Rasagiline: a review of its use in the treatment of idiopathic Parkinson's disease.
Rasagiline (Azilect®), a selective, irreversible, monoamine oxidase-B inhibitor, is available in the EU, the US and in several other countries worldwide, including Canada and Israel. It is indicated for the treatment of idiopathic Parkinson's disease as monotherapy or as adjunctive therapy to levodopa in patients [corrected]with end-of-dose fluctuations in the EU and for the treatment of adult patients with the signs and symptoms of idiopathic Parkinson's disease in the US. This article reviews the pharmacological properties, therapeutic efficacy and tolerability of rasagiline as monotherapy or as adjunctive therapy to levodopa in patients with Parkinson's disease. Oral rasagiline as monotherapy or as adjunctive therapy to levodopa was effective in the symptomatic treatment of adult patients with Parkinson's disease participating in double-blind, placebo-controlled, multinational studies. In patients with early Parkinson's disease, monotherapy with rasagiline 1 mg/day (recommended dosage) significantly slowed the rate of worsening (i.e. an increase in the Unified Parkinson's Disease Rating Scale [UPDRS] score) in the ADAGIO and TEMPO studies, with the results from the ADAGIO study for rasagiline 1 mg/day suggesting a slowing of clinical progression. However, at the higher dosage of 2 mg/day, rasagiline met the primary endpoint in the TEMPO study and the first, but not the second, of three hierarchical primary endpoints in the ADAGIO study. Compared with delayed-start rasagiline monotherapy, early initiation was associated with a slower long-term progression of the clinical signs and symptoms of Parkinson's disease in the TEMPO study. As adjunctive therapy to levodopa in the LARGO and PRESTO studies, rasagiline 0.5 and/or 1 mg/day significantly reduced the total daily 'off' time (primary efficacy endpoint) and significantly improved the Clinical Global Impression score, the UPDRS activities of daily living subscale score during 'off' time and the UPDRS motor subscale score during 'on' time compared with placebo in patients with advanced Parkinson's disease. Although rasagiline showed neuroprotective properties both in vitro and in vivo, identifying its potential to slow clinical progression in the clinical setting has been elusive to date and was not definitively demonstrated in the studies discussed in this article. Additional rasagiline studies specifically designed to assess the clinical progression of Parkinson's disease while addressing the potentially confounding factors of the delayed-start study design would therefore be of interest. As monotherapy or as adjunctive therapy to levodopa, rasagiline was generally well tolerated, with the frequency and nature of treatment-emergent adverse events generally similar across clinical studies and between rasagiline and placebo groups. Therapy with rasagiline appears to be associated with a low incidence of cognitive and behavioural adverse events. Thus, oral rasagiline as monotherapy or as adjunctive therapy to levodopa provides a useful option in the treatment of adult patients with Parkinson's disease.


Experimental, clinical, and metabolic results of side-to-side portacaval shunt for intractable cirrhotic ascites.
Intractable ascites, refractory to medical therapy, occurs in approximately 10 percent of patients with ascites from cirrhosis and is almost always fatal. Sinusoidal hypertension resulting from hepatic venous outflow obstruction plays a primary role in the pathogenesis of cirrhotic ascites and provides the rationale for decompression of the liver by side-to-side portacaval shunt in treatment of intractable ascites. This report presents the experimental basis for the use of side-to-side shunt and long-term results of a prospective study in 34 selected patients with intractable cirrhotic ascites. In the experimental studies, hepatic venous outflow obstruction and massive ascites were produced in dogs by ligation of the hepatic veins, and the effect of portacaval shunts on ascites, thoracic duct lymph flow, and aldosterone secretion were measured. In the clinical study, 34 carefully selected patients with cirrhosis (91 percent alcoholic) and truly intractable ascites (failure of medical therapy for 5 to 24 months) underwent side-to-side portacaval shunt. The effects on ascites, survival, metabolic abnormalities, and quality of life were studied prospectively during follow-up that was longer than 5 years in all but two patients. Quantitative Child's risk classes in percent of patients were A in 0, B in 68, and C in 32. In the experimental studies, side-to-side portacaval shunt permanently relieved severe ascites, reduced the 13-fold increase in thoracic duct lymph flow rate to almost normal, and abolished the aldosterone hypersecretory response to minimal hepatic venous outflow obstruction. End-to-side portacaval shunt was much less effective. In the clinical study, side-to-side portacaval shunt reduced mean portal vein-inferior vena cava pressure gradient from 282 mm saline to 4 mm and permanently relieved all patients of ascites without subsequent requirement of diuretic therapy. Two patients who died of hepatoma, and one who died of heart failure developed terminal ascites. Thirty-day mortality rate was 6 percent, and long-term survival rates at 5, 10, and 15 years were 75 percent, 74 percent, and 73 percent. In metabolic studies, side-to-side shunt produced marked diuresis and natriuresis and abolished hypersecretion of aldosterone. Quality of life was generally improved as a result of a low incidence of recurrent portal-systemic encephalopathy (6 percent), abstinence from alcohol in 91 percent, improvement in liver function in 81 percent, and improvement in Child's risk class. The portacaval anastomosis remained permanently patent in every patient. Side-to-side portacaval shunt is very effective treatment of intractable ascites from cirrhosis. Our results are attributable to careful selection of patients, an organized system of care, and a program of rigorous, lifelong follow-up that emphasizes abstinence from alcohol and dietary protein restriction.


On the structural differences between markers and genomic AC microsatellites.
AC microsatellites have proved particularly useful as genetic markers. For some purposes, such as in population biology, the inferences drawn depend on the quantitative values of their mutation rates. This, together with intrinsic biological interest, has led to widespread study of microsatellite mutational mechanisms. Now, however, inconsistencies are appearing in the results of marker-based versus non-marker-based studies of mutational mechanisms. The reasons for this have not been investigated, but one possibility, pursued here, is that the differences result from structural differences between markers and genomic microsatellites. Here we report a comparison between the CEPH AC marker microsatellites and the global population of AC microsatellites in the human genome. AC marker microsatellites are longer than the global average. Controlling for length, marker microsatellites contain on average fewer interruptions, and have longer segments, than their genomic counterparts. Related to this, marker microsatellites show a greater tendency to concentrate the majority of their repeats into one segment. These differences plausibly result from scientists selecting markers for their high polymorphism. In addition to the structural differences, there are differences in the base composition of flanking sequences, marker flanking regions being richer in C and G and poorer in A and T. Our results indicate that there are profound differences between marker and genomic microsatellites that almost certainly affect their mutation rates. There is a need for a unified model of mutational mechanisms that accounts for both marker-derived and genomic observations. A suggestion is made as to how this might be done.


Toward atomic resolution diffractive imaging of isolated molecules with X-ray free-electron lasers.
We give a detailed account of the theoretical analysis and the experimental results of an X-ray-diffraction experiment on quantum-state selected and strongly laser-aligned gas-phase ensembles of the prototypical large asymmetric rotor molecule 2,5-diiodobenzonitrile, performed at the Linac Coherent Light Source [Phys. Rev. Lett.112, 083002 (2014)]. This experiment is the first step toward coherent diffractive imaging of structures and structural dynamics of isolated molecules at atomic resolution, i.e., picometers and femtoseconds, using X-ray free-electron lasers.


Preparation of Poly(vinyl Alcohol) Microparticles for Freeze Protection of Sensitive Fruit Crops.
Poly(vinyl alcohol) (PVA) displays ice recrystallization inhibition (IRI) properties as many antifreeze proteins found in cold tolerant organisms. The molecular architecture and composition (molecular weight and distribution of pendant OH and acetate groups) have been studied to improve the antifreezing properties of PVA, suggesting that the molecular architecture of PVA plays an important role in IRI activity. The present work deals with the preparation of PVA microparticles using an alkaline treatment. The effect of PVA molecular weight on the morphology and antifreezeing properties of PVA microparticles was investigated. The antifreezeing property of PVA microparticles on the susceptibility of flower bud tissues to freeze damage was also evaluated. The alkaline treatment of an aqueous PVA solution produced stable polymer chain aggregates with spherical shapes. The average size of the PVA microparticles increased significantly with the increasing molecular weight of the PVA macromolecule precursor. The PVA microparticles inhibited the growth of ice crystals and blocked ice growth at concentrations as low as 0.01 % w/v. The effect of impeding ice crystal growth by preventing the joining of adjacent ice crystals is attributed to the larger size of the PVA particles adsorbed on the ice surface compared to the aggregated PVA macromolecules in saline solution. The thermal hysteresis activity of PVA macromolecules and microparticles was not detected by differential scanning calorimetry analysis. The PVA microparticles reduced the incidence of freeze injuries in flower bud tissues by 55% and their application, considering the low toxicity of PVA, has a high potential for freeze protection in fruit crops.


Therapy in prion diseases.
In the last two decades, knowledge of the neurobiology of prion diseases or transmissible spongiform encephalopathies (TSE) has significantly advanced, but a successful therapy to stop or delay the progression of these disorders remains one of the most challenging goals of biomedical research. Several obstacles to this achievement are in common with other neurodegenerative disorders: difficulties to move from experimental level to clinical stage; appropriate timing of intervention; correct set up of clinical trial. Also in terms of molecular bases of disease, TSE and the other neurodegenerative disorders associated with protein misfolding such as Alzheimer, Parkinson and Huntington diseases, share a central pathogenic role of soluble small aggregates, named oligomers, considered the culprit of neuronal dysfunction: accordingly, these disorders could by termed oligomeropathies. However, the rapid progression of TSE, together with their clinical and molecular heterogeneity, make the therapeutic approach particularly problematic. The main target of the antiprion strategy has been the pathological form of the cellular prion protein (PrP(C)) termed PrP(Sc), invariably associated with the diseases. Several compounds have been found to affect PrP(Sc) formation or enhance its clearance in in vitro models, and prolong survival in experimental animals. However, few of them such as quinacrine and pentosan polysulfate have reached the clinical evaluation; more recently, we have conducted a clinical trial with doxycycline in patients with Creutzfeldt-Jakob disease without satisfactory results. In experimental conditions, active and passive immunization with antibodies against PrP and mucosal vaccination have shown to protect from peripheral infection. Other studies have proposed new potentially effective molecules targeting PrP oligomers. Furthermore, the possibility to interfere with PrP(C) to PrP(Sc) conversion by an active control of PrP(C) is another interesting approach emerging from experimental studies. However, in common with the other oligomeropathies, early diagnosis allowing to treat at risk population in a preclinical stage represent the more realistic perspective for efficient TSE therapy.


Transient Two-Dimensional Electronic Spectroscopy: Coherent Dynamics at Arbitrary Times along the Reaction Coordinate.
Recent advances in two-dimensional electronic spectroscopy (2DES) have enabled identification of fragile quantum coherences in condensed-phase systems near the equilibrium molecular geometry. In general, traditional 2DES cannot measure such coherences associated with photophysical processes that occur at times significantly after the initially prepared state has dephased, such as the evolution of the initial excited state into a charge transfer state. We demonstrate the use of transient two-dimensional electronic spectroscopy (t-2DES) to probe coherences in an electron donor-acceptor dyad consisting of a perylenediimide (PDI) acceptor and a perylene (Per) donor. An actinic pump pulse prepares the lowest excited singlet state of PDI followed by formation of the PDI•--Per•+ ion pair, which is probed at different times following the actinic pulse using 2DES. Analysis of the observed coherences provides information about electronic, vibronic, and vibrational interactions at any time along the reaction coordinate for ion pair formation.


Ischaemia is linked to inflammation and induction of angiogenesis in pancreatic islets.
Beta-cell replacement is the only way to restore euglycaemia in patients with type-1 diabetes. Pancreatic tissue, processed for subsequent clinical islet transplantation, is exposed to ischaemia causing injury and death in a large number of islets before and after transplantation. In this review we summarize what is known on the sources of environmental stress for pancreatic islets, such as insufficient oxygen supply during pancreas procurement and in culture prior to intraportal transplantation, nutritional and oxygen deprivation during the isolation process, and the consequences of hyperglycaemia. An increasingly recognized role in the modulation of beta-cell function and these environmental stress factors plays the vascular network of the pancreatic islets. Islet revascularization by angiogenesis is relevant for the survival of the graft subsequent to transplantation. Potential strategies offered by therapeutic induction of revascularization to ameliorate the detrimental impact of these factors on the quality of islet transplants are discussed.


Female gender may predict response to FOLFIRINOX in patients with unresectable pancreatic cancer: a single institution retrospective review.
FOLFIRINOX is a highly active regimen for the treatment of patients with unresectable pancreatic cancer. However, treatment with FOLFIRINOX is associated with relevant toxicity and predictors for response to therapy are warranted. We retrospectively analyzed 49 patients with unresectable pancreatic cancer treated with FOLFIRINOX in order to evaluate a possible predictive role of clinical parameters and tumor characteristics for response to chemotherapy. Tumor samples were characterized histopathologically before treatment and expression of p53 and Ki67 was analyzed using automated immunohistochemistry. Overall survival (OS) and progression-free survival (PFS) were estimated by the Kaplan-Meier method. The overall objective response rate was 55.1%, the disease control rate was 70.6%. Female gender was associated with a significantly higher disease control rate of 91.7 compared to 48.0% in male patients (p=0.001) which reached 100% in female patients when primarily treated compared to treatment after surgical resection and relapse (77.8%, p=0.057). For all patients median PFS was 3.5 months (95% CI, 2.7-4.3 months) and median OS was 13 months (95% CI, 9.4-16.6 months). Female patients showed a tendency towards a longer median PFS (5.0 months, 95% CI, 3.6-6.4 months) compared to males (3.0 months, 95% CI, 2.4-3.6 months) (p=0.099). Serum levels of CA19.9 and CEA were significantly higher in female patients compared to male patients (p=0.037, p=0.05). Tumors of patients with response to FOLFIRINOX showed a higher expression level of p53 and Ki67 as well as higher serum levels of CA19.9 compared to non-responders, which was statistically not significant. Our study indicates that female gender is a positive predictor for therapy response to FOLFIRINOX in patients with unresectable pancreatic cancer. Female gender in turn was associated with increased levels of tumor markers CEA and CA19.9 and patients with higher serum levels of CA19.9 were more responsive to FOLFIRINOX.


Recruitment of circulating breast cancer cells is stimulated by radiotherapy.
Radiotherapy (RT) is a localized therapy that is highly effective in killing primary tumor cells located within the field of the radiation beam. We present evidence that irradiation of breast tumors can attract migrating breast cancer cells. Granulocyte-macrophage colony stimulating factor (GM-CSF) produced by tumor cells in response to radiation stimulates the recruitment of migrating tumor cells to irradiated tumors, suggesting a mechanism of tumor recurrence after radiation facilitated by transit of unirradiated, viable circulating tumor cells to irradiated tumors. Data supporting this hypothesis are presented through in vitro invasion assays and in vivo orthotopic models of breast cancer. Our work provides a mechanism for tumor recurrence in which RT attracts cells outside the radiation field to migrate to the site of treatment.


Simultaneous comparisons of treatments at multiple time points: Combined marginal models versus joint modeling.
We discuss several aspects of multiple inference in longitudinal settings, focusing on many-to-one and all-pairwise comparisons of (a) treatment groups simultaneously at several points in time, or (b) time points simultaneously for several treatments. We assume a continuous endpoint that is measured repeatedly over time and contrast two basic modeling strategies: fitting a joint model across all occasions (with random effects and/or some residual covariance structure to account for heteroscedasticity and serial dependence), and a novel approach combining a set of simple marginal, i.e. occasion-specific models. Upon parameter and covariance estimation with either modeling approach, we employ a variant of multiple contrast tests that acknowledges correlation between time points and test statistics. This method provides simultaneous confidence intervals and adjusted p-values for elementary hypotheses as well as a global test decision. We compare via simulation the powers of multiple contrast tests based on a joint model and multiple marginal models, respectively, and quantify the benefit of incorporating longitudinal correlation, i.e. the advantage over Bonferroni. Practical application is illustrated with data from a clinical trial on bradykinin receptor antagonism.


Taxonomy and evolution of bacteriochlorophyll a-containing members of the OM60/NOR5 clade of marine gammaproteobacteria: description of Luminiphilus syltensis gen. nov., sp. nov., reclassification of Haliea rubra as Pseudohaliea rubra gen. nov., comb. nov., and emendation of Chromatocurvus halotolerans.
Aerobic gammaproteobacteria affiliated to the OM60/NOR5 clade are widespread in saline environments and of ecological importance in several marine ecosystems, especially the euphotic zone of coastal areas. Within this group a close relationship between aerobic anoxygenic photoheterotrophs and non-phototrophic members has been found. Several strains of aerobic red-pigmented bacteria affiliated to the OM60/NOR5 clade were obtained from tidal flat sediment samples at the island of Sylt (North Sea, Germany). Two of the novel isolates, Rap1red and Ivo14(T), were chosen for an analysis in detail. Strain Rap1red shared a 16S rRNA sequence identity of 99% with the type strain of Congregibacter litoralis and was genome-sequenced to reveal the extent of genetic microheterogeneity among closely related strains within this clade. In addition, a draft genome sequence was obtained from the isolate Ivo14(T), which belongs to the environmental important NOR5-1 lineage that contains so far no cultured representative with a comprehensive description. Strain Ivo14(T) was characterized using a polyphasic approach and compared with other red-pigmented members of the OM60/NOR5 clade, including Congregibacter litoralis DSM 17192(T), Haliea rubra DSM 19751(T) and Chromatocurvus halotolerans DSM 23344(T). All analyzed strains contained bacteriochlorophyll a and spirilloxanthin as photosynthetic pigments. Besides a detailed phenotypic characterization including physiological and chemotaxonomic traits, sequence information based on protein-coding genes and a comparison of draft genome data sets were used to identify possible features characteristic for distinct taxa within this clade. Comparative sequence analyses of the pufLM genes of genome-sequenced representatives of the OM60/NOR5 clade indicated that the photosynthetic apparatus of these species was derived from a common ancestor and not acquired by multiple horizontal gene transfer from phylogenetically distant species. An affiliation of the characterized bacteriochlorophyll a-containing strains to different genera was indicated by significant phenotypic differences and pufLM nucleotide sequence identity values below 82%. The revealed high genotypic and phenotypic diversity of closely related strains within this phylogenetic group reflects a rapid evolution and frequent niche separation in the OM60/NOR5 clade, which is possibly driven by the necessities of an adaptation to oligotrophic marine habitats.


The extraordinary healing properties of the pediatric tympanic membrane: a study of atelectasis in the pediatric ear.
Traumatic perforations of the eardrum are generally treated conservatively as they tend to heal spontaneously. Retraction pockets of the eardrum, leading to atelectasis of the middle ear and often ultimately though unpredictably, to cholesteatoma formation, are treated in a variety of ways including complex grafting procedures. On the premise that a surgically resected retraction pocket is in essence a traumatic perforation, we examined the results of spontaneous healing of the resected retraction pocket. Retrospective case note study. A group of 86 ears in 62 patients who had undergone simple excision of the atrophic segment of the eardrum and who had no grafting procedure of the resultant defect, were followed at 6 days, 6-7 weeks and then every 6 months to determine the incidence of spontaneous healing of the eardrum. Audiograms were taken preoperatively, at 7 weeks and between 7 and 15 months postoperative. 94.2% of the eardrums (81 out of 86 ears) healed spontaneously without any form of grafting within 7 weeks. Air bone gap showed improvement in 53 patients. Only one of the four ears that failed to achieve closure had postoperative ear discharge. There were no other complications. In 17 ears (19.8%) there was a recurrence of the atelectasis. Excision of retraction pockets in pediatric ear atelectasis, in principle does not require grafting, as the great majority will heal spontaneously. The spontaneous healing in this study is comparable to reported studies of spontaneous healing of traumatic perforations. Irrespective of the size of the atrophic part of the membrane, recurrence rates for the atelectasis after simple excision is similar to that described in other studies. Retraction pockets may be excised and allowed to heal without risk to the hearing.


Treatment and outcomes for patients with bacteremic pneumococcal pneumonia.
Delayed time to antibiotic administration has been linked with higher mortality for patients with community-acquired pneumonia, but the impact of antibiotic resistance on clinical outcomes has been controversial. In the current study we assess the combined impact of antibiotic resistance and antibiotic timing on outcomes, including inhospital mortality, complications, length of stay, and time to stability, for patients hospitalized with community-acquired bacteremic pneumococcal pneumonia. We conducted a retrospective cohort study in 43 hospitals in the Southeastern Pennsylvania region from 2001 to 2004. Eligible adult patients had pneumococcal bacteremia and radiographic evidence of pneumonia. Outcomes were assessed based on medical record review. Multivariable regression was used to adjust for severity of illness and sequentially assess the impact of antibiotic resistance and time to active antibiotic therapy. The overall inhospital mortality was 10%. Overall, levels of macrolide, cephalosporin, and fluoroquinolone resistance were low and did not adversely impact the time to administration of active antibiotic therapy. Receipt of at least 1 active antibiotic within 4 hours was associated with reduced mortality (odds ratio [OR], 0.47; 95% confidence interval [CI], 0.2-1.0) and shortened length of stay (OR, 0.77; CI, 0.6-1.0) but did not reduce the risk of other adverse outcomes. We conclude that early antibiotic administration reduces the risks of mortality in patients with bacteremic pneumococcal pneumonia. Current patterns of drug resistance did not lead to delays in administration of active antimicrobial therapy.


Developmental regulation of heterochromatin-mediated gene silencing in Drosophila.
The roles of differentiation, mitotic activity and intrinsic promoter strength in the maintenance of heterochromatic silencing were investigated during development using an inducible lacZ gene as an in vivo probe. Heterochromatic silencing is initiated at the onset of gastrulation, approximately 1 hour after heterochromatin is first visible cytologically. A high degree of silencing is maintained in the mitotically active imaginal cells from mid-embryogenesis until early third instar larval stage, and extensive relaxation of silencing is tightly associated with the onset of differentiation. Relaxation of silencing can be triggered in vitro by ecdysone. In contrast, timing and extent of silencing at both the initiation and relaxation stages are insensitive to changes in cell cycle activity, and intrinsic promoter strength also does not influence the extent of silencing by heterochromatin. These data suggest that the silencing activity of heterochromatin is developmentally programmed.


Campylobacter jejuni colonization is associated with a dysbiosis in the cecal microbiota of mice in the absence of prominent inflammation.
Campylobacter jejuni causes enterocolitis in humans, but does not incite disease in asymptomatic carrier animals. To survive in the intestine, C. jejuni must successfully compete with the microbiota and overcome the host immune defense. Campylobacter jejuni colonization success varies considerably amongst individual mice, and we examined the degree to which the intestinal microbiota was affected in mice (i.e. a model carrier animal) colonized by C. jejuni at high relative to low densities. Mice were inoculated with C. jejuni or buffer, and pathogen shedding and intestinal colonization were measured. Histopathologic scoring and quantification of mRNA expression for α-defensins, toll-like receptors, and cytokine genes were conducted. Mucosa-associated bacterial communities were characterized by two approaches: multiplexed barcoded pyrosequencing and terminal restriction fragment length polymorphism analysis. Two C. jejuni treatments were established based on the degree of cecal and colonic colonization; C. jejuni Group A animals were colonized at high cell densities, and C. jejuni Group B animals were colonized at lower cell densities. Histological examination of cecal and colonic tissues indicated that C. jejuni did not incite visible pathologic changes. Although there was no significant difference among treatments in expression of mRNA for α-defensins, toll-like receptors, or cytokine genes, a trend for increased expression of toll-like receptors and cytokine genes was observed for C. jejuni Group A. The results of the two methods to characterize bacterial communities indicated that the composition of the cecal microbiota of C. jejuni Group A mice differed significantly from C. jejuni Group B and Control mice. This difference was due to a reduction in load, diversity and richness of bacteria associated with the cecal mucosa of C. jejuni Group A mice. High density colonization by C. jejuni is associated with a dysbiosis in the cecal microbiota independent of prominent inflammation.


[Emerging viral zoonoses: hantavirus infections].
Hantaviruses are rodent-borne agents belonging to the Bunyaviridae family. These viruses, which are found throughout Europe, Asia and the Americas, are maintained by different species of rodents, in which they produce chronic, inapparent infections. Humans become infected through contact with urine, saliva or faeces from infected rodents, mainly via the aerosol route. In humans, clinical disease occurs in the form of two major syndromes: haemorrhagic fever with renal syndrome (HFRS) or hantavirus pulmonary syndrome (HPS). Haemorrhagic fever with renal syndrome mainly occurs in Europe and Asia and HPS has only ever been reported in the Americas. Person-to-person transmission of hantaviruses, although uncommon, was described during an outbreak of HPS in southern Argentina. Most epidemics of HFRS and HPS occur in areas with large populations of rodents that have a relatively high prevalence of infection.


The measurement and analysis of age-related changes in Caenorhabditis elegans.
Aging is characterized by progressive degenerative changes in tissue organization and function that increase the probability of mortality. Major goals of aging research include elucidating the series of events that cause degenerative changes and analyzing environmental and genetic factors that modulate these changes. The basis for mechanistic studies of aging are accurate and precise descriptions of age-related changes, since these descriptions define the aging phenotype. Here we review studies that describe age-related changes in C. elegans including measurements of integrated functions such as behavior, microscopic analyses of tissue organization, and biochemical studies of macromolecules. Genetic and environmental factors that influence these changes are described, and studies that analyze the relationships between different age-related changes are discussed. Together these studies provide fundamental insights into aging in C. elegans that may be relevant to aging in other animals.


Developmental and temporal characteristics of clonal sperm mosaicism.
Throughout development and aging, human cells accumulate mutations resulting in genomic mosaicism and genetic diversity at the cellular level. Mosaic mutations present in the gonads can affect both the individual and the offspring and subsequent generations. Here, we explore patterns and temporal stability of clonal mosaic mutations in male gonads by sequencing ejaculated sperm. Through 300× whole-genome sequencing of blood and sperm from healthy men, we find each ejaculate carries on average 33.3 ± 12.1 (mean ± SD) clonal mosaic variants, nearly all of which are detected in serial sampling, with the majority absent from sampled somal tissues. Their temporal stability and mutational signature suggest origins during embryonic development from a largely immutable stem cell niche. Clonal mosaicism likely contributes a transmissible, predicted pathogenic exonic variant for 1 in 15 men, representing a life-long threat of transmission for these individuals and a significant burden on human population health.


Structural-thermodynamic relationships of interactions in the N-terminal ATP-binding domain of Hsp90.
Despite its importance as a target in anti-cancer therapeutics and the numerous rational-based inhibitor design efforts aimed at it, there are only limited data available on structural-thermodynamic relationships of interactions of the N-terminal ATP-binding domain of Hsp90 (N-Hsp90). Here, we redress this by presenting an investigation of binding of nucleotides and ansamycin compounds to this domain. Interactions of nucleotides with N-Hsp90 are relatively weak (>10 microM) and are strongly enthalpy driven over the temperature range 10-25 degrees C. Geldanamycin (GA) and its analogues 17-AAG [17-(allylamino)-17-demethoxy-GA] and 17-DMAG (17-N,N-dimethylaminoethylamino-17-demethoxy-GA) bind more strongly and have a dominant favourable enthalpic contribution over the temperature range investigated. We investigated the temperature dependence of the enthalpic contribution to binding. We found that while the ansamycin compounds have the commonly observed negative value, the nucleotides show a negligible or even a positive DeltaC(p) of binding. These data represent the first observation of a single binding site for which interactions with different ligands result in both negative and positive DeltaC(p) values. By addressing the likely impact of the potential contributions from protein-ligand interactions, we are able to attribute the anomalous DeltaC(p) for the nucleotides largely to a change in the conformation of the domain structure and local motion in the lid region of N-Hsp90 with the concomitant exposure of hydrophobic amino acid side chains.


Possible human-avian transmission of Mycobacterium tuberculosis in a green-winged macaw (Ara chloroptera).
This report describes a case of Mycobacterium tuberculosis infection in a green-winged macaw (Ara chloroptera), confirmed by microbiologic and pathologic diagnostics, and notes a possible human-avian transmission. Clinical signs included cutaneous swellings, profound leukocytosis, and signs of osteomyelitis in the long bones. Proliferation consisted of several nodules with small greenish-caseous foci in cross-section and revealed a severe granulomatous inflammation with intralesional acid-fast rods. Mycobacterium tuberculosis was isolated from subcutaneous nodules and biochemically confirmed. The disease in avian species is of zoonotic importance.


Linkage disequilibrium and modification of risk for Huntington disease.
The major limitation in performing predictive testing for Huntington disease (HD) is the unavailability of DNA from crucial family members. In our program approximately 20% (36/183) of persons have been excluded from predictive testing because of this reason. The major aim of this study was to examine whether data derived from linkage disequilibrium could modify risk analysis for persons at risk for HD. As a first step, we assessed whether the previously reported linkage disequilibrium between alleles recognized by probe pBS674E-D at locus D4S95 remained significant in a much larger data set. A total of 1,150 chromosomes from 622 individuals--200 affected and 422 unaffected--from 118 families were assessed. Significant haplotype association was detected with AccI and MboI RFLPs at the locus D4S95, with all the families (P = .00003), as well as for a subset from the United Kingdom (P = .0037). Data derived from linkage disequilibrium studies using D4S95 modifies the risk for HD, especially in persons of U.K. descent. Utilization of this approach for risk modification of HD awaits both validation of these data and additional information concerning ethnic-specific alleles at the D4S95 locus.


Productivity, embryo and eggshell characteristics, and contaminants in bald eagles from the Great Lakes, USA, 1986 to 2000.
Chlorinated hydrocarbon concentrations in eggs of fish-eating birds from contaminated environments such as the Great Lakes of North America tend to be highly intercorrelated, making it difficult to elucidate mechanisms causing reproductive impairment, and to ascribe cause to specific chemicals. An information- theoretic approach was used on data from 197 salvaged bald eagle (Haliaeetus leucocephalus) eggs (159 clutches) that failed to hatch in Michigan and Ohio, USA (1986-2000). Contaminant levels declined over time while eggshell thickness increased, and by 2000 was at pre-1946 levels. The number of occupied territories and productivity increased during 1981 to 2004. For both the entire dataset and a subset of nests along the Great Lakes shoreline, polychlorinated biphenyls (SigmaPCBs, fresh wet wt) were generally included in the most parsimonious models (lowest-Akaike's information criterion [AICs]) describing productivity, with significant declines in productivity observed above 26 microg/g SigmaPCBs (fresh wet wt). Of 73 eggs with a visible embryo, eight (11%) were abnormal, including three with skewed bills, but they were not associated with known teratogens, including SigmaPCBs. Eggs with visible embryos had greater concentrations of all measured contaminants than eggs without visible embryos; the most parsimonious models describing the presence of visible embryos incorporated dieldrin equivalents and dichlorodiphenyldichloroethylene (DDE). There were significant negative correlations between eggshell thickness and all contaminants, with SigmaPCBs included in the most parsimonious models. There were, however, no relationships between productivity and eggshell thickness or Ratcliffe's index. The SigmaPCBs and DDE were negatively associated with nest success of bald eagles in the Great Lakes watersheds, but the mechanism does not appear to be via shell quality effects, at least at current contaminant levels, while it is not clear what other mechanisms were involved.


Gene and protein kinase expression profiling of reactive oxygen species-associated lipotoxicity in the pancreatic beta-cell line MIN6.
Oligonucleotide microarrays were used to define oleic acid (OA)-regulated gene expression and proteomic technology to screen protein kinases in MIN6 insulinoma cells. The effects of oxidative stress caused by OA and potential protective effects of N-acetyl-L-cysteine (NAC), a scavenger of reactive oxygen species (ROS), on global gene expression and beta-cell function were investigated. Long-term exposure of MIN6 cells to OA led to a threefold increase in basal insulin secretion, a 50% decrease in insulin content, an inhibition of glucose-stimulated insulin secretion (GSIS), and a twofold increase in the level of ROS. The addition of NAC normalized both the OA-induced insulin content and ROS elevation, but it failed to restore GSIS. Microarray studies and subsequent quantitative PCR analysis showed that OA consistently regulated the expression of 45 genes involved in metabolism, cell growth, signal transduction, transcription, and protein processing. The addition of NAC largely normalized the expression of the OA-regulated genes involved in cell growth and differentiation but not other functions. A protein kinase screen showed that OA regulated the expression and/or phosphorylation levels of kinases involved in stress-response mitogen-activated protein kinase, phosphatidylinositol 3-kinase, and cell cycle control pathways. Importantly, these findings indicate that chronic OA exposure can impair beta-cell function through ROS-dependent and -independent mechanisms.


Effects of dipeptide insertions between codons 69 and 70 of human immunodeficiency virus type 1 reverse transcriptase on primer unblocking, deoxynucleoside triphosphate inhibition, and DNA chain elongation.
Finger insertion mutations of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) (T69S mutations followed by various dipeptide insertions) have a multinucleoside resistance phenotype that can be explained by decreased sensitivity to deoxynucleoside triphosphate (dNTP) inhibition of the nucleotide-dependent unblocking activity of RT. We show that RTs with SG or AG (but not SS) insertions have three- to fourfold-increased unblocking activity and that all three finger insertion mutations have threefold-decreased sensitivity to dNTP inhibition. The additional presence of M41L and T215Y mutations increased unblocking activity for all three insertions, greatly reduced the sensitivity to dNTP inhibition, and resulted in defects in in vitro DNA chain elongation. The DNA chain elongation defects were partially repaired by additional mutations at positions 210, 211, and 214. These results suggest that structural communication between the regions of RT defined by these mutations plays a role in the multinucleoside resistance phenotype.


Vitamin D deficiency rickets mimicking pseudohypoparathyroidism.
Vitamin D deficiency rickets (VDDR) is a disorder biochemically characterized by elevated serum alkaline phosphatase (ALP) activity, normal or decreased serum calcium (Ca) and inorganic phosphate concentrations, secondary hyperparathyroidism and decreased serum 25-hydroxyvitamin D (25(OH)D) levels. In stage 1 VDDR, urinary amino acid and phosphate excretion are normal with minimal or no findings of rickets on radiographs. Pseudohypoparathyroidism (PHP) is an inherited disorder characterized by end-organ resistance to parathormone (PTH). VDDR occasionally resembles PHP type 2 in clinical presentation and biochemical features, creating difficulties in the differential diagnosis of these two entities. Here we report an infant diagnosed with VDDR. In addition to inadequate vitamin D intake, usage of antiepileptic drugs (AED) may have led to the worsening of the vitamin D deficiency. The patient presented with a history of febrile convulsions, for which he received phenobarbital treatment. The initial findings of hypocalcemia, hyperphosphatemia and normal tubular reabsorption of phosphate, mimicking PHP 2, responded well to vitamin D and oral Ca treatment with normalization of serum Ca, phosphorus (P), ALP and PTH levels.


Adherence to COVID-19 Preventive Measures in Mozambique: Two Consecutive Online Surveys.
We assessed adherence to government recommendations implemented shortly after the introduction of COVID-19 in Mozambique in March 2020, through two online cross-sectional surveys in April and June 2020. We quantified adherence to preventive measures by a composite score comprising of five measures: physical distancing, face mask use, hand hygiene, cough hygiene, and avoidance of touching the face. 3770 and 1115 persons participated in the first and second round respectively. Wearing face masks, regular handwashing and cough hygiene all reached compliance rates of over 90% while physical distancing and avoiding to touch the face reached a compliance rate of 80-90%. A multivariable model investigating factors associated with adherence found that being older, more educated, and belonging to the healthcare sector increased the odds for higher adherence. Private workers and retired people, respondents receiving COVID-19 information through social media, and those who reported flu-like symptoms were less likely to adhere. 6% of respondents reported flu-like symptoms which aligned with the WHO clinical definition of COVID-19, suggesting low level community transmission. In conclusion, most respondents in this online survey in Mozambique complied well with strategies to prevent COVID-19. Whether the good preventive behaviour explains the low grade COVID-19 transmission requires further study.


Blastococcus atacamensis sp. nov., a novel strain adapted to life in the Yungay core region of the Atacama Desert.
A polyphasic study was undertaken to establish the taxonomic status of a Blastococcus strain isolated from an extreme hyper-arid Atacama Desert soil. The isolate, strain P6T, was found to have chemotaxonomic and morphological properties consistent with its classification in the genus Blastococcus. It was shown to form a well-supported branch in the Blastococcus 16S rRNA gene tree together with the type strains of Blastococcus capsensis and Blastococcus saxobsidens and was distinguished from the latter, its close phylogenetic neighbour, by a broad range of phenotypic properties. The draft genome sequence of isolate P6T showed 84.6 % average nucleotide identity, 83.0 % average amino acid identity and a digital DNA-DNA hybridisation value of 27.8 % in comparison with the genome sequence of B. saxobsidens DSM 44509T, values consistent with its assignment to a separate species. Based on these data it is proposed that isolate P6T (NCIMB 15090T=NRRL B-65468T) be assigned to the genus Blastococcus as Blastococcus atacamensis sp. nov. Analysis of the whole genome sequence of B. atacamensis P6T, with 3778 open reading frames and a genome size of 3.9 Mb showed the presence of genes and gene clusters that encode for properties that reflect its adaptation to the extreme environmental conditions that prevail in Atacama Desert soils.


Chemical composition, cytotoxicity effect and antimicrobial activity of Ceratonia siliqua essential oil with preservative effects against Listeria inoculated in minced beef meat.
The present study describes the phytochemical profile and the protective effects of Ceratonia siliqua pods essential oil (CsEO), a food and medicinal plant widely distributed in Tunisia. Twenty five different components were identified in the CsEO. Among them, the major detected components were: Nonadecane, Heneicosane , Naphthalene, 1,2-Benzenedicarboxylic acid dibutylester, Heptadecane, Hexadecanoic acid, Octadecanoic acid, 1,2-Benzenedicarboxylic acid, Phenyl ethyl tiglate, Eicosene, Farnesol 3, Camphor, Nerolidol and n-Eicosane. The antimicrobial activity of CsEO was evaluated against a panel of 13 bacteria and 8 fungal strains using agar diffusion and broth microdilution methods. Results have shown that CsEO exhibited moderate to strong antimicrobial activity against the tested species. In addition, the inhibitory effect of this CsEO was evaluated in vivo against a foodborne pathogens Listeria monocytogenes, experimentally inoculated in minced beef meat (2×10(2) CFU/g of meat) amended with different concentrations of the CsEO and stored at 7 °C for 10 days. The antibacterial activity of CsEO in minced beef meat was clearly evident and its presence led to a strong inhibitory effect against the pathogens at 7 °C. On the other hand, the cytotoxic effects of the essential oil against two tumoral human cell lines HeLa and MCF-7 were examined by MTT assay. The CsEO showed an inhibition of both cell lines with significantly stronger activity against HeLa cells. The IC(50) values were 210 and 800 μg/ml for HeLa and MCF-7 cells, respectively. Overall, results presented here suggest that the EO of C. siliqua possesses antimicrobial and cytotoxic properties, and is therefore a potential source of active ingredients for food and pharmaceutical industry.


Failure mechanisms in denture adhesives.
The mechanical properties of bio adhesives in oral care application are expected to be critical in defining the stability and release of devices such as dentures from the oral tissue. A multiscale experimental mechanical approach is used to evaluate the performance of denture adhesive materials. The inherent mechanical behavior of denture fixatives was examined by separating adhesive material from a representative polymethyl methacrylate (PMMA) surface using atomic force microscopy (AFM) approaches and compared to macroscopic mechanical testing. Failure of denture adhesive material was found to be critically dependent on the formation of fibrillar structures within the adhesive. Small scale mechanical testing provided evidence for the mechanical properties of the fibrillar structures formed within the adhesive in macroscopic mechanical testing and indicated the importance of the forces required to fail the adhesive at these small length scales in controlling both the maximum forces sustained by the bulk material as well as the ease of separating the adhesive from PMMA surfaces. Our results are important in defining the performance of denture fixative materials and their control of adhesive behavior, allowing the potential to tune properties required in the adhesion and removal of dentures.


Helicobacter pylori attenuates lipopolysaccharide-induced nitric oxide production by murine macrophages.
Intragastric growth of Helicobacter pylori and non-Helicobacter microorganisms is thought to be associated with elevated levels of pro-inflammatory cytokines and the production of NO these effects can lead to chronic inflammation. Microorganisms can activate the expression of iNOS and the production of NO by macrophages through stimulation with bacterial LPS. Helicobacter pylori can evade these vigorous immune responses, but the underlying mechanism remains unknown. In this study, we used a murine model of macrophage infection to demonstrate that H. pylori inhibits LPS-induced expression of iNOS and production of NO by macrophages. Suppression of LPS-induced NO production by macrophages led to elevated survival of H. pylori in a trans-well system. This effect was abrogated in macrophages from iNOS(-/-) mice. Analysis of iNOS mRNA and protein levels revealed that H. pylori inhibits iNOS expression at both transcriptional and post-transcriptional levels, and that these effects occurred with live bacteria. Furthermore, the effect of H. pylori involved down-regulation of the mitogen-activated protein kinase pathway and the translocation of active NF-κB into the nucleus. Taken together, our results reveal a new mechanism by which H. pylori modulates the innate immune responses of the host and maintains a persistent infection within the stomach.


Beyond the black letter of the law: an empirical study of an individual judge's decision process for civil commitment hearings.
To study the role of parens patriae and "police powers" considerations in an individual judge's civil commitment decisions, the judge's reports of the impact of various characteristics of the patient were analyzed. The validity of this methodology was tested by comparing it to an alternative technique based on objective statistical analysis of the dependence of the judge's decisions upon patient characteristics. A probate court judge filled out a questionnaire after each civil commitment hearing over which he presided during a seven-month study. For each of 26 decisions, the judge rated the patient on 26 features and indicated the impact of each feature on the decision. The judge's responses were analyzed to measure the role of various statutory and nonstatutory considerations (expressed as patient characteristics) in the judge's decisions. Results using self-reported impacts are compared with an objective, statistical characterization of the judge's decision-making policy. As in previous studies, the parens patriae model more closely described the individual judge's decision process than the "police powers" model. Contextual variables (e.g., the patient's family favoring commitment) also were influential. Results with the two methods were similar. The methodology developed here can be used not only in further research on judicial commitment decisions but also to educate judges and other decision-makers individually faced with potentially tragic choices as to their personal implicit decision-making strategies.


Can textured insoles improve ankle proprioception and performance in dancers?
With the aim of determining both the acute and the chronic effects of textured insoles on the ankle discrimination and performance ability of dancers, 60 ballet dancers from the Australian Ballet School, aged 14-19 years, were divided into three groups (two intervention groups and a control group), age- and level-matched. In the first 5 weeks (weeks 1 to 5), the first intervention group (GRP1) was asked to wear textured insoles in their ballet shoes and the second intervention group (GRP2) was not given textured insoles to wear. In the next 5 weeks (weeks 6 to 10), GRP2 was asked to wear the same type of textured insoles and GRP1 did not wear the textured insoles. The control group (CTRL) did not wear textured insoles during the whole 10 weeks. All participants were tested preintervention, after 5 weeks and after 10 weeks for ankle discrimination score (AUC scores). Dance performance was assessed by 5-7 dance teachers. Pre-to-post change in AUC scores was significantly greater for the groups wearing insoles than for the controls (P = .046) and the size of pre-to-post changes did not differ between the two intervention groups (P = .834). Significant correlation was found between ankle discrimination score and performance scores, using the textured insoles (r = .412; P = .024). In conclusion, the stimulation to the proprioceptive system arising from textured insoles worn for five weeks was sufficient to improve the proprioceptive ability and performance ability of ballet dancers.


Baseline Body Mass Predicts Average Depressive Symptoms over the Next Two Decades for White but Not Black Older Adults.
Although obesity and depression have a bidirectional association, this link may vary based on race. The current study tested racial variation in bidirectional links between depressive symptoms and body mass index (BMI) over 24 years of follow-up in older adults over the age of 50 in the United States. We hypothesized weaker bidirectional links in Blacks compared to Whites. Data came from waves 1 to 12 (1990 to 2014) of the Health and Retirement Study (HRS), an ongoing state-of-the-art national cohort. The study followed a representative sample of Americans (n = 15,194; 2,200 Blacks and 12,994 Whites) over the age of 50. Dependent variables were average depressive symptoms and BMI over 24 years, based on measurements every other year, from 1990 to 2014. Independent variables included baseline depressive symptoms and BMI. Covariates included age, gender, marital status, veteran status, and activities of daily living. Structural equation models were fitted to the data for data analysis. In the pooled sample, bidirectional associations were found between BMI and depressive symptoms as baseline BMI predicted average depressive symptoms over time and baseline depressive symptoms predicted average BMI over 24 years. Racial differences were found in the bidirectional association between BMI and depressive symptoms, with both directions of the associations being absent for Blacks. For Whites, baseline BMI predicted average depressive symptoms over the next 24 years. Reciprocal associations between BMI and depressive symptoms over a 24-year period among individuals over the age of 50 vary for Blacks and Whites. As these associations are stronger for Whites than Blacks, clinical and public health programs that simultaneously target comorbid obesity and depression may be more appropriate for Whites than Blacks.


Intestinal barrier regulates immune responses in the liver via IL-10-producing macrophages.
The gut-liver axis is of clinical importance as a potential therapeutic target in a wide range of liver diseases; however, the mechanisms underlying interactions between microbial products and immune responses in the liver remain unknown. In this study, we demonstrated that IL-10-producing macrophages contribute to immune tolerance in the inflamed liver under intestinal barrier disruption in a murine tandem model of dextran sulfate sodium (DSS) colitis and concanavalin A (Con A) hepatitis. Intestinal barrier disruption protected mice from subsequent liver injury, and the severity of colitis directly affected susceptibility to such injury. The protective effect of DSS-Con A was canceled in gut-sterilized mice, suggesting that gut microbiota play a substantial role in this process. Altered gut microbiota and their metabolites, along with a disrupted intestinal barrier, directly gave rise to immunological permissiveness in the inflamed liver. We identified 1-methylnicotinamide (1-MNA) as a candidate metabolite capable of suppressing liver injury with the potential to induce IL-10-producing macrophages. Consistently, expression of nicotinamide N-methyltransferase, which converts nicotinamide to 1-MNA, was upregulated in the liver of DSS-Con A mice, and this effect was abrogated by gut sterilization. Collectively, our results provide a mechanistic insight into the regulation of immunological balance in the liver via the gut-liver axis.


Cargo Release from Myosin V Requires the Convergence of Parallel Pathways that Phosphorylate and Ubiquitylate the Cargo Adaptor.
Cellular function requires molecular motors to transport cargoes to their correct intracellular locations. The regulated assembly and disassembly of motor-adaptor complexes ensures that cargoes are loaded at their origin and unloaded at their destination. In Saccharomyces cerevisiae, early in the cell cycle, a portion of the vacuole is transported into the emerging bud. This transport requires a myosin V motor, Myo2, which attaches to the vacuole via Vac17, the vacuole-specific adaptor protein. Vac17 also binds to Vac8, a vacuolar membrane protein. Once the vacuole is brought to the bud cortex via the Myo2-Vac17-Vac8 complex, Vac17 is degraded and the vacuole is released from Myo2. However, mechanisms governing dissociation of the Myo2-Vac17-Vac8 complex are not well understood. Ubiquitylation of the Vac17 adaptor at the bud cortex provides spatial regulation of vacuole release. Here, we report that ubiquitylation alone is not sufficient for cargo release. We find that a parallel pathway, which initiates on the vacuole, converges with ubiquitylation to release the vacuole from Myo2. Specifically, we show that Yck3 and Vps41, independent of their known roles in homotypic fusion and protein sorting (HOPS)-mediated vesicle tethering, are required for the phosphorylation of Vac17 in its Myo2 binding domain. These phosphorylation events allow ubiquitylated Vac17 to be released from Myo2 and Vac8. Our data suggest that Vps41 is regulating the phosphorylation of Vac17 via Yck3, a casein kinase I, and likely another unknown kinase. That parallel pathways are required to release the vacuole from Myo2 suggests that multiple signals are integrated to terminate organelle inheritance.


A novel method for the filterless preconcentration of iron.
A novel method of analysis of iron by filterless preconcentration is presented. This is the first example of efficient preconcentration of a refractory transition metal where coprecipitation and columns were omitted. The method applies a manifold of flow injection analysis (FIA) to iron species that are preconcentrated on the inner walls of a tubular reactor. It was found that the adsorption of iron species to the walls was particularly pronounced in reactors of nylon material and enrichment factors of 30-35 could be attained, as dependent on the length of the reactor and on the time of preconcentration. In the preconcentration step of the FIA accessory, the optimum efficacy was obtained when the acidity of the samples was adjusted by HCl to pH = 2.5 whereas the ammonia preconcentration buffer should be kept alkaline at pH = 9.8. After being preconcentrated on the tubular reactor, the iron species were eluted by hydrochloric acid and analysed by flame atomic absorption spectrometry (FAAS). An unprecedented low limit of detection (LOD, 3sigma) of 1.8 microg L(-1) was thus obtained for the analysis of iron by FAAS, and the calibration line was linear up to 100 microg L(-1). A high sampling frequency of 40 per hour was obtained and the protocol allowed analysis of small amounts of iron in drinking water, in digested samples and in saline water. The major influence of interferences originated from ligands that are known to complex strongly to iron species. The method thus developed was uncomplicated in operation and it exhibited an excellent long-term stability and a high selectivity. Further, it was environmentally safe owing to production of non-toxic waste disposals. Equally high enrichment factors (EF) were obtained for Fe(ii) and Fe(iii), which is explained by the very low solubility product of both species.


Targeted removal of ant colonies in ecological experiments, using hot water.
Ecological experiments on fire ants cannot, or should not, use poison baits to eliminate the fire ants because such baits are not specific to fire ants, or even to ants. Hot water is an extremely effective and specific killing agent for fire ant colonies, but producing large amounts of hot water in the field, and making the production apparatus mobile have been problematical. The construction and use of a charcoal-fired kiln made from a 55-gal. oil drum lined with a sand-fireclay mixture is described. An automobile heater fan powered from a 12-v battery provided a draft. Dual bilge pumps pumped water from a large tank through a long coil of copper tubing within the kiln to produce 4 to 5 l. of hot water per min. The hot water was collected in 20 l. buckets and poured into fire ant nests previously opened by piercing with a stick. The entire assembly was transported in and operated from the back of a pickup truck. Five experimental plots containing 32 to 38 colonies of the fire ant, Solenopsis invicta, Buren (Hymenoptera: Formicidae), were treated with hot water over a period of two years. All colonies on the treatment plots were treated twice with hot water early in 2004, reducing their numbers to zero. However new colonies were formed, and mature colonies expanded into the plots. A third treatment was made in the spring of 2005, after which fire ant populations were suppressed for over a year. Whereas the 5 control plots contained a total of 166 mostly large colonies, the 5 treatment plots contained no live colonies at all. Averaged over a two-year period, a 70% reduction in total number of colonies was achieved (P < 0.001) on the treatment plots, and a 93% reduction of large, mature colonies. Over this same time span, the number of colonies in control plots remained stable. The reduction in colony numbers on the treatment plots was reflected in the pitfall trap samples that recorded a 60% reduction in fire ants.


DamID as an approach to studying long-distance chromatin interactions.
How transcription is controlled by distally located cis-regulatory elements is an active area of research in biology. As such, there have been many techniques developed to probe these long-distance chromatin interactions. Here, we focus on one such method, called DamID (van Steensel and Henikoff, Nat Biotechnol 18(4):424-428, 2000). While other methods like 3C (Dekker et al., Science 295(5558):1306-1311, 2002), 4C (Simonis et al., Nat Genet 38(11):1348-1354, 2006; Zhao et al., Nat Genet 38(11):1341-1347, 2006), and 5C (Dostie et al., Genome Res 16(10):1299-1309, 2006) are undoubtedly powerful, the DamID method can offer some advantages over these methods if the genetic locus can be easily modified. The lack of tissue fixation, the low amounts of starting material required to perform the experiment, and the relatively modest hardware requirements make DamID experiments an interesting alternative to consider when examining long-distance chromatin interactions.


Impaired binding of ZASP/Cypher with phosphoglucomutase 1 is associated with dilated cardiomyopathy.
Z-band alternatively spliced PDZ-motif protein (ZASP)/Cypher is a Z-disc component of which several dilated cardiomyopathy (DCM)-associated mutations have been reported. Most of the mutations were found in exons 4 and 10 of ZASP/Cypher gene LDB3 and both exons were expressed preferentially in the heart. The aim of this study was to investigate the functional alteration of ZASP/Cypher caused by the DCM-associated mutations. The yeast-two-hybrid method was used to identify the protein bound to a domain encoded by exon 4 of LDB3. Interaction of ZASP/Cypher with the binding protein was investigated in relation to the functional alterations caused by LDB3 mutations. Localization of the ZASP/Cypher-binding protein was examined at the cellular level in rat cardiomyocytes. Phosphoglucomutase 1 (PGM1), a metabolic enzyme involved in glycolysis and gluconeogenesis, was identified as a protein interacting with ZASP/Cypher. PGM1 bound to ZASP/Cypher at the domains encoded by exons 4 and 10. Two LDB3 mutations in exon 4 (Ser189Leu and Thr206Ile) and another mutation in exon 10 (Ile345Met) reduced the binding to PGM1. PGM1 showed diffuse localization in the cytoplasm of rat cardiomyocytes under standard culture conditions, and distribution at the Z-discs was observed under stressed culture conditions. Binding of endogenous PGM1 and ZASP/Cypher was found to be enhanced by stress in rat cardiomyocytes. ZASP/Cypher anchors PGM1 to Z-disc under conditions of stress. The impaired binding of PGM1 to ZASP/Cypher might be involved in the pathogenesis of DCM.


Norwegian midwives' perception of the labour admission test.
To explore what information and knowledge the labour admission test is perceived to provide and what meaning the test carries in the daily work of practising midwives. In-depth interviews transcribed verbatim and analysed using the grounded theory technique. Four different labour wards in Norway. A theoretical sample of 12 practising midwives. The core category "experiencing contradictions" was identified during the analyses, indicating that the midwives found conflicting interests within themselves, or between themselves and others when using the labour admission test. They experienced contradictions between professional identity and the increasing use of technology, between feeling safe and feeling unsafe and between having power and being powerless. The labour admission traces could be difficult to interpret, especially for newly qualified midwives. Some midwives thought that a labour admission trace could protect them in case of litigation. The hierarchy of power in the labour ward influences the use and interpretation of the labour admission test. Some midwives felt their professional identity threatened and that midwives in general are losing their traditional skills because of the increasing use of obstetric technology. The findings of the present study should be taken into consideration when changing practice to not routinely perform the labour admission test. There is also a need for further research on what effect the increasing use of obstetric technology has on traditional midwifery skills.


Age-related differences in laterality of cortical activations in swallowing.
The present study examined age differences in neural lateralization patterns during swallowing and three related tasks, using functional magnetic resonance imaging (fMRI). Ten healthy right-handed young adults (mean age = 21.7 years, SD = 2.1 years) and nine healthy elders (mean age = 70.2 years, SD = 3.9 years) were scanned in a 3-T MRI head scanner. Participants were visually cued to "prepare to swallow," "swallow," "tap your tongue," and "clear your throat" in randomized order. Laterality preference for each task was examined within and between groups using region-of-interest (ROI) analyses in seven areas of the left and right primary sensorimotor and premotor cortices. Results of the within-group comparisons verified a more active role of the left premotor cortex in motor-cognitive planning of deglutition in both young and older adults and a more active role of selected areas of the right hemisphere during swallowing in young adults. Greater variability was seen during tongue tapping and throat clearing in both groups. Finally, as people age the cortical hemispheric control of swallowing seems to start becoming more symmetrical/bilateral, which may indicate neural compensatory mechanisms of the aging brain commonly seen for other motor and cognitive functions.


Age-adjusted baseline data for women with hallux valgus undergoing corrective surgery.
Functional and health-related quality-of-life data on a population of patients with hallux valgus are lacking. Similarly, the correlation of the severity of the deformity with these measures is unknown. Two hundred and eighty-five women with an average age of forty-nine years who were scheduled for bunion surgery were enrolled in the study. The patients completed a baseline American Academy of Orthopaedic Surgeons (AAOS) foot and ankle outcomes questionnaire, which includes the Short Form-36 (SF-36) and a specific lower-extremity section on the foot and ankle. Preoperative radiographic data with regard to the hallux valgus angle and the intermetatarsal angle were stratified into groups according to the severity of the deformity (mild, moderate, or severe). The data were then stratified into age-groups consistent with those reported for the SF-36, and the results were compared with the SF-36 scores for the general population. The global foot and ankle score and the shoe comfort score were compared with general population scores that were published previously. The severity of the preoperative deformity was correlated with the baseline scores. General health scores were noted to be relatively stable throughout the age-groups for patients with bunions, with the older groups demonstrating better scores than the general population. Bodily pain scores were consistently worse for patients with a bunion through all age-groups compared with the general population. The average global foot and ankle score and the shoe comfort score were significantly lower (p < 0.001 for both) for the patients with a bunion than for the general population. The severity of the preoperative deformity did not correlate with any of the outcome scores. The bodily pain score from the SF-36 appears to be a sensitive measure of problems experienced by patients undergoing bunion surgery. Surprisingly, the severity of the deformity as measured radiographically did not correlate with any of the fifteen scores measured. These data may serve as a baseline for clinical hallux valgus studies with use of the SF-36 or the AAOS outcomes questionnaire.


Dangerousness commitments: indices of future violence potential?
This study examines the accuracy of long-term clinical predictions of dangerousness among psychiatric inpatients and explores factors influencing the levels of such accuracy. Hospital and state criminal history records of all psychiatric patients (N = 31) for whom, during a four-year period, treatment staff pursued extended civil commitments based on dangerousness under the Postcertification for the imminently Dangerous statute (California Welfare and Institutions Code section 5300) were reviewed. A matched control group consisted of 31 patients who had been placed on 14-Day Certifications for Dangerousness to Others, but who were not subsequently placed on 180-Day Postcertifications. Sixty-one percent of patients in the postcertification group engaged in documented physically assaultive behavior during the extended one- to five-year follow-up period, compared with 26 percent of patients in the matched control group, suggesting that inclusion in the extended commitment group was indicative of greater long-term potential for assault. Differences in assaultiveness did not emerge during the first year of followup, but became clear and significant over subsequent years. Accuracy of prediction differed as a function of patient ethnic group.


Through bleaching and tsunami: Coral reef recovery in the Maldives.
Coral reefs are degrading worldwide, but little information exists on their previous conditions for most regions of the world. Since 1989, we have been studying the Maldives, collecting data before, during and after the bleaching and mass mortality event of 1998. As early as 1999, many newly settled colonies were recorded. Recruits shifted from a dominance of massive and encrusting corals in the early stages of recolonisation towards a dominance of Acropora and Pocillopora by 2009. Coral cover, which dropped to less than 10% after the bleaching, returned to pre-bleaching values of around 50% by 2013. The 2004 tsunami had comparatively little effect. In 2014, the coral community was similar to that existing before the bleaching. According to descriptors and metrics adopted, recovery of Maldivian coral reefs took between 6 and 15years, or may even be considered unachieved, as there are species that had not come back yet.


Combining wavelet analysis and Bayesian networks for the classification of auditory brainstem response.
The auditory brainstem response (ABR) has become a routine clinical tool for hearing and neurological assessment. In order to pick out the ABR from the background EEG activity that obscures it, stimulus-synchronized averaging of many repeated trials is necessary, typically requiring up to 2000 repetitions. This number of repetitions can be very difficult, time consuming and uncomfortable for some subjects. In this study, a method combining wavelet analysis and Bayesian networks is introduced to reduce the required number of repetitions, which could offer a great advantage in the clinical situation. 314 ABRs with 64 repetitions and 155 ABRs with 128 repetitions recorded from eight subjects are used here. A wavelet transform is applied to each of the ABRs, and the important features of the ABRs are extracted by thresholding and matching the wavelet coefficients. The significant wavelet coefficients that represent the extracted features of the ABRs are then used as the variables to build the Bayesian network for classification of the ABRs. In order to estimate the performance of this approach, stratified ten-fold cross-validation is used.


Biodegradable zinc oxide composite scaffolds promote osteochondral differentiation of mesenchymal stem cells.
Osteoarthritis (OA) involves the degeneration of articular cartilage and subchondral bone. The capacity of articular cartilage to repair and regenerate is limited. A biodegradable, fibrous scaffold containing zinc oxide (ZnO) was fabricated and evaluated for osteochondral tissue engineering applications. ZnO has shown promise for a variety of biomedical applications but has had limited use in tissue engineering. Composite scaffolds consisted of ZnO nanoparticles embedded in slow degrading, polycaprolactone to allow for dissolution of zinc ions over time. Zinc has well-known insulin-mimetic properties and can be beneficial for cartilage and bone regeneration. Fibrous ZnO composite scaffolds, having varying concentrations of 1-10 wt.% ZnO, were fabricated using the electrospinning technique and evaluated for human mesenchymal stem cell (MSC) differentiation along chondrocyte and osteoblast lineages. Slow release of the zinc was observed for all ZnO composite scaffolds. MSC chondrogenic differentiation was promoted on low percentage ZnO composite scaffolds as indicated by the highest collagen type II production and expression of cartilage-specific genes, while osteogenic differentiation was promoted on high percentage ZnO composite scaffolds as indicated by the highest alkaline phosphatase activity, collagen production, and expression of bone-specific genes. This study demonstrates the feasibility of ZnO-containing composites as a potential scaffold for osteochondral tissue engineering.


Energetic costs of mange in wolves estimated from infrared thermography.
Parasites, by definition, extract energy from their hosts and thus affect trophic and food web dynamics even when the parasite may have limited effects on host population size. We studied the energetic costs of mange (Sarcoptes scabiei) in wolves (Canis lupus) using thermal cameras to estimate heat losses associated with compromised insulation during the winter. We combined the field data of known, naturally infected wolves with a data set on captive wolves with shaved patches of fur as a positive control to simulate mange-induced hair loss. We predict that during the winter in Montana, more severe mange infection increases heat loss by around 5.2-12 MJ per night (1,240-2,850 kcal, or a 65-78% increase) for small and large wolves, respectively, accounting for wind effects. To maintain body temperature would require a significant proportion of a healthy wolf's total daily energy demands (18-22 MJ/day). We also predict how these thermal costs may increase in colder climates by comparing our predictions in Bozeman, Montana to those from a place with lower ambient temperatures (Fairbanks, Alaska). Contrary to our expectations, the 14°C differential between these regions was not as important as the potential differences in wind speed. These large increases in energetic demands can be mitigated by either increasing consumption rates or decreasing other energy demands. Data from GPS-collared wolves indicated that healthy wolves move, on average, 17 km per day, which was reduced by 1.5, 1.8, and 6.5 km for light, medium, and severe hair loss. In addition, the wolf with the most hair loss was less active at night and more active during the day, which is the converse of the movement patterns of healthy wolves. At the individual level, mange infections create significant energy demands and altered behavioral patterns, this may have cascading effects on prey consumption rates, food web dynamics, predator-prey interactions, and scavenger communities.


Flavonoids in baby spinach (Spinacia oleracea L.): changes during plant growth and storage.
The variation in flavonoid concentration and composition was investigated in baby spinach (Spinacia oleracea L.) cv. Emilia sown on three occasions, each harvested at three growth stages at 6-day intervals. After harvest, leaves were stored in polypropylene bags at 2 or 10 degrees C. Flavonoids were analyzed by reversed phase HPLC. Twelve flavonoid peaks were detected. The main flavonoid, making up on average 43% of the total flavonoid concentration, was identified as 5,3',4'-trihydroxy-3-methoxy-6:7-methylenedioxyflavone-4'-glucuronide. Four other flavonoids each contributed 7-12% of the total flavonoid content. Total flavonoid content was relatively stable during normal retail storage conditions, although some of the individual flavonoid compounds showed considerable variation. The youngest plants had the highest flavonoid concentration, indicating that by harvesting the baby spinach a few days earlier than the current commercial stage of harvest, the flavonoid concentration in the product may be increased and the content of potentially health-promoting compounds enhanced.


Medical treatment of recurrent temporomandibular joint dislocation using botulinum toxin A.
This paper describes a new technique for prophylactic treatment of recurrent mandibular dislocation using injection of botulinum toxin A (BtA) into the lateral pterygoid muscles. BtA temporarily weakens muscles by blocking acetylcholine release, and thus operates through a principle different from established treatments such as joint sclerosant therapy, eminectomy or Dautry's procedure. The patient suffered recurrent mandibular dislocations caused by tardive dystonia. We injected 75 mu BtA percutaneously into each lateral pterygoid muscle under electromyographic guidance. No further dislocations occurred over the subsequent 10 months, and follow-up continues. There were no immediate or delayed side effects. More experience is required before this becomes an established treatment. BtA is usually given in outpatients, and is less invasive or destructive than previous options. It may not be suitable if dislocation is due to lax ligaments or weak muscles. Operators must be aware that other BtA preparations require a different dose.


Quantifying the Impact of Trainee Providers on Outpatient Clinic Workflow using Secondary EHR Data.
Providers today face productivity challenges including increased patient loads, increased clerical burdens from new government regulations and workflow impacts of electronic health records (EHR). Given these factors, methods to study and improve clinical workflow continue to grow in importance. Despite the ubiquitous presence of trainees in academic outpatient clinics, little is known about the impact of trainees on academic workflow. The purpose of this study is to demonstrate that secondary EHR data can be used to quantify that impact, with potentially important results for clinic efficiency and provider reimbursement models. Key findings from this study are that (1) Secondary EHR data can be used to reflect in clinic trainee activity, (2) presence of trainees, particularly in high-volume clinic sessions, is associated with longer session lengths, and (3) The timing of trainee appointments within clinic sessions impacts the session length.


Large dense-core vesicle exocytosis in pancreatic beta-cells monitored by capacitance measurements.
This article discusses the currently used methodologies for monitoring exocytosis as changes in cell capacitance. Details are given on composition of solutions, experimental protocols, and how the observed responses can be interpreted physiologically. The concepts are illustrated by examples from our own work on insulin-releasing pancreatic beta-cells. Finally, we consider the feasibility of applying capacitance measurements to endocrine cells in intact pancreatic islets, where the cells are electrically coupled to each other.


Hallucinations in the months after a trauma: An investigation of the role of cognitive processing of a physical assault in the occurrence of hallucinatory experiences.
The role that cognitive processing of a recent trauma has in the occurrence of hallucinations has not been examined longitudinally. This study investigated trauma-related cognitive predictors of hallucinations in the months following an interpersonal assault. Four weeks after treatment at an emergency department for interpersonal assault injuries, 106 participants were assessed for peri-traumatic cognitive processing, cognitive responses to trauma memories, negative beliefs about the self, Posttraumatic-stress disorder (PTSD), and hallucinatory experiences. Hallucinatory experiences were reassessed six months later. Cognitive processing during trauma (lack of self-referential processing, and dissociation), beliefs about permanent negative change, self-vulnerability, and self-blame and cognitive response styles (thought suppression, rumination, and numbing) were significant predictors of later hallucinations. The way in which trauma is processed may partly determine the occurrence of hallucinations.


Stakeholders' participation in planning and priority setting in the context of a decentralised health care system: the case of prevention of mother to child transmission of HIV programme in Tanzania.
In Tanzania, decentralisation processes and reforms in the health sector aimed at improving planning and accountability in the sector. As a result, districts were given authority to undertake local planning and set priorities as well as allocate resources fairly to promote the health of a population with varied needs. Nevertheless, priority setting in the health care service has remained a challenge. The study assessed the priority setting processes in the planning of the prevention of mother to child transmission of HIV (PMTCT) programme at the district level in Tanzania. This qualitative study was conducted in Mbarali district, south-western Tanzania. The study applied in-depth interviews and focus group discussions in the data collection. Informants included members of the Council Health Management Team, regional PMTCT managers and health facility providers. Two plans were reported where PMTCT activities could be accommodated; the Comprehensive Council Health Plan and the Regional PMTCT Plan that was donor funded. As donors had their own globally defined priorities, it proved difficult for district and regional managers to accommodate locally defined PMTCT priorities in these plans. As a result few of these were funded. Guidelines and main priority areas of the Ministry of Health and Social Welfare (MoHSW) also impacted on the ability of the districts and regions to act, undermining the effectiveness of the decentralisation policy in the health sector. The challenges in the priority setting processes revealed within the PMTCT initiative indicate substantial weaknesses in implementing the Tanzania decentralisation policy. There is an urgent need to revive the strategies and aims of the decentralisation policy at all levels of the health care system with a view to improving health service delivery.


Multi-Factor Analysis of Single-Center Asthma Control in Xiamen, China.
We evaluated the effects of air pollutants, age, allergic history, family allergic history, treatment, treatment steps, and compliance on uncontrolled childhood asthma in Xiamen, China. The clinical data of children with asthma in the pediatric outpatient department of the First Affiliated Hospital of Xiamen University from January 2016 to June 2018 were analyzed retrospectively. According to the assessment of the patients' outcome including well-controlled, partly-controlled and uncontrolled, 7,211 cases of 3,268 patients were selected. Rank sum test and ordered multi-class logistic regression analysis were used. In the rank sum test, age, allergic history, family allergic history, season, treatment, treatment steps and compliance were found associated with uncontrolled rate (all P < 0.001). Logistic regression analysis showed that PM10, NO2, and SO2 raised uncontrolled-asthma rate (aOR 1.311, aOR 1.281, aOR 1.252, respectively). Older children had lower uncontrolled rate (OR = 0.849, 95% CI: 0.758-0.950), children with higher treatment steps had higher uncontrolled rate (OR = 1.227, 95%CI: 1.040-1.448), and children with better treatment compliance have lower uncontrolled rate (OR = 0.374 95% CI: 0.331-0.424). The order of the uncontrolled rate of asthma from high to low was winter, spring, autumn, and summer. PM10, NO2, SO2, age, season, treatment steps, and treatment compliance have significance for predicting the control rate of childhood asthma in Xiamen, China.


Awareness of metabolic concerns and perceived impact of pharmacotherapy in patients with bipolar disorder: a survey of 500 US psychiatrists.
An online survey was conducted to assess psychiatrists' familiarity with the metabolic syndrome and its components in patients with bipolar disorder, and characterize their perspectives and practices regarding its impact on patient management. Participants were US psychiatrists from a random sample of those in the AMA database. Qualified respondents practiced 4-30 years, spent > or = 50% of their time in direct patient care, and treated > or = 25 bipolar patients in the last month. Results were collected and tabulated by Harris Interactive from Nov-Dec 2005. Five hundred psychiatrists qualified and completed the survey, and 50 respondents also participated in follow-up interviews. Most respondents (94%) viewed metabolic syndrome as a significant health risk requiring monitoring and treatment. While 76% have diagnosed it, only 28% correctly identified the five NCEP diagnostic criteria. Medication adverse effects of greatest concern were weight gain, glucose intolerance, and dyslipidemia. During treatment, 78% of respondents reported monitoring weight, 69% glucose, 61% lipids, and 52% blood pressure. Most respondents (92%) reported referring patients to specialists or primary care for metabolic abnormalities. Changes in metabolic profile were reported to prompt many psychiatrists (85%) to stop or switch bipolar medications, especially those who treat a large number of bipolar patients (89%). The follow-up interviews supported a change in practice patterns over the last 5 years. Nearly all respondents have metabolic concerns with medical therapies used to treat bipolar disorder. Many now routinely monitor weight and other metabolic parameters. Most have referred patients for medical management and adjusted bipolar therapies due to metabolic abnormalities.


Genetics and Genomics of Longitudinal Lung Function Patterns in Individuals with Asthma.
Patterns of longitudinal lung function growth and decline in childhood asthma have been shown to be important in determining risk for future respiratory ailments including chronic airway obstruction and chronic obstructive pulmonary disease. To determine the genetic underpinnings of lung function patterns in subjects with childhood asthma. We performed a genome-wide association study of 581 non-Hispanic white individuals with asthma that were previously classified by patterns of lung function growth and decline (normal growth, normal growth with early decline, reduced growth, and reduced growth with early decline). The strongest association was also measured in two additional cohorts: a small asthma cohort and a large chronic obstructive pulmonary disease metaanalysis cohort. Interaction between the genomic region encompassing the most strongly associated single-nucleotide polymorphism and nearby genes was assessed by two chromosome conformation capture assays. An intergenic single-nucleotide polymorphism (rs4445257) on chromosome 8 was strongly associated with the normal growth with early decline pattern compared with all other pattern groups (P = 6.7 × 10-9; odds ratio, 2.8; 95% confidence interval, 2.0-4.0); replication analysis suggested this variant had opposite effects in normal growth with early decline and reduced growth with early decline pattern groups. Chromosome conformation capture experiments indicated a chromatin interaction between rs4445257 and the promoter of the distal CSMD3 gene. Early decline in lung function after normal growth is associated with a genetic polymorphism that may also protect against early decline in reduced growth groups. Clinical trial registered with www.clinicaltrials.gov (NCT00000575).


Diferric transferrin regulates transferrin receptor 2 protein stability.
Transferrin receptor 2 (TfR2) is a type 2 transmembrane protein expressed in hepatocytes that binds iron-bound transferrin (Tf). Mutations in TfR2 cause one form of hereditary hemochromatosis, a disease in which excessive absorption of dietary iron can lead to liver cirrhosis, diabetes, arthritis, and heart failure. The function of TfR2 in iron homeostasis is unknown. We have studied the regulation of TfR2 in HepG2 cells. Western blot analysis shows that TfR2 increases in a time- and dose-dependent manner after diferric Tf is added to the culture medium. In cells exposed to diferric Tf, the amount of TfR2 returns to control levels within 8 hours after the removal of diferric Tf from the medium. However, TfR2 does not increase when non-Tf-bound iron (FeNTA) or apo Tf is added to the medium. The response to diferric Tf appears to be hepatocyte specific. Real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis shows that TfR2 mRNA levels do not change in cells exposed to diferric Tf. Rather, the increase in TfR2 is attributed to an increase in the half-life of TfR2 protein in cells exposed to diferric Tf. Our results support a role for TfR2 in monitoring iron levels by sensing changes in the concentration of diferric Tf.


Role of block copolymer-micelle nanocomposites in dye-bovine serum albumin binding: a combined experimental and molecular docking study.
The role of a nanocomposite (NC), composed of intercalation of the diblock copolymer polyethylene-b-polyethylene glycol (PE-b-PEG) with the anionic surfactant sodium dodecyl sulphate (SDS), on the binding characteristics of bovine serum albumin (BSA) with a dye (1,8-naphthalimide, NAPMD) compared to the interaction between the same players in aqueous solution has been examined comprehensively in this paper. Static quenching due to complex formation in both NC medium and in buffer solution has been inferred on the basis of considerable changes in the absorption spectra of BSA on addition of NAPMD, of which the interaction is found to be stronger in NC medium. Temperature dependent fluorescence data also confirm an effective static quenching and stronger binding of NAPMD with BSA in NC medium. Peptide chain unfolding and denaturing of BSA in NC medium have been confirmed from steady state and time-resolved emission and circular dichroism data. This exposes both the tyrosine and tryptophan moieties as a unique case. Increased energy transfer between NAPMD and the tryptophan residue in the unfolded form of BSA helps in the appearance of tyrosine fluorescence in NC medium by quenching the tryptophan band. Ionization of the hydroxyl group in the aromatic ring of the tyrosine residue by the PEG group present in the NC medium produces a downshift of the tyrosine fluorescence band. The use of site selective markers confirms that NAPMD is near tryptophan in Sudlow's site I in NC medium and in buffer solution it is away from tryptophan in Sudlow's site II. The theoretical docking studies also vindicate the results of binding of NAPMD with BSA in site I or site II in NC and buffer media, as observed from different emission experiments including the site selective markers study.


Expression of MLL-AF4 or AF4-MLL fusions does not impact the efficiency of DNA damage repair.
The most frequent rearrangement of the human MLL gene fuses MLL to AF4 resulting in high-risk infant B-cell acute lymphoblastic leukemia (B-ALL). MLL fusions are also hallmark oncogenic events in secondary acute myeloid leukemia. They are a direct consequence of mis-repaired DNA double strand breaks (DNA-DSBs) due to defects in the DNA damage response associated with exposure to topoisomerase-II poisons such as etoposide. It has been suggested that MLL fusions render cells susceptible to additional chromosomal damage upon exposure to etoposide. Conversely, the genome-wide mutational landscape in MLL-rearranged infant B-ALL has been reported silent. Thus, whether MLL fusions compromise the recognition and/or repair of DNA damage remains unanswered. Here, the fusion proteins MLL-AF4 (MA4) and AF4-MLL (A4M) were CRISPR/Cas9-genome edited in the AAVS1 locus of HEK293 cells as a model to study MLL fusion-mediated DNA-DSB formation/repair. Repair kinetics of etoposide- and ionizing radiation-induced DSBs was identical in WT, MA4- and A4M-expressing cells, as revealed by flow cytometry, by immunoblot for γH2AX and by comet assay. Accordingly, no differences were observed between WT, MA4- and A4M-expressing cells in the presence of master proteins involved in non-homologous end-joining (NHEJ; i.e.KU86, KU70), alternative-NHEJ (Alt-NHEJ; i.e.LigIIIa, WRN and PARP1), and homologous recombination (HR, i.e.RAD51). Moreover, functional assays revealed identical NHEJ and HR efficiency irrespective of the genotype. Treatment with etoposide consistently induced cell cycle arrest in S/G2/M independent of MA4/A4M expression, revealing a proper activation of the DNA damage checkpoints. Collectively, expression of MA4 or A4M does neither influence DNA signaling nor DNA-DSB repair.


Questionnaire survey of acute and delayed adverse reactions to ferumoxides.
The purpose of this study was to assess acute and delayed adverse reactions to ferumoxides. 473 consecutive patients who underwent ferumoxides-enhanced MRI were enrolled in this study. Following non-contrast-enhanced MRI, patients were interviewed about their allergic history, and after ferumoxides-enhanced MRI, they were asked to fill out a questionnaire sheet to record any symptoms within one week. 345 of 473 questionnaires (72.9%) were returned. Fifty-eight of 345 patients (16.8%) showed some adverse reactions considered to be related to ferumoxides: 25 (7.2%) during administration, 25 (7.2%) later on the day of ferumoxides administration, and 17 (4.9%) within one week. Thirty-three patients (9.6%) showed only delayed adverse reactions. Delayed adverse reactions were more frequent in women than in men (p<0.01), and in patients with any allergic history than in those without such history (p<0.05). Acute adverse reactions tended to be more frequent in patients who had some adverse events to any contrast medium than in those with no such history (p=0.069). By using the questionnaire, the incidence of adverse reactions to ferumoxides (16.8%) was higher than previously reported from Japanese clinical trials.


The Effects of Quantitative Trait Architecture on Detection Power in Short-Term Artificial Selection Experiments.
Evolve and resequence (E&R) experiments, in which artificial selection is imposed on organisms in a controlled environment, are becoming an increasingly accessible tool for studying the genetic basis of adaptation. Previous work has assessed how different experimental design parameters affect the power to detect the quantitative trait loci (QTL) that underlie adaptive responses in such experiments, but so far there has been little exploration of how this power varies with the genetic architecture of the evolving traits. In this study, we use forward simulation to build a more realistic model of an E&R experiment in which a quantitative polygenic trait experiences a short, but strong, episode of truncation selection. We study the expected power for QTL detection in such an experiment and how this power is influenced by different aspects of trait architecture, including the number of QTL affecting the trait, their starting frequencies, effect sizes, clustering along a chromosome, dominance, and epistasis patterns. We show that all of these parameters can affect allele frequency dynamics at the QTL and linked loci in complex and often unintuitive ways, and thus influence our power to detect them. One consequence of this is that existing detection methods based on models of independent selective sweeps at individual QTL often have lower detection power than a simple measurement of allele frequency differences before and after selection. Our findings highlight the importance of taking trait architecture into account when designing and interpreting studies of molecular adaptation with temporal data. We provide a customizable modeling framework that will enable researchers to easily simulate E&R experiments with different trait architectures and parameters tuned to their specific study system, allowing for assessment of expected detection power and optimization of experimental design.