Standards and guidelines for the interpretation of sequence variants: a joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology.
The American College of Medical Genetics and Genomics (ACMG) previously developed guidance for the interpretation of sequence variants.(1) In the past decade, sequencing technology has evolved rapidly with the advent of high-throughput next-generation sequencing. By adopting and leveraging next-generation sequencing, clinical laboratories are now performing an ever-increasing catalogue of genetic testing spanning genotyping, single genes, gene panels, exomes, genomes, transcriptomes, and epigenetic assays for genetic disorders. By virtue of increased complexity, this shift in genetic testing has been accompanied by new challenges in sequence interpretation. In this context the ACMG convened a workgroup in 2013 comprising representatives from the ACMG, the Association for Molecular Pathology (AMP), and the College of American Pathologists to revisit and revise the standards and guidelines for the interpretation of sequence variants. The group consisted of clinical laboratory directors and clinicians. This report represents expert opinion of the workgroup with input from ACMG, AMP, and College of American Pathologists stakeholders. These recommendations primarily apply to the breadth of genetic tests used in clinical laboratories, including genotyping, single genes, panels, exomes, and genomes. This report recommends the use of specific standard terminology-"pathogenic," "likely pathogenic," "uncertain significance," "likely benign," and "benign"-to describe variants identified in genes that cause Mendelian disorders. Moreover, this recommendation describes a process for classifying variants into these five categories based on criteria using typical types of variant evidence (e.g., population data, computational data, functional data, segregation data). Because of the increased complexity of analysis and interpretation of clinical genetic testing described in this report, the ACMG strongly recommends that clinical molecular genetic testing should be performed in a Clinical Laboratory Improvement Amendments-approved laboratory, with results interpreted by a board-certified clinical molecular geneticist or molecular genetic pathologist or the equivalent.


Application of the COOP/WONCA charts to aged patients with chronic obstructive pulmonary disease: a comparison between Japanese and Chinese populations.
The prevalence of chronic obstructive pulmonary disease (COPD) is similar in Japan and China and is increasing due to high rates of smoking in these countries. Reducing COPD is an important public health issue. The goals of this study were to verify the reliability and validity of the Japanese version of the COOP/WONCA charts, a tool for measuring health status, and to examine the qualitative differences in health status between Japanese and Chinese patients with COPD and between these patients and healthy subjects. From 2008 to 2011, we examined the factors affecting the health status of Japanese and Chinese populations living in six cities. Participants were patients with COPD staged according to the Global Initiative for Chronic Obstructive Lung Disease (GOLD) criteria (140 Japanese, 201 Chinese) and healthy subjects (243 Japanese, 199 Chinese), all 50 to 79 years old. Health status was measured by using the COOP/WONCA charts, and basic information such as smoking status and medical history was reported by the participants. The Japanese and Chinese versions of the COOP/WONCA charts were shown to be reliable and valid by test-retest, comparison with the SF-36 and respiratory symptoms, and correlation of results obtained from patients and their physicians. Stepwise multiple regression analyses demonstrated that "Physical fitness", "Daily activities", and "Social activities" were predicted by COPD status and/or respiratory symptoms; "Feelings" by nationality and respiratory symptoms; "Pain" by sex and respiratory symptoms; and "Overall health" by nationality. When the COOP/WONCA scores were stratified by nationality, age, sex, and COPD status, the difference of each score between the patients and healthy subjects was larger for the Chinese subjects than for the Japanese. The physical, psychosocial activities, and pain scores increased significantly as COPD status worsened in Chinese subjects, whereas these scores were not affected by sex, age, or COPD status for Japanese subjects. Brinkman index and use of smoky fuel indoors affected the COOP/WONCA scores in Chinese patients but not in Japanese patients. The Japanese COOP/WONCA charts are reliable and valid. COPD more severely affected the health status of Chinese participants than of Japanese participants. These results suggest that countermeasures against insufficient health care and smoky environments may improve the health status of Chinese patients with COPD.


Sexual harassment and victimization of students: a case study of a higher education institution in South Africa.
Sexual harassment has been identified as a major public health problem that is hidden in most institutions/organizations. This study assessed sexual harassment and victimization of students in a higher institution in South Africa. A quantitative, cross-sectional descriptive design was used in this study. The target population was registered students of the higher education institution and the sample size was 342. Questionnaire was used for data collection and data were analyzed using Statistical Package for Social Sciences (SPSS) Version 23.0 program. The basic principles of ethics were duly observed and the ethical clearance certificate was obtained prior to data collection. The findings revealed that 27 (17.3%) of the male and 47 (25.5%) of the female respondents (P = 0.047) had personally experienced unwanted touching. Two (1.3%) male and 5 (2.7%) female students admitted that they have been raped. Seventeen (10.8%) of the males and 19 (10.2%) of the females had been coerced to comply with a sexual relationship on campus. This study shows that both male and female students on campus are experiencing different forms of sexual harassment.


How do interprofessional student teams interact in a primary care clinic? A qualitative analysis using activity theory.
Practice based interprofessional education opportunities are proposed as a mechanism for health professionals to learn teamwork skills and gain an understanding of the roles of others. Primary care is an area of practice that offers a promising option for interprofessional student learning. In this study, we investigated what and how students from differing professions learn together. Our findings inform the design of future interprofessional education initiatives. Using activity theory, we conducted an ethnographic investigation of interprofessional education in primary care. During a 5 months period, we observed 14 clinic sessions involving mixed discipline student teams who interviewed people with chronic disease. Teams were comprised of senior medicine, nursing, occupational therapy, pharmacy and physiotherapy entry level students. Semi-structured interviews were also conducted with seven clinical educators. Data were analysed to ascertain the objectives, tools, rules and division of labour. Two integrated activity systems were identified: (1) student teams gathering information to determine patients' health care needs and (2) patients either as health consumers or student educators. Unwritten rules regarding 'shared contribution', 'patient as key information source' and 'time constraints' were identified. Both the significance of software literacy on team leadership, and a pre-determined structure of enquiry, highlighted the importance of careful consideration of the tools used in interprofessional education, and the way they can influence practice. The systems of practice identified provide evidence of differing priorities and values, and multiple perspectives of how to manage health. The work reinforced the value of the patients' voice in clinical and education processes.


Evolution of protein families: is it possible to distinguish between domains of life?
Understanding evolutionary relationships between species can shed new light into the rooting of the tree of life and the origin of eukaryotes, thus, resulting in a long standing interest in accurately assessing evolutionary parameters at time scales on the order of a billion of years. Prior work suggests large variability in molecular substitution rates, however, we still do not know whether such variability is due to species-specific trends at a genomic scale, or whether it can be attributed to the fluctuations inherent in any stochastic process. Here, we study the statistical properties of gene and protein-family sizes in order to quantify the long time scale evolutionary differences and similarities across species. We first determine the protein families of 209 species of bacteria and 20 species of archaea. We find that we are unable to reject the null hypothesis that the protein-family sizes of these species are drawn from the same distribution. In addition, we find that for species classified in the same phylogenetic branch or in the same lifestyle group, family size distributions are not significantly more similar than for species in different branches. These two findings can be accounted for in terms of a dynamical birth, death, and innovation model that assumes identical protein-family evolutionary rates for all species. Our theoretical and empirical results thus strongly suggest that the variability empirically observed in protein-family size distributions is compatible with the expected stochastic fluctuations for an evolutionary process with identical genomic evolutionary rates. Our findings hold special importance for the plausibility of some theories of the origin of eukaryotes which require drastic changes in evolutionary rates for some period during the last 2 billion years.


Outcome of untreated meningiomas.
Nowadays meningiomas are frequently detected incidentally. Their natural history has not yet been established because it is difficult to predict the growth pattern. Therefore, the management, after the radiological diagnosis, is still controversial. To evaluate the clinical outcome and growth rate of conservatively treated meningiomas at our tertiary center, identify prognostic factors of tumor growth, and suggest guidelines based on the available data and our experience. We reviewed the clinical records of 56 patients with 63 untreated meningiomas. Most were diagniosed incidentally. Clinical features and imaging findings at diagnosis and during follow-up were compared between growing and non-growing tumors. Potential patient- and tumor-related predictive factors for growth were analyzed. The study group included 46 women (52 meningiomas) and 10 men (11 meningiomas) aged 39-83 years. Mean tumor size was 18 +/- 11 mm (range 3-70 mm) at diagnosis and 22 +/- 11 mm (range 8-70 mm) at last follow-up; mean follow-up time was 65 +/- 34 months (range 15-152 months). During follow-up 24 tumors (38%) grew ata rate of 4 mm per year; none became symptomatic. Only two prognostic factors were statistically significantly associated with low growth rate: older age and tumor calcifications. Given our finding of a low growth incidence of meningiomas in the elderly, we support conservative management in patients aged 70 years or older. Calcifications into the meningioma are also indicative of slow growth, suggesting a conservative strategy. Surgery is recommended in younger patients in whom tumor growth occurs more often and a longer follow-up is necessary.


Permeability of normal versus carious dentin.
Although a number of reports have been published demonstrating that carious dentin is less permeable than normal dentin, these reports have been qualitative rather than quantitative. The purpose of this in vitro study was to apply a quantitative technique to the study of the permeability of carious human teeth before and after excavation, before and after removal of the smear layer and before and after preparation of a control cavity of similar size and depth in normal dentin subjected to the same measurements, for comparative purposes. Dentin permeability was measured as a hydraulic conductance. The permeability values measured at each step in the protocol were expressed as a percent of the maximum permeability of both cavities, permitting each tooth the serve as its own control. Carious lesions exhibited a slight degree of permeability (2.3 +/- 0.6% of controls) which remained unchanged after excavation of the lesions. Removal of the smear layer in the excavated carious lesions increased the permeability significantly to 6.9 +/- 3.2%. Preparation of a control cavity of the same area and depth increased the permeability slightly. Removal of its smear layer increased the permeability of the dentin 91%. These results confirm previous qualitative studies that carious dentin, even after excavation and removal of the smear layer has a very low permeability.


Initiation of repair of DNA-polypeptide cross-links by the UvrABC nuclease.
Although the biochemical pathways that repair DNA-protein cross-links have not been clearly elucidated, it has been proposed that the partial proteolysis of cross-linked proteins into smaller oligopeptides constitutes an initial step in removal of these lesions by nucleotide excision repair (NER). To test the validity of this repair model, several site-specific DNA-peptide and DNA-protein cross-links were engineered via linkage at (1) an acrolein-derived gamma-hydroxypropanodeoxyguanosine adduct and (2) an apurinic/apyrimidinic site, and the initiation of repair was examined in vitro using recombinant proteins UvrA and UvrB from Bacillus caldotenax and UvrC from Thermotoga maritima. The polypeptides cross-linked to DNA were Lys-Trp-Lys-Lys, Lys-Phe-His-Glu-Lys-His-His-Ser-His-Arg-Gly-Tyr, and the 16 kDa protein, T4 pyrimidine dimer glycosylase/apurinic/apyrimidinic site lyase. For the substrates examined, DNA incision required the coordinated action of all three proteins and occurred at the eighth phosphodiester bond 5' to the lesion. The incision rates for DNA-peptide cross-links were comparable to or greater than that measured on fluorescein-adducted DNA, an excellent substrate for UvrABC. Incision rates were dependent on both the site of covalent attachment on the DNA and the size of the bound peptide. Importantly, incision of a DNA-protein cross-link occurred at a rate approximately 3.5-8-fold slower than the rates observed for DNA-peptide cross-links. Thus, direct evidence has been obtained indicating that (1) DNA-peptide cross-links can be efficiently incised by the NER proteins and (2) DNA-peptide cross-links are preferable substrates for this system relative to DNA-protein cross-links. These data suggest that proteolytic degradation of DNA-protein cross-links may be an important processing step in facilitating NER.


Shotgun proteomics data from multiple organisms reveals remarkable quantitative conservation of the eukaryotic core proteome.
Genome-wide, absolute quantification of expressed proteins is not yet within reach for most eukaryotes. However, large numbers of MS-based protein identifications have been deposited in databases, together with information on the observation frequencies of each peptide spectrum ("spectral counts"). We have conducted a meta-analysis using several million peptide observations from five model eukaryotes, establishing a consistent, semi-quantitative analysis pipeline. By inferring and comparing protein abundances across orthologs, we observe: (i) the accuracy of spectral counting predictions increases with sampling depth and can rival that of direct biochemical measurements, (ii) the quantitative makeup of the consistently observed core proteome in eukaryotes is remarkably stable, with abundance correlations exceeding R(S)=0.7 at an evolutionary distance greater than 1000 million years, and (iii) some groups of proteins are more constrained than others. We argue that our observations reveal stabilizing selection: central parts of the eukaryotic proteome appear to be expressed at well-balanced, near-optimal abundance levels. This is consistent with our further observations that essential proteins show lower abundance variations than non-essential proteins, and that gene families that tend to undergo gene duplications are less well constrained than families that keep a single-copy status.


Protection against woodchuck hepatitis virus (WHV) infection by gene gun coimmunization with WHV core and interleukin-12.
Woodchuck hepatitis virus (WHV) and hepatitis B virus (HBV) are closely similar with respect to genomic organization, host antiviral responses, and pathobiology of the infection. T-cell immunity against viral nucleocapsid (HBcAg or WHcAg) has been shown to play a critical role in viral clearance and protection against infection. Here we show that vaccination of healthy woodchucks by gene gun bombardment with a plasmid coding for WHcAg (pCw) stimulates proliferation of WHcAg-specific T cells but that these cells do not produce significant levels of gamma interferon (IFN-gamma) upon antigen stimulation. In addition, animals vaccinated with pCw alone were not protected against WHV inoculation. In order to induce a Th1 cytokine response, another group of woodchucks was immunized with pCw together with another plasmid coding for woodchuck interleukin-12 (IL-12). These animals exhibited WHcAg-specific T-cell proliferation with high IFN-gamma production and were protected against challenge with WHV, showing no viremia or low-level transient viremia after WHV inoculation. In conclusion, gene gun immunization with WHV core generates a non-Th1 type of response which does not protect against experimental infection. However, steering the immune response to a Th1 cytokine profile by IL-12 coadministration achieves protective immunity. These data demonstrate a crucial role of Th1 responses in the control of hepadnavirus replication and suggest new approaches to inducing protection against HBV infection.


Real-time optoacoustic tomography of indocyanine green perfusion and oxygenation parameters in human finger vasculature.
We interrogated whether optoacoustic tomography could be employed to study blood functional parameters and biodistribution of injected fluorescent agents in humans. Using a multichannel scanner at a frame rate of 10 images per second, we obtained cross-sectional images of the human finger in real time, before and after the administration of indocyanine green. We demonstrated that multispectral optoacoustic tomography can sense fast flow kinetics and resolve spatiotemporal characteristics of a common fluorochrome in human vasculature at clinically relevant concentrations. We further register ICG images with oxygen saturation maps and anatomical views of the proximal interphalangeal joint of a healthy volunteer.


RNA-seq reveals the RNA binding proteins, Hfq and RsmA, play various roles in virulence, antibiotic production and genomic flux in Serratia sp. ATCC 39006.
Serratia sp. ATCC 39006 (S39006) is a Gram-negative enterobacterium that is virulent in plant and animal models. It produces a red-pigmented trypyrrole secondary metabolite, prodigiosin (Pig), and a carbapenem antibiotic (Car), as well as the exoenzymes, pectate lyase and cellulase. Secondary metabolite production in this strain is controlled by a complex regulatory network involving quorum sensing (QS). Hfq and RsmA (two RNA binding proteins and major post-transcriptional regulators of gene expression) play opposing roles in the regulation of several key phenotypes within S39006. Prodigiosin and carbapenem production was abolished, and virulence attenuated, in an S39006 ∆hfq mutant, while the converse was observed in an S39006 rsmA transposon insertion mutant. In order to define the complete regulon of Hfq and RsmA, deep sequencing of cDNA libraries (RNA-seq) was used to analyse the whole transcriptome of S39006 ∆hfq and rsmA::Tn mutants. Moreover, we investigated global changes in the proteome using an LC-MS/MS approach. Analysis of differential gene expression showed that Hfq and RsmA directly or indirectly regulate (at the level of RNA) 4% and 19% of the genome, respectively, with some correlation between RNA and protein expression. Pathways affected include those involved in antibiotic regulation, virulence, flagella synthesis, and surfactant production. Although Hfq and RsmA are reported to activate flagellum production in E. coli and an adherent-invasive E. coli hfq mutant was shown to have no flagella by electron microscopy, we found that flagellar production was increased in the S39006 rsmA and hfq mutants. Additionally, deletion of rsmA resulted in greater genomic flux with increased activity of two mobile genetic elements. This was confirmed by qPCR and analysis of rsmA culture supernatant revealed the presence of prophage DNA and phage particles. Finally, expression of a hypothetical protein containing DUF364 increased prodigiosin production and was controlled by a putative 5' cis-acting regulatory RNA element. Using a combination of transcriptomics and proteomics this study provides a systems-level understanding of Hfq and RsmA regulation and identifies similarities and differences in the regulons of two major regulators. Additionally our study indicates that RsmA regulates both core and variable genome regions and contributes to genome stability.


Mixed Invasive Ductal and Lobular Carcinoma of the Breast: Prognosis and the Importance of Histologic Grade.
The diagnosis of mixed invasive ductal and lobular carcinoma (IDC-L) in clinical practice is often associated with uncertainty related to its prognosis and response to systemic therapies. With the increasing recognition of invasive lobular carcinoma (ILC) as a distinct disease subtype, questions surrounding IDC-L become even more relevant. In this study, we took advantage of a detailed clinical database to compare IDC-L and ILC regarding clinicopathologic and treatment characteristics, prognostic power of histologic grade, and survival outcomes. In this retrospective cohort study, we identified 811 patients diagnosed with early-stage breast cancer with IDC-L or ILC. Descriptive statistics were performed to compare baseline clinicopathologic characteristics and treatments. Survival rates were subsequently analyzed using the Kaplan-Meier method and compared using the Cox proportional hazards model. Patients with ILC had more commonly multifocal disease, low to intermediate histologic grade, and HER2-negative disease. Histologic grade was prognostic for patients with IDC-L but had no significant discriminatory power in patients with ILC. Among postmenopausal women, those with IDC-L had significantly better outcomes when compared with those with ILC: disease-free survival (DFS) and overall survival (OS; adjusted hazard ratio [HR], 0.54; 95% confidence interval [CI] 0.31-0.95). Finally, postmenopausal women treated with an aromatase inhibitor had more favorable DFS and OS than those treated with tamoxifen only (OS adjusted HR, 0.50; 95% CI, 0.29-0.87), which was similar for both histologic types (p = .212). IDC-L tumors have a better prognosis than ILC tumors, particularly among postmenopausal women. Histologic grade is an important prognostic factor in IDC-L but not in ILC. This study compared mixed invasive ductal and lobular carcinoma (IDC-L) with invasive lobular carcinomas (ILCs) to assess the overall prognosis, the prognostic role of histologic grade, and response to systemic therapy. It was found that patients with IDC-L tumors have a better prognosis than ILC, particularly among postmenopausal women, which may impact follow-up strategies. Moreover, although histologic grade failed to stratify the risk of ILC, it showed an important prognostic power in IDC-L, thus highlighting its clinical utility to guide treatment decisions of IDC-L. Finally, the disease-free survival advantage of adjuvant aromatase inhibitors over tamoxifen in ILC was consistent in IDC-L.


Electrically controlling cell adhesion, growth and migration.
We have developed a neurochip to control the adhesion and outgrowth of individual neurons by electrochemical removal of protein repellent molecules from transparent electrodes. The neurochip architecture is based on three parallel indium-tin-oxide (ITO) electrodes on a SiO(2) substrate and a photoresist structure forming a landing spot for the neuron soma and two lateral outgrowth pathways for the neurites. The whole surface was turned protein and cell repellent with poly(ethylene glycol) grafted-poly(L-lysine) (PLL-g-PEG) before enabling neuron soma adhesion by selective PLL-g-PEG removal. After the neuron has settled down a potential was applied to the pathway electrodes to permit the neurite outgrowth along pathways formed by the SU8 structure. We also show the possibility to control cell migration by small pulsed currents. Myoblasts were therefore seeded on a chemical pattern of cell adhesive PLL and cell resistant PLL-g-PEG. The PLL-g-PEG was then removed electrochemically from the electrodes to permit migration onto the cell free electrodes. Electrodes without applied current were confluently overgrown within 24 h but a small pulsed current was able to inhibit cell growth on the bare ITO electrode for more than 72 h. With both techniques, cell adhesion, growth and migration can be controlled dynamically after the cells started to grow on the substrate. This opens new possibilities: we believe the key to control the development of topologically controlled neuron networks or more complex co-cultures is the combination of passive surface modifications and active control over the surface properties at any time of the experiment.


Local tenomodulin absence, angiogenesis, and matrix metalloproteinase activation are associated with the rupture of the chordae tendineae cordis.
Rupture of the chordae tendineae cordis (CTC) is a well-known cause of mitral regurgitation. Despite its importance, the mechanisms by which the CTC is protected and the cause of its rupture remain unknown. CTC is an avascular tissue. We investigated the molecular mechanisms underlying the avascularity of CTC and the correlation between avascularity and CTC rupture. We found that tenomodulin, which is a recently isolated antiangiogenic factor, was expressed abundantly in the elastin-rich subendothelial outer layer of normal rodent, porcine, canine, and human CTC. Conditioned medium from cultured CTC interstitial cells strongly inhibited tube formation and mobilization of endothelial cells; these effects were partially inhibited by small-interfering RNA against tenomodulin. The immunohistochemical analysis was performed on 12 normal and 16 ruptured CTC obtained from the autopsy or surgical specimen. Interestingly, tenomodulin was locally absent in the ruptured areas of CTC, where abnormal vessel formation, strong expression of vascular endothelial growth factor-A and matrix metalloproteinases, and infiltration of inflammatory cells were observed, but not in the normal or nonruptured area. In anesthetized open-chest dogs, the tenomodulin layer of tricuspid CTC was surgically filed, and immunohistological analysis was performed after several months. This intervention gradually caused angiogenesis and expression of vascular endothelial growth factor-A and matrix metalloproteinases in the core collagen layer in a time-dependent manner. These findings provide evidence that tenomodulin is expressed universally in normal CTC in a concentric pattern and that local absence of tenomodulin, angiogenesis, and matrix metalloproteinase activation are associated with CTC rupture.


Modification of Tobacco rattle virus RNA1 to serve as a VIGS vector reveals that the 29K movement protein is an RNA silencing suppressor of the virus.
Tobacco rattle virus (TRV) has a bipartite, positive-sense single-stranded RNA genome and is widely used for virus-induced gene silencing (VIGS) in plants. RNA1 of TRV that lacks the gene for the cysteine-rich 16K silencing-suppression protein infects plants systemically in the absence of RNA2. Here, we attempted to engineer RNA1 for use as a VIGS vector by inserting heterologous gene fragments to replace 16K. The RNA1 vector systemically silenced the phytoene desaturase (PDS) gene, although less efficiently than when the original VIGS vector system was used, which consists of wild-type RNA1 and engineered RNA2 carrying the heterologous gene. Infectious RNA1 mutants with a dysfunctional 16K suppressed silencing and enhanced transgene expression in green fluorescent protein-transgenic Nicotiana benthamiana following inoculation by agroinfiltration, unlike mutants that also lacked 29K, a movement protein (MP) gene. The 30K MP gene of Tobacco mosaic virus complemented in cis the movement defect but not the silencing suppression functions of TRV 29K. Silencing suppression by 29K occurred in the context of RNA1 replication but not in an agroinfiltration assay which tested 29K alone for suppression of sense-mediated silencing. Both 29K and 16K were needed to avoid necrotic symptoms in RNA1-infected N. benthamiana. The results shed new light on virulence factors of TRV.


The geriatric floating interdisciplinary transition team.
Older adults often receive suboptimal care during hospitalizations and transitions to postacute settings. Inpatient geriatric services have been shown to increase care quality but have not improved patient outcomes consistently. Acute Care for the Elderly units improve patient outcomes but are resource intensive. Transitional care has been shown to reduce hospital readmissions and healthcare costs. This article describes the Geriatric Floating Interdisciplinary Transition Team (Geri-FITT), a model that combines the strengths of inpatient geriatric evaluation and comanagement and transitional care models by creating an inpatient comanagement service that also delivers transitional care. The Geri-FITT model is designed to improve the hospital care of older adults and their transitions to postacute settings. In Geri-FITT, a geriatrician-geriatric nurse practitioner team assesses patients, comanages geriatric syndromes, provides staff education, encourages patient self-management, communicates with primary care providers, and follows up with patients soon after discharge. This pilot cohort study of Geri-FITT included hospitalized patients aged 70 and older on four general medicine services (two Geri-FITT, two usual care) at an academic medical center (N=717). The study assessed the effect of Geri-FITT on patients' care transition quality (Care Transitions Measure) and their satisfaction with hospital care (four questions). The results indicate that Geri-FITT is associated with slightly higher, though not statistically significantly so, quality care transitions and greater patient satisfaction with inpatient care. Geri-FITT may be a feasible approach to enhancing inpatient management and transitional care for older adults. Further study of its effect on these and other outcomes in other healthcare settings seems warranted.


Primary stability and stiffness in ankle arthrodes-crossed screws versus anterior plating.
Ankle arthrodesis is commonly used for the treatment of osteoarthritis or failed arthroplasty. Screw fixation is the predominant technique to perform ankle arthrodesis. Due to a considerable frequency of failures research suggests the use of an anatomically shaped anterior double plate system as a reliable method for isolated tibiotalar arthrodesis. The purpose of the present biomechanical study was to compare two groups of ankle fusion constructs - three screw fixation and an anterior double plate system - in terms of primary stability and stiffness. Six matched-pairs human cadaveric lower legs (Thiel fixated) were used in this study. One specimen from each pair was randomly assigned to be stabilized with the anterior double plate system and the other with the three-screw technique. The different arthrodesis methods were tested by dorsiflexing the foot until failure of the system, defined as rotation of the talus relative to the tibia in the sagittal plane. Experiments were performed on a universal materials testing machine. The force required to make arthrodesis fail was documented. For calculation of the stiffness, a linear regression was fitted to the force-displacement curve in the linear portion of the curve and its slope taken as the stiffness. For the anatomically shaped double-plate system a mean load of 967N was needed (range from 570N to 1400N) to make arthrodesis fail. The three-screw fixation method resisted a mean load of 190N (range from 100N to 280N) (p=0.005). In terms of stiffness a mean of 56N/mm (range from 35N/mm to 79N/mm) was achieved for the anatomically shaped double-plate system whereas a mean of 10N/mm (range from 6N/mm to 18N/mm) was achieved for the three-screw fixation method (p=0.004). Our biomechanical data demonstrates that the anterior double-plate system is significantly superior to the three-screw fixation technique for ankle arthrodesis in terms of primary stability and stiffness.


Changes to SNAP-authorized retailer stocking requirements and the supply of foods and beverages in low-income communities in seven U.S. states.
Low-income communities often lack access to supermarkets and healthy foods. Enhanced stocking requirements for staple foods for Supplemental Nutrition Assistance Program (SNAP)-authorized retailers may increase availability of healthy foods in smaller stores which are prevalent in low-income areas. This study aimed to evaluate the extent that small food stores located in low-income areas met the U.S. Department of Agriculture's 2016 final rule on SNAP-authorized retailer stocking requirements, which increased the minimum number of required staple food varieties from three to seven for each staple food category, required a depth of stock of three units of each variety, and increased the required number of categories with perishables from two to three. A multisite research project was conducted in 2017. Nine research teams located in seven U.S. states audited the availability of perishable and nonperishable staple foods and beverages in 351 small food stores in low-income areas. Analyses determined the extent to which stores met all or part of the stocking requirements and tested differences by store type. 30.2% of stores met all of the 2016 final rule requirements; 86.3% met the requirements for fruits and vegetables, whereas only 30.5% met requirements for dairy. 53.1% of non-chain small grocery stores met all requirements compared to 17.1% of convenience stores (p < .0001). Less than one half of the food stores audited met the U.S. Department of Agriculture's 2016 final rule that would expand SNAP-authorized retailer stocking requirements suggesting that, if implemented, the rule may generate increased offerings of staple foods in small stores in low-income areas.


Diminished error processing in smokers during smoking cue exposure.
Deficits in error processing may contribute to the continuation of impulsive behaviors such as smoking. Previous studies show deficits in error processing among substance abuse patients. However, these studies were all conducted during affectively neutral conditions. Deficits in error processing in smokers may become more pronounced under affectively challenging conditions, such as during smoking cue exposure. The aim of the present study was to investigate whether smokers showed initial error processing deficits, as measured with the error-related negativity (ERN), and decreased motivational significance attributed to an error, as measured with the error positivity (Pe) when exposed to smoking cues. Additionally, we examined the nature of the ERN and Pe amplitudes in more detail by investigating their associations with trait impulsivity, nicotine dependence levels and cigarette craving. Event-related potentials were measured during a modified Erikson flanker task in both smokers and non-smoking controls. Smokers showed reduced ERN and Pe amplitudes after making an error, accompanied by diminished post-error slowing of reaction times. These results suggest that initial error processing and motivational significance attributed to an error are affected in smokers during smoking cue exposure. Furthermore, individual variation in impulsivity and nicotine dependence was associated with reduced ERN amplitudes.


Cardiovascular depression and stabilization by central vasopressin in rats.
The role of endogenous vasopressin in cardiovascular homeostasis was examined using vasopressin-deficient rats (Brattleboro) (n = 194) and their parent strain, Long-Evans rats (n = 181). Mean arterial pressure (blood pressure) and heart rate were measured every 4 seconds with or without infusion of drug solution for 21 hours, and mean values and their standard deviations (lability) were calculated. Blood pressure in Brattleboro rats (116 +/- 1.1 mm Hg, mean +/- SEM) was significantly higher than that in Long-Evans rats (96 +/- 0.7 mm Hg, p less than 0.001), whereas heart rates (381 +/- 3.3 and 375 +/- 2.9 beats/min, respectively) were similar. The lability of blood pressure and heart rate in Brattleboro rats (9.2 +/- 0.1 mm Hg and 42.3 +/- 0.7 beats/min) was also greater than that in Long-Evans rats (6.7 +/- 0.1 mm Hg, p less than 0.001 and 38.4 +/- 0.8 beats/min, p less than 0.01, respectively). In Brattleboro rats, intravenous vasopressin (0.1 ng/kg/min or 0.6 ng/kg/min) did not affect blood pressure, although it did reduce heart rate and decreased lability of blood pressure and heart rate. Intracerebroventricular (central) infusion of vasopressin (2 pg/kg/min) in Brattleboro rats induced initial hypertension and tachycardia followed by long-lasting hypotension and bradycardia, whereas in Long-Evans rats it induced only hypertension and tachycardia. In both strains, central vasopressin dramatically decreased the lability of blood pressure and heart rate. Neither intravenous (0.2 ng/kg/min) nor central desmopressin (2 pg/kg/min or 0.2 ng/kg/min), a V2 renal receptor agonist, changed any of these parameters in Brattleboro rats, although both diminished urinary volume. Neither intravenous (50 ng/kg/min) nor central (3.3 pg/kg/min) d(CH2)5-Tyr(Me)-arginine vasopressin, a vasopressin V1 receptor antagonist, modulated any of these parameters in Long-Evans rats. These results suggest that endogenous as well as exogenous vasopressin acts centrally as a cardiovascular inhibitor and stabilizer through a receptor mechanism other than V1 or V2 receptor mechanisms.


Radiological Examination of Postoperative Cervical Alignment and Stability in Patients with Dialysis-Associated Spondylosis Excluding Destructive Spondyloarthropathy: Comparison with Patients with Cervical Spondylotic Myelopathy.
Several reports have demonstrated the surgical treatment strategy for patients with dialysis-associated spondylosis in the cervical spine (CDAS) with destructive spondyloarthropathy (DSA). However, studies focusing on the clinical outcome of patients with CDAS without DSA remain scarce. We aimed to review the treatment strategy of patients with CDAS but without DSA. The clinical data and surgical records of consecutive patients with CDAS without DSA (n = 9; D-group) and cervical spondylotic myelopathy (CSM) (n = 30; C-group) who underwent modified double-door laminoplasty(DDL) were reviewed retrospectively. We investigated four radiologic factors in the pre-and postoperative periods that have been reported to be the risk factors for worsening of clinical symptoms in various studies and examined statistical comparison between the D and C groups. In the D group, the pre- versus postoperative C2-C7 sagittal angles were not significantly different, and only two patients (22%) had kyphosis postoperatively. There was a significant difference in the pre- and postoperative C2-C7 angles in the two groups (P = 0.031). Regarding the change in segmental alignment, the local open angle increased at the C4/C5 level in the D group. Also there was a significant difference in the local angles between the two groups at C4/5 and C5/6 (P = 0.00038, and 0.037), suggesting that postoperative segmental mobility at C4/5 and C5/6 was higher in the D group than in the C group. In the present study, DDL in patients with CDAS without DSA did not adversely affect the postoperative alignment and stability compared with CSM patients with CSM. However, patients in the D group may have a chance to develop DSA change at the C4/5 level in the future, and careful long-term follow-up is warranted.


Cancer occurrence among workers exposed to acrylonitrile.
A MEDLINE search identified 12 published epidemiologic studies that have reported incidence or mortality experience among workers exposed to acrylonitrile. Many of the studies contain scanty descriptions of subject ascertainment, and most do not have good information on exposure assessment. Many also may have suffered from incomplete follow-up, as evinced by an overall deficit in the number of deaths observed, compared with the number expected from general population mortality rates. Such problems are not unique to studies on acrylonitrile, and to some extent they reflect the difficulties of conducting retrospective cohort studies. Despite these drawbacks, a simplified meta-analysis of the mortality experience reported for these cohorts revealed little evidence for carcinogenicity. Approximately the same number of cancer deaths was observed as was expected according to general population mortality rates (standardized mortality ratio 1.03, 90% confidence interval 0.92-1.15). The combined information from these studies is insufficient to support confidence about a lack of carcinogenicity at all sites. Nevertheless, despite the flaws in some of the individual studies, the summarized findings offer reassurance that workers exposed to acrylonitrile face no striking increases in mortality for all cancers or for respiratory cancer.


Asymmetric collapse in biomimetic complex coacervates revealed by local polymer and water dynamics.
Complex coacervation is a phenomenon characterized by the association of oppositely charged polyelectrolytes into micrometer-scale liquid condensates. This process is the purported first step in the formation of underwater adhesives by sessile marine organisms, as well as the process harnessed for the formation of new synthetic and protein-based contemporary materials. Efforts to understand the physical nature of complex coacervates are important for developing robust adhesives, injectable materials, or novel drug delivery vehicles for biomedical applications; however, their internal fluidity necessitates the use of in situ characterization strategies of their local dynamic properties, capabilities not offered by conventional techniques such as X-ray scattering, microscopy, or bulk rheological measurements. Herein, we employ the novel magnetic resonance technique Overhauser dynamic nuclear polarization enhanced nuclear magnetic resonance (DNP), together with electron paramagnetic resonance (EPR) line shape analysis, to concurrently quantify local molecular and hydration dynamics, with species- and site-specificity. We observe striking differences in the structure and dynamics of the protein-based biomimetic complex coacervates from their synthetic analogues, which is an asymmetric collapse of the polyelectrolyte constituents. From this study we suggest charge heterogeneity within a given polyelectrolyte chain to be an important parameter by which the internal structure of complex coacervates may be tuned. Acquiring molecular-level insight to the internal structure and dynamics of dynamic polymer complexes in water through the in situ characterization of site- and species-specific local polymer and hydration dynamics should be a promising general approach that has not been widely employed for materials characterization.


Vincristine-Induced Peripheral Neuropathy in Pediatric Oncology: A Randomized Controlled Trial Comparing Push Injections with One-Hour Infusions (The VINCA Trial).
Vincristine (VCR) is a frequently used chemotherapeutic agent. However, it can lead to VCR-induced peripheral neuropathy (VIPN). In this study we investigated if one-hour infusions of VCR instead of push-injections reduces VIPN in pediatric oncology patients. We conducted a multicenter randomized controlled trial in which participants received all VCR administrations through push injections or one-hour infusions. VIPN was measured at baseline and 1-5 times during treatment using Common Terminology Criteria of Adverse Events (CTCAE) and pediatric-modified Total Neuropathy Score. Moreover, data on co-medication, such as azole antifungals, were collected. Overall, results showed no effect of administration duration on total CTCAE score or ped-mTNS score. However, total CTCAE score was significantly lower in patients receiving one-hour infusions concurrently treated with azole antifungal therapy (β = -1.58; p = 0.04). In conclusion, generally VCR administration through one-hour infusions does not lead to less VIPN compared to VCR push injections in pediatric oncology patients. However, one-hour infusions lead to less severe VIPN compared to push-injections when azole therapy is administered concurrently with VCR. These results indicate that in children treated with VCR and requiring concurrent azole therapy, one-hour infusions might be beneficial over push injections, although larger trials are needed to confirm this association.


Bacterial swarms recruit cargo bacteria to pave the way in toxic environments.
Swarming bacteria are challenged by the need to invade hostile environments. Swarms of the flagellated bacterium Paenibacillus vortex can collectively transport other microorganisms. Here we show that P. vortex can invade toxic environments by carrying antibiotic-degrading bacteria; this transport is mediated by a specialized, phenotypic subpopulation utilizing a process not dependent on cargo motility. Swarms of beta-lactam antibiotic (BLA)-sensitive P. vortex used beta-lactamase-producing, resistant, cargo bacteria to detoxify BLAs in their path. In the presence of BLAs, both transporter and cargo bacteria gained from this temporary cooperation; there was a positive correlation between BLA resistance and dispersal. P. vortex transported only the most beneficial antibiotic-resistant cargo (including environmental and clinical isolates) in a sustained way. P. vortex displayed a bet-hedging strategy that promoted the colonization of nontoxic niches by P. vortex alone; when detoxifying cargo bacteria were not needed, they were lost. This work has relevance for the dispersal of antibiotic-resistant microorganisms and for strategies for asymmetric cooperation with agricultural and medical implications. Antibiotic resistance is a major health threat. We show a novel mechanism for the local spread of antibiotic resistance. This involves interactions between different bacteria: one species provides an enzyme that detoxifies the antibiotic (a sessile cargo bacterium carrying a resistance gene), while the other (Paenibacillus vortex) moves itself and transports the cargo. P. vortex used a bet-hedging strategy, colonizing new environments alone when the cargo added no benefit, but cooperating when the cargo was needed. This work is of interest in an evolutionary context and sheds light on fundamental questions, such as how environmental antibiotic resistance may lead to clinical resistance and also microbial social organization, as well as the costs, benefits, and risks of dispersal in the environment.


Lateralization of motor excitability during observation of bimanual signs.
Viewing another person's hand actions enhances excitability in an observer's left and right primary motor (M1) cortex. We aimed to determine whether viewing communicative hand actions alters this bilateral sensorimotor resonance. Using single-pulse transcranial magnetic stimulation (TMS), we measured excitability in the left and right M1 while right-handed non-signing participants observed bimanual communicative hand actions, i.e., meaningful signs in British Sign Language. TMS-induced motor evoked potentials were recorded from hand muscles during sign observation before and after teaching the participants to associate meanings with half of the signs. Before this teaching, when participants did not know that the presented hand actions were signs, excitability of left and right M1 was modulated equally. After learning the meanings of half the signs, excitability of the left, but not right, M1 was significantly enhanced. This left-lateralized enhancement of M1 excitability occurred during observation of signs with known and unknown meanings. The findings suggest that awareness of the communicative nature of another person's hand actions strengthens sensorimotor resonance in the left M1 cortex and alters hemispheric balance during action observation.


p27(Kip1) deficiency promotes prostate carcinogenesis but does not affect the efficacy of retinoids in suppressing the neoplastic process.
p27 is a cell cycle suppressor gene, whose protein is a negative regulator of cyclin/cdk complexes. p27 is also a potential target of retinoids in cancer prevention studies. In benign prostate hyperplasia (BPH), and in most carcinomas, p27(Kip1) is down-regulated, suggesting its potential resistance to retinoids. To test this hypothesis, we examined the efficacy of 9-cis retinoic acid (9cRA) to suppress prostate cell proliferation (PECP) and carcinogenesis in p27(Kip1) deficient mice. p27(Kip1) deficient (-/-), heterozygous (+/-) and homozygous (+/+) mice were treated for 7 days with testosterone, 9cRA, or with both, and cell proliferation in dorsolateral prostate (DLP) was determined by BrdU labeling. Prostate carcinogenesis was induced by N-Methyl-N-Nitrosourea (MNU) and hormone stimulation. PECP in DLP of two-month-old mice of all genotypes was similar but significantly increased in old p27-/- mice only. Testosterone treatment increased PECP in all three p27 genotypes with the highest values in p27-/- mice. p27(Kip1) deficiency did not affect the response of PEC to 9cRA and to 9cRA+testosterone. The decrease of p27(Kip1) in p27+/- and p27-/- mice progressively increased the incidence and frequency of PIN and tumors. 9cRA suppressed PIN in all three p27 genotypes and this was associated with decreased PECP and increased cellular senescence. This data indicates that p27(Kip1) deficiency promotes prostate cell proliferation and carcinogenesis but does not affect 9cRA's potential to suppress prostate carcinogenesis, suggesting that patients with PIN and carcinomas lacking or having a low level of p27(Kip1) expression may also benefit from clinical trials with retinoids.


The redox state of recombinant human serum albumin and its optimal concentration for mouse embryo culture.
Albumin has multiple physiological functions in embryo culture, such as a chelator of heavy metals, free radical scavenger, pH and osmotic regulator, a stabilizer, growth factor carrier, a surfactant, and a nutrient. However, the commercially available human serum albumin (HSA) products may not be completely safe since they could be contaminated with viruses and prions. Recombinant human serum albumin (rHSA) has been reported to be as efficient as commercial HSA for fertilization and embryo development. Despite the possible benefits of rHSA, it has not been widely used for embryo culture due to its high cost of production. Our objective was to analyze the redox state of different types of HSA products and rHSA to define oxidative status batch variations of HSA and rHSA and to evaluate the optimal concentration of rHSA for mouse embryo culture. The redox state of the HSA and rHSA used in embryo culture media was found to vary widely. Redox variations were found among different HSA batches as well as among rHSA batches. The highest blastocyst development and hatching rates were obtained with rHSA used at a concentration of 0.05 mg/mL. We showed that very low concentrations of rHSA were most favorable for advanced mouse embryo development in culture.


Genome-wide association mapping identifies the genetic basis of discrete and quantitative variation in sexual weaponry in a wild sheep population.
Understanding the genetic architecture of phenotypic variation in natural populations is a fundamental goal of evolutionary genetics. Wild Soay sheep (Ovis aries) have an inherited polymorphism for horn morphology in both sexes, controlled by a single autosomal locus, Horns. The majority of males have large normal horns, but a small number have vestigial, deformed horns, known as scurs; females have either normal horns, scurs or no horns (polled). Given that scurred males and polled females have reduced fitness within each sex, it is counterintuitive that the polymorphism persists within the population. Therefore, identifying the genetic basis of horn type will provide a vital foundation for understanding why the different morphs are maintained in the face of natural selection. We conducted a genome-wide association study using ∼36000 single nucleotide polymorphisms (SNPs) and determined the main candidate for Horns as RXFP2, an autosomal gene with a known involvement in determining primary sex characters in humans and mice. Evidence from additional SNPs in and around RXFP2 supports a new model of horn-type inheritance in Soay sheep, and for the first time, sheep with the same horn phenotype but different underlying genotypes can be identified. In addition, RXFP2 was shown to be an additive quantitative trait locus (QTL) for horn size in normal-horned males, accounting for up to 76% of additive genetic variation in this trait. This finding contrasts markedly from genome-wide association studies of quantitative traits in humans and some model species, where it is often observed that mapped loci only explain a modest proportion of the overall genetic variation.


Intracellular carbonic anhydrase activity sensitizes cancer cell pH signaling to dynamic changes in CO2 partial pressure.
Carbonic anhydrase (CA) enzymes catalyze the chemical equilibration among CO2, HCO3(-) and H(+). Intracellular CA (CAi) isoforms are present in certain types of cancer, and growing evidence suggests that low levels correlate with disease severity. However, their physiological role remains unclear. Cancer cell CAi activity, measured as cytoplasmic CO2 hydration rate (kf), ranged from high in colorectal HCT116 (∼2 s(-1)), bladder RT112 and colorectal HT29, moderate in fibrosarcoma HT1080 to negligible (i.e. spontaneous kf = 0.18 s(-1)) in cervical HeLa and breast MDA-MB-468 cells. CAi activity in cells correlated with CAII immunoreactivity and enzymatic activity in membrane-free lysates, suggesting that soluble CAII is an important intracellular isoform. CAi catalysis was not obligatory for supporting acid extrusion by H(+) efflux or HCO3(-) influx, nor for maintaining intracellular pH (pHi) uniformity. However, in the absence of CAi activity, acid loading from a highly alkaline pHi was rate-limited by HCO3(-) supply from spontaneous CO2 hydration. In solid tumors, time-dependence of blood flow can result in fluctuations of CO2 partial pressure (pCO2) that disturb cytoplasmic CO2-HCO3(-)-H(+) equilibrium. In cancer cells with high CAi activity, extracellular pCO2 fluctuations evoked faster and larger pHi oscillations. Functionally, these resulted in larger pH-dependent intracellular [Ca(2+)] oscillations and stronger inhibition of the mTORC1 pathway reported by S6 kinase phosphorylation. In contrast, the pHi of cells with low CAi activity was less responsive to pCO2 fluctuations. Such low pass filtering would "buffer" cancer cell pHi from non-steady-state extracellular pCO2. Thus, CAi activity determines the coupling between pCO2 (a function of tumor perfusion) and pHi (a potent modulator of cancer cell physiology).


Rapid detection of cocaine, benzoylecgonine and methylecgonine in fingerprints using surface mass spectrometry.
Latent fingerprints provide a potential route to the secure, high throughput and non-invasive detection of drugs of abuse. In this study we show for the first time that the excreted metabolites of drugs of abuse can be detected in fingerprints using ambient mass spectrometry. Fingerprints and oral fluid were taken from patients attending a drug and alcohol treatment service. Gas chromatography mass spectrometry (GC-MS) was used to test the oral fluid of patients for the presence of cocaine and benzoylecgonine. The corresponding fingerprints were analysed using Desorption Electrospray Ionization (DESI) which operates under ambient conditions and Ion Mobility Tandem Mass Spectrometry Matrix Assisted Laser Desorption Ionization (MALDI-IMS-MS/MS) and Secondary Ion Mass Spectrometry (SIMS). The detection of cocaine, benzoylecgonine (BZE) and methylecgonine (EME) in latent fingerprints using both DESI and MALDI showed good correlation with oral fluid testing. The sensitivity of SIMS was found to be insufficient for this application. These results provide exciting opportunities for the use of fingerprints as a new sampling medium for secure, non-invasive drug detection. The mass spectrometry techniques used here offer a high level of selectivity and consume only a small area of a single fingerprint, allowing repeat and high throughput analyses of a single sample.


Carbon fiber epoxy composites for both strengthening and health monitoring of structures.
This paper presents a study of the electrical and mechanical behavior of several continuous carbon fibers epoxy composites for both strengthening and monitoring of structures. In these composites, the arrangement of fibers was deliberately diversified to test and understand the ability of the composites for self-sensing low strains. Composites with different arrangements of fibers and textile weaves, mainly unidirectional continuous carbon reinforced composites, were tested at the dynamometer. A two-probe method was considered to measure the relative electrical resistance of these composites during loading. The measured relative electrical resistance includes volume and contact electrical resistances. For all tested specimens, it increases with an increase in tensile strain, at low strain values. This is explained by the improved alignment of fibers and resulting reduction of the number of possible contacts between fibers during loading, increasing as a consequence the contact electrical resistance of the composite. Laboratory tests on strengthening of structural elements were also performed, making hand-made composites by the "wet process", which is commonly used in civil engineering for the strengthening of all types of structures in-situ. Results show that the woven epoxy composite, used for strengthening of concrete elements is also able to sense low deformations, below 1%. Moreover, results clearly show that this textile sensor also improves the mechanical work of the strengthened structural elements, increasing their bearing capacity. Finally, the set of obtained results supports the concept of a textile fabric capable of both structural upgrade and self-monitoring of structures, especially large structures of difficult access and needing constant, sometimes very expensive, health monitoring.


Occurrence of Erysiphe necator chasmothecia and their natural parasitism by Ampelomyces quisqualis.
In northern Italy, Erysiphe necator overwinters almost exclusively as chasmothecia. From 2004 to 2008, we investigated the occurrence of natural parasitism of grapevine powdery mildew chasmothecia by Ampelomyces quisqualis in the Trentino-Alto Adige region, in northern Italy. The survey was conducted in 18 vineyards in autumn 2004 and 2005 and in 45 vineyards in autumn 2006 and 2007. The incidence of powdery mildew signs (white powdery mycelia and conidia), the number of chasmothecia and their development pattern, and the incidence of parasitism by A. quisqualis were assessed. The production of E. necator chasmothecia on leaves is related to the incidence and severity of the disease on leaves at the end of the season and is not correlated with the elevation of the vineyard, which is inversely related to the temperature. A limited amount of natural parasitism of E. necator by Ampelomyces spp. (0.17 to 3.51%) was observed in all of the years of the survey. Pycnidia and conidia of Ampelomyces spp. parasitizing E. necator chasmothecia were found in conventional, organically grown, and untreated vineyards. Some of the isolated Ampelomyces strains have conidia that are shaped differently than those of the commercial A. quisqualis strain (AQ10) and are phylogenetically different from AQ10.


Seasonal variation of tungiasis in an endemic community.
Tungiasis (caused by the sand flea Tunga penetrans) is hyperendemic in many resource-poor communities in Brazil. To understand transmission dynamics of this parasitic skin disease in a typical endemic area, a longitudinal study was carried out in a slum in Fortaleza in northeastern Brazil. In a door-to-door survey, the population of a randomly selected area (n = 1,460) was examined on four occasions for the presence of embedded sand fleas. Prevalence rates were 33.6% in March (rainy season), 23.8% in June (end of the rainy season), 54.4% in September (peak of the dry season), and 16.8% in January (begin of the rainy season). Tungiasis was more common in males than in females. The intensity of infestation was correlated with the prevalence. The study shows that prevalence of tungiasis and parasite burden vary significantly during the year with a peak in the dry season. These findings have important consequences for the design of control measures.


Genetic dissection of systemic lupus erythematosus pathogenesis: partial functional complementation between Sle1 and Sle3/5 demonstrates requirement for intracellular coexpression for full phenotypic expression of lupus.
Sle1 on chromosome 1 and Sle3/5 on chromosome 7 are two of the most critical lupus susceptibility loci of the New Zealand Black/White-derived NZM2410 mouse strain. In contrast to C57BL/6 mice congenic for either Sle1 (B6.Sle1) or Sle3/5 (B6.Sle3/5), strains that express only a modest lupus-related phenotype, the bicongenic B6.Sle1.Sle3/5 strain has a robust phenotype, suggesting a critical role for epistatic interactions in lupus pathogenesis. Mixed chimera experiments indicated that the two loci are functionally expressed by different cell populations and predicted that phenotypic expression of the phenotypic features of the B6.Sle1.Sle3/5 strain could be fully reproduced with a combination of B6.Sle1 and B6.Sle3/5 bone marrow. Contrary to our expectations, there was only a partial functional complementation in these mixed chimeras. Spleen enlargement, CD4:CD8 ratio elevation, and epitope spreading of autoantibodies were fully developed in B6+B6.Sle1.Sle3/5 but not in B6.Sle1+B6.Sle3/5 mixed chimeras. This study is the first to present evidence that the pathways mediated by two critical lupus susceptibility loci derived from the New Zealand White strain must be integrated intracellularly for epistatic interactions to occur. Our mixed chimera approach continues to provide novel insights into the functional genetic pathways underlying this important murine model of systemic autoimmunity.


Structural and functional plasticity of subcellular tethering, targeting and processing of RPGRIP1 by RPGR isoforms.
Mutations affecting the retinitis pigmentosa GTPase regulator-interacting protein 1 (RPGRIP1) interactome cause syndromic retinal dystrophies. RPGRIP1 interacts with the retinitis pigmentosa GTPase regulator (RPGR) through a domain homologous to RCC1 (RHD), a nucleotide exchange factor of Ran GTPase. However, functional relationships between RPGR and RPGRIP1 and their subcellular roles are lacking. We show by molecular modeling and analyses of RPGR disease-mutations that the RPGR-interacting domain (RID) of RPGRIP1 embraces multivalently the shared RHD of RPGR(1-19) and RPGR(ORF15) isoforms and the mutations are non-overlapping with the interface found between RCC1 and Ran GTPase. RPGR disease-mutations grouped into six classes based on their structural locations and differential impairment with RPGRIP1 interaction. RPGRIP1α(1) expression alone causes its profuse self-aggregation, an effect suppressed by co-expression of either RPGR isoform before and after RPGRIP1α(1) self-aggregation ensue. RPGR(1-19) localizes to the endoplasmic reticulum, whereas RPGR(ORF15) presents cytosolic distribution and they determine uniquely the subcellular co-localization of RPGRIP1α(1). Disease mutations in RPGR(1) (-19), RPGR(ORF15), or RID of RPGRIP1α(1), singly or in combination, exert distinct effects on the subcellular targeting, co-localization or tethering of RPGRIP1α(1) with RPGR(1-19) or RPGR(ORF15) in kidney, photoreceptor and hepatocyte cell lines. Additionally, RPGR(ORF15), but not RPGR(1-19), protects the RID of RPGRIP1α(1) from limited proteolysis. These studies define RPGR- and cell-type-dependent targeting pathways with structural and functional plasticity modulating the expression of mutations in RPGR and RPGRIP1. Further, RPGR isoforms distinctively determine the subcellular targeting of RPGRIP1α(1,) with deficits in RPGR(ORF15)-dependent intracellular localization of RPGRIP1α(1) contributing to pathomechanisms shared by etiologically distinct syndromic retinal dystrophies.


Assessment of anti-diabetic activity of the aqueous extract of leaves of Astilboides tabularis.
Astilboides tabularis has a long history of usage as the functional food with the effect of adjunctive therapy of diabetic in northeast of China. The present study was undertaken to assess anti-diabetic activity of the aqueous extract of leaves of A. tabularis and provided experimental evidence for the clinical usage of A. tabularis in the treatment of diabetes mellitus. The aqueous extracts of leaves of A. tabularis (EAT) were orally administered at 300 and 600mg/kg dose to epinephrine-induced and alloxan-induced diabetes mice. Hypoglycemic effects, change in body weight, water intake, food intake, lipid profile and lipid peroxidant were assessed. Furthermore, the main chemical components of EAT were isolated and high performance liquid chromatography (HPLC) analysis was employed to identify the phytochemical composition of the plant extract. The finally, total flavonoids content and total phenolic content were quantified by colorimetric assay. EAT showed a significant reduction in blood glucose level at both 300 and 600mg/kg (ig). In addition, EAT also exhibited a profound antioxidant effect for increasing SOD and GSH-px activity and decreasing TBARS activity at both low and high dose. Furthermore, EAT also could significantly decrease the contents of TC and TG. The 8 main constituents of EAT are also isolated and characterized and the content of total flavonoid and total phenolic are 22% and 25% in EAT respectively. The present investigation revealed that EAT possess potent anti-diabetic activity and polyphenol compounds may be the main active ingredients.


Impact of Cash Transfers on the Use of Oral Health Services and Oral Health Outcomes: A Systematic Review.
The objective of the present study was to summarize the evidence available on the impact of cash transfers on the use of oral health services and oral health outcomes. 9 databases were searched for studies on cash transfers and oral health: PubMed, Embase, The Cochrane Library, CINAHL, EconLit, Dentistry and Oral Sciences Source, Scopus, Web of Science, and LILACS. We conducted a systematic review of studies that evaluated the impact of cash transfers on the use of oral health services and other oral health outcomes. Three studies with more than 13,000 participants conducted in Brazil and Argentina were included. One study from Brazil found that participants in the Bolsa Familia Program (BFP) were more likely not to use oral health services (aPR: 6.18; 95% CI: 3.07-12.45; P < 0.001) and had a higher probability of presenting dental caries (aPR: 2.00; 95% CI: 1.47-2.69) and severe caries (aRR: 1.53; 95% CI: 1.18-2.00). Another study conducted in Brazil found that the BFP was associated with fewer dental caries among those enrolled in the first 2 years of the BFP as well as after six years. On the other hand, the Argentina study found that the Universal Child Allowance program did not have a statistically significant average treatment effect (ATE = -0.05; P > 0.05) on the use of dental health services. There is a lack of evidence about the impact of CT on the use of oral health services and oral health outcomes. The evidence suggests that cash transfers might not have a positive impact on the use of oral health services. The results regarding the impact of receiving cash transfers on dental caries are contradictory. However, more evidence is needed to draw stronger conclusions for policy taking. The protocol was registered in PROSPERO (CRD42021268234).


Position-Independent Decoding of Movement Intention for Proportional Myoelectric Interfaces.
In this decade, myoelectric interfaces based on pattern recognition have gained considerable attention thanks to their naturalness enabling human intentions to be conveyed to and in control of a machine. However, the high variations of electromyogram signal patterns caused by arm position changes prohibit application to the real world. In this paper, we propose a novel method of decoding movement intentions robust to arm position changes towards proportional myoelectric interfaces. Specifically, we devise the position-independent decoding that estimates the likelihood of different arm positions, which we predefine during a training step, and also decodes the movement intention in a unified framework. The proposed method has an advantage that could be used to decode the movement intentions on untrained arm positions in a realistic scenario. Our experimental results showed that the proposed method could successfully decode the continuous movement intentions (e.g., flexion/extension and radial/ulnar deviation) on both trained and untrained arm positions. Our study also proved the effectiveness of the proposed method by comparing the existing methods in terms of the decoded trajectories as movement intentions in untrained arm positions.


Missing clinical information in NHS hospital outpatient clinics: prevalence, causes and effects on patient care.
In Britain over 39,000 reports were received by the National Patient Safety Agency relating to failures in documentation in 2007 and the UK Health Services Journal estimated in 2008 that over a million hospital outpatient visits each year might take place without the full record available. Despite these high numbers, the impact of missing clinical information has not been investigated for hospital outpatients in the UK.Studies in primary care in the USA have found 13.6% of patient consultations have missing clinical information, with this adversely affecting care in about half of cases, and in Australia 1.8% of medical errors were found to be due to the unavailability of clinical information.Our objectives were to assess the frequency, nature and potential impact on patient care of missing clinical information in NHS hospital outpatients and to assess the principal causes. This is the first study to present such figures for the UK and the first to look at how clinicians respond, including the associated impact on patient care. Prospective descriptive study of missing information reported by surgeons, supplemented by interviews on the causes.Data were collected by surgeons in general, gastrointestinal, colorectal and vascular surgical clinics in three teaching hospitals across the UK for over a thousand outpatient appointments. Fifteen interviews were conducted with those involved in collating clinical information for these clinics.The study had ethics approval (Hammersmith and Queen Charlotte's & Chelsea Research Ethics Committee), reference number (09/H0707/27). Participants involved in the interviews signed a consent form and were offered the opportunity to review and agree the transcript of their interview before analysis. No patients were involved in this research. In 15% of outpatient consultations key items of clinical information were missing. Of these patients, 32% experienced a delay or disruption to their care and 20% had a risk of harm. In over half of cases the doctor relied on the patient for the information, making a clinical decision despite the information being missing in 20% of cases. Hospital mergers, temporary staff and non-integrated IT systems were contributing factors. If these findings are replicated across the NHS then almost 10 million outpatients are seen each year without key clinical information, creating over a million unnecessary appointments, and putting nearly 2 million patients at risk of harm. There is a need for a systematic, regular audit of the prevalence of missing clinical information. Only then will we know the impact on clinical decision making and patient care of new technology, service reorganisations and, crucially given the present financial climate, temporary or reduced staffing levels. Further research is needed to assess the relationship between missing clinical information and diagnostic errors; to examine the issue in primary care; and to consider the patients perspective.


Identification of ion channel genes in the Acyrthosiphon pisum genome.
Aphids are major pests of crops, causing hundreds of millions of dollars worth of damage annually. Ion channel proteins are often the targets of modern insecticides and mutations in ion channel genes can lead to resistance to many leading classes of insecticides. The sequencing of the pea aphid, Acyrthosiphon pisum, genome has now allowed detailed in silico analysis of the aphid ion channels. The study has revealed significant differences in the composition of the ion channel families between the aphid and other insects. For example A. pisum does not appear to contain a homologue of the nACh receptor alpha 5 gene whilst the calcium channel beta subunit has been duplicated. These variations could result in differences in function or sensitivity to insecticides. The genome sequence will allow the study of aphid ion channels to be accelerated, leading to a better understanding of the function of these economically important channels. The potential for identifying novel insecticide targets within the aphid is now a step closer.


Recent advances in ginseng as cancer therapeutics: a functional and mechanistic overview.
Cancer is one of the leading causes of death worldwide. Ginseng, a key ingredient in traditional Chinese medicine, shows great promise as a new treatment option. As listed by the U.S. National Institutes of Health as a complementary and alternative medicine, its anti-cancer functions are being increasingly recognized. This review covers the mechanisms of action of ginsenosides and their metabolites, which can modulate signaling pathways associated with inflammation, oxidative stress, angiogenesis, metastasis, and stem/progenitor-like properties of cancer cells. The emerging use of structurally modified ginsenosides and recent clinical studies on the use of ginseng either alone or in combination with other herbs or Western medicines which are exploited as novel therapeutic strategies will also be explored.


Inflammation and coronary artery calcification in South Asians: The Mediators of Atherosclerosis in South Asians Living in America (MASALA) study.
Inflammatory biomarkers and adipocytokines (IBA) may contribute to atherosclerosis by promoting vascular inflammation. The association between IBA and coronary artery calcium (CAC), a marker of subclinical atherosclerosis, is not well defined in South Asians (SA). We hypothesized that IBA (high sensitivity C-reactive protein [hsCRP], tumor necrosis factor alpha [TNF-α], adiponectin, and leptin) were independently associated with and improved discrimination of CAC among SA. We analyzed IBA and CAC among participants in the prospective Mediators of Atherosclerosis in South Asians Living in America (MASALA) study. We used logistic regression models to examine cross-sectional associations of IBA with CAC presence (CAC >0) and severity (CAC >100), and C-statistics to assess the incremental contribution of each IBA to traditional risk factors (TRF) from the AHA/ACC Pooled Cohort Equations (PCE) for discrimination of CAC. Among 906 participants in the MASALA study, women (n = 420) had significantly higher levels of hsCRP, adiponectin, and leptin but lower levels of TNF-α than men (p < .01 for all). There was no significant association between any of the four IBA and either CAC category in multivariable-adjusted models, respectively. Lastly, none of the four IBA improved discrimination of CAC presence or severity when added to elements of the PCE. IBA were not associated with CAC presence or severity in the MASALA population. IBA did not help identify SA at risk of subclinical atherosclerosis, although associations with ASCVD events remain unclear. In SA, CAC may have a distinct pathophysiology independent of inflammation as measured by IBA.


A Bacillus-based Coxsackie virus A16 mucosal vaccine induces strong neutralizing antibody responses.
The purpose of this study was to construct a Coxsackie virus A16 (CA16) mucosal vaccine and evaluate its ability to induce immune response. VP1 gene of CA16 was inserted into the genome of Bacillus subtilis via recombination and displayed on the surface of the spores. This Bacillus-based vaccine was used for intranasal immunization of mice and the serum antibody titer was determined by enzyme-linked immunosorbent assay (ELISA). Neutralization activity of the serum from immunized mice was analyzed by an in vitro neutralizing test. VP1 gene was successfully integrated into the genome of Bacillus subtilis and was expressed on the surface of Bacillus spores. Intranasal immunization of mice with this vaccine induced a higher level of VP1 specific IgA and IgG than in mice of the control group (p < 0.05). The neutralizing antibody titer in the spore immunization group was 1 : 169, which was higher than that in the control group (p < 0.05). We concluded that vaccine prepared by displaying CA16 VP1 protein on the surface of Bacillus subtilis spores can stimulate mice to produce protective neutralizing antibodies, which provides foundations for the development of CA16 mucosal vaccine.


Complexity of MRI induced heating on metallic leads: experimental measurements of 374 configurations.
MRI induced heating on PM leads is a very complex issue. The widely varying results described in literature suggest that there are many factors that influence the degree of heating and that not always are adequately addressed by existing testing methods. We present a wide database of experimental measurements of the heating of metallic wires and PM leads in a 1.5 T RF coil. The aim of these measurements is to systematically quantify the contribution of some potential factors involved in the MRI induced heating: the length and the geometric structure of the lead; the implant location within the body and the lead path; the shape of the phantom used to simulate the human trunk and its relative position inside the RF coil. We found that the several factors are the primary influence on heating at the tip. Closer locations of the leads to the edge of the phantom and to the edge of the coil produce maximum heating. The lead length is the other crucial factor, whereas the implant area does not seem to have a major role in the induced temperature increase. Also the lead structure and the geometry of the phantom revealed to be elements that can significantly modify the amount of heating. Our findings highlight the factors that have significant effects on MRI induced heating of implanted wires and leads. These factors must be taken into account by those who plan to study or model MRI heating of implants. Also our data should help those who wish to develop guidelines for defining safe medical implants for MRI patients. In addition, our database of the entire set of measurements can help those who wish to validate their numerical models of implants that may be exposed to MRI systems.


Mutually regulated expression of Pax6 and Six3 and its implications for the Pax6 haploinsufficient lens phenotype.
Pax6 is a key regulator of eye development in vertebrates and invertebrates, and heterozygous loss-of-function mutations of the mouse Pax6 gene result in the Small eye phenotype, in which a small lens is a constant feature. To provide an understanding of the mechanisms underlying this haploinsufficient phenotype, we evaluated in Pax6 heterozygous mice the effects of reduced Pax6 gene dosage on the activity of other transcription factors regulating eye formation. We found that Six3 expression was specifically reduced in lenses of Pax6 heterozygous mouse embryos. Interactions between orthologous genes from the Pax and Six families have been identified in Drosophila and vertebrate species, and we examined the control of Pax6 and Six3 gene expression in the developing mouse lens. Using in vitro and transgenic approaches, we found that either transcription factor binds regulatory sequences from the counterpart gene and that both genes mutually activate their expression. These studies define a functional relationship in the lens in which Six3 expression is dosage-dependent on Pax6 and where, conversely, Six3 activates Pax6. Accordingly, we show a rescue of the Pax6 haploinsufficient lens phenotype after lens-specific expression of Six3 in transgenic mice. This phenotypic rescue was accompanied by cell proliferation and activation of the platelet-derived growth factor alpha-R/cyclin D1 signaling pathway. Our findings thus provide a mechanism implicating gene regulatory interactions between Pax6 and Six3 in the tissue-specific defects found in Pax6 heterozygous mice.


The Relationship Between CGM-Derived Metrics, A1C, and Risk of Hypoglycemia in Older Adults With Type 1 Diabetes.
Continuous glucose monitoring (CGM) is now commonly used in the management of type 1 diabetes (T1D). The CGM-derived coefficient of variation (CV) measures glucose variability, and the glucose management indicator (GMI) measures mean glycemia (previously called estimated A1C). However, their relationship with laboratory-measured A1C and the risk of hypoglycemia in older adults with T1D is not well studied. In a single-center study, older adults (age ≥65 years) with T1D wore a CGM device for 14 days. The CV (%) and GMI were calculated, and A1C and clinical and demographic information were collected. We evaluated 130 older adults (age 71 ± 5 years), of whom 55% were women, 97% were White, diabetes duration was 39 ± 17 years, and A1C was 7.3 ± 0.6% (56 ± 15 mmol/mol). Participants were stratified by high CV (>36%; n = 77) and low CV (≤36%; n = 53). Although there was no difference in A1C levels between the groups with high and low CV (7.3% [56 mmol/mol] vs. 7.3% [53 mmol/mol], P = 0.4), the high CV group spent more time in hypoglycemia (<70 mg/dL and ≤54 mg/dL) compared with the group with low CV (median 31 vs. 84 min/day, P < 0.0001; 8 vs. 46 min/day, P < 0.001, respectively). An absolute difference between A1C and GMI of ≥0.5% was observed in 46% of the cohort. When the A1C was higher than the GMI by ≥0.5%, a higher duration of hypoglycemia was observed (P = 0.02). In older adults with T1D, the use of CGM-derived CV and GMI can better identify individuals at higher risk for hypoglycemia compared with A1C alone. These measures should be combined with A1C for better diabetes management in older adults with T1D.


Acylation of the Incretin Peptide Exendin-4 Directly Impacts Glucagon-Like Peptide-1 Receptor Signaling and Trafficking.
The glucagon-like peptide-1 receptor (GLP-1R) is a class B G protein-coupled receptor and mainstay therapeutic target for the treatment of type 2 diabetes and obesity. Recent reports have highlighted how biased agonism at the GLP-1R affects sustained glucose-stimulated insulin secretion through avoidance of desensitization and downregulation. A number of GLP-1R agonists (GLP-1RAs) feature a fatty acid moiety to prolong their pharmacokinetics via increased albumin binding, but the potential for these chemical changes to influence GLP-1R function has rarely been investigated beyond potency assessments for cAMP. Here, we directly compare the prototypical GLP-1RA exendin-4 with its C-terminally acylated analog, exendin-4-C16. We examine relative propensities of each ligand to recruit and activate G proteins and β-arrestins, endocytic and postendocytic trafficking profiles, and interactions with model and cellular membranes in HEK293 and HEK293T cells. Both ligands had similar cAMP potency, but exendin-4-C16 showed ∼2.5-fold bias toward G protein recruitment and a ∼60% reduction in β-arrestin-2 recruitment efficacy compared with exendin-4, as well as reduced GLP-1R endocytosis and preferential targeting toward recycling pathways. These effects were associated with reduced movement of the GLP-1R extracellular domain measured using a conformational biosensor approach and a ∼70% increase in insulin secretion in INS-1 832/3 cells. Interactions with plasma membrane lipids were enhanced by the acyl chain. Exendin-4-C16 showed extensive albumin binding and was highly effective for lowering of blood glucose in mice over at least 72 hours. Our study highlights the importance of a broad approach to the evaluation of GLP-1RA pharmacology. SIGNIFICANCE STATEMENT: Acylation is a common strategy to enhance the pharmacokinetics of peptide-based drugs. This work shows how acylation can also affect various other pharmacological parameters, including biased agonism, receptor trafficking, and interactions with the plasma membrane, which may be therapeutically important.


Breast cancer screening in older women.
There is currently an epidemic of breast cancer in women 65 years of age and older. The purposes of this paper are to explore the breast cancer screening behaviors of older women and to identify some of the determinants of screening in these women. Data were analyzed from the 1987 National Health Interview Survey, a continuous nationwide household interview survey of the U.S. civilian, noninstitutionalized population. As in other studies, the utilization of breast cancer screening by older women was less in older women than in younger women. This was true for both mammography and clinical breast examination. A number of determinants of screening in older women were identified here. Women with a usual source of care and/or no activity limitation, as well as high school graduates, were the ones most likely to have received a screening mammogram and/or a screening clinical breast exam during the past year. The failure of older women to receive adequate breast cancer screening is an important concern which should be reevaluated, given the breast cancer epidemic in this population. This study identified a number of determinants of breast cancer screening in older women. For the most part, these determinants point to the primary care physician as the key to breast cancer screening in these women. Therefore, the primary care physician must be informed of, and encouraged to follow, the recommendations for periodic breast cancer screening in older women.


Following translation by single ribosomes one codon at a time.
We have followed individual ribosomes as they translate single messenger RNA hairpins tethered by the ends to optical tweezers. Here we reveal that translation occurs through successive translocation--and-pause cycles. The distribution of pause lengths, with a median of 2.8 s, indicates that at least two rate-determining processes control each pause. Each translocation step measures three bases--one codon-and occurs in less than 0.1 s. Analysis of the times required for translocation reveals, surprisingly, that there are three substeps in each step. Pause lengths, and thus the overall rate of translation, depend on the secondary structure of the mRNA; the applied force destabilizes secondary structure and decreases pause durations, but does not affect translocation times. Translocation and RNA unwinding are strictly coupled ribosomal functions.


Unconventional cytokine profiles and development of T cell memory in long-term survivors after cancer vaccination.
Cancer vaccine trials frequently report on immunological responses, without any clinical benefit. This paradox may reflect the challenge of discriminating between effective and pointless immune responses and sparse knowledge on their long-term development. Here, we have analyzed T cell responses in long-term survivors after peptide vaccination. There were three main study aims: (1) to characterize the immune response in patients with a possible clinical benefit. (2) To analyze the long-term development of responses and effects of booster vaccination. (3) To investigate whether the Th1/Th2-delineation applies to cancer vaccine responses. T cell clones were generated from all nine patients studied. We find that surviving patients harbor durable tumor-specific responses against vaccine antigens from telomerase, RAS or TGFbeta receptor II. Analyses of consecutive samples suggest that booster vaccination is required to induce robust T cell memory. The responses exhibit several features of possible clinical advantage, including combined T-helper and cytotoxic functionality, recognition of naturally processed antigens and diverse HLA-restriction and fine-specificity. CD4(-)CD8(-) T cell clones display unconventional cytotoxicity and specifically kill tumor cells expressing mutated TGFbeta receptor II. Cytokine profiling on the long-term survivors demonstrates high IFN gamma/IL10-ratios, favoring immunity over tolerance, and secretion of multiple chemokines likely to mobilize the innate and adaptive immune system. Interestingly, these pro-inflammatory cytokine profiles do not follow a Th1/Th2-delineation. Most IFN gamma(high)/IL4(low)/IL10(low) cultures include high concentrations of hallmark Th2-cytokines IL-5 and IL-13. This does not reflect a mixture of Th1- and Th2-clones, but applies to 19/20 T cell clones confirmed to be monoclonal through TCR clonotype mapping. The present study identifies several factors that may promote clinical efficacy and suggests that cytokine profiling should not rely on the Th1/Th2-paradigm, but assess the overall inflammatory milieu and the balance between key cytokines.


Automatic detection of small bowel contraction frequencies in motility plots using lomb-scargle periodogram and sinus-fitting method--initial experience.
Contraction frequencies are an important parameter for the analysis of bowel motility in MRI. The contraction curve can be rather noisy and the frequency-evaluation might be difficult. The aim was to evaluate manual calculations of small bowel contraction frequency in comparison with automatic calculations using two mathematically established methods. The institutional-review-board approved study consisted of 48 segmental small-bowel motility-plots out of a previous study with 25 patients (15 men/10 women; mean 39 years, standard deviation ± 14.67) undergoing MRI (1.5 T, GE-Medical Systems; two-dimensional-Fiesta, pulse repetition time 2.91, echo time 1.25, flip-angle 45°, matrix 256 × 256, slice thickness 10 mm) were evaluated. Calculations of contraction frequency was performed either manually or using Lomb-Scargle-periodograms and Sinus-Fitting method. The results were compared using intraclass correlation coefficient, Friedman's-test and Wilcoxon-matched-paired-signed-rank-test (P < 0.05 considered statistically significant). Mean contraction frequency was 5.69 (standard deviation ± 1.99) for manual calculations showing moderate interreader reliability (intraclass correlation coefficient 0.72; 95% confidence-interval: 0.59;0.82). No significant difference (P = 0.776) was found to Lomb-Scargle-periodograms (5.34 contractions-per-minute [cpm], standard deviation ± 2.47; P = 0.174) and to the sinus-fitting-method (5.47 cpm; standard deviation ± 2.57; P = 0.0779). For the latter calculations failed in 3/48 plots, while manual and Lomb-Scargle-periodograms could be performed successfully in all 48/48 plots. Herein, we could show that Lomb-Scargle-periodograms and Sinus-Fitting are fast and reliable methods to automatically measure small-bowel contraction frequencies even in noisy small bowel motility plots.


Thermally stable and uniform DNA amplification with picosecond laser ablated graphene rapid thermal cycling device.
Rapid thermal cycling (RTC) in an on-chip device can perform DNA amplification in vitro through precise thermal control at each step of the polymerase chain reaction (PCR). This study reports a straightforward fabrication technique for patterning an on-chip graphene-based device with hole arrays, in which the mechanism of surface structures can achieve stable and uniform thermal control for the amplification of DNA fragments. A thin-film based PCR device was fabricated using picosecond laser (PS-laser) ablation of the multilayer graphene (MLG). Under the optimal fluence of 4.72 J/cm2 with a pulse overlap of 66%, the MLG can be patterned with arrays of 250 μm2 hole surface structures. A 354-bp DNA fragment of VP1, an effective marker for diagnosing the BK virus, was amplified on an on-chip device in less than 60 min. A thin-film electrode with the aforementioned MLG as the heater was demonstrated to significantly enhance temperature stability for each stage of the thermal cycle. The temperature control of the heater was performed by means of a developed programmable PCR apparatus. Our results demonstrated that the proposed integration of a graphene-based device and a laser-pulse ablation process to form a thin-film PCR device has cost benefits in a small-volume reagent and holds great promise for practical medical use of DNA amplification.


DNA demethylases: a new epigenetic frontier in drug discovery.
DNA methylation is one of the most extensively studied, and one of the most stable, of all epigenetic modifications. Two drugs that target DNA methyltransferase enzymes are licensed for clinical use in oncology but relatively little attention has focused on the enzymatic pathways by which DNA methylation can be reversed. Recent breakthroughs have identified at least two classes of enzymes that can achieve functional reversal. This review discusses the significance of DNA demethylation in a range of human diseases, the candidate proteins that mediate the demethylation and the opportunities and challenges in targeting these candidates to develop new therapeutics.


Diverse laboratory colonies of Aedes aegypti harbor the same adult midgut bacterial microbiome.
Host-associated microbes, collectively known as the microbiota, play an important role in the biology of multicellular organisms. In mosquito vectors of human pathogens, the gut bacterial microbiota influences vectorial capacity and has become the subject of intense study. In laboratory studies of vector biology, genetic effects are often inferred from differences between geographically and genetically diverse colonies of mosquitoes that are reared in the same insectary. It is unclear, however, to what extent genetic effects can be confounded by uncontrolled differences in the microbiota composition among mosquito colonies. To address this question, we used 16S metagenomics to compare the midgut bacterial microbiome of six laboratory colonies of Aedes aegypti recently derived from wild populations representing the geographical range and genetic diversity of the species. We found that the diversity, abundance, and community structure of the midgut bacterial microbiome was remarkably similar among the six different colonies of Ae. aegypti, regardless of their geographical origin. We also confirmed the relatively low complexity of bacterial communities inhabiting the mosquito midgut. Our finding that geographically diverse colonies of Ae. aegypti reared in the same insectary harbor a similar gut bacterial microbiome supports the conclusion that the gut microbiota of adult mosquitoes is environmentally determined regardless of the host genotype. Thus, uncontrolled differences in microbiota composition are unlikely to represent a significant confounding factor in genetic studies of vector biology.


The tuning of human visual cortex to variations in the 1/fα amplitude spectra and fractal properties of synthetic noise images.
Natural scenes share a consistent distribution of energy across spatial frequencies (SF) known as the 1/fα amplitude spectrum (α≈0.8-1.5, mean 1.2). This distribution is scale-invariant, which is a fractal characteristic of natural scenes with statistically similar structure at different spatial scales. While the sensitivity of the visual system to the 1/f properties of natural scenes has been studied extensively using psychophysics, relatively little is known about the tuning of cortical responses to these properties. Here, we use fMRI and retinotopic mapping techniques to measure and analyze BOLD responses in early visual cortex (V1, V2, and V3) to synthetic noise images that vary in their 1/fα amplitude spectra (α=0.25 to 2.25, step size: 0.50) and contrast levels (10% and 30%) (Experiment 1). To compare the dependence of the BOLD response between the photometric (intensity based) and geometric (fractal) properties of our stimuli, in Experiment 2 we compared grayscale noise images to their binary (thresholded) counterparts, which contain only black and white regions. In both experiments, early visual cortex responded maximally to stimuli generated to have an input 1/f slope corresponding to natural 1/fα amplitude spectra, and lower BOLD responses were found for steeper or shallower 1/f slopes (peak modulation: 0.59% for 1.25 vs. 0.31% for 2.25). To control for changing receptive field sizes, responses were also analyzed across multiple eccentricity bands in cortical surface space. For most eccentricity bands, BOLD responses were maximal for natural 1/fα amplitude spectra, but importantly there was no difference in the BOLD response to grayscale stimuli and their corresponding thresholded counterparts. Since the thresholding of an image changes its measured 1/f slope (α) but not its fractal characteristics, this suggests that neuronal responses in early visual cortex are not strictly driven by spectral slope values (photometric properties) but rather their embedded geometric, fractal-like scaling properties.