% of patients in the second cohort. Compared with patients without follow-up care within 30 days, the adjusted odds of filling an evidence-based antihypertensive medication prescription 1 year later in the incident group were 2.36 (95% confidence interval [CI] 1.86-2.99) for those who received early care, and 2.00 (95% CI 1.55-2.58) for those who received basic care. The adjusted odds in the second cohort were 2.12 (95% CI 1.84-2.43) and 1.96 (95% CI 1.69-2.27), respectively. Early follow-up care after leaving an emergency department with a diagnosis of hypertension was associated with improved long-term use of evidence-based antihypertensive medication. As patients increasingly present to the emergency department for hypertension, a formal, timely follow-up care system could improve patient use of evidence-based antihypertensive medication.


Gut Microbiota and Brain Function: An Evolving Field in Neuroscience.
There is a growing appreciation of the importance of gut microbiota to health and disease. This has been driven by advances in sequencing technology and recent findings demonstrating the important role of microbiota in common health disorders such as obesity. Moreover, the potential role of gut microbiota in influencing brain function, behavior, and mental health has attracted the attention of neuroscientists and psychiatrists. At the 29(th) International College of Neuropsychopharmacology (CINP) World Congress held in Vancouver, Canada, in June 2014, a group of experts presented the symposium, "Gut microbiota and brain function: Relevance to psychiatric disorders" to review the latest findings in how gut microbiota may play a role in brain function, behavior, and disease. The symposium covered a broad range of topics, including gut microbiota and neuroendocrine function, the influence of gut microbiota on behavior, probiotics as regulators of brain and behavior, and imaging the gut-brain axis in humans. This report provides an overview of these presentations.


Monitoring reversal of MET-mediated resistance to EGFR tyrosine kinase inhibitors in non-small cell lung cancer using 3'-deoxy-3'-[18F]-fluorothymidine positron emission tomography.
MET amplification is one of the mechanisms underlying acquired resistance to EGFR tyrosine kinase inhibitors (TKI) in non-small cell lung cancer (NSCLC). Here, we tested whether 3'-deoxy-3'-[(18)F]-fluorothymidine ([(18)F]FLT) positron emission tomography/computerized tomography (PET/CT) can detect MET-mediated resistance to EGFR TKIs and monitor the effects of MET inhibitors in NSCLC. H1993 and H820 NSCLC cells with high and low levels of MET amplification, respectively, and HCC827-expressing MET, but without gene amplification, were tested for the effects of MET inhibitors on the EGFR pathway and proliferation both in vitro and in vivo. Nude mice bearing NSCLCs with and without MET amplification were subjected to [(18)F]FLT PET/CT before and after treatment with crizotinib or erlotinib (50 mg/kg and 100 mg/kg p.o. for 3 days). H1993 cells showed high responsiveness to MET inhibitors and were resistant to erlotinib. Conversely, HCC827 cells showed high sensitivity to erlotinib and were resistant to MET inhibitors. Accordingly, H1993 tumors bearing MET amplification showed a mean reduction in [(18)F]FLT uptake of 28% and 41% after low- and high-dose treatment with crizotinib for 3 days, whereas no posttherapy changes of [(18)F]FLT uptake were observed in HCC827 tumors lacking MET amplification. Furthermore, a persistently high [(18)F]FLT uptake was observed in H1993 tumors after treatment with erlotinib, whereas HCC827 tumors showed up to 39% reduction of [(18)F]FLT uptake following erlotinib treatment. Imaging findings were confirmed by Ki67 immunostaining of tumor sections. [(18)F]FLT PET/CT can detect MET-mediated resistance to EGFR TKIs and its reversal by MET inhibitors in NSCLC.


Contribution of chronic diseases to the disability burden in a population 15 years and older, Belgium, 1997-2008.
Age-associated disability reduces quality of life in older populations and leads to wide-range implications for social and health policy. The identification of diseases that contribute to the disability burden is crucial to the development of prevention and intervention strategies to reduce disability. In this study, we assessed the contribution of chronic diseases to the prevalence of disability in Belgium. Data from 35,837 individuals aged 15 years or older who participated in the 1997, 2001, 2004, or 2008 Belgian Health Interview Surveys were used. Disability was defined as difficulties in doing at least one of six activities of daily living (transfer in and out of bed, transfer in and out of chair, dressing, washing hands and face, feeding, and going to the toilet) and/or mobility limitations (ability to walk without stopping less than 200 m). Multiple additive regression models were fitted separately for men and women to estimate the age-specific background disability rate (experienced by everyone, independent of the presence of specific diseases) and disease-specific disability rates (disability rate in subjects who reported selected chronic diseases). Musculoskeletal, cardiovascular, and respiratory diseases were the main contributors to the disability burden in Belgium. Musculoskeletal diseases were the most prevalent diseases in men and women in all age groups. Neurological diseases and stroke were the most disabling diseases, i.e. caused the highest level of disability among the diseased individuals, in all age groups for men and women, respectively. Back pain was the main cause of disability in men aged 15 to 64 years, while heart attack was the major contributor to the disability prevalence in men aged 65 or older. Likewise, arthritis was the main cause of disability among women across all age groups. Depression was also an important contributor in young subjects (15-54 years). Cancer was not an important contributor to the disability prevalence in Belgium. To reduce the burden of disability in Belgium, interventions should target musculoskeletal, cardiovascular and respiratory diseases especially among elderly. Furthermore, attention should also be given to depression in young individuals.


A SERRS/MRI multimodal contrast agent based on naked Au nanoparticles functionalized with a Gd(iii) loaded PEG polymer for tumor imaging and localized hyperthermia.
Multimodal contrast agents offer new interesting diagnostic possibilities, summing the benefits of multiple imaging techniques. Magnetic resonance and optical imaging are complementary techniques. The first allows total body screening, even though it suffers from low spatial resolution and needs high loadings, whereas the second shows lower penetration, but bright signals, and a higher spatial resolution and needs lower loadings. We present a plasmonic nanosystem as a MRI (magnetic resonance imaging) and SERRS (surface enhanced resonance Raman scattering) multimodal contrast agent. Naked gold nanoparticles, obtained by laser ablation synthesis in solution, are organized as a highly efficient SERRS substrate with a naphthalocyanine reporter and functionalized with a MRI contrast agent with a newly synthesized 3DOTA-PEG polymer, with a high GdIII loading. As a proof of concept, in vivo and ex vivo MRI and SERRS experiments are also performed. The plasmonic property of the nanosystem is then exploited to show its usefulness for localized hyperthermia.


Insulin signaling in osteoblasts integrates bone remodeling and energy metabolism.
The broad expression of the insulin receptor suggests that the spectrum of insulin function has not been fully described. A cell type expressing this receptor is the osteoblast, a bone-specific cell favoring glucose metabolism through a hormone, osteocalcin, that becomes active once uncarboxylated. We show here that insulin signaling in osteoblasts is necessary for whole-body glucose homeostasis because it increases osteocalcin activity. To achieve this function insulin signaling in osteoblasts takes advantage of the regulation of osteoclastic bone resorption exerted by osteoblasts. Indeed, since bone resorption occurs at a pH acidic enough to decarboxylate proteins, osteoclasts determine the carboxylation status and function of osteocalcin. Accordingly, increasing or decreasing insulin signaling in osteoblasts promotes or hampers glucose metabolism in a bone resorption-dependent manner in mice and humans. Hence, in a feed-forward loop, insulin signals in osteoblasts activate a hormone, osteocalcin, that promotes glucose metabolism.


Bladder leiomyoma: case report.
Leiomyoma is a benign lesion which represents 0.04-0.5% of bladder tumors. It is more common in females and its peak incidence is between 4th and 5th decades. Surgery is the treatment of choice and adequate results have been previously reported. 38 years old male patient who consulted for chronic pelvic pain syndrome. CT scan showed a 2 cm diameter exophitic lesion at the anterior left lateral bladder wall, which protruded into the perivesical fat. We performed a laparoscopic partial cistectomy locating the tumor and resecting it with simultaneous cystoscopic control, obtaining negative margins. The operative time was 70 minutes with an intraoperative blood loss of 50 ml. Postoperative period was uneventful. Final pathology reported: Bladder wall leiomyoma, without mitosis or atypia. Immunohistochemistry was positive for Actine and Vimentine stablishing diagnosis. Cd 1 17 (c-kit) was negative and ruled out a Gastrointestinal Stromal Tumor. Leiomyoma is bladder's most common benign non epithelial tumor. It represents 35-46% of these lesions with a 2:5 male/female ratio. It origins from the smooth muscle bundles and at the urinary tract the most common localizations are kidney and bladder. Clinical presentation depends on tumor size and localization. Ultrasound is the most useful diagnostic tool and the pathological diagnosis is mandatory. Surgery is the treatment of choice and technique depends on tumor size and localization. The laparoscopic approach seems to be an effective alternative in this group of tumors. Prognosis is good and recurrence is rare.


Asymmetric coevolutionary networks facilitate biodiversity maintenance.
The mutualistic interactions between plants and their pollinators or seed dispersers have played a major role in the maintenance of Earth's biodiversity. To investigate how coevolutionary interactions are shaped within species-rich communities, we characterized the architecture of an array of quantitative, mutualistic networks spanning a broad geographic range. These coevolutionary networks are highly asymmetric, so that if a plant species depends strongly on an animal species, the animal depends weakly on the plant. By using a simple dynamical model, we showed that asymmetries inherent in coevolutionary networks may enhance long-term coexistence and facilitate biodiversity maintenance.


Achieving new levels of recall in consent to research by combining remedial and motivational techniques.
Research supports the efficacy of both a remedial consent procedure (corrected feedback (CF)) and a motivational consent procedure (incentives) for improving recall of informed consent to research. Although these strategies were statistically superior to standard consent, effects were modest and not clinically significant. This study examines a combined incentivised consent and CF procedure that simplifies the cognitive task and increases motivation to learn consent information. We randomly assigned 104 individuals consenting to an unrelated host study to a consent as usual (CAU) condition (n=52) or an incentivised CF (ICF) condition (n=52). All participants were told they would be quizzed on their consent recall following their baseline assessment and at 4 monthly follow-ups. ICF participants were also informed that they would earn $5 for each correct answer and receive CF as needed. Quiz scores in the two conditions did not differ at the first administration (p=0.39, d=0.2); however, ICF scores were significantly higher at each subsequent administration (second: p=0.003, Cohen's d=0.6; third: p<0.0001, d=1.4; fourth: p<0.0001, d=1.6; fifth: p<0.0001, d=1.8). The ICF procedure increased consent recall from 72% to 83%, compared with the CAU condition in which recall decreased from 69% to 59%. This supports the statistical and clinical utility of a combined remedial and motivational consent procedure for enhancing recall of study information and human research protections.


Establishment of a predictive model of serum glucose changes under different exercise intensities and durations among patients with type 2 diabetes mellitus.
Regular exercise is regarded as one of necessary elements in treating diabetes mellitus (DM). The purpose of this study was to investigate the influence of different exercise intensities and durations on serum glucose changes after exercise in type 2 DM patients and to establish a predictive model of changes in serum glucose under different exercise intensities and durations. Thirty-seven type 2 DM patients were recruited from four teaching hospitals. A total of 12 exercise sessions were scheduled according to the results of a graded treadmill exercise test. The 12 exercise sessions were designed on the basis of different exercise intensities (40%, 60%, and 80% maximal workload) and exercise durations (10, 20, 30, and 40 min). Serum glucose level was measured before and after exercise. The findings indicate that the main effect of exercise intensity and duration was significant, but there was no interaction effect. All four variables, including exercise intensity, exercise duration, pre-exercise serum glucose levels, and gender, explained 37% of the variance in serum glucose changes after exercise. In conclusion, a dose-response relationship between exercise amount and serum glucose changes was demonstrated. This is helpful for health professionals to teach type 2 DM patients how to predict serum glucose response in different exercise situations.


Serum Jo-1 Autoantibody and Isolated Arthritis in the Antisynthetase Syndrome: Review of the Literature and Report of the Experience of AENEAS Collaborative Group.
Anti-Jo-1 is the most frequently detectable antibody in the antisynthetase syndrome (ASSD), an autoimmune disease characterized by the occurrence of arthritis, myositis, and interstitial lung disease (ILD). Recently, we organized an international collaborative group called American and European NEtwork of Antisynthetase Syndrome (AENEAS) for the study of this rare and fascinating disease. The group collected and published one of the largest series of ASSD patients ever described and with one of the longer follow-up ever reported. The number of participating centers is steadily increasing, as well as the available cohort. In the first paper, we showed that arthritis, myositis, and ILD may be frequently the only feature at disease onset, raising problems to reach a correct diagnosis of this syndrome. Nevertheless, we first observed that the ex novo appearance of further manifestations is common during the follow-up, strengthening the importance of a correct diagnosis. In our cohort, the 24 % of the 243 patients up to now collected had isolated arthritis as a presenting feature. These patients represent the most intriguing group in terms of differential diagnosis and clinical time course. Furthermore, data on this aspect are scanty, the reason that lead us to evaluate these aspects in our cohort of patients, reviewing also available literature. In fact, the most relevant aspect is that ASSD is rarely suspected in this setting of patients, in particular in case of poliarticular involvement, positive rheumatoid factor (RF), or anti-cyclic citrullinated peptide antibodies (ACPA) or evidence of joint erosions at plain radiographs. These findings were not rare in our cohort, and they have been also described in other series. Furthermore, manifestations such as Raynaud's phenomenon, mechanic's hands, and fever that may lead to the suspect of ASSD are observed only in a third of cases. If we consider the high rate of clinical picture progression in these patients, we feel that ASSD should be carefully considered in all patients presenting with isolated arthritis, even in those with erosive, RF, and ACPA-positive arthritis.


Differences in childhood physical abuse reporting and the association between CPA and alcohol use disorder in European American and African American women.
The goal of the current study was to examine whether the magnitude of the association between childhood physical abuse (CPA) and alcohol use disorder (AUD) varies by type of CPA assessment and race of the respondents. Data are from the Missouri adolescent female twins study and the Missouri family study (N = 4508) where 21.2% identified as African American (AA) and 78.8% as European American (EA); mean age = 23.8. Data were collected using a structured comprehensive interview which assessed CPA experiences using behavioral questions about specific abusive behaviors and trauma checklist items. Cox proportional hazards regression analyses were conducted, adjusting for additional risk factors associated with AUD, including co-occurring psychiatric disorders (defined as time-varying) and parental alcohol misuse. Overall, CPA reporting patterns were highly correlated (tetrachoric ρ = 0.73); although, only 25.8% of women who endorsed behaviorally defined CPA also endorsed checklist items whereas 72.2% of women who endorsed checklist items also endorsed behavioral questions. Racial disparities were evident, with behaviorally defined CPA increasing the hazard for AUD in EA but not AA women. Additional racial disparities in the risk for AUD were observed: increased hazard for AUD were associated with major depressive disorder in AA, and cannabis dependence and paternal alcohol problems in EA, women. Results demonstrate the relevance of the type of CPA measure in assessing CPA in studies of alcohol-related problems-behavioral items may be more inclusive of CPA exposure and more predictive of AUD- and highlight racial distinctions of AUD etiology in women. (PsycINFO Database Record


Optimizing Egg Recovery From Wild Northern Corn Rootworm Beetles (Coleoptera: Chrysomelidae).
The northern corn rootworm, Diabrotica barberi Smith & Lawrence (Coleoptera: Chrysomelidae), is one of the most important insect pests in the U.S. Corn Belt. Efforts to obtain eggs from wild northern corn rootworm populations using techniques developed for other rootworm species have been unsuccessful due to lack of oviposition. In 2016, we evaluated four oviposition media in choice tests within each of three female densities in 30.5 × 30.5 × 30.5 cm BugDorm cages. The number of eggs laid per female was significantly affected by female density and the interaction of female density × oviposition media, but oviposition was relatively poor in all oviposition media (1.2 eggs per female when averaging the three female densities and all oviposition media). Single females were also evaluated in nonchoice assays in 6 cm × 6 cm × 8 cm clear plastic boxes and averaged up to 108 eggs per female depending on the oviposition media. In 2017, the cumulative number of eggs laid per female in boxes with one female was not significantly different from the number of eggs laid per female in boxes with 3 females. In 2018, the cumulative number of eggs laid per female was not significantly different between female densities of 1, 3, 5, or 10 females per box. Total egg production per box therefore increased as female density increased. More than 27,000 wild northern corn rootworm eggs were collected from just 190 females when collected relatively early in the field season. We now have an efficient and robust system for obtaining eggs from wild northern corn rootworm females.


Osteoporosis medication and reduced mortality risk in elderly women and men.
Osteoporotic fractures are associated with premature mortality. Antiresorptive treatment reduces refracture but mortality reduction is unclear. The objective of the study was to examine the effect of osteoporosis treatment [bisphosphonates (BP), hormone therapy (HT), and calcium ± vitamin D only (CaD)] on mortality risk. This was a prospective cohort study (April 1989 to May 2007). The study was conducted with community-dwelling elderly (aged 60+ yr) subjects in Dubbo, a semiurban city, Australia. Subjects included 1223 and 819 women and men in the Dubbo Osteoporosis Epidemiology Study. Mortality according to treatment group was recorded. There were 325 (BP, n = 106; HT, n = 77; CaD, n = 142) women and 37 men (BP, n = 15; CaD, n = 22) on treatment. In women, mortality rates were lower with BP 0.8/100 person-years (0.4, 1.4) and HT 1.2/100 person-years (0.7, 2.1) but not CaD 3.2/100 person-years (2.5, 4.1) vs. no treatment 3.5/100 person-years (3.1, 3.8). Accounting for age, fracture occurrence, comorbidities, quadriceps strength, and bone mineral density, mortality risk remained lower for women on BP [hazard ratio (HR) 0.3 (0.2, 0.6)] but not HT [HR 0.8 (0.4, 1.8)]. For 429 women with fractures, mortality risk was still reduced in the BP group [adjusted HR 0.3 (0.2, 0.7)], not accounted for by a reduction in subsequent fractures. In men, lower mortality rates were observed with BP but not CaD [BP 1.0/100 person-years (0.3, 3.9) and CaD 3.1/100 person-years (1.5, 6.6) vs. no treatment 4.3/100 person-years (3.9, 4.8)]. After adjustment, mortality was similar, although not significant [HR 0.5 (0.1, 2.0)]. Osteoporosis therapy appears to reduce mortality risk in women and possibly men.


Breastfed at Tiffany's.
The importance of breast milk for the growing infant is undisputed; breastfeeding decreases infantile mortality by tenfold and decreases the incidence of infectious diseases. Despite its recognized benefits, the structural richness of breast milk has also impeded the characterization of the multiple effects of milk components on infant physiology. However, the important roles of some components of breast milk are beginning to be dissected. For instance, molecules such as immunoglobulin A (IgA) and milk oligosaccharides protect from gastrointestinal infections and influence the development of the gut microbiota. Deciphering the complex composition of breast milk brings to light multifaceted contributions that combine to make breast milk the ultimate personalized medicine.


Cyclin specificity in the phosphorylation of cyclin-dependent kinase substrates.
Cell-cycle events are controlled by cyclin-dependent kinases (CDKs), whose periodic activation is driven by cyclins. Different cyclins promote distinct cell-cycle events, but the molecular basis for these differences remains unclear. Here we compare the specificity of two budding yeast cyclins, the S-phase cyclin Clb5 and the M-phase cyclin Clb2, in the phosphorylation of 150 Cdk1 (Cdc28) substrates. About 24% of these proteins were phosphorylated more efficiently by Clb5-Cdk1 than Clb2-Cdk1. The Clb5-specific targets include several proteins (Sld2, Cdc6, Orc6, Mcm3 and Cdh1) involved in early S-phase events. Clb5 specificity depended on an interaction between a hydrophobic patch in Clb5 and a short sequence in the substrate (the RXL or Cy motif). Phosphorylation of Clb5-specific targets during S phase was reduced by replacing Clb5 with Clb2 or by mutating the substrate RXL motif, confirming the importance of Clb5 specificity in vivo. Although we did not identify any highly Clb2-specific substrates, we found that Clb2-Cdk1 possessed higher intrinsic kinase activity than Clb5-Cdk1, enabling efficient phosphorylation of a broad range of mitotic Cdk1 targets. Thus, Clb5 and Clb2 use distinct mechanisms to enhance the phosphorylation of S-phase and M-phase substrates.


Altered cytokine responses of dengue-specific CD4+ T cells to heterologous serotypes.
The interplay of different inflammatory cytokines induced during a dengue (DEN) virus infection plays a role in either protection or increased disease severity. We measured the frequencies and characterized the cytokine responses of DEN virus-specific memory CD4+ T cells in PBMC of six volunteers who received experimental live attenuated monovalent DEN vaccines. IFN-gamma and TNF-alpha responses to inactivated DEN Ags were detected in up to 0.54 and 1.17% of total circulating CD4+ T cells, respectively. Ags from the homologous serotype elicited the highest IFN-gamma response. The ratio of TNF-alpha- to IFN-gamma-producing CD4+ T cells was higher after stimulation with Ags from heterologous DEN serotypes. Peptide-specific CD4+ T cell frequencies of up to 0.089% were detected by direct staining using HLA class II tetramers. IFN-gamma and TNF-alpha responses to individual HLA class II-restricted peptide epitopes were detected in up to 0.05 and 0.27% of CD4+ T cells, respectively. Peptide sequences from the homologous serotype elicited a variety of cytokine response patterns. TNF-alpha- to IFN-gamma-positive CD4+ T cell ratios varied between peptides, but the ratio of the sum of responses was highest against heterologous serotypes. These results demonstrate epitope sequence-specific differences in T cell effector function. These patterns of effector responses may play a role in the immunopathogenesis of DEN hemorrhagic fever.


Formation of H2 on graphene using Eley-Rideal and Langmuir-Hinshelwood processes.
A hydrogen atom can either physisorb or chemisorb onto a graphene surface. To describe the interaction of H with graphene, we trained the C-C, H-H, and C-H interactions of the ReaxFF CHO bond order potential to reproduce Density Functional Theory (DFT) generated values of graphene cohesive energy and lattice constant, H2 dissociation energy, H on graphene adsorption potentials, and H2 formation on graphene using the Eley-Rideal (ER) and Langmuir-Hinshelwood (LH) processes. The results, generated from the trained H-graphene potentials, are in close agreement with the corresponding results from DFT. The advantage of using optimized CH potentials is, for example, the inclusion of physisorption interactions and quantum mechanical features of chemical bonding in the functional forms of the potentials. The trained CH potentials are utilized to study the energetics of formation of an H2 molecule on graphene using the Eley-Rideal and Langmuir-Hinshelwood processes. Potential energy surfaces for the formation of H2 through ER are generated for the collinear and oblique approach of the second hydrogen atom. Energetics of the formation of H2 through LH is studied for a variety of cases such as when hydrogen atoms are chemisorbed or physisorbed and when hydrogen occupies ortho, meta, or para chemisorption sites. The likelihood of H2 formation through LH for various configurations is discussed. Furthermore, the tunneling probability of an atom through a continuous symmetric/asymmetric barrier is calculated and applied to an adsorbed hydrogen atom on graphene.


Aromatic ionene topology and counterion-tuned gelation of acidic aqueous solutions.
Unusual gelation of acidic solutions was achieved using polycations bearing quaternary ammonium moieties. These ionene polymers are based on a disubstituted phenylene dibenzamide core, which allows the construction of different topomers (i.e. ortho-1, meta-2 and para-3). The topology of the polymers was found to play a key role on their aggregation behaviour both in pure water and in a variety of aqueous acidic solutions leading to the formation of stable acidic gels. Specifically, ortho-1 showed superior gelation ability than the analogues meta-2 and para-3 in numerous solutions of different pH and ionic strengths. Lower critical gelation concentrations, higher gel-to-sol transition temperatures and faster gelation were usually observed for ortho-1 regardless the solvent system. Detailed computational molecular dynamic simulations revealed a major role of the counterion (Cl-) and specific polymerpolymer interactions. In particular, hydrogen bonds, N-Hπ interactions and intramolecular π-π stacking networks are distinctive in ortho-1. In addition, counterions located at internal hydration regions also affect to such polymerpolymer interactions, acting as binders and, therefore, providing additional stability.


Immediate effects of UV radiation on the skin: modification by an antioxidant complex containing carotenoids.
The ultraviolet (UV) portion of sunlight is involved in the induction and development of skin cancers against which a limited photoprotection may be provided by reduced time of exposure, clothing, and sunscreen applications. The concept of an effective, safe, systemic photoprotection will circumvent many of the shortcomings. The UV-induced oxidative stress is a cause of DNA damage and a few publications have shown, in humans, minimal benefits, if any, of the oral intake of antioxidant complex, contrasting with the large literature showing beneficial effects in vitro or in animal models. We investigated, in 25 healthy individuals, the capacity of an antioxidant complex (AOC) - vitamins (lycopene, beta-carotene, alpha-tocopherol), selenium - to reduce UV-induced damages. The AOC was administered orally, daily during 7 weeks. Before and after irradiations, before and after the intake of the product, six parameters were studied: skin color by chromametry, minimal erythemal dose and, on skin biopsies, sunburn cells (SBCs), p53 detected by immunohistochemistry, pigmentation index, and levels of lipoperoxides (thiobarbituric acid reaction). After the oral intake of AOC, we observed an elevation of the actinic erythema threshold (+20%, P=0.01) and a general reduction of the UV-induced erythemas, a reduction of the UV-induced p53 expression (P<0.05) and of SBCs (P<0.01), and a parallel reduction of the lipoperoxide levels (P<0.01). The pigmentation was increased (P<0.01). After the oral intake of an antioxidant complex, many parameters of the epidermal defense against UV-induced damages are significantly improved. The oral intake of AOC could provide a safe, daylong and efficient complement to photo-protective measures provided by topical and physical agents and may contribute to reduce the DNA damages leading to skin aging and skin cancers.


Cloning of the neurodegeneration gene drop-dead and characterization of additional phenotypes of its mutation.
Mutations in the Drosophila gene drop-dead (drd) result in early adult lethality and neurodegeneration, but the molecular identity of the drd gene and its mechanism of action are not known. This paper describes the characterization of a new X-linked recessive adult-lethal mutation, originally called lot's wife (lwf(1)) but subsequently identified as an allele of drd (drd(lwf)); drd(lwf) mutants die within two weeks of eclosion. Through mapping and complementation, the drd gene has been identified as CG33968, which encodes a putative integral membrane protein of unknown function. The drd(lwf) allele is associated with a nonsense mutation that eliminates nearly 80% of the CG33968 gene product; mutations in the same gene were also found in two previously described drd alleles. Characterization of drd (lwf) flies revealed additional phenotypes of drd, most notably, defects in food processing by the digestive system and in oogenesis. Mutant flies store significantly more food in their crops and defecate less than wild-type flies, suggesting that normal transfer of ingested food from the crop into the midgut is dependent upon the DRD gene product. The defect in oogenesis results in the sterility of homozygous mutant females and is associated with a reduction in the number of vitellogenic egg chambers. The disruption in vitellogenesis is far more severe than that seen in starved flies and so is unlikely to be a secondary consequence of the digestive phenotype. This study demonstrates that mutation of the drd gene CG33968 results in a complex phenotype affecting multiple physiological systems within the fly.


Neuroprotection from tissue inhibitor of metalloproteinase-1 and its nanoparticles.
Matrix metalloproteinases (MMPs) are family of zinc dependent endopeptidases, which cleave extracellular matrix proteins, and play an important role in tissue remodelling in physiological and pathological processes. There is enhanced expression of MMPs, in particular MMP-9, during numerous pathological conditions, including epilepsy and ischemic stroke. Therefore, inhibition of MMP-9 is considered as a potential therapeutic target. Tissue Inhibitor of Matrix Metalloproteinase-1 (TIMP-1) is a 28kDa endogenous inhibitor of MMP-9. In this study we examined recombinant mouse TIMP-1 for its in-vitro neuroprotective effects, against Kainic Acid (KA) induced excitotoxicity in organotypic hippocampal slice culture (OHC) model. We also studied, sustained release effects of TIMP-1 in OHC by using poly lactic-co-glycolic acid (PLGA) nanoparticles (NPs). TIMP-1 and TIMP-1 PLGA NPs were added to the slice cultures at different time points, i.e., 30min before treatment with KA and 6h after KA treatment. Propidium iodide staining was used to reveal cell toxicity in the cultures. In addition, neurotoxicity was assessed using standard lactate dehydrogenase (LDH) release assay. Gelatinolytic activity in conditioned cultured medium of OHC was accessed by a fluorescent substrate assay. Briefly, our result show that TIMP-1 provided significant level of neuroprotection, especially when given before 30min of KA and released from the NPs. Since gelatinolytic activity assay showed a decrease in MMP-9 activity, it can be suggested that this neuroprotection might be mediated by the gelatinase inhibition.


Strategies to manage hepatitis C virus (HCV) disease burden.
The number of hepatitis C virus (HCV) infections is projected to decline while those with advanced liver disease will increase. A modeling approach was used to forecast two treatment scenarios: (i) the impact of increased treatment efficacy while keeping the number of treated patients constant and (ii) increasing efficacy and treatment rate. This analysis suggests that successful diagnosis and treatment of a small proportion of patients can contribute significantly to the reduction of disease burden in the countries studied. The largest reduction in HCV-related morbidity and mortality occurs when increased treatment is combined with higher efficacy therapies, generally in combination with increased diagnosis. With a treatment rate of approximately 10%, this analysis suggests it is possible to achieve elimination of HCV (defined as a >90% decline in total infections by 2030). However, for most countries presented, this will require a 3-5 fold increase in diagnosis and/or treatment. Thus, building the public health and clinical provider capacity for improved diagnosis and treatment will be critical.


Phase I study of Rituximab-CHOP regimen in combination with granulocyte colony-stimulating factor in patients with follicular lymphoma.
Rituximab is an anti-CD20 monoclonal antibody, and it is used to treat B-cell lymphomas. Antibody-dependent cellular cytotoxicity (ADCC) is considered one of the mechanisms through which rituximab exerts its effects. Granulocyte colony-stimulating factor (G-CSF) enhances the cytotoxicity of neutrophils through ADCC, and it can be speculated that a combination of rituximab and G-CSF may augment the treatment efficacy of rituximab. We administered rituximab with CHOP (cyclophosphamide, doxorubicin, vincristine, prednisone) treatment with G-CSF to 15 patients with follicular lymphoma, and we investigated the safety and efficacy of this regimen. We investigated ADCC activity in neutrophils and the expression of cell surface antigens including Fcgamma receptor type I [FcgammaRI (CD64)] on neutrophils to determine the optimal dose of G-CSF. Adverse reactions occurred in 14 of 15 patients and consisted mainly of grade 3/4 hematological toxicity. The response rate was 100%, with complete remission in 12 patients (80%) and partial remission in 3 patients (20%). At 14 months, the median length of the observation period, 2 of 12 patients had relapsed. G-CSF administration increased both FcgammaRI expression and ADCC activity. There were no significant differences in the levels of FcgammaRI expression or ADCC activity between the 2 microg/kg G-CSF and 5 microg/kg G-CSF groups, indicating that the optimal dose of G-CSF was 2 microg/kg. We conclude that the combination of rituximab-CHOP and G-CSF is well tolerated. We plan to carry out a randomized trial to compare efficacy between rituximab-CHOP treatment and treatment with a combination of rituximab-CHOP and G-CSF.


Fleas (Siphonaptera) are Cretaceous, and evolved with Theria.
Fleas (order Siphonaptera) are highly-specialized, diverse blood-feeding ectoparasites of mammals and birds with an enigmatic evolutionary history and obscure origin. We here present a molecular phylogenetic study based on a comprehensive taxon sampling of 259 flea taxa, representing 16 of the 18 extant families of this order. A Bayesian phylogenetic tree with strong nodal support was recovered, consisting of seven sequentially derived lineages with Macropsyllidae as the earliest divergence, followed by Stephanocircidae. Divergence times of flea lineages were estimated based on fossil records and host specific associations to bats (Chiroptera), suggesting that the common ancestor of extant Siphonaptera diversified during the Cretaceous. However, most of the intraordinal divergence into extant lineages took place after the K-Pg boundary. Ancestral states of host association and biogeographical distribution were reconstructed, suggesting with high likelihood that fleas originated in the southern continents (Gondwana) and migrated from South America to their extant distributions in a relatively short time frame. Theria (placental mammals and marsupials) represent the most likely ancestral host group of extant Siphonaptera, with marsupials occupying a more important role than previously assumed. Major extant flea families evolved in connection to post K-Pg diversification of Placentalia. The association of fleas with monotremes and birds is likely due to later secondary host association. These results suggest caution in casually interpreting recently discovered Mesozoic fossil "dinosaur fleas" of Northeast Asia as part of what we currently consider Siphonaptera.


Peripheral vascular resistance and angiotensin II levels during pulsatile and non-pulsatile cardiopulmonary bypass.
The effects of pulsatile and non-pulsatile cardiopulmonary bypass (CPB) on levels of peripheral vascular resistance and plasma angiotensin II (AII) have been studied in 24 patients submitted to elective cardiac surgical procedures. Twelve patients had conventional non-pulsatile perfusion throughout the period of CPB (non-pulsatile group), while 12 had pulsatile perfusion during the central period of total CPB, using the Stockert pulsatile pump system (pulsatile group). There were no significant differences between the groups in respect of age, weight, bypass time, cross-clamp time, or in mean pump flow or mean perfusion pressure at the onset of CPB. Peripheral vascular resistance index (PVRI) and plasma AII levels were measured at the onset of total CPB and at the end of total CPB. In the non-pulsatile group PVRI rose from 19.6 units to 29.96 units during perfusion. In the pulsatile group PVRI showed little change from 20.89 units to 21.45 units during perfusion (P less than 0.001). Plasma AII levels (normal less than 35 pg/ml) rose during perfusion from 49 pg/ml to 226 pg/ml in the non-pulsatile group. The rise in the pulsatile group from 44 pg/ml to 98 pg/ml was significantly smaller than that in the non-pulsatile group (P less than 0.01). These results indicate that pulsatile cardiopulmonary bypass prevents the rise in PVRI associated with non-pulsatile perfusion, and that this effect may be achieved by preventing excessive activation of the renin-angiotensin system, thus producing significantly lower plasma concentrations of the vasoconstrictor angiotensin II.


Concentration, pH, and surface charge effects on cadmium and lead sorption in three tropical soils.
Reactions of heavy metals with soil are important in determining metal fates in the environment. Sorption characteristics of two heavy metals, Cd and Pb, in three tropical soils (Mollisol, Oxisol, and Ultisol) from Puerto Rico were assessed at varying metal concentrations (0 to 1.2 mM) and pH values (approximately 2 to 7). All soils sorbed more Pb than Cd. Sorption maxima were obtained for each metal for the Oxisol and Ultisol soils, but not the Mollisol. Sorption appeared to depend more on soil mineralogy than organic matter content. Sorption isotherms were linear within the sorption envelope with similar slopes for each soil-metal curve, when plotting metal sorption as a function of pH. Cadmium and Pb isotherms yielded average slopes of approximately 36+/-1 and 28+/-1 units (percent increase in metal sorption per 1-unit increase in pH), respectively. Metal sorption depended more on metal type than soil composition. Cadmium sorption displayed a greater pH dependence than Pb. Cadmium sorption was less than or equal to the amount of negative surface charge except at pH values greater than the point of zero net charge (PZNC). This suggests that Cd was probably sorbed via electrostatic surface reactions and/or possible inner-sphere complexation at pH > 3.7. However, the amount of Pb sorbed by the Oxisol was greater than the amount of negative surface charge, suggesting that Pb participates in inner-sphere surface reactions. Lead was sorbed more strongly than Cd in our soils and poses less of a threat to underlying ground water systems due to its lower mobility and availability.


Deficient E-cadherin adhesion in C57BL/6J-Min/+ mice is associated with increased tyrosine kinase activity and RhoA-dependent actomyosin contractility.
The Min/+ mouse is a model for APC-dependent colorectal cancer (CRC). We showed that tumorigenesis in this animal was associated with decreased E-cadherin adhesion and increased epidermal growth factor receptor (Egfr) activity in the non-tumor intestinal mucosa. Here, we tested whether these abnormalities correlated with changes in the actin cytoskeleton due to increased Rho-ROCK signaling. We treated Apc+/+ (WT) littermate small intestine with EGTA, an inhibitor of E-cadherin, and with LPA, an RhoA activator; both induced effects on adhesion and kinase activity that mimicked the Min/+ phenotype. GTP-bound Rho was increased in Min/+ enterocytes relative to WT. Since RhoA activity is associated with actomyosin contractility, markers of this signaling cascade were assessed including phosphorylated myosin light chain (MLC), cofilin, Pyk2, Src, and MAPK kinases. The increased actomyosin contractility characterizing Min/+ intestinal tissue was suppressed by the ROCK inhibitor, Y27632, but was inducible in the WT by EGTA or LPA. Finally, ultrastructural imaging revealed changes consistent with actomyosin contractility in Min/+ enterocytes. Thus, the positive regulation of E-cadherin adhesion provided by Apc+ in vivo allows proper negative regulation of Egfr, Src, Pyk2, and MAPK, as well as RhoA activities.


Influence of a novel calcium-phosphate coating on the mechanical properties of highly porous collagen scaffolds for bone repair.
Lyophilised collagen scaffolds have shown enormous potential in tissue engineering in a number of areas due to their excellent biological performance. However, they are limited for use in bone tissue engineering due to poor mechanical properties. This paper discusses the development of a calcium-phosphate coating for collagen scaffolds in order to improve their mechanical properties for bone tissue engineering. Pure collagen scaffolds produced in a lyophilization process were coated by immersing them in sodium ammonium hydrogen phosphate (NaNH(4)HPO(4)) followed by calcium chloride (CaCl(2)). The optimal immersing sequence, duration, as well as the optimal solution concentration which facilitated improved mechanical properties of the scaffolds was investigated. The influence of the coating on composition, structural and material properties was analysed. This investigation successfully developed a novel collagen/calcium-phosphate composite scaffold. An increase in the mechanical properties of the scaffolds from 0.3 kPa to up to 90 kPa was found relative to a pure collagen scaffold, while the porosity was maintained as high as 92%, indicating the potential of the scaffold for bone tissue engineering or as a bone graft substitute.


Integrating Social Services and Home-Based Primary Care for High-Risk Patients.
There is a consensus that our current hospital-intensive approach to care is deeply flawed. This review article describes the research evidence for developing a better system of care for high-cost, high-risk patients. It reviews the evidence that home-centered care and integration of health care with social services are the cornerstones of a more humane and efficient system. The article describes the strengths and weaknesses of research evaluating the effects of social services in addressing social determinants of health, and how social support is critical to successful acute care transition programs. It reviews the history of incorporating social services into care management, and the prospects that recent payment reforms and regulatory initiatives can succeed in stimulating the financial integration of social services into new care coordination initiatives. The article reviews the literature on home-based primary care for the chronically ill and disabled, and suggests that it is the emergence of this care modality that holds the greatest promise for delivery system reform. In the hope of stimulating further discussion and debate, the authors summarize existing viewpoints on how a home-centered system, which integrates social and medical services, might emerge in the next few years.


Consolidated criteria for strengthening reporting of health research involving indigenous peoples: the CONSIDER statement.
Research reporting guidelines are increasingly commonplace and shown to improve the quality of published health research and health outcomes. Despite severe health inequities among Indigenous Peoples and the potential for research to address the causes, there is an extended legacy of health research exploiting Indigenous Peoples. This paper describes the development of the CONSolIDated critERtia for strengthening the reporting of health research involving Indigenous Peoples (CONSIDER) statement. A collaborative prioritization process was conducted based on national and international statements and guidelines about Indigenous health research from the following nations (Peoples): Australia (Aboriginal and Torres Strait Islanders), Canada (First Nations Peoples, Métis), Hawaii (Native Hawaiian), New Zealand (Māori), Taiwan (Taiwan Indigenous Tribes), United States of America (First Nations Peoples) and Northern Scandinavian countries (Sami). A review of seven research guidelines was completed, and meta-synthesis was used to construct a reporting guideline checklist for transparent and comprehensive reporting of research involving Indigenous Peoples. A list of 88 possible checklist items was generated, reconciled, and categorized. Eight research domains and 17 criteria for the reporting of research involving Indigenous Peoples were identified. The research reporting domains were: (i) governance; (ii) relationships; (iii) prioritization; (iv) methodologies; (v) participation; (vi) capacity; (vii) analysis and findings; and (viii) dissemination. The CONSIDER statement is a collaborative synthesis and prioritization of national and international research statements and guidelines. The CONSIDER statement provides a checklist for the reporting of health research involving Indigenous peoples to strengthen research praxis and advance Indigenous health outcomes.


How I treat malignant pleural mesothelioma.
Malignant pleural mesothelioma is a rare and aggressive malignancy mostly associated with occupational asbestos exposure. Prognosis is poor and only highly selected patients may benefit from aggressive surgical management, also as part of a multimodal approach. In advanced disease, the combination of pemetrexed and platinum remains the only established treatment, while efficacy evidence of second line chemotherapy is lacking. Thus, a deeper knowledge of biology of the disease and more effective treatments are urgently needed. Refer to specialised centres with multidisciplinary expertise is mandatory, as well as inclusion of patients in clinical trials is advisable whenever possible. In all stages of disease focus on symptoms control is paramount.


Recombinant silk-elastinlike protein polymer displays elasticity comparable to elastin.
We evaluated the mechanical properties of the genetically engineered, recombinant silk-elastinlike protein copolymer, SELP-47K. In tensile stress-strain analysis, methanol-treated non-cross-linked SELP-47K films exceeded the properties of native aortic elastin, attaining an ultimate tensile strength of 2.5 +/- 0.4 MPa, an elastic modulus of 1.7 +/- 0.4 MPa, an extensibility of 190 +/- 60%, and a resilience of 86 +/- 4% after 10 cycles of mechanical preconditioning. Stress-relaxation and creep analysis showed that films substantially maintained their elastic properties under sustained deformation. Chemical cross-linking of SELP-47K films doubled the elastic modulus and ultimate tensile strength and enhanced the extensibility and resilience. The underlying conformational and microstructural features of the films were examined. Raman spectroscopy revealed that the silklike blocks of SELP-47K existed in antiparallel beta-sheet crystals in the films, likely responsible for the robust physical cross-links. Scanning electron microscopy (SEM) revealed that the various processing treatments and the mechanical deformation of the films induced changes in their surface microstructure consistent with the coagulation and alignment of polymer chains. These results demonstrate that films with excellent elasticity, comparable to native aortic elastin, are obtainable from SELP-47K, a protein copolymer combining both silk- and elastin-derived sequences in a single polymer chain.


Identification and management of bed bug infestations in austere environments.
Military forces have missions that send them all over the globe. With the reemergence of bed bugs worldwide, the possibility of Servicemembers encountering them has increased. Special Operations Forces are often sent to locations that may not have integrated pest management support. Knowing how to identify and manage a bed bug infestation, with and without proper equipment and supplies, may become necessary in the very near future. It is also important that Servicemembers are aware of how bed bugs travel, to prevent their dispersal back to the United States and into their barracks and homes.


Acute elevation of intra-abdominal pressure contributes to extravascular shift of fluid and proteins in an experimental porcine model.
Intra-abdominal hypertension and abdominal compartment syndrome contribute significantly to increased morbidity and mortality in critically ill patients. This study describes pathophysiologic effects of the acutely elevated intra-abdominal pressure on microvascular fluid exchange and microcirculation. The resulting changes could contribute to development of organ dysfunction or failure. 16 pigs were randomly allocated to a control-group (C-group) or an interventional group (P-group). After 60 min of stabilization, intra-abdominal pressure of the P-group animals was elevated to 15 mmHg by Helium insufflation and after 120 min to a level of 30 mmHg for two more hours. The C-group animals were observed without insufflation of gas. Laboratory and hemodynamic parameters, plasma volume, plasma colloid osmotic pressure, total tissue water content, tissue perfusion, markers of inflammation and cerebral energy metabolism were measured and net fluid balance and fluid extravasation rates calculated. Analysis of variance for repeated measurements with post-tests were used to evaluate the results with respect to differences within or between the groups. In the C-group hematocrit, net fluid balance, plasma volume and the fluid extravasation rate remained essentially unchanged throughout the study as opposed to the increase in hematocrit (P < 0.001), fluid extravasation rate (P < 0.05) and decrease in plasma volume (P < 0.001) of the P-group. Hemodynamic parameters remained stable or were slightly elevated in the C-group while the P-group demonstrated an increase in femoral venous pressure (P < 0.001), right atrial pressure (P < 0.001), pulmonary capillary wedge pressure (P < 0.01) and mean pulmonary arterial pressure (P < 0.001). The protein mass decreased in both study groups but was significantly lower in the P-group as compared with the C-group, after 240 min of intervention. The increased intra-abdominal pressure was associated with elevated intracranial pressure and reduced tissue perfusion of the pancreas and the gastric- and intestinal mucosa. Elevation of intra-abdominal pressure has an immediate impact on microvascular fluid extravasation leading to plasma volume contraction, reduced cardiac output and deranged perfusion of abdominal organs.


Vascular muscle membrane cation mechanisms and total peripheral resistance.
Passive and active carrier-mediated transport of sodium across vascular muscle membranes has been suggested to be more important in the increased total peripheral resistance found in genetic hypertension. Using manipulations of ion gradients and recordings of ion currents, membrane potentials, and tension, I have found evidence for calcium regulation as the central pathophysiological mechanism in spontaneously hypertensive rats. Increased sodium pump activity, which may be a partial compensation for the increased sodium influx in hypertension, may thus be secondary to altered calcium channel regulation in hypertension. The calcium channel, and the membrane potentials governing it, seem to be the most immediately important membrane mechanisms for hypertension research.


Requirement of heparan sulfate for bFGF-mediated fibroblast growth and myoblast differentiation.
Basic fibroblast growth factor (bFGF) binds to heparan sulfate proteoglycans at the cell surface and to receptors with tyrosine kinase activity. Prevention of binding between cell surface heparan sulfate and bFGF (i) substantially reduces binding of fibroblast growth factor to its cell-surface receptors, (ii) blocks the ability of bFGF to support the growth of Swiss 3T3 fibroblasts, and (iii) induces terminal differentiation of MM14 skeletal muscle cells, which is normally repressed by fibroblast growth factor. These results indicate that cell surface heparan sulfate is directly involved in bFGF cell signaling.


Quantum reference frames and their applications to thermodynamics.
We construct a quantum reference frame, which can be used to approximately implement arbitrary unitary transformations on a system in the presence of any number of extensive conserved quantities, by absorbing any back action provided by the conservation laws. Thus, the reference frame at the same time acts as a battery for the conserved quantities. Our construction features a physically intuitive, clear and implementation-friendly realization. Indeed, the reference system is composed of the same types of subsystems as the original system and is finite for any desired accuracy. In addition, the interaction with the reference frame can be broken down into two-body terms coupling the system to one of the reference frame subsystems at a time. We apply this construction to quantum thermodynamic set-ups with multiple, possibly non-commuting conserved quantities, which allows for the definition of explicit batteries in such cases.This article is part of a discussion meeting issue 'Foundations of quantum mechanics and their impact on contemporary society'.


Hippocampal sharp wave bursts coincide with neocortical "up-state" transitions.
The sleeping neocortex shows nested oscillatory activity in different frequency ranges, characterized by fluctuations between "up-states" and "down-states." High-density neuronal ensemble recordings in rats now reveal the interaction between synchronized activity in the hippocampus and neocortex: Electroencephalographic sharp waves in the hippocampus were more probable during down-states than during up-states, and tended to coincide with transitions from down-states to up-states. The form of cortical activity fluctuations and their interactions with sharp waves depend on sleep depth: In deeper sleep stages, characterized by strong neocortical oscillation in the delta range or slower (approximately 0.8-4 Hz), sharp-wave-triggered peri-event time histograms (PETH) are consistent with a longer duration for down-states than for up-states. In lighter sleep, the sharp-wave-triggered PETH suggested longer up-states than down-states. These results highlight the interplay in the hippocampal/neocortical loop: Decreased neocortical input during down-states may be a factor in generation of sharp waves. In turn, sharp waves may facilitate down-to-up transitions. This interplay may reflect joint memory trace reactivation in the hippocampus and in the neocortex, possibly contributing to consolidation of long-term memory: Off-line reactivation of recent neural activity patterns in the hippocampus occurs during 50-100-msec electroencephalographic sharp waves, corresponding to pyramidal-cell population bursts. The neocortical up-states starting in correspondence with sharp waves may be influenced by the reactivated information carried by the hippocampal sharp wave.


The influence of surface oxide on the growth of metal/semiconductor nanowires.
We report the critical effects of oxide on the growth of nanostructures through silicide formation. Under an in situ ultrahigh vacuum transmission electron microscope, it is observed from the conversion of Si nanowires into the metallic PtSi grains epitaxially through controlled reactions between lithographically defined Pt pads and Si nanowires. With oxide, instead of contact area, single crystal PtSi grains start forming either near the center between two adjacent pads or from the ends of Si nanowires, resulting in the heterostructure formation of Si/PtSi/Si. Without oxide, transformation from Si into PtSi begins at the contact area between them, resulting in the heterostructure formation of PtSi/Si/PtSi. The nanowire heterostructures have an atomically sharp interface with epitaxial relationships of Si(20-2)//PtSi(10-1) and Si[111]//PtSi[111]. Additionally, it has been observed that the existence of oxide significantly affects not only the growth position but also the growth behavior and the growth rate by two orders of magnitude. Molecular dynamics simulations have been performed to support our experimental results and the proposed growth mechanisms. In addition to fundamental science, the significance of the study matters for future processing techniques in nanotechnology and related applications as well.


The role of gap states in the energy level alignment at the organic-organic heterojunction interfaces.
The interface properties of organic-organic heterojunctions (OOHs), such as interface energy level alignment (ELA), interfacial charge transfer, interface nanostructuring, molecular orientation and so on, play an essential role in determining the device performance for some technologically important organic electronic devices, encompassing organic solar cells, bipolar organic field-effect-transistors, and organic light-emitting-diodes. The aim of this article is to provide a balanced assessment on the understanding of the ELA at the small-molecule based OOH interfaces with well-defined molecular orientation, with particular emphasis on the role of gap states in organic thin films. A generalized picture of gap states determined ELA at the OOH interfaces is provided and their implications in relevant organic electronic devices have been discussed.


The genetic architecture of seed composition in soybean is refined by genome-wide association scans across multiple populations.
Soybean oil and meal are major contributors to world-wide food production. Consequently, the genetic basis for soybean seed composition has been intensely studied using family-based mapping. Population-based mapping approaches, in the form of genome-wide association (GWA) scans, have been able to resolve loci controlling moderately complex quantitative traits (QTL) in numerous crop species. Yet, it is still unclear how soybean's unique population history will affect GWA scans. Using one of the populations in this study, we simulated phenotypes resulting from a range of genetic architectures. We found that with a heritability of 0.5, ∼100% and ∼33% of the 4 and 20 simulated QTL can be recovered, respectively, with a false-positive rate of less than ∼6×10(-5) per marker tested. Additionally, we demonstrated that combining information from multi-locus mixed models and compressed linear-mixed models improves QTL identification and interpretation. We applied these insights to exploring seed composition in soybean, refining the linkage group I (chromosome 20) protein QTL and identifying additional oil QTL that may allow some decoupling of highly correlated oil and protein phenotypes. Because the value of protein meal is closely related to its essential amino acid profile, we attempted to identify QTL underlying methionine, threonine, cysteine, and lysine content. Multiple QTL were found that have not been observed in family-based mapping studies, and each trait exhibited associations across multiple populations. Chromosomes 1 and 8 contain strong candidate alleles for essential amino acid increases. Overall, we present these and additional data that will be useful in determining breeding strategies for the continued improvement of soybean's nutrient portfolio.


Transatlantic discovery of Notocotylus atlanticus (Digenea: Notocotylidae) based on life cycle data.
Digenean parasites feature a series of stages with a distinct appearance, reproduction mode, and lifestyle that together constitute their well-known, complex life cycle. Species descriptions of Digenea have always been based on one of these stages-the marita, or sexually reproducing adult in the final host. However, in some cases, data on the life cycle are essential for the differential diagnosis of closely related species. Here, we present the case of Notocotylus atlanticus, where different stages of its life cycle were discovered for the first time since the species description, and across the Atlantic. We used a material from a naturally infected intertidal marine snail, Ecrobia ventrosa, and several waterfowl species and also carried out infection experiments. For morphological studies, we employed light microscopy, SEM, and CLSM; molecular data obtained include sequences of ITS1 and 28S rRNA gene. We demonstrate that N. atlanticus adult worm morphology is barely sufficient to distinguish it from several other species. Cercariae morphology and identity of the first intermediate hosts provide crucial additional information. According to our preliminary phylogenetic reconstructions, two notocotylid lineages are associated with two major gastropod lineages-the Caenogastropoda and the Heterobranchia. The traditional character to identify notocotylid genera (structure of ventral organs) fails to explain the phylogeny and thus requires reassessment. Further reliable morphological, life cycle and molecular data on other species are likely to reveal more patterns in notocotylid systematics, host specificity, and evolution.


The effect of 6-aminohexanoic acid and fucoidan on the activation of glutamic plasminogen by streptokinase.
Studies were conducted on the effect of 6-aminohexanoic acid (6-AH) or fucoidan on the activation of glutamic plasminogen (glu-plg) by streptokinase using 0.05 mol/l Tris buffer containing a physiological concentration of NaCl. In contrast to the earlier reports where no NaCl was added to the buffer solution, addition of 6-AH enhanced the initial rate while the inhibition by fucoidan was not affected. Double reciprocal plots of the activation of glu-plg by streptokinase in the presence of 6-AH showed an increase in Vmax, but no change in Km. However, the addition of fucoidan showed a decrease in Vmax, but no change in Km. To determine whether the stimulatory effect of 6-AH was specifically directed towards glu-plg or streptokinase, the ratios of the initial rate of plasmin generation in the presence of 6-AH over the controls were plotted against the inverse of the volume fraction of glu-plg or streptokinase after serial dilutions. The results indicated that the dilutions of glu-plg, but not of streptokinase, influenced the ratios, suggesting an interaction of 6-AH with glu-plg. Similar experiments were conducted to determine the mechanism of inhibition of streptokinase by fucoidan. The results indicated that fucoidan was interacting with streptokinase, but not with glu-plg. Circular dichroism studies of glu-plg in the near-ultraviolet spectra (250-308 nm) showed that addition of 6-AH enhanced the spectra in the region around certain chromophores, which reflected conformational changes. On the contrary, the far-ultraviolet spectra were almost identical.


Properties of fiber optics for application in astronomical interferometry.
We tested such properties of single-mode and multimode fiber optics that are relevant for use in a long-baseline astronomical interferometer, and we give quantitative values for modulation, transmission, and stability of the fiber optics as measured in a Mach-Zehnder interferometer configuration. Only polarization-maintaining single-mode fibers gave satisfying results. Good preservation of modulation and ease of beam combining are strong features of these fibers, whereas the light losses that are due to imperfect coupling may present a serious problem.


The simultaneous use of electrocochleogram, brainstem auditory evoked potential and facial muscle EMG in cerebellopontine angle tumor removal.
In six cases of acoustic neurilemmoma, electrocochleogram (ECOchG), brainstem auditory evoked potentials (BAEP) and facial muscle electromyograms (EMG) were recorded to monitor facial nerve and brainstem function. Under isoflurane and nitrous oxide anesthesia, we recorded ECOchG from the tympanic membrane, BAEP from the scalp needle, and facial muscle EMG from the mentalis muscle. During surgery, the body temperature was kept above 36.5 degrees C, and PaCO2 above 30 mmHg. In all cases, the peak N1 of ECOchG and wave I of BAEP had identical latencies throughout the monitoring period. The response was faster and the amplitude was higher in the ECOchG recordings. For calculation of the I-III or I-V interpeak latency of BAEP, the wave I of BAEP could be confirmed more quickly and precisely by the peak N1 of ECOchG. During tumor removal, the embedded facial nerve pathway in the tumor was identified by electric stimulation of the intracranial facial nerve, followed by evoked facial muscle EMG. Facial nerve function was confirmed by nerve traction or direct electric stimulation after total removal of the tumor. No facial palsy or other neurologic sequelae was found after the operations.


Microsaccadic sampling of moving image information provides Drosophila hyperacute vision.
Small fly eyes should not see fine image details. Because flies exhibit saccadic visual behaviors and their compound eyes have relatively few ommatidia (sampling points), their photoreceptors would be expected to generate blurry and coarse retinal images of the world. Here we demonstrate that Drosophila see the world far better than predicted from the classic theories. By using electrophysiological, optical and behavioral assays, we found that R1-R6 photoreceptors' encoding capacity in time is maximized to fast high-contrast bursts, which resemble their light input during saccadic behaviors. Whilst over space, R1-R6s resolve moving objects at saccadic speeds beyond the predicted motion-blur-limit. Our results show how refractory phototransduction and rapid photomechanical photoreceptor contractions jointly sharpen retinal images of moving objects in space-time, enabling hyperacute vision, and explain how such microsaccadic information sampling exceeds the compound eyes' optical limits. These discoveries elucidate how acuity depends upon photoreceptor function and eye movements.


Gene-expression profiling discriminates between cerebral malaria (CM)-susceptible mice and CM-resistant mice.
The development of cerebral malaria (CM) in mice with Plasmodium berghei ANKA infection is under genetic control. Brain gene-expression patterns were investigated in well-defined genetically CM-resistant (CM-R; BALB/c and DBA/2) and CM-susceptible (CM-S; C57BL/6 and CBA/J) mice by use of cDNA microarrays. By combining transcriptional profiling with rigorous statistical methods and cluster analysis, we identified a set of 69 genes that perfectly discriminated between mouse strains and between CM-R and CM-S mice. The analysis of gene ontological terms revealed that the genes that clustered and were related to susceptibility to CM preferentially belonged to some biological process classes, such as those pertaining to immune responses. Using a false discovery rate of 5% and the Welch t test, we identified 31 genes with consistent differential expression between CM-R and CM-S mice. These data indicate that microarray analysis may be useful for identification of candidate genes that are potentially responsible for resistance or susceptibility to mouse CM and suggest that candidate genes identified in mice could be specifically tested in humans for an association with disease severity.


Eosinophilic pustular folliculitis (Ofuji's disease). Immunohistochemical analysis.
Eosinophilic pustular folliculitis is a distinctive dermatosis that was first described in Japan. Although the histopathologic feature of eosinophilic pustular folliculitis is characterized by follicular infiltrates with numerous eosinophils, its pathophysiology remains unclear. The lesional skin of five patients with eosinophilic pustular folliculitis was examined using several monoclonal antibodies including a variety of anti-leukocyte adhesion molecules: endothelial cell adhesion molecule 1, vascular cell adhesion molecule 1, and intercellular adhesion molecule 1, by means of immunohistochemical techniques. Intercellular adhesion molecule 1 expression by keratinocytes was observed on follicular epithelium, but not on epidermis. The migration of eosinophils and lymphocytes, which were intensely positive for antilymphocyte function-associated antigen 1, was limited to follicular epithelium. Endothelial cell adhesion molecule 1 and vascular cell adhesion molecule 1 expression of vascular endothelium was more often observed around hair follicles. There was no reactivity for interleukin 8 in follicular epithelium. These findings may explain the selective migration of eosinophils and lymphocytes to the hair follicles in eosinophilic pustular folliculitis.


Resilience to capsaicin-induced mitochondrial damage in trigeminal ganglion neurons.
Capsaicin is an agonist of transient receptor potential cation channel subfamily V member 1 (TRPV1). Strong TRPV1 stimulation with capsaicin causes mitochondrial damage in primary sensory neurons. However, the effect of repetitive and moderate exposure to capsaicin on the integrity of neuronal mitochondria remains largely unknown. Our electron microscopic analysis revealed that repetitive stimulation of the facial skin of mice with 10 mM capsaicin induced short-term damage to the mitochondria in small-sized trigeminal ganglion neurons. Further, capsaicin-treated mice exhibited decreased sensitivity to noxious heat stimulation, indicating TRPV1 dysfunction, in parallel with the mitochondrial damage in the trigeminal ganglion neurons. To analyze the capsaicin-induced mitochondrial damage and its relevant cellular events in detail, we performed cell-based assays using TRPV1-expressing PC12 cells. Dose-dependent capsaicin-mediated mitochondrial toxicity was observed. High doses of capsaicin caused rapid destruction of mitochondrial internal structure, while low doses induced mitochondrial swelling. Further, capsaicin induced a dose-dependent loss of mitochondria and autophagy-mediated degradation of mitochondria (mitophagy). Concomitantly, transcriptional upregulation of mitochondrial proteins, cytochrome c oxidase subunit IV, Mic60/Mitofilin, and voltage-dependent anion channel 1 was observed, which implied induction of mitochondrial biogenesis to compensate for the loss of mitochondria. Collectively, although trigeminal ganglion neurons transiently exhibit mitochondrial damage and TRPV1 dysfunction following moderate capsaicin exposure, they appear to be resilient to such a challenge. Our in vitro data show a dose-response relationship in capsaicin-mediated mitochondrial toxicity. We postulate that induction of mitophagy and mitochondrial biogenesis in response to capsaicin stimulation play important roles in repairing the damaged mitochondrial system.


Long-term evolution of brainstem electrical evoked responses to sound after restricted ablation of the auditory cortex.
This study aimed to assess the top-down control of sound processing in the auditory brainstem of rats. Short latency evoked responses were analyzed after unilateral or bilateral ablation of auditory cortex. This experimental paradigm was also used towards analyzing the long-term evolution of post-lesion plasticity in the auditory system and its ability to self-repair. Auditory cortex lesions were performed in rats by stereotactically guided fine-needle aspiration of the cerebrocortical surface. Auditory Brainstem Responses (ABR) were recorded at post-surgery day (PSD) 1, 7, 15 and 30. Recordings were performed under closed-field conditions, using click trains at different sound intensity levels, followed by statistical analysis of threshold values and ABR amplitude and latency variables. Subsequently, brains were sectioned and immunostained for GAD and parvalbumin to assess the location and extent of lesions accurately. Alterations in ABR variables depended on the type of lesion and post-surgery time of ABR recordings. Accordingly, bilateral ablations caused a statistically significant increase in thresholds at PSD1 and 7 and a decrease in waves amplitudes at PSD1 that recover at PSD7. No effects on latency were noted at PSD1 and 7, whilst recordings at PSD15 and 30 showed statistically significant decreases in latency. Conversely, unilateral ablations had no effect on auditory thresholds or latencies, while wave amplitudes only decreased at PSD1 strictly in the ipsilateral ear. Post-lesion plasticity in the auditory system acts in two time periods: short-term period of decreased sound sensitivity (until PSD7), most likely resulting from axonal degeneration; and a long-term period (up to PSD7), with changes in latency responses and recovery of thresholds and amplitudes values. The cerebral cortex may have a net positive gain on the auditory pathway response to sound.


The contracting round: achieving health gain or financial balance?
In the 1991 National Health Service reforms, health authorities became responsible for the health of their resident population, and they contract for health services from NHS providers - trusts and primary care services. A case study in Camden and Islington, an inner London health district, during 1996-1997 shows that contracting was directed more towards achieving financial balance than health objectives. Reasons include the inflationary effect of competition within an internal market, the power of administrators in decision-making within the health authority, and lack of adequate financial accounting in the NHS to relate costs to health outcomes. The introduction of programme budgets for districts would provide more cost-effective use of the nation's resources.


Net changes in antibiotic concentrations downstream from an effluent discharge.
Many studies have shown the occurrence of antibiotics and degradation products in streams; however, relatively little work has applied a functional perspective to antibiotic transport and uptake. This study examined net changes in antibiotic concentrations downstream from a wastewater treatment plant (WWTP) effluent discharge and estimated net uptake length (Snet), net uptake velocity (v(f-net)), and net areal uptake rate (Unet) of antibiotics over a 3-km stream reach at Mud Creek, northwest Arkansas, USA, during June, September, and December 2006. Ten antibiotics and one degradation product (azithromycin, ciprofloxacin, erythromycin, erythromycin-H2O, ofloxacin, sulfachloropyridazine, sulfadimethoxine, sulfamethoxazole, tetracycline, oxytetracycline, and trimethoprim) were found at least once at Mud Creek downstream from the effluent discharge. All chemicals persisted in measurable concentrations in the water column over the 3-km stream reach, and we observed significant net retention of some antibiotics and one degradation product across the sampling events. Antibiotics that were significantly retained traveled kilometer-scale distances (Snet: 1.8 to 51.5 km) with relatively low uptake velocities (v(f-net): 1.6 to 33.9 x 10(-6) m s(-1)) and rates (Unet: 0.01 to 38.4 x 10(-6) microg m(-2) s(-1)). This study illustrates that some antibiotics do not travel conservatively in streams and that uptake processes occur over the scale of kilometers, linking upstream effluent sources to downstream processing over large spatial scales.


Early developmental exposure to volatile anesthetics causes behavioral defects in Caenorhabditis elegans.
Mounting evidence from animal studies shows that anesthetic exposure in early life leads to apoptosis in the developing nervous system. This loss of neurons has functional consequences in adulthood. Clinical retrospective reviews have suggested that multiple anesthetic exposures in early childhood are associated with learning disabilities later in life as well. Despite much concern about this phenomenon, little is known about the mechanism by which anesthetics initiate neuronal cell death. Caenorhabditis elegans, a powerful genetic animal model, with precisely characterized neural development and cell death pathways, affords an excellent opportunity to study anesthetic-induced neurotoxicity. We hypothesized that exposing the nematode to volatile anesthetics early in life would induce neuron cell death, producing a behavioral defect that would be manifested in adulthood. After synchronization and hatching, larval worms were exposed to volatile anesthetics at their 95% effective concentration for 4 hours. On day 4 of life, exposed and control worms were tested for their ability to sense and move to an attractant (i.e., to chemotax). We determined the rate of successful chemotaxis using a standardized chemotaxis index. Wild-type nematodes demonstrated striking deficits in chemotaxis indices after exposure to isoflurane (ISO) or sevoflurane (SEVO) in the first larval stage (chemotaxis index: untreated, 85 ± 2; ISO, 52 ± 2; SEVO, 47 ± 2; P < 0.05 for both exposures). The mitochondrial mutant gas-1 had a heightened effect from the anesthetic exposure (chemotaxis index: untreated, 71 ± 2; ISO, 29 ± 12; SEVO, 24 ± 13; P < 0.05 for both exposures). In contrast, animals unable to undergo apoptosis because of a mutation in the pathway that mediates programmed cell death (ced-3) retained their ability to sense and move toward an attractant (chemotaxis index: untreated, 76 ± 10; ISO, 73 ± 9; SEVO, 76 ± 10). Furthermore, we discovered that the window of greatest susceptibility to anesthetic neurotoxicity in nematodes occurs in the first larval stage after hatching (L1). This coincides with a period of neurogenesis in this model. All values are means ± SD. These data indicate that anesthetics affect neurobehavior in nematodes, extending the range of phyla in which early exposure to volatile anesthetics has been shown to cause functional neurological deficits. This implies that anesthetic-induced neurotoxicity occurs via an ancient underlying mechanism. C elegans is a tractable model organism with which to survey an entire genome for molecules that mediate the toxic effects of volatile anesthetics on the developing nervous system.


High-sensitivity cardiac troponin t concentrations below the limit of detection to exclude acute myocardial infarction: a prospective evaluation.
Initial reports suggest that concentrations of high-sensitivity cardiac troponin T (hs-cTnT) (Roche Diagnostics Elecsys(®)) below the limit of blank (LoB) (3 ng/L) or limit of detection (LoD) (5 ng/L) of the assay have almost 100% negative predictive value (NPV) for acute myocardial infarction (AMI), particularly among patients without electrocardiograph (ECG) evidence of ischemia. We aimed to prospectively validate those findings. We included adults presenting to the emergency department with suspected cardiac chest pain. Standard troponin T (cTnT) and hs-cTnT (both Roche Elecsys) were tested in samples drawn on arrival. The primary outcome was AMI, adjudicated by 2 investigators on the basis of clinical data and ≥12-h cTnT testing. We also evaluated diagnostic performance when AMI was readjudicated on the basis of hs-cTnT (≥12-h) concentrations. Of 463 patients included, 79 (17.1%) had AMI. Twenty-four patients (5.2%) had hs-cTnT concentrations below the LoB, although none had AMI. Ninety-six patients (20.7%) had hs-cTnT concentrations below the LoD, 1 of whom had AMI. Thus, diagnostic sensitivity was 98.7% (95% CI 87.5%-98.6%) and NPV was 99.0% (95% CI 94.3%-100.0%). Of the 17.3% (n = 80) patients with hs-cTnT below the LoD and no ECG ischemia, none had AMI. Thus, diagnostic sensitivity was 100.0% (95% CI 95.4%-100.0%) and NPV was 100.0% (95% CI 95.5%-100.0%). Sensitivity and NPV were maintained when AMI was readjudicated on the basis of hs-cTnT. Our findings confirm that patients with nonischemic ECG and undetectable hs-cTnT at presentation have a very low probability of AMI, although the proportion of patients affected was smaller than in previous research.


Improving the quality of survivorship for older adults with cancer.
In May 2015, the Cancer and Aging Research Group, in collaboration with the National Cancer Institute and the National Institute on Aging through a U13 grant, convened a conference to identify research priorities to help design and implement intervention studies to improve the quality of life and survivorship of older, frailer adults with cancer. Conference attendees included researchers with multidisciplinary expertise and advocates. It was concluded that future intervention trials for older adults with cancer should: 1) rigorously test interventions to prevent the decline of or improve health status, especially interventions focused on optimizing physical performance, nutritional status, and cognition while undergoing cancer treatment; 2) use standardized care plans based on geriatric assessment findings to guide targeted interventions; and 3) incorporate the principles of geriatrics into survivorship care plans. Also highlighted was the need to integrate the expertise of interdisciplinary team members into geriatric oncology research, improve funding mechanisms to support geriatric oncology research, and disseminate high-impact results to the research and clinical community. In conjunction with the 2 prior U13 meetings, this conference provided the framework for future research to improve the evidence base for the clinical care of older adults with cancer. Cancer 2016;122:2459-68. © 2016 American Cancer Society.


Insights on the identities of sharks of the Rhizoprionodon acutus (Elasmobranchii: Carcharhiniformes) species complex based on three new species of Phoreiobothrium (Cestoda: Onchoproteocephalidea).
Recent molecular work on milk sharks (Rhizoprionodon acutus [Rüppell]) suggests that, rather than a single widely distributed species, R. acutus represents a complex of four narrowly distributed cryptic species. Examination of the cestodes in three of the four members of that complex globally led to the discovery and description of three new species in the onchoproteocephalidean genus Phoreiobothrium Linton, 1889. The host associations and geographic distributions of the new species are fully congruent with the geographic distributions and species boundaries inferred for the sharks from molecular data: Phoreiobothrium jahki n. sp. parasitizes Rhizoprionodon cf. acutus 3 off Borneo, P. nadiae n. sp. parasitizes R. cf. acutus 1 off Senegal, and P. swaki n. sp. parasitizes R. cf. acutus 2 off northern Australia. The new cestodes differ from one another and from their 11 valid congeners in morphological features such as sublocular configuration and number, hook size, and testis number. Given the notoriously oioxenous nature of elasmobranch-hosted onchoproteocephalidean cestodes, these results provide further support for recognition of the milk shark species complex. This work also raises questions about the Phoreiobothrium species reported in cursory descriptions from India; further examination of these cestodes is key because they are potentially hosted by the fourth member of the R. acutus complex. To encourage future taxonomic work on the morphology of sharks in this complex, comparative photographs of representatives of the four potential host species are provided.


Enrichment of in vitro and in vivo immunologic activity of purified fractions of calf thymic hormone.
Thymus humoral factor (THF), a thymus hormone which participates in the processes leading to acquisition of immunocompetence of lymphoid cells has been isolated in our laboratory by a stepwise gel filtration through various Sephadex columns. THF so isolated appears to be a polypeptide of 3,000 mol wt which contains approximately 30 amino acid residues. Here we have tested the biological activity of THF fractions of successive degrees of purity upon lymphoid cells from both intact and neonatally thymectomized mice. The lymphoid cell populations were treated with the various THF fractions by in vitro incubation for a short time and by repeated injection in vivo. The treated cells evidenced increased ability to react in the graft-versus-host assay in vivo and in mixed lymphocyte cultures in vitro concomitantly with the rise of intracellular cAMP. On the other hand no activity whatsoever was shown by any of the control materials tested. These bioassays permitted isolation of fractions progressively more active than the original crude dialyzate of thymus extract tested. Thus the active peptide component of THF eluted from DEAE Sephadex A-25 column was estimated to be 2 X 10(4)-fold more active than the crude dialyzate of thymus extract which served as a starting material.


Adenosine deaminase enhances the immunogenicity of human dendritic cells from healthy and HIV-infected individuals.
ADA is an enzyme implicated in purine metabolism, and is critical to ensure normal immune function. Its congenital deficit leads to severe combined immunodeficiency (SCID). ADA binding to adenosine receptors on dendritic cell surface enables T-cell costimulation through CD26 crosslinking, which enhances T-cell activation and proliferation. Despite a large body of work on the actions of the ecto-enzyme ADA on T-cell activation, questions arise on whether ADA can also modulate dendritic cell maturation. To this end we investigated the effects of ADA on human monocyte derived dendritic cell biology. Our results show that both the enzymatic and non-enzymatic activities of ADA are implicated in the enhancement of CD80, CD83, CD86, CD40 and CCR7 expression on immature dendritic cells from healthy and HIV-infected individuals. These ADA-mediated increases in CD83 and costimulatory molecule expression is concomitant to an enhanced IL-12, IL-6, TNF-α, CXCL8(IL-8), CCL3(MIP1-α), CCL4(MIP-1β) and CCL5(RANTES) cytokine/chemokine secretion both in healthy and HIV-infected individuals and to an altered apoptotic death in cells from HIV-infected individuals. Consistently, ADA-mediated actions on iDCs are able to enhance allogeneic CD4 and CD8-T-cell proliferation, globally yielding increased iDC immunogenicity. Taken together, these findings suggest that ADA would promote enhanced and correctly polarized T-cell responses in strategies targeting asymptomatic HIV-infected individuals.


Developing therapeutic rapport: a training validation study.
Patient-centred consulting is at the heart of modern medicine, and training programmes now routinely highlight a range of communication skills that enhance the doctor-patient relationship. The present pilot study focuses on the central role played by empathic skills in developing rapport and shared understanding, as captured in a new model developed by the authors. The specific aim of the study was to evaluate the potential of a short-term training programme using this model to improve rapport-related behaviour in general practitioner (GP) trainees. In a quasi-experimental study, 37 GP trainees from schemes in central England (the training group) participated in a training programme based around a new model of therapeutic rapport, with pre- and post-training analysis of relevant knowledge-based, affective and behavioural outcomes. A control group of ten GP trainees, from schemes in a comparable UK medical deanery, took no part in the training programme but were involved in all pre- and post-training analyses. The training group demonstrated significant increases in rapport-related knowledge and all three affective dimensions (attitudes, confidence and motivation); there was a similar finding in terms of 'positive engagement' and all expert-rated aspects of rapport-related behaviours. The control group showed no comparable improvement in any area, and recorded a significant drop in demonstration of positive engagement behaviours. The significant improvement in rapport-related knowledge, attitudes and behaviour after training, and the lack of any comparable change in the control group, gives some support to the suggestion that the module (and the model that underpinned it) may have triggered such an impact independent of ongoing experience or other educational activities.


A pilot experimental lateral ridge augmentation study using bone morphogenetic protein 2 in dogs.
The loss of multiple teeth in the posterior mandible often results in deficient ridge width for prosthetic tooth rehabilitation. Multiple approaches have been used to regenerate lost bone; however, the outcomes are highly variable. Several approaches use combination therapy to make the procedure more predictable. In this experimental study in dogs, a chronic defect was created and then treated with one of four therapies: (1) autograft, (2) recombinant human bone morphogenetic protein 2 (rhBMP-2) in absorbable collagen sponge (ACS), (3) ACS wrapped around a collagen-ceramic matrix, and (4) rhBMP-2 in ACS around the collagen-ceramic matrix. Two metal screws were used for space maintenance. After 2 months, the metal pins were removed and an endosseous dental implant was placed in the regenerated bone. Two months later, the animals were sacrificed and specimens prepared for histologic evaluation. Only five animals were used, with each condition evaluated in each animal. With this low number of animals and with the observed variability, no quantitative differences were found between each of the four conditions evaluated. All conditions resulted in new bone growth. Dense native bone was found in the autograft sites and the sites treated with rhBMP-2 and ACS. Sites treated with the collagen-ceramic matrix with and without rhBMP-2 in the ACS had residual ceramic and large porous areas. Bone was found in varying degrees along the implant surfaces. These results suggest that multiple approaches can be used to augment bone horizontally in the posterior mandible of dogs. Interestingly, rhBMP-2 combined with a non-space maintaining collagen carrier yielded new bone densities similar to the autograft in this model.


Facility and area variation affecting the use of physical restraints in nursing homes.
Various individual characteristics increase a nursing home resident's likelihood of being mechanically restrained. However, there is not good information on the role played by facility characteristics and geographic area in determining the likelihood of physical restraint use. This study investigates the importance of individual, facility, and area indicators in determining the likelihood of restraint use. This research uses data on 2,000 nursing home residents receiving care in more than 250 nursing homes in 10 different areas. Resident-level data were collected in 1990 by research nurses using the Minimum Data Set for Nursing Home Resident Assessment and Care Screening. Facility-level data were gathered by telephone interviews with facility staff. The relations among the variables were investigated using contingency tables and multivariate logistic regression models. Controlling for residents' physical and cognitive function, facility characteristics and location had a significant impact on restraint practices. Those facilities with low nurse staffing were more likely to restrain residents. Furthermore, controlling for these factors, restraint practices varied significantly across the geographic areas in the study. Like hospitals, nursing homes exhibit significant interfacility and regional variation in their clinical practices. Future analyses of nursing home care might benefit from considering both factors in much finer detail.