A proposed architecture and method of operation for improving the protection of privacy and confidentiality in disease registers.
Disease registers aim to collect information about all instances of a disease or condition in a defined population of individuals. Traditionally methods of operating disease registers have required that notifications of cases be identified by unique identifiers such as social security number or national identification number, or by ensembles of non-unique identifying data items, such as name, sex and date of birth. However, growing concern over the privacy and confidentiality aspects of disease registers may hinder their future operation. Technical solutions to these legitimate concerns are needed. An alternative method of operation is proposed which involves splitting the personal identifiers from the medical details at the source of notification, and separately encrypting each part using asymmetrical (public key) cryptographic methods. The identifying information is sent to a single Population Register, and the medical details to the relevant disease register. The Population Register uses probabilistic record linkage to assign a unique personal identification (UPI) number to each person notified to it, although not necessarily everyone in the entire population. This UPI is shared only with a single trusted third party whose sole function is to translate between this UPI and separate series of personal identification numbers which are specific to each disease register. The system proposed would significantly improve the protection of privacy and confidentiality, while still allowing the efficient linkage of records between disease registers, under the control and supervision of the trusted third party and independent ethics committees. The proposed architecture could accommodate genetic databases and tissue banks as well as a wide range of other health and social data collections. It is important that proposals such as this are subject to widespread scrutiny by information security experts, researchers and interested members of the general public, alike.


Pupil diameter reflects uncertainty in attentional selection during visual search.
Pupil diameter has long been used as a metric of cognitive processing. However, recent advances suggest that the cognitive sources of change in pupil size may reflect LC-NE function and the calculation of unexpected uncertainty in decision processes (Aston-Jones and Cohen, 2005; Yu and Dayan, 2005). In the current experiments, we explored the role of uncertainty in attentional selection on task-evoked changes in pupil diameter during visual search. We found that task-evoked changes in pupil diameter were related to uncertainty during attentional selection as measured by reaction time (RT) and performance accuracy (Experiments 1-2). Control analyses demonstrated that the results are unlikely to be due to error monitoring or response uncertainty. Our results suggest that pupil diameter can be used as an implicit metric of uncertainty in ongoing attentional selection requiring effortful control processes.


Metagenomic insights into strategies of aerobic and anaerobic carbon and nitrogen transformation in boreal lakes.
Thousands of net-heterotrophic and strongly stratifying lakes dominate the boreal landscape. Besides their central role as emitters of greenhouse gases, we have only recently begun to understand the microbial systems driving the metabolic processes and elemental cycles in these lakes. Using shotgun metagenomics, we show that the functional potential differs among lake types, with humic lakes being particularly enriched in carbon degradation genes. Most of the metabolic pathways exhibit oxygen- and temperature-dependent stratification over depth, coinciding with shifts in bacterial community composition, implying that stratification is a major factor controlling lake metabolism. In the bottom waters, rare and poorly characterized taxa, such as ε-Proteobacteria, but also autotrophs, such as photolithotrophic Chlorobia were abundant. These oxygen-depleted layers exhibited high genetic potential for mineralization, but also for fixation of carbon and nitrogen, and genetic markers for both methane production and oxidation were present. Our study provides a first glimpse of the genetic versatility of freshwater anoxic zones, and demonstrates the potential for complete turnover of carbon compounds within the water column.


Variation in personality and behavioural plasticity across four populations of the great tit Parus major.
1. Interest in the evolutionary origin and maintenance of individual behavioural variation and behavioural plasticity has increased in recent years. 2. Consistent individual behavioural differences imply limited behavioural plasticity, but the proximate causes and wider consequences of this potential constraint remain poorly understood. To date, few attempts have been made to explore whether individual variation in behavioural plasticity exists, either within or between populations. 3. We assayed 'exploration behaviour' among wild-caught individual great tits Parus major when exposed to a novel environment room in four populations across Europe. We quantified levels of individual variation within and between populations in average behaviour, and in behavioural plasticity with respect to (i) repeated exposure to the room (test sequence), (ii) the time of year in which the assays were conducted and (iii) the interval between successive tests, all of which indicate habituation to novelty and are therefore of functional significance. 4. Consistent individual differences ('I') in behaviour were present in all populations; repeatability (range: 0.34-0.42) did not vary between populations. Exploration behaviour was also plastic, increasing with test sequence - but less so when the interval between subsequent tests was relatively large - and time of year; populations differed in the magnitude of plasticity with respect to time of year and test interval. Finally, the between-individual variance in exploration behaviour increased significantly from first to repeat tests in all populations. Individuals with high initial scores showed greater increases in exploration score than individuals with low initial scores; individual by environment interaction ('I × E') with respect to test sequence did not vary between populations. 5. Our findings imply that individual variation in both average level of behaviour and behavioural plasticity may generally characterize wild great tit populations and may largely be shaped by mechanisms acting within populations. Experimental approaches are now needed to confirm that individual differences in behavioural plasticity (habituation) - not other hidden biological factors - caused the observed patterns of I × E. Establishing the evolutionary causes and consequences of this variation in habituation to novelty constitutes an exciting future challenge.


Experimental study of the effect of IL-6 on cancer cachexia.
Several cytokines, including IL-1, TNF, LIF and IL-6 have recently been proposed as cachexia inducers. We experimentally examined the participation of cytokines, particularly, IL-6, in cancer cachexia using the human digestive cancer cell lines MKN 28, MKN 45, MKN 74, Kato-III, OCUM-2M (gastric cancer), SW1990, Panc-1 (pancreatic cancer), and OCUG (gallbladder cancer). A high level of IL-6 was detected in the OCUG culture medium. Nude mice bearing OCUG tumor had reduced body weight even when the tumor was relatively small. Loss of both muscle and adipose tissue, anemia, hypoglycemia, and a high serum level of human IL-6 were observed in these mice. However, body weight recovered rapidly to the level of that of nontumor-bearing mice after resection of OCUG tumor. Antihuman IL-6 but not anti-murine IL-6 receptor antibodies significantly suppressed the development of cachexia as measured by various indicators of cachexia including loss of both muscle and adipose tissue, anemia and hypoglycemia, as well as weight loss. These results suggest that OCUG-bearing mice exhibited cancer cachexia mediated by IL-6, and that of OCUG cell line might be useful as a human digestive cancer cachexia model.


Serum perfluoroalkyl substances in children exposed to the world trade center disaster.
The World Trade Center (WTC) disaster released large amounts of various chemical substances into the environment, including perfluoroalkyl substances (PFASs). Yet, no studies have examined exposures in children living or attending schools near the disaster site. We measured serum PFASs in WTC Health Registry (WTCHR) respondents who were ≤8 years of age on September 11, 2001 and a sociodemographically-matched comparison group. We also examined the relationship of PFASs levels with dust cloud exposure; home dust exposure, and with traumatic exposure, the latter to take into account differences related to possible mental health consequences and associated behavioral problems. Serum samples, collected between 2014 and 2016, were analyzed from 123 WTCHR participants and from 185 participants in the comparison group. In the WTCHR group, median perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) levels were 1.81ng/mL and 3.72ng/mL, respectively. Controlling for sex, caloric intake, race/ethnicity, and date of birth, significant increases among WTCHR participants compared with the matched comparison group were detected for perfluorohexanesulfonate (0.23ng/mL increase or 0.24log unit increase, p=0.006); PFOS (0.86ng/mL increase or 0.16log unit increase, p=0.011); PFOA (0.35ng/mL increase or 0.18log unit increase, p<0.001); perfluorononanoic acid (0.12ng/mL increase or 0.17log unit increase, p=0.003); perfluorodecanoic acid (0.06ng/mL increase or 0.42log unit increase, p<0.001); and perfluoroundecanoic acid (0.03ng/mL increase or 0.32log unit increase, p=0.019). Stronger associations were identified for home dust exposures and traumatic exposures than dust cloud. These findings highlight the importance of conducting longitudinal studies in this population to assess possible cardiometabolic and renal consequences related to these exposures.


Quasinormal-Mode Perturbation Theory for Dissipative and Dispersive Optomechanics.
Despite the several novel features arising from the dissipative optomechanical coupling, such effect remains vastly unexplored due to the lack of a simple formalism that captures non-Hermiticity in the engineering of optomechanical systems. In this Letter, we show that quasinormal-mode-based perturbation theory is capable of correctly predicting both dispersive and dissipative optomechanical couplings. We validate our model through simulations and also by comparison with experimental results reported in the literature. Finally, we apply this formalism to plasmonic systems, used for molecular optomechanics, where strong dissipative coupling signatures in the amplification of vibrational modes could be observed.


Implementation of Pharmacogenetic Testing Within the Veterans Health Administration From 2011 to 2013.
We assessed implementation of precision medicine within the Veterans Health Administration. We analyzed the use of interleukin-28B (IL28B) pharmacogenetic test, which predicts interferon-α treatment response in patients with hepatitis C. Patients with favorable CC genotype exhibit a two-fold higher response than patients with less favorable genotypes (CT and TT). Linking IL28B tests to Veterans Health Administration clinical data, we analyzed test use, concordance with guidelines, subsequent interferon-α treatment, and site variations. From January 2011 until December 2013, 3,529 Veterans underwent IL28B testing. There were 2,988 (85%) tests linked to electronic health record data. Demographics in those with clinical data: 97% male, mean age 59 years (SD 6.8), race: White 50%, Black 43%, Hispanic 3%, and other 3%. In Whites, the favorable CC genotype was three-fold more prevalent (541, 36%) than in Blacks (151, 12%). Analysis of IL28B timing revealed 2,373 (79%) Veterans were appropriately tested before interferon-α, 49 (2%) tested concurrent with interferon-α, and 566 (19%) tested post interferon-α treatment. Of the 630 treatment-naïve Veterans with CC genotype, only 144 (23%) initiated interferon-α treatment post-testing. Although 35% of IL28B tests overall did not influence care, the majority of tests were guideline concordant and clinically useful. IL28B testing varied substantially by site and state.


Efficient Generation and Selection of Virtual Populations in Quantitative Systems Pharmacology Models.
Quantitative systems pharmacology models mechanistically describe a biological system and the effect of drug treatment on system behavior. Because these models rarely are identifiable from the available data, the uncertainty in physiological parameters may be sampled to create alternative parameterizations of the model, sometimes termed "virtual patients." In order to reproduce the statistics of a clinical population, virtual patients are often weighted to form a virtual population that reflects the baseline characteristics of the clinical cohort. Here we introduce a novel technique to efficiently generate virtual patients and, from this ensemble, demonstrate how to select a virtual population that matches the observed data without the need for weighting. This approach improves confidence in model predictions by mitigating the risk that spurious virtual patients become overrepresented in virtual populations.


Investigation into the effect of hepatitis B virus on apoliprotein A1 expression and its mechanism.
Hepatitis B virus (HBV) infection poses a serious threat to human health, with China being one of the highly affected countries. However, the pathogenesis of chronic hepatitis B (CHB) is still unclear. Apolipoprotein A1 (ApoA1) which represents the major protein component of high-density lipoprotein is normally secreted by hepatocytes. When hepatocytes are infected with HBV may lead to the disruption of ApoA1 secretion. In this study, we investigated the effect of HBV on ApoA1 expression and preliminarily explored its molecular mechanism of regulation for revealing the pathogenesis of CHB. The expression of mRNA and protein of ApoA1 in Human HepG2 hepatoblastoma cells and subline HepG2.2.15 cells were performed by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blot. The serum ApoA1, by the immune turbidimetric test, and high-density lipoprotein cholesterol (HDL-C) in CHB patients and healthy controls, based on the enzymatic method, were measured with autobiochemical analyzer. The statistical difference was analyzed by SPSS 13.0. HBV infectious clone, pHBV1.3, and ApoA1 gene promoter were co-transfected into HepG2, and the luciferase activity was determined. The changes of ApoA1 mRNA and protein expression were detected by RT-PCR and Western-blot method, after HepG2 cells were transfected with pHBV1.3. The expression of ApoA1 mRNA and protein in HepG2.2.15 were lower than those in HepG2, and when compared with healthy controls, serum levels of ApoA1 and HDL-C in CHB patients were lower (P < 0.05). pHBV1.3 in HepG2 cells restrained the activity of ApoA1 promoter, mRNA and protein expression. HBV could inhibit the expression of ApoA1 in vitro and in vivo.


Identification of vigilance lapses using EEG/EOG by expert human raters.
It is critically important for certain occupational groups to remain highly alert throughout their working day. For safety reasons, it would be useful to automatically detect lapses in performance using EEG/EOG. Automating the detection process could be simplified considerably if we could mimic human experts. Surprisingly, it is unclear to what extent human EEG raters are able to detect lapses. Consequently, we undertook a study in which 4 expert EEG raters assessed the level of alertness of 10 air traffic controllers by observing a combination of their EEG and EOG while they performed a 10 min psychomotor vigilance task (PVT). They were specifically required to identify lapses or sleep episodes that might lead to a lapse in PVT performance. A reaction time .. 500 ms was defined as a PVT lapse. There was a total of 101 lapses (mean duration = 1.00 s). Of these, only 6 lapses were detected by one or more raters and all of these were marked as ;sleep'. Overall the human expert raters were unable to reliably identify lapses based only on EEG and EOG. This poor performance suggests an automated system would need to identify subtle features not overtly visible in the EEG.


Physical interactions among Mcm proteins and effects of Mcm dosage on DNA replication in Saccharomyces cerevisiae.
Mcm2, Mcm3, and Mcm5/Cdc46 are conserved proteins essential for the initiation of DNA synthesis at replication origins in Saccharomyces cerevisiae. The accumulation of these proteins in the nucleus before the onset of DNA synthesis suggests that they play a role in restricting DNA synthesis to once per cell cycle. In this work, we show that Mcm2, Mcm3, and Mcm5 self-interact and interact with one another to form complexes. Mcm2 and Mcm3 are abundant proteins, present in approximately 4 X 10(4) and 2 X 10(5) copies per cell, respectively. Reducing the dosage of Mcm2 by half results in diminished usage of specific replication origins. These results together suggest that a significant molar excess of Mcm proteins relative to replication origins is required for the proper initiation of all replication origins.


Symptom burden and infection occurrence among individuals with extremity lymphedema.
Currently, there is a lack of data related to differences in symptoms and infection across different types and anatomical sites of lymphedema. The objective of this study was to examine differences in symptoms and infection status among individuals with lymphedema of the upper or lower extremities. The National Lymphedema Network initiated an online survey of self-report lymphedema data from March 2006 through January 2010. Descriptive statistics, Mann-Whitney tests, and Chi-square tests were used to analyze data. 723 individuals with upper extremity lymphedema and 1114 individuals with lower extremity lymphedema completed the survey. Individuals with extremity lymphedema experienced high symptom burden and infectious complications. Compared with individuals with upper extremity lymphedema, individuals with lower extremity lymphedema experienced more frequent and more severe symptoms (p<.001), infection episodes (p<.001), and infection-related hospitalizations (p<.001). No statistically significant differences of symptom burden and infection status were identified between individuals with lower extremity primary and secondary lymphedema. Individuals with extremity lymphedema experience substantial symptom burden and infectious complications; however, those with lower extremity lymphedema have more severe symptoms and more infections than those with upper extremity lymphedema.


[Design and validation of scales to measure adolescent attitude toward eating and toward physical activity].
Different authors suggest that attitude is a mediator in behavior change, so it is a predictor of behavior practice. The main of this study was to design and to validate two scales for measure adolescent attitude toward healthy eating and adolescent attitude toward healthy physical activity. Scales were design based on a literature review. After, they were validated using an on-line Delphi Panel with eighteen experts, a pretest, and a pilot test with a sample of 188 high school students. Comprehensibility, content validity, adequacy, as well as the reliability (alpha of Cronbach test), and construct validity (exploratory factor analysis) of scales were tested. Scales validated by experts were considered appropriate in the pretest. In the pilot test, the ten-item Attitude to Eating Scale obtained α=0.72. The eight-item Attitude to Physical Activity Scale obtained α=0.86. They showed evidence of one-dimensional interpretation after factor analysis, a) all items got weights r>0.30 in first factor before rotations, b) the first factor explained a significant proportion of variance before rotations, and c) the total variance explained by the main factors extracted was greater than 50%. The Scales showed their reliability and validity. They could be employed to assess attitude to these priority intervention areas in Spanish adolescents, and to evaluate this intermediate result of health interventions and health programs.


Prostatic relapse of an undifferentiated teratoma 24 years after orchidectomy.
Non-seminomatous germ cell tumours make up about 40 % of all germ cell tumours, which in turn are the most common tumours in men aged 15-44 years. Low risk stage I non-seminomatous germ cell tumours, which are confined to the testes, are commonly treated by orchiectomy and surveillance. Up to 20 % of patients with this diagnosis relapse, usually within 1-2 years of follow up, but very rarely after more than 5 years. The most common sites of relapse are the retroperitoneal lymph nodes, the mediastinum, and the lungs. We describe a case of relapse in the prostate over 20 years after initial diagnosis, which has not been described in the literature so far. This report presents a 49-year-old white British man with relapsed testicular non-seminomatous germ cell tumour 22 years after initial treatment with orchidectomy only. He relapsed with a prostatic mass, haematospermia and back pain. His prostate specific antigen levels were within normal range. Alpha feto-protein and lactate dehydrogenase levels were elevated, and his human chorionic gonadotrophin levels were normal. A biopsy confirmed undifferentiated malignant tumour, shown immunohistochemically to be a yolk sac tumour. The patient was initially treated with bleomycin, etoposide and cisplatin chemotherapy, but developed bleomycin-related pulmonary side effects after two cycles. His treatment was changed and he completed four cycles of chemotherapy by receiving two cycles of etoposide, ifosfamide, and cisplatin. Post treatment blood tumour markers were normal, but a follow up computed tomography showed a mass in the base of the prostate, the trigone and the left distal ureter which was surgically resected. The histology from the surgical resection was of necrotic tissue. The patient is now in follow up at 3 years after treatment with no evidence of residual disease on computed tomography. His Alpha feto-protein, beta human chorionic gonadotrophin and lactate dehydrogenase levels are normal. Very late relapse in stage I non-seminomatous germ cell tumours is extremely rare and the prostate is a highly unusual site of relapsed disease. For diagnosis of late relapse, this case confirms the value of serum biomarkers in germ cell tumours, in particular non-seminomatous germ cell tumours.


Targeting osteopontin, the silent partner of Na+/H+ exchanger isoform 1 in cardiac remodeling.
Cardiac hypertrophy (CH), characterized by the enlargement of cardiomyocytes, fibrosis and apoptosis, contributes to cardiac remodeling, which if left unresolved results in heart failure. Understanding the signaling pathways underlying CH is necessary to identify potential therapeutic targets. The Na(+) /H(+) -exchanger isoform I (NHE1), a ubiquitously expressed glycoprotein and cardiac specific isoform, regulates intracellular pH. Recent studies have demonstrated that enhanced expression/activity of NHE1 contributes to cardiac remodeling and CH. Inhibition of NHE1 in both in vitro and in vivo models have suggested that inhibition of NHE1 protects against hypertrophy. However, clinical trials using NHE1 inhibitors have proven to be unsuccessful, suggesting that additional factors maybe contributing to cardiac remodeling. Recent studies have indicated that the upregulation of NHE1 is associated with enhanced levels of osteopontin (OPN) in the setting of CH. OPN has been demonstrated to be upregulated in left ventricular hypertrophy, dilated cardiomyopathy and in diabetic cardiomyopathy. The cellular interplay between OPN and NHE1 in the setting of CH remains unknown. This review focuses on the role of NHE1 and OPN in cardiac remodeling and emphasizes the signaling pathways implicating OPN in the NHE1-induced hypertrophic response.


PEGylated Chitosan Nanoparticles Encapsulating Ascorbic Acid and Oxaliplatin Exhibit Dramatic Apoptotic Effects against Breast Cancer Cells.
This study aims to design a pH-responsive dual-loaded nanosystem based on PEGylated chitosan nanoparticles loaded with ascorbic acid (AA) and oxaliplatin (OX) for the effective treatment of breast cancer. In this regard, non-PEGylated and PEGylated chitosan nanoparticles (CS NPs) loaded with either ascorbic acid (AA), oxaliplatin (OX), or dual-loaded with AA-OX were fabricated using the ionotropic gelation method. The hydrodynamic diameters of the fabricated AA/CS NPs, OX/CS NPs, and AA-OX/CS NPs were 157.20 ± 2.40, 188.10 ± 9.70, and 261.10 ± 9.19 nm, respectively. While the hydrodynamic diameters of the designed AA/PEG-CS NPs, OX/PEG-CS NPs, and AA-OX/PEG-CS NPs were 152.20 ± 2.40, 156.60 ± 4.82, and 176.00 ± 4.21 nm, respectively. The ζ-potential of the prepared nanoparticles demonstrated high positive surface charges of +22.02 ± 1.50, +22.58 ± 1.85 and +40.4 ± 2.71 mV for AA/CS NPs, OX/CS NPs, and AA-OX/CS NPs, respectively. The ζ-potential of the PEGylated CS NPs was reduced owing to the shielding of the positive charges by the PEG chains. Additionally, all the prepared nanoparticles exhibited high entrapment efficiencies (EE%) and spherical-shaped morphology. The chemical features of the prepared nanoparticles were investigated using Fourier transform infrared (FTIR) spectroscopy. Release studies showed the capability of the prepared non-PEGylated and PEGylated chitosan NPs to release their cargo in the acidic environment of cancer tissue (pH 5.5). Furthermore, the AA/CS NPs, AA/PEG-CS NPs, OX/CS NPs, OX/PEG-CS NPs, AA-OX/CS NPs and AA-OX/PEG-CS NPs exhibited remarkable cytotoxic activities against breast adenocarcinoma (MCF-7) cells with IC50 values of 44.87 ± 11.49, 23.3 ± 3.73, 23.88 ± 6.29, 17.98 ± 3.99, 18.69 ± 2.22, and 7.5 ± 0.69 µg/mL, respectively; as compared to free AA and OX (IC50 of 150.80 ± 26.50 and 147.70 ± 63.91 µg/mL, respectively). Additionally, treatment of MCF-7 cells with IC50 concentrations of AA, AA/CS NPs, AA/PEG-CS NPs, OX, OX/CS NPs, OX/PEG-CS NPs, AA-OX/CS NPs or AA-OX/PEG-CS NPs increased the percentages of early apoptotic cells to 5.28%, 9.53%, 11.20%, 5.27%, 13.80%, 8.43%, 2.32%, and 10.10%, respectively, and increased the percentages of late apoptotic cells to 0.98%, 0.37%, 2.41%, 2.06%, 0.97%, 9.66%, 56%, and 81.50%, respectively. These results clearly indicate that PEGylation enhances the apoptotic effect of AA and OX alone, in addition to potentiating the apoptotic effect of AA and OX when combined on MCF-7 cells. In conclusion, PEGylated chitosan nanoparticles encapsulating AA, OX, or AA and OX represent an effective formula for induction of apoptosis in MCF-7 cells.


[Longitudinal study of the metabolic syndrome after kidney transplantation].
The occurence of the metabolic syndrome (MS) between the renal receptors is one of the major complications after transplantation and is associated with an increased risk of graft failure and high rates of obesity and diabetes new appearance. This study aims to investigate the prevalence and risk factors associated with the development of the MS and to evaluate the association between the same with the allograft dysfunction. The samples consisted of 138 renal transplant patients, 83 men and 55 women, kidney transplant, which was attended by over five years for the transplant consultation. Were analyzed as potential risk factors for MS: age, sex, body mass index (BMI), weight, hypertension, diabetes, LDL, HDL, triglycerides in serum and immunosuppressive therapy (cyclosporine, tacrolimus, mycophenolate mofetil), was also assessed the prevalence of acute rejection episodes and renal function. The prevalence of MS was 39.85 %. As statistically significant risk factors were obtained the BMI, overweight, HDL cholesterol levels, triglycerides and LDL as well as hypertension and diabetes. There were high rates of acute rejection and differences in story to the glomerular filtration rate. There is a high prevalence of the MS that severely compromised renal function and graft survival in renal transplant patients, it is very important the control and strict monitoring of all risk factors identified.


miR-125a-5p/miR-125b-5p contributes to pathological activation of angiotensin II-AT1R in mouse distal convoluted tubule cells by the suppression of Atrap.
The renin-angiotensin system plays a crucial role in the regulation of blood pressure. Activation of the angiotensin II (Ang II)-Ang II type 1 receptor (AT1R) signaling pathway contributes to the pathogenesis of hypertension and subsequent organ damage. AT1R-associated protein (ATRAP) has been identified as an endogenous inhibitory protein of the AT1R pathological activation. We have shown that mouse Atrap (Atrap) represses various Ang II-AT1R-mediated pathologies, including hypertension in mice. The expression of human ATRAP (ATRAP)/Atrap can be altered in various pathological states in humans and mice, such as Ang II stimulation and serum starvation. However, the regulatory mechanisms of ATRAP/Atrap are not yet fully elucidated. miRNAs are 21 to 23 nucleotides of small RNAs that post-transcriptionally repress gene expression. Single miRNA can act on hundreds of target mRNAs, and numerous miRNAs have been identified as the Ang II-AT1R signaling-associated disease phenotype modulator, but nothing is known about the regulation of ATRAP/Atrap. In the present study, we identified miR-125a-5p/miR-125b-5p as the evolutionarily conserved miRNAs that potentially act on ATRAP/Atrap mRNA. Further analysis revealed that miR-125a-5p/miR-125b-5p can directly repress both ATRAP and Atrap. In addition, the inhibition of miR-125a-5p/miR-125b-5p resulted in the suppression of the Ang II-AT1R signaling in mouse distal convoluted tubule cells. Taken together, miR-125a-5p/miR-125b-5p activates Ang II-AT1R signaling by the suppression of ATRAP/Atrap. Our results provide new insights into the potential approaches for achieving the organ-protective effects by the repression of the miR-125 family associated with the enhancement of ATRAP/Atrap expression.


Mutual coherence function of a finite optical beam and application to coherent detection.
The mutual coherence function of a finite laser beam propagating through a random medium is calculated from the extended Huygens principle. Formulas are presented for the transverse coherence from an arbitrarily truncated focused Gaussian beam. Expressions are derived for a modified Kolmogorov spectrum for the refractive index fluctuations and for the case of nonuniform turbulence. The formalism is used to derive the conditions under which instantaneous reciprocity exists between a transmitter and receiver in a coherent detection system. It is also shown that a maximum useful transmitter (as well as receiver) diameter exists for computing the average normalized signal-to-noise ratio of this system. This maximum dimension is the smaller of the turbulence-induced coherence length and a Fresnel zone.


Search filters for finding prognostic and diagnostic prediction studies in Medline to enhance systematic reviews.
The interest in prognostic reviews is increasing, but to properly review existing evidence an accurate search filer for finding prediction research is needed. The aim of this paper was to validate and update two previously introduced search filters for finding prediction research in Medline: the Ingui filter and the Haynes Broad filter. Based on a hand search of 6 general journals in 2008 we constructed two sets of papers. Set 1 consisted of prediction research papers (n = 71), and set 2 consisted of the remaining papers (n = 1133). Both search filters were validated in two ways, using diagnostic accuracy measures as performance measures. First, we compared studies in set 1 (reference) with studies retrieved by the search strategies as applied in Medline. Second, we compared studies from 4 published systematic reviews (reference) with studies retrieved by the search filter as applied in Medline. Next--using word frequency methods--we constructed an additional search string for finding prediction research. Both search filters were good in identifying clinical prediction models: sensitivity ranged from 0.94 to 1.0 using our hand search as reference, and 0.78 to 0.89 using the systematic reviews as reference. This latter performance measure even increased to around 0.95 (range 0.90 to 0.97) when either search filter was combined with the additional string that we developed. Retrieval rate of explorative prediction research was poor, both using our hand search or our systematic review as reference, and even combined with our additional search string: sensitivity ranged from 0.44 to 0.85. Explorative prediction research is difficult to find in Medline, using any of the currently available search filters. Yet, application of either the Ingui filter or the Haynes broad filter results in a very low number missed clinical prediction model studies.


Evaluation of a cancer survivorship protocol: transitioning patients to survivors.
This article describes the implementation and evaluation of a survivorship protocol for cancer survivors to improve knowledge about their disease and decrease anxiety. The study included a convenience sample of 30 cancer survivors at an outpatient community cancer center in the southeastern United States following completion of acute oncology treatment. One month after the survivorship protocol was delivered, knowledge about diagnosis, treatments, recommended follow-up, signs of recurrence, and late side effects increased. Anxiety scores were lower one month after the intervention, and satisfaction with the protocol was high. The results demonstrated that the survivorship protocol is a feasible method of educating cancer survivors, supporting the Institute of Medicine's recommendation that strategies for delivering education to cancer survivors are important. Survivors have additional needs that must be addressed following treatment, and a survivorship protocol can provide the knowledge survivors need to participate in their own health care.


Under what conditions is it ethical to offer incentives to encourage drug-using women to use long-acting forms of contraception?
To stimulate debate by examining ethical issues raised by Project Prevention, a US-based organization that offers $US300 to addicted individuals who agree to either undergo surgical sterilization or use long-acting forms of contraception. An analysis of key ethical questions raised by Project Prevention. The important issues for debate are: (i) what are the reproductive rights of drug-using women; (ii) does a substantial cash incentive undermine the ability of addicted women to make free and informed decisions about long-term contraception; and (iii) how can we best assist addicted women to access good reproductive health care and obtain treatment for their addiction? We need more research on ways in which small non-cash incentives for reversible methods of contraception could be used in a morally acceptable and effective way to promote the sexual, reproductive and general health of addicted women.


Long-Term Outcomes of Patients with Tracheoesophageal Fistula/Esophageal Atresia: Survey Results from Tracheoesophageal Fistula/Esophageal Atresia Online Communities.
Introduction Outcome studies of tracheoesophageal fistula (TEF) and/or esophageal atresia (EA) are limited to retrospective chart reviews. This study surveyed TEF/EA patients/parents engaged in social media communities to determine long-term outcomes. Materials and Methods A 50-point survey was designed to study presentation, interventions, and ongoing symptoms after repair in patients with TEF/EA. It was validated using a test population and made available on TEF/EA online communities. Results In this study, 445 subjects completed the survey during a 2-month period. Mean age of patients when surveyed was 8.7 years (0-61 years) and 56% were male. Eighty-nine percent of surveys were completed by the parent of the patient. Sixty-two percent of patients underwent repair in the first 7 days of life. Standard open repair was most common (56%), followed by primary esophageal replacement (13%) and thoracoscopic repair (13%). Out of 405, 106 (26%) patients had postoperative leak. Postoperative leak was least likely in primary esophageal replacement (18%) and standard open repair (19%). Leak occurred in 32% of patients who had thoracoscopic repair; 31% (128/413) reported long-gap atresia, which was significantly associated with increased risk of postoperative leak (54/128, 42%) when compared with standard short-gap atresia (odds ratio, 3.5; p = 0.001). Out of 409, 221 (54%) patients reported dysphagia after repair, with only 77/221 (34.8%) reporting resolution by age 5. Out of 381, 290 (76%) patients reported symptoms of gastroesophageal reflux disease (GERD). There was no difference in dysphagia rates or GERD symptoms based on type of initial repair. Antireflux surgery was required in 63/290, 22% of patients with GERD (15% of all patients) and 27% of these patients who had surgery required more than one procedure antireflux procedure. The most common was Nissen fundoplication (73%), followed by partial wrap (14%). Reflux recurred in 32% of patients after antireflux surgery. Conclusion TEF/EA patients have long-term dysphagia and GERD that may be under reported. Retrospective studies of outcomes after TEF/EA repair may underestimate long-term esophageal dysmotility, dysphagia, GERD, and strictures that occur regardless of the type of repair and adversely affect quality of life. Fifteen percent of all TEF/EA patients surveyed required an antireflux procedure during childhood, and more than one-quarter of those required repeat surgery. These data demonstrate the need for long-term follow-up as pediatric patients transition to adult care.


Risk factors for and knowledge of Mycobacterium tuberculosis infection among drug users in substance abuse treatment.
To determine the prevalence, incidence and risk factors for Mycobacterium tuberculosis infection, as well as to assess TB knowledge and attitudes, among a group of known drug users in a city with low TB incidence (11.3 per 100,000 in 1995). Patients of an urban drug treatment facility enrolled in opioid substitution, opioid antagonist and other drug treatment programs were screened for TB, including tuberculin skin testing and standardized data collection on TB risk factors. A subsample of clients was interviewed about TB knowledge and attitudes. Between 1 June 1995 and 31 May 1996, 1055 individuals were screened. The prevalence of infection was 15.7% (CI: 13.2-18.2%). PPD positivity was associated with older age (per annum, OR = 1.08, CI: 1.05-1.11), non-white race (OR = 2.81, CI: 1.72-4.60), foreign birth (OR = 4.24, CI: 2.35-7.62) and a history of injecting drug use (OR = 1.89, CI: 1.14, 3.12). The incidence of infection was 2.9 per 100 person-years (CI: 1.8-4.7). Thirty-two per cent of 79 drug users interviewed about TB knowledge and attitudes thought TB could be prevented by bleaching or not sharing needles/syringes. Fifty-one per cent thought anyone with a positive TB skin test was contagious. M. tuberculosis infection was common in this population and associated with injecting drugs and several demographic factors. The incidence of new infection was relatively low. In this non-endemic environment, the detection and treatment of latent infection are important aspects of TB control. Misconceptions about TB transmission were also widespread in this population. Drug treatment programs can play a key role by undertaking screening programs that educate about TB and identify infected subjects who would benefit from preventive therapy.


Incorporating alignment uncertainty into Felsenstein's phylogenetic bootstrap to improve its reliability.
Most evolutionary analyses are based on pre-estimated multiple sequence alignment. Wong et al. established the existence of an uncertainty induced by multiple sequence alignment when reconstructing phylogenies. They were able to show that in many cases different aligners produce different phylogenies, with no simple objective criterion sufficient to distinguish among these alternatives. We demonstrate that incorporating MSA induced uncertainty into bootstrap sampling can significantly increase correlation between clade correctness and its corresponding bootstrap value. Our procedure involves concatenating several alternative multiple sequence alignments of the same sequences, produced using different commonly used aligners. We then draw bootstrap replicates while favoring columns of the more unique aligner among the concatenated aligners. We named this concatenation and bootstrapping method, Weighted Partial Super Bootstrap (wpSBOOT). We show on three simulated datasets of 16, 32 and 64 tips that our method improves the predictive power of bootstrap values. We also used as a benchmark an empirical collection of 853 one to one orthologous genes from seven yeast species and found wpSBOOT to significantly improve discrimination capacity between topologically correct and incorrect trees. Bootstrap values of wpSBOOT are comparable to similar readouts estimated using a single method. However, for reduced trees by 50 and 95% bootstrap thresholds, wpSBOOT comes out the lowest Type I error (less FP). The automated generation of replicates has been implemented in the T-Coffee package, which is available as open source freeware available from www.tcoffee.org. Supplementary data are available at Bioinformatics online.


Exploring the potential of halophilic archaea for the decolorization of azo dyes.
Azo dyes are being extensively used in textile industries, so finding a proper solution to decolorize them is of high importance. In order to find azo dye decolorizing strains among haloarchaea, which are well known for their tolerance to harsh environmental conditions, fifteen haloarchaeal strains were screened. Halogeometricum sp. strain A and Haloferax sp. strain B with the highest decolorization ability (95% and 91% for Remazol black B; both about 60% for Acid blue 161, respectively) were selected for further studies. It was shown that both strains were able to grow and decolorize the dye in a medium containing up to 5 M NaCl, with optimum decolorization activity at 2.5-3.4 M, pH 7, and a wide temperature range between 30 to 45 °C. Moreover, both strains were able to tolerate and decolorize up to 1,000 mg l-1 Remazol black B. Also, they were able to survive in 5,000 mg l-1 of the dye after 20 days' incubation. Glucose and yeast extract were found to be the best carbon and nitrogen sources in the decolorization medium for both strains. This is the first report studying decolorization of azo dyes using halophilic archaea.


Neoangiogenesis-related genes are hallmarks of fast-growing hepatocellular carcinomas and worst survival. Results from a prospective study.
The biological heterogeneity of hepatocellular carcinoma (HCC) makes prognosis difficult. We translate the results of a genome-wide high-throughput analysis into a tool that accurately predicts at presentation tumour growth and survival of patients with HCC. Ultrasound surveillance identified HCC in 78 (training set) and 54 (validation set) consecutive patients with cirrhosis. Patients underwent two CT scans 6 weeks apart (no treatment in-between) to determine tumour volumes (V0 and V1) and calculate HCC doubling time. Baseline-paired HCC and surrounding tissue biopsies for microarray study (Agilent Whole Human Genome Oligo Microarrays) were also obtained. Predictors of survival were assessed by multivariate Cox model. Calculated tumour doubling times ranged from 30 to 621 days (mean, 107±91 days; median, 83 days) and were divided into quartiles: ≤53 days (n=19), 54-82 days (n=20), 83-110 days (n=20) and ≥111 days (n=19). Median survival according to doubling time was significantly lower for the first quartile versus the others (11 vs 41 months, 42, and 47 months, respectively) (p<0.0001). A five-gene transcriptomic hepatic signature including angiopoietin-2 (ANGPT2), delta-like ligand 4 (DLL4), neuropilin (NRP)/tolloid (TLL)-like 2 (NETO2), endothelial cell-specific molecule-1 (ESM1), and nuclear receptor subfamily 4, group A, member 1 (NR4A1) was found to accurately identify rapidly growing HCCs of the first quartile (ROC AUC: 0.961; 95% CI 0.919 to 1.000; p<0.0001) and to be an independent factor for mortality (HR: 3.987; 95% CI 1.941 to 8.193, p<0.0001). The hepatic five-gene signature was able to predict HCC growth in individual patient and the consequent risk of death. This implies a role of this molecular tool in the future therapeutic management of patients with HCC. ClinicalTrials.gov Identifier: NCT01657695.


Bone marrow-derived mesenchymal stem cells ameliorate chronic high glucose-induced β-cell injury through modulation of autophagy.
Chronic hyperglycemia causes a progressive decrease of β-cell function and mass in type 2 diabetic patients. Growing evidence suggests that augment of autophagy may be an effective approach to protect β cells against various extra-/intracellular stimuli. In this study, we thus investigated whether bone marrow-derived mesenchymal stem cells (BM-MSCs) could ameliorate chronic high glucose (HG)-induced β-cell injury through modulation of autophagy. Prolonged exposure to HG decreased cell viability, increased cell apoptosis and impaired basal insulin secretion and glucose-stimulated insulin secretion of INS-1 cells, but BM-MSC treatment significantly alleviated these glucotoxic alternations. In addition, western blotting displayed upregulated expression of Beclin1 and LC3-II in INS-1 cells co-cultured with BM-MSCs. Results from immunofluorescence staining and transmission electronic microscope analysis also revealed that BM-MSCs promoted autophagosomes and autolysosomes formation in HG-treated INS-1 cells. However, it should be noted that inhibition of autophagy significantly diminished the protective effects of BM-MSCs on HG-treated INS-1 cells, suggesting that the improvement of β-cell function and survival induced by BM-MSCs was mediated through autophagy. Furthermore, our results showed that BM-MSCs improved mitochondrial function and reduced reactive oxygen species production in HG-treated INS-1 cells, largely owing to autophagic clearance of impaired mitochondria. In vivo study was performed in rats with type 2 diabetes (T2D). BM-MSC infusion not only ameliorated hyperglycemia, but also promoted restoration of pancreatic β cells in T2D rats. Meanwhile, BM-MSC infusion upregulated LAMP2 expression and enhanced formation of autophagosomes and autolysosomes, combined with reduced β-cell apoptosis and increased number of insulin granules. These findings together indicated that BM-MSCs could protect β cells against chronic HG-induced injury through modulation of autophagy in vitro and in vivo. This study unveiled novel evidence of BM-MSCs as an ideal strategy to enhance autophagy for treatment of T2D mellitus.


Naturally-Acquired Immune Response against Plasmodium vivax Rhoptry-Associated Membrane Antigen.
Rhoptry-associated membrane antigen (RAMA) is an abundant glycophosphatidylinositol (GPI)-anchored protein that is embedded within the lipid bilayer and is implicated in parasite invasion. Antibody responses against rhoptry proteins are produced by individuals living in a malaria-endemic area, suggesting the immunogenicity of Plasmodium vivax RAMA (PvRAMA) for induction of immune responses during P. vivax infection. To determine whether PvRAMA contributes to the acquisition of immunity to malaria and could be a rational candidate for a vaccine, the presence of memory T cells and the stability of the antibody response against PvRAMA were evaluated in P. vivax-exposed individuals. The immunogenicity of PvRAMA for the induction of T cell responses was evaluated by in vitro stimulation of peripheral blood mononuclear cells (PBMCs). High levels of interferon (IFN)-γ and interleukin (IL)-10 cytokines were detected in the culture supernatant of PBMCs, and the CD4+ T cells predominantly produced IL-10 cytokine. The levels of total anti-PvRAMA immunoglobulin G (IgG) antibody were significantly elevated, and these antibodies persisted over the 12 months of the study. Interestingly, IgG1, IgG2 and IgG3 were the major antibody subtypes in the response to PvRAMA. The frequency of IgG3 in specific to PvRAMA antigen maintained over 12 months. These data could explain the immunogenicity of PvRAMA antigen in induction of both cell-mediated and antibody-mediated immunity in natural P. vivax infection, in which IFN-γ helps antibody class switching toward the IgG1, IgG2 and IgG3 isotypes and IL-10 supports PvRAMA-specific antibody production.


The discriminative ability of FRAX, the WHO algorithm, to identify women with prevalent asymptomatic vertebral fractures: a cross-sectional study.
A Moroccan model for the FRAX tool to determine the absolute risk of osteoporotic fracture at 10 years has been established recently. The study aimed to assess the discriminative capacity of FRAX in identifying women with prevalent asymptomatic vertebral fractures (VFs). We enrolled in this cross-sectional study 908 post-menopausal women with a mean age of 60.9 years ± 7.7 (50 to 91) with no prior known diagnosis of osteoporosis. Subjects were recruited from asymptomatic women selected from the general population. Lateral VFA images and scans of the lumbar spine and proximal femur were obtained using a GE Healthcare Lunar Prodigy densitometer. VFs were defined using a combination of Genantsemiquantitative (SQ) approach and morphometry. We calculated the absolute risk of major fracture and hip fracture with and without bone mineral density (BMD)using the FRAX website.The overall discriminative value of the different risk scores was assessed by calculating the areas under the ROC curve (AUC). VFA images showed that 179 of the participants (19.7%) had at least one grade 2/3 VF. The group of women with VFs had a statistically significant higher FRAX scores for major and hip fractures with and without BMD, and lower weight, height, and lumbar spine and hip BMD and T-scores than those without a VFA-identified VF. The AUC ROC of FRAX for major fracture without BMD was 0.757 (CI 95%; 0.718-0.797) and 0.736 (CI 95%; 0.695-0.777) with BMD, being 0.756 (CI 95%; 0.716-0.796) and 0.747 (CI 95%; 0.709-0.785), respectively for FRAX hip fracture without and with BMD. The AUC ROC of lumbar spine T-score and femoral neck T-score were 0.660 (CI 95%; 0.611-0.708) and 0.707 (CI 95%; 0.664-0.751) respectively. In asymptomatic post-menopausal women, the FRAX risk for major fracture without BMD had a better discriminative capacity in identifying the women with prevalent VFs than lumbar spine and femoral neck T-scores suggesting its usefulness in identifying women in whom VFA could be indicated.


Practical Synthesis and Properties of 2,5-Diarylarsoles.
2,5-Diarylarsoles were easily synthesized from nonvolatile arsenic precursors. Diiodoarsine was generated in situ and reacted with titanacyclopentadienes to give 2,5-diarylarsoles. The structures and optical properties were studied in comparison with those of 2,5-diarylphosphole. It was found that the arsoles were much more stable in the air than the phosphole. Single crystal X-ray diffraction revealed the arsenic atoms adopted a trigonal pyramidal structure, reflecting on the s-character of the lone pair. The obtained 2,5-diarylarsoles and 2,5-diarylphosphole showed intense emission in solutions and solid state. In addition, the optical properties were controlled by transition-metal coordination.


Preoperative sCD40L levels predict risk of atrial fibrillation after off-pump coronary artery bypass graft surgery.
The risk of atrial fibrillation (AF) after coronary bypass surgery has been related to redox state, inflammation, and ischemia. Platelet activation is common to all of these pathways. We investigated the relation between AF and preoperative soluble CD40 ligand (sCD40L), a proinflammatory marker released by activated platelets. Furthermore, we studied the role of inflammation, endothelial function, and redox state in this relation. sCD40L levels were measured in 144 patients in sinus rhythm the day before off-pump coronary artery surgery. Systemic inflammation was assessed from levels of C-reactive protein and soluble intercellular adhesion molecule-1, and endothelial function was assessed from the brachial artery flow-mediated dilatation response. Graft samples were collected during surgery to assess vascular redox state. AF occurred in 33% of patients after surgery, with 3% still in AF after 6 weeks. Preoperative sCD40L levels were significantly higher in those who developed in-hospital AF (odds ratio for a 1-SD increase in log[sCD40L]=1.97; 95% CI, 1.21 to 3.22; P=0.007; after adjustment for age, sex, Euroscore, and total duration of operation). sCD40L and vascular superoxide levels were higher in patients still in AF at 6 weeks, and endothelial function was lower, although the small number of events precluded statistical analysis in this group. Systemic endothelial function, redox state, and preoperative markers of systemic inflammation were not associated with in-hospital postoperative AF. Preoperative platelet activation, as assessed by sCD40L levels, is a novel predictor of postoperative AF, independent of systemic endothelial function, vascular redox state, and systemic inflammation.


Genetic Diversity and Virulence Determinants of Escherichia coli Strains Isolated from Patients with Crohn's Disease in Spain and Chile.
Adherent-invasive Escherichia coli (AIEC) strains are genetically variable and virulence factors for AIEC are non-specific. FimH is the most studied pathogenicity-related protein, and there have been few studies on other proteins, such as Serine Protease Autotransporters of Enterobacteriacea (SPATEs). The goal of this study is to characterize E. coli strains isolated from patients with Crohn's disease (CD) in Chile and Spain, and identify genetic differences between strains associated with virulence markers and clonality. We characterized virulence factors and genetic variability by pulse field electrophoresis (PFGE) in 50 E. coli strains isolated from Chilean and Spanish patients with CD, and also determined which of these strains presented an AIEC phenotype. Twenty-six E. coli strains from control patients were also included. PFGE patterns were heterogeneous and we also observed a highly diverse profile of virulence genes among all E. coli strains obtained from patients with CD, including those strains defined as AIEC. Two iron transporter genes chuA, and irp2, were detected in various combinations in 68-84% of CD strains. We found that the most significant individual E. coli genetic marker associated with CD E. coli strains was chuA. In addition, patho-adaptative fimH mutations were absent in some of the highly adherent and invasive strains. The fimH adhesin, the iron transporter irp2, and Class-2 SPATEs did not show a significant association with CD strains. The V27A fimH mutation was detected in the most CD strains. This study highlights the genetic variability of E. coli CD strains from two distinct geographic origins, most of them affiliated with the B2 or D E. coli phylogroups and also reveals that nearly 40% of Chilean and Spanish CD patients are colonized with E.coli with a characteristic AIEC phenotype.


Sensitive and accurate identification of protein-DNA binding events in ChIP-chip assays using higher order derivative analysis.
Immuno-precipitation of protein-DNA complexes followed by microarray hybridization is a powerful and cost-effective technology for discovering protein-DNA binding events at the genome scale. It is still an unresolved challenge to comprehensively, accurately and sensitively extract binding event information from the produced data. We have developed a novel strategy composed of an information-preserving signal-smoothing procedure, higher order derivative analysis and application of the principle of maximum entropy to address this challenge. Importantly, our method does not require any input parameters to be specified by the user. Using genome-scale binding data of two Escherichia coli global transcription regulators for which a relatively large number of experimentally supported sites are known, we show that ∼90% of known sites were resolved to within four probes, or ∼88 bp. Over half of the sites were resolved to within two probes, or ∼38 bp. Furthermore, we demonstrate that our strategy delivers significant quantitative and qualitative performance gains over available methods. Such accurate and sensitive binding site resolution has important consequences for accurately reconstructing transcriptional regulatory networks, for motif discovery, for furthering our understanding of local and non-local factors in protein-DNA interactions and for extending the usefulness horizon of the ChIP-chip platform.


NCHS urban-rural classification scheme for counties.
This report details the National Center for Health Statistics' (NCHS) development of the 2006 NCHS Urban-Rural Classification Scheme for Counties and provides some examples of how the scheme can be used to describe differences in health measures by urbanization level. The 2006 NCHS urban-rural classification scheme classifies all U.S. counties and county-equivalents into six levels--four for metropolitan counties and two for nonmetropolitan counties. The Office of Management and Budget's delineation of metropolitan and nonmetropolitan counties forms the foundation of the scheme. The NCHS scheme also uses the cut points of the U.S. Department of Agriculture Rural-Urban Continuum Codes to subdivide the metropolitan counties based on the population of their metropolitan statistical area (MSA): large, for MSA population of 1 million or more; medium, for MSA population of 250,000-999,999; and small, for MSA population below 250,000. Large metro counties were further separated into large central and large fringe metro categories using classification rules developed by NCHS. Nonmetropolitan counties were assigned to two levels based on the Office of Management and Budget's designated micropolitan or noncore status. The 2006 scheme was applied to data from the National Vital Statistics System (NVSS) and the National Health Interview Survey (NHIS) to illustrate its ability to capture health differences by urbanization level. Application of the 2006 NCHS scheme to NVSS and NHIS data shows that it identifies important health disparities among communities, most notably those for inner city and suburban communities. The design of the NCHS Urban-Rural Classification Scheme for Counties makes it particularly well-suited for assessing and monitoring health differences across the full urbanization continuum.


Interactions between interlimb and intralimb coordination during the performance of bimanual multijoint movements.
The simultaneous performance of movements involving different effectors gives rise to neural and biomechanical interactions between and within limbs. The present study addressed the role of interlimb and intralimb constraints during the control of bimanual multijoint movements. Thirteen participants performed eight tasks involving the bilateral elbows and wrists under different coordination conditions. With respect to interlimb coordination, coordination patterns referred to the in-phase and anti-phase coordination modes, involving the simultaneous timing of homologous versus non-homologous muscles, respectively. With respect to inter-segmental (intralimb) coordination, the isodirectional mode referred to simultaneous flexions and extensions in the ipsilateral wrist and elbow joints, whereas the non-isodirectional mode involved simultaneous flexion in one joint together with extension in the other joint, or vice versa. The analysis of the data focused upon measures of relative phasing between proximal and distal joints within a limb as well as between the homologous joints of both limbs. With respect to interlimb coordination, findings revealed that adoption of the in-phase mode resulted in a higher quality of interlimb coordination than the anti-phase mode. However, the mode adopted in the distal joints had a larger impact on the quality of interlimb coordination than the mode adopted in the proximal joints. More specifically, in-phase coordination of the distal joints had a positive, and anti-phase coordination a negative, influence on the global coordinative behavior of the system. Minor effects of intralimb coordination modes on interlimb coordination were observed. With respect to intralimb coordination between the ipsilateral elbow and wrist, the isodirectional mode was performed with higher stability than the non-isodirectional mode. The mode of interlimb coordination also affected the quality of intralimb coordination, such that generating anti-phase coordination patterns in the distal joints had a negative influence on the accuracy and stability of intralimb coordination. Taken together, the present findings suggest a hierarchical structure whereby interlimb coordination constraints have a stronger impact on the global coordinative behavior of the system than intralimb coordination constraints. Moreover, the global coordinative state of the system is more affected by the coordination between the distal than between the proximal joints. Overall, the findings suggest that the mirror-image symmetry constraint has a powerful influence on bimanual multijoint coordination.


Role of microRNA in metabolic shift during heart failure.
Heart failure (HF) is an end point resulting from a number of disease states. The prognosis for HF patients is poor with survival rates precipitously low. Energy metabolism is centrally linked to the development of HF, and it involves the proteomic remodeling of numerous pathways, many of which are targeted to the mitochondrion. microRNAs (miRNA) are noncoding RNAs that influence posttranscriptional gene regulation. miRNA have garnered considerable attention for their ability to orchestrate changes to the transcriptome, and ultimately the proteome, during HF. Recently, interest in the role played by miRNA in the regulation of energy metabolism at the mitochondrion has emerged. Cardiac proteome remodeling during HF includes axes impacting hypertrophy, oxidative stress, calcium homeostasis, and metabolic fuel transition. Although it is established that the pathological environment of hypoxia and hemodynamic stress significantly contribute to the HF phenotype, it remains unclear as to the mechanistic underpinnings driving proteome remodeling. The aim of this review is to present evidence highlighting the role played by miRNA in these processes as a means for linking pathological stimuli with proteomic alteration. The differential expression of proteins of substrate transport, glycolysis, β-oxidation, ketone metabolism, the citric acid cycle (CAC), and the electron transport chain (ETC) are paralleled by the differential expression of miRNA species that modulate these processes. Identification of miRNAs that translocate to cardiomyocyte mitochondria (miR-181c, miR-378) influencing the expression of the mitochondrial genome-encoded transcripts as well as suggested import modulators are discussed. Current insights, applications, and challenges of miRNA-based therapeutics are also described.


Quantitation of human genomic DNA through amplification of the amelogenin locus.
An alternate method for quantitation of human genomic DNA is presented. Quantitative template amplification technology (abbreviated "Q-TAT") estimates the quantity of human DNA present in an extract by comparing fluorescence in X and Y amplicons produced from unknowns with fluorescence in a standard curve amplified from known quantities of reference DNA. Q-TAT utilizes PCR and electrophoresis with fluorescent detection/quantitation, precluding the need for new instrumentation, methodology, or quality assurance associated with slot-blot or real-time PCR. In a comparison study incorporating shared samples, Q-TAT was found to be more sensitive than widely used slot-blot methods but somewhat less sensitive than real-time PCR. Among samples containing DNA concentrations ranging from 100 pg/microL to 2-4 ng/microL, Q-TAT produced DNA concentration estimates that agreed reasonably well with either Quantiblot or real-time PCR. Q-TAT was reproducible with a typical coincidence of variation of about 35%. Quantitation of human DNA in this study involved summing fluorescence in X and Y amplicons in unknowns and quantitation standards. However, analyzing fluorescence in X and Y amplicons individually could allow estimates of male and female DNA present in mixtures to be made. Moreover, since X and Y amplicons exhibit sizes of 210 and 216 bp, respectively, the integrity as well as the concentration of the genomic DNA template can be assessed. Q-TAT represents an alternate method useful for the quantitation of human genomic DNA prior to amplification of STR loci used for identity testing purposes. The method uses existing equipment and procedures in conjunction with a well-characterized DNA standard to produce concentration estimates for unknowns that reliably produce STR profiles suitable for analysis.


Post-translational modification of the deubiquitinating enzyme otubain 1 modulates active RhoA levels and susceptibility to Yersinia invasion.
Microbial pathogens exploit the ubiquitin system to facilitate infection and manipulate the immune responses of the host. In this study, susceptibility to Yersinia enterocolitica and Yersinia pseudotuberculosis invasion was found to be increased upon overexpression of the deubiquitinating enzyme otubain 1 (OTUB1), a member of the ovarian tumour domain-containing protein family. Conversely, OTUB1 knockdown interfered with Yersinia invasion in HEK293T cells as well as in primary monocytes. This effect was attributed to a modulation of bacterial uptake. We demonstrate that the Yersinia-encoded virulence factor YpkA (YopO) kinase interacts with a post-translationally modified form of OTUB1 that contains multiple phosphorylation sites. OTUB1, YpkA and the small GTPase ras homologue gene family member A (RhoA) were found to be part of the same protein complex, suggesting that RhoA levels are modulated by OTUB1. Our results show that OTUB1 is able to stabilize active RhoA prior to invasion, which is concomitant with an increase in bacterial uptake. This effect is modulated by post-translational modifications of OTUB1, suggesting a new entry point for manipulating Yersinia interactions with the host.


Formation of Atomically Flat Silver Films on GaAs with a "Silver Mean" Quasi Periodicity
A flat epitaxial silver film on a gallium arsenide [GaAs(110)] surface was synthesized in a two-step process. Deposition of a critical thickness of silver at low temperature led to the formation of a dense nanocluster film. Upon annealing, all atoms rearranged themselves into an atomically flat film. This silver film has a close-packed (111) structure modulated by a "silver mean" quasi-periodic sequence. The ability to grow such epitaxial overlayers of metals on semiconductors enables the testing of theoretical models and provides a connection between metal and semiconductor technologies.


Central serotonin agonist actions of LY 165163, 1-(m-trifluoromethylphenyl)-4-(p-aminophenylethyl) piperazine, in rats.
1-(m-Trifluoromethylphenyl)-4-(p-aminophenylethyl)piperazine (LY 156163), reported previously to have selective affinity for the 5-HT1A subtype of serotonin receptor in vitro, was studied at doses of 1.25 to 20 mg/kg i.p. in rats to determine if it had properties characteristic of centrally acting serotonin agonists. LY 165163 decreased whole brain concentrations of the serotonin metabolite, 5-hydroxyindoleacetic acid, but not of serotonin itself, decreased the rate of accumulation of 5-hydroxyindoleacetic acid after probenecid administration to block its efflux from brain, decreased the rate of decline in serotonin concentration after inhibition of serotonin synthesis with alpha-propyldopacetamide and decreased the accumulation of 5-hydroxytryptophan after decarboxylase inhibition by m-hydroxybenzylhydrazine. LY 165163 also decreased 5-hydroxyindoleacetic acid concentrations in two specific brain regions, striatum and hypothalamus. Serum concentrations of corticosterone and prolactin were increased by doses of LY 165163 that reduced serotonin turnover. These effects are all consistent with evidence from other studies that LY 165163 is a centrally acting serotonin agonist. LY 165163 also increased the concentrations of two dopamine metabolites, 3,4-dihydroxyphenylacetic acid and homovanillic acid, measured in whole brain as well as in striatum and hypothalamus, but did not alter dopamine concentration. The accumulation of dopa after decarboxylase inhibition was accelerated by LY 165163. The increases in hormone concentrations in serum and of dopamine metabolite concentrations in brain were not antagonized by pretreatment with metergoline, a serotonin antagonist. The mechanisms of those effects require further study.


Response evaluation in mesothelioma: Beyond RECIST.
Malignant pleural mesothelioma (MPM) typically demonstrates a non-spherical growth pattern, so it is often difficult to accurately categorize change in tumour burden using size-based tumour response criteria (e.g., WHO (World Health Organisation), RECIST (Response Evaluation Criteria in Solid Tumours) and modified RECIST). Functional imaging techniques are applied to derive quantitative measurements of tumours, which reflect particular aspects of the tumour pathophysiology. By quantifying how these measurements change with treatment, it is possible to observe treatment effects. In this review, we survey the existing roles of CT and MRI for the management of MPM, including the currently applied size measurement criteria for the assessment of treatment response. New functional imaging techniques, such as positron emission tomography (PET), diffusion-weighted MRI (DWI) and dynamic contrast-enhanced MRI (DCE-MRI) that may potentially improve the assessment of treatment response will be highlighted and discussed.


Acute respiratory distress syndrome: new definition, current and future therapeutic options.
Since acute respiratory distress syndrome (ARDS) was first described in 1967 there has been large number of studies addressing its pathogenesis and therapies. Despite this intense research activity, there are very few effective therapies for ARDS other than the use of lung protection strategies. This lack of therapeutic modalities is not only related to the complex pathogenesis of this syndrome but also the insensitive and nonspecific diagnostic criteria to diagnose ARDS. This review article will summarize the key features of the new definition of ARDS, and provide a brief overview of innovative therapeutic options that are being assessed in the management of ARDS.


The value of pathogen information in treating clinical mastitis.
The objective of this study was to determine the economic value of obtaining timely and more accurate clinical mastitis (CM) test results for optimal treatment of cows. Typically CM is first identified when the farmer observes recognisable outward signs. Further information of whether the pathogen causing CM is Gram-positive, Gram-negative or other (including no growth) can be determined by using on-farm culture methods. The most detailed level of information for mastitis diagnostics is obtainable by sending milk samples for culture to an external laboratory. Knowing the exact pathogen permits the treatment method to be specifically targeted to the causation pathogen, resulting in less discarded milk. The disadvantages are the additional waiting time to receive test results, which delays treating cows, and the cost of the culture test. Net returns per year (NR) for various levels of information were estimated using a dynamic programming model. The Value of Information (VOI) was then calculated as the difference in NR using a specific level of information as compared to more detailed information on the CM causative agent. The highest VOI was observed where the farmer assumed the pathogen causing CM was the one with the highest incidence in the herd and no pathogen specific CM information was obtained. The VOI of pathogen specific information, compared with non-optimal treatment of Staphylococcus aureus where recurrence and spread occurred due to lack of treatment efficacy, was $20.43 when the same incorrect treatment was applied to recurrent cases, and $30.52 when recurrent cases were assumed to be the next highest incidence pathogen and treated accordingly. This indicates that negative consequences associated with choosing the wrong CM treatment can make additional information cost-effective if pathogen identification is assessed at the generic information level and if the pathogen can spread to other cows if not treated appropriately.


Concurrent repeatability and reproducibility analyses of four marker placement protocols for the foot-ankle complex.
Multi-segment models of the foot have been proposed in the past years to overcome limitations imposed by oversimplified traditional approaches used to describe foot kinematics, but they have been only partially validated and never compared. This paper presents a unique comparative assessment of the four most widely adopted foot kinematic models and aims to provide a guidance for the clinical interpretation of their results. Sensitivity of the models to differences between treadmill and overground walking was tested in nine young healthy adults using a 1D paired t-test. Repeatability was assessed by investigating the joint kinematics obtained when the same operator placed the markers on thirteen young healthy adults in two occasions. Reproducibility was then assessed using data from three randomly selected participants, asking three operators to repeat the marker placement three times. The analyses were performed on sagittal kinematics using curve similarity and correlation indices (Linear Fit Method) and absolute differences between selected points. Differences between treadmill and overground gait were highlighted by all the investigated models. The two most repeatable and reproducible investigated models had average correlations higher than 0.70, with the lowest values (0.56) obtained for the midfoot. Averaged correlations were always higher than 0.74 for the former and 0.70 for the latter, with the lowest obtained for the midfoot (0.64 and 0.51). For all investigated models, foot kinematics generally showed low repeatability: normative bands must be adopted with caution when used for comparison with patient data.


Neuronal nitric oxide synthase contributes to the regulation of hematopoiesis.
Nitric oxide (NO) signaling is important for the regulation of hematopoiesis. However, the role of individual NO synthase (NOS) isoforms is unclear. Our results indicate that the neuronal NOS isoform (nNOS) regulates hematopoiesis in vitro and in vivo. nNOS is expressed in adult bone marrow and fetal liver and is enriched in stromal cells. There is a strong correlation between expression of nNOS in a panel of stromal cell lines established from bone marrow and fetal liver and the ability of these cell lines to support hematopoietic stem cells; furthermore, NO donor can further increase this ability. The number of colonies generated in vitro from the bone marrow and spleen of nNOS-null mutants is increased relative to wild-type or inducible- or endothelial NOS knockout mice. These results describe a new role for nNOS beyond its action in the brain and muscle and suggest a model where nNOS, expressed in stromal cells, produces NO which acts as a paracrine regulator of hematopoietic stem cells.


Economic evaluations of implemented vaccination programmes: key methodological challenges in retrospective analyses.
Post-implementation evaluation should play an important role in assessing the success of public health programmes; however, the value for money achieved by vaccine programmes after introduction has received relatively little attention to date. In this article we explore the methodological challenges in these analyses and offer direction for future evaluations in the area. We identify alternative approaches to addressing these challenges, which include the estimation of disease changes attributable to vaccination efforts, the hypothetical no vaccination comparator scenario and the full benefit achieved by implemented vaccination programmes. We also outline other important considerations such as the evolution of prices over time. Further work needs to be done to explore these issues and to determine how the application of different approaches may impact on the results of evaluations in various circumstances. As retrospective analyses are likely to become more frequent and influential, it is important that both the benefits and the limitations of post-implementation evaluations are recognised and understood. We argue that it would be useful to establish a methodological framework to provide standards and guidance on how to undertake such analyses in the future.


Large-scale gene discovery in the pea aphid Acyrthosiphon pisum (Hemiptera).
Aphids are the leading pests in agricultural crops. A large-scale sequencing of 40,904 ESTs from the pea aphid Acyrthosiphon pisum was carried out to define a catalog of 12,082 unique transcripts. A strong AT bias was found, indicating a compositional shift between Drosophila melanogaster and A. pisum. An in silico profiling analysis characterized 135 transcripts specific to pea-aphid tissues (relating to bacteriocytes and parthenogenetic embryos). This project is the first to address the genetics of the Hemiptera and of a hemimetabolous insect.


Retention in an antiretroviral therapy programme during an era of decreasing drug cost in Limbe, Cameroon.
In 2002, Cameroon initiated scale up of antiretroviral therapy (ART); on 1 October 2004, a substantial reduction in ART cost occurred. We assessed the impact of this event and other factors on enrolment and retention in care among HIV-infected patients initiating ART from February 2002 to December 2005 at the single ART clinic serving the Southwest Region in Limbe, Cameroon. We retrospectively analyzed clinical and pharmacy payment records of HIV-infected patients initiating ART according to national guidelines. We compared two cohorts of patients, enrolled before and after 1 October 2004, to determine if price reduction was associated with enhanced enrolment. We assessed factors associated with retention and survival by Cox proportional hazards models. Retention in care implied patients who had contact with the healthcare system as of 31 December 2005 (including those who were transferred to continue care in other ART centres), although these patients may have interrupted therapy at some time. A patient who was not retained in care may have dropped out (lost to follow up) or died. Mean enrolment rates for 2920 patients who initiated ART before and after the price reduction were 46.5 and 95.5 persons/month, respectively (p < 0.001). The probabilities of remaining alive and in care were 0.66 (95% CI 0.64-0.68) at six months, 0.58 (95% CI 0.56-0.60) at one year, 0.47 (95% CI 0.45-0.49) at two years and 0.35 (95% CI 0.32-0.38) at three years; they were not significantly different between the two cohorts of patients enrolled before and after the price reduction over the first 15 months of comparable follow up (hazard ratio 1.1; 95% CI 0.9-1.2, p = 0.27). In multivariable analysis using multiple imputations to compensate for missing values, factors associated with dropping out of care or dying were male gender (HR 1.33 [1.18-1.50], p = 0.003), treatment paid by self, family or partly by other (HR 3.05 [1.99-4.67], p < 0.001), and, compared with residents of Limbe, living more than 150 km from Limbe (HR 1.41 [1.18-1.69], p < 0.001), or being residents of Douala (HR 1.51 [1.16-1.98], p < 0.001). Reducing the cost of ART increased enrolment of clients in the programme, but did not change retention in care. In a system where most clients pay for ART, an accessible clinic location may be more important than the cost of medication for retention in care. Decentralizing ART clinics might improve retention and survival among patients on ART.


Functional Thin Films on Surfaces.
The motion of a thin viscous film of fluid on a curved surface exhibits many intricate visual phenomena, which are challenging to simulate using existing techniques. A possible alternative is to use a reduced model, involving only the temporal evolution of the mass density of the film on the surface. However, in this model, the motion is governed by a fourth-order nonlinear PDE, which involves geometric quantities such as the curvature of the underlying surface, and is therefore difficult to discretize. Inspired by a recent variational formulation for this problem on smooth surfaces, we present a corresponding model for triangle meshes. We provide a discretization for the curvature and advection operators which leads to an efficient and stable numerical scheme, requires a single sparse linear solve per time step, and exactly preserves the total volume of the fluid. We validate our method by qualitatively comparing to known results from the literature, and demonstrate various intricate effects achievable by our method, such as droplet formation, evaporation, droplets interaction and viscous fingering. Finally, we extend our method to incorporate non-linear van der Waals forcing terms which stabilize the motion of the film and allow additional effects such as pearling.


GSTM1 Deletion Exaggerates Kidney Injury in Experimental Mouse Models and Confers the Protective Effect of Cruciferous Vegetables in Mice and Humans.
GSTM1 encodes glutathione S-transferase μ-1 (GSTM1), which belongs to a superfamily of phase 2 antioxidant enzymes. The highly prevalent GSTM1 deletion variant is associated with kidney disease progression in human cohorts: the African American Study of Kidney Disease and Hypertension and the Atherosclerosis Risk in Communities (ARIC) Study. We generated a Gstm1 knockout mouse line to study its role in a CKD model (involving subtotal nephrectomy) and a hypertension model (induced by angiotensin II). We examined the effect of intake of cruciferous vegetables and GSTM1 genotypes on kidney disease in mice as well as in human ARIC study participants. We also examined the importance of superoxide in the mediating pathways and of hematopoietic GSTM1 on renal inflammation. Gstm1 knockout mice displayed increased oxidative stress, kidney injury, and inflammation in both models. The central mechanism for kidney injury is likely mediated by oxidative stress, because treatment with Tempol, an superoxide dismutase mimetic, rescued kidney injury in knockout mice without lowering BP. Bone marrow crosstransplantation revealed that Gstm1 deletion in the parenchyma, and not in bone marrow-derived cells, drives renal inflammation. Furthermore, supplementation with cruciferous broccoli powder rich in the precursor to antioxidant-activating sulforaphane significantly ameliorated kidney injury in Gstm1 knockout, but not wild-type mice. Similarly, among humans (ARIC study participants), high consumption of cruciferous vegetables was associated with fewer kidney failure events compared with low consumption, but this association was observed primarily in participants homozygous for the GSTM1 deletion variant. Our data support a role for the GSTM1 enzyme in the modulation of oxidative stress, inflammation, and protective metabolites in CKD.


Effect of organic fouling on micro-pollutant rejection in membrane bioreactor treating municipal solid waste landfill leachate.
Effect of membrane fouling on the removal of micro-pollutants from municipal solid waste landfill leachate, i.e. 4-methyl-2,6-di-tert-butylphenol (BHT), bisphenol A (BPA), and bis(2-ethylhexyl)phthalate (DEHP), in membrane bioreactor (MBR) was investigated. Modifications of membrane surface properties were analyzed to determine their relationship with their removals. Membrane fouling was simulated with foulants of different particle sizes on cellulose acetate (CA) microfiltration membrane to investigate the effect of foulant characteristics on BHT, BPA, and DEHP retention in the filtration experiment. The rejection efficiencies of the organic micro-pollutants in the MBR were 82-97% by fouled membrane, and 70-90% by cleaned membrane. The fouled membrane provided higher rejection of micro-pollutants from about 5% for BPA and BHT to 19% for DEHP. These improvements were due to the modification of membrane surface characteristics in terms of surface morphology, and contact angle after membrane fouling. The degree of rejection was found to be dependent upon the characteristics of foulant deposited on CA membrane surface. Increasing foulant particle size and its density shifted the mechanism of micro-pollutant rejection from membrane pore narrowing to pore blocking and cake formation while increasing pollutant adsorption capacity onto the foulant layer.


Meta-Analysis of NOS3 G894T Polymorphisms with Air Pollution on the Risk of Ischemic Heart Disease Worldwide.
The purpose of this updated meta-analysis was to investigate the effect of nitric oxide synthase-3 (NOS3) G894T polymorphisms, air pollution and their interaction on ischemic heart disease (IHD) risk across populations worldwide. Recursive partition trees, nonlinear association curve fit and geographic information system maps were incorporated to verify results of conventional pooled analyses for sources of heterogeneity. Results from 61 studies (16,219 cases, 12,222 controls) revealed a significant increased relative risk (RR) of IHD associated with NOS3 894 polymorphisms TT (RR = 1.44) and GT (RR = 1.37). Subgroup analysis revealed that the TT polymorphism genotype had significantly increased risk of IHD in Caucasian, East Asian, South Asian, and Middle Eastern populations (all p < 0.05). It is important to point out that many countries demonstrated an average risk of greater than two, which identifies the NOS3 894 TT polymorphism as a potential causal factor and biological marker of IHD, based on criteria for strong evidence used in international consensus panels. These 10 countries include Ukraine, the United Kingdom, Brazil, Chile, Japan, South Korea, India, Iran, Egypt and Morocco. For these countries with elevated risk (RR > 2) from the NOS3 894 TT polymorphism, meta-predictive analysis demonstrated an increasing trend in air pollution association with increased NOS3 894 polymorphisms. Further studies are needed to explore the complexity of the associations among NOS3 gene polymorphisms per population stratifications within countries, detailed air pollution data for added specificity for geographic location across time, and disease risk.


The knockdown of c-myc expression by RNAi inhibits cell proliferation in human colon cancer HT-29 cells in vitro and in vivo.
We investigated the effects of RNA interference-mediated silencing of the c-myc gene on celluar proliferation and apoptosis in human colon cancer HT-29 cells in vitro and in vivo. A small interfering RNA (siRNA) targeting c-myc was designed, the DNA template was synthesized, and the siRNA was obtained by in vitro transcription. After siRNA transfection into HT-29 and human neuroblastoma IMR-32 cells with Lipofectamine 2000, the proliferation of the HT-29 and IMR-32 cells was assessed via 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) colorimetry, and Hoechst 33258 staining was used to observe cell apoptosis. Following gene transfer to HT-29 cells, the expression of c-myc mRNA was examined via reverse transcription polymerase chain reaction, and the level of the protein via Western blot assay. Growth curves were constructed and in vivo experiments were performed on nude mice to assess the effects of c-myc silencing on tumor growth. The c-myc expression in the tumor tissue was measured by reverse transcription polymerase chain reaction and subsequently by immunohistochemistry. Our paper demonstrates that the delivery of siRNA directed against c-myc not only efficiently down-regulated the expression of c-myc, inhibited the proliferation of HT-29 cells and induced apoptosis in vitro, but also suppressed the growth of colon cancer cells in vivo.


Biology of upper-body and lower-body adipose tissue--link to whole-body phenotypes.
The distribution of adipose tissue in the body has wide-ranging and reproducible associations with health and disease. Accumulation of adipose tissue in the upper body (abdominal obesity) is associated with the development of cardiovascular disease, insulin resistance, type 2 diabetes mellitus and even all-cause mortality. Conversely, accumulation of fat in the lower body (gluteofemoral obesity) shows opposite associations with cardiovascular disease and type 2 diabetes mellitus when adjusted for overall fat mass. The abdominal depots are characterized by rapid uptake of predominantly diet-derived fat and a high lipid turnover that is easily stimulated by adrenergic receptor activation. The lower-body fat stores have a reduced lipid turnover with a capacity to accommodate fat undergoing redistribution. Lower-body adipose tissue also seems to retain the capacity to recruit additional adipocytes as a result of weight gain and demonstrates fewer signs of inflammatory insult. New data suggest that the profound functional differences between the upper-body and lower-body tissues are controlled by site-specific sets of developmental genes, such as HOXA6, HOXA5, HOXA3, IRX2 and TBX5 in subcutaneous abdominal adipose tissue and HOTAIR, SHOX2 and HOXC11 in gluteofemoral adipose tissue, which are under epigenetic control. This Review discusses the developmental and functional differences between upper-body and lower-body fat depots and provides mechanistic insight into the disease-protective effects of lower-body fat.


Novel, Synergistic Antifungal Combinations that Target Translation Fidelity.
There is an unmet need for new antifungal or fungicide treatments, as resistance to existing treatments grows. Combination treatments help to combat resistance. Here we develop a novel, effective target for combination antifungal therapy. Different aminoglycoside antibiotics combined with different sulphate-transport inhibitors produced strong, synergistic growth-inhibition of several fungi. Combinations decreased the respective MICs by ≥8-fold. Synergy was suppressed in yeast mutants resistant to effects of sulphate-mimetics (like chromate or molybdate) on sulphate transport. By different mechanisms, aminoglycosides and inhibition of sulphate transport cause errors in mRNA translation. The mistranslation rate was stimulated up to 10-fold when the agents were used in combination, consistent with this being the mode of synergistic action. A range of undesirable fungi were susceptible to synergistic inhibition by the combinations, including the human pathogens Candida albicans, C. glabrata and Cryptococcus neoformans, the food spoilage organism Zygosaccharomyces bailii and the phytopathogens Rhizoctonia solani and Zymoseptoria tritici. There was some specificity as certain fungi were unaffected. There was no synergy against bacterial or mammalian cells. The results indicate that translation fidelity is a promising new target for combinatorial treatment of undesirable fungi, the combinations requiring substantially decreased doses of active components compared to each agent alone.


Ets2 determines the inflammatory state of endothelial cells in advanced atherosclerotic lesions.
Neovascularization is required for embryonic development and plays a central role in diseases in adults. In atherosclerosis, the role of neovascularization remains to be elucidated. In a genome-wide microarray-screen of Flk1+ angioblasts during murine embryogenesis, the v-ets erythroblastosis virus E26 oncogene homolog 2 (Ets2) transcription factor was identified as a potential angiogenic factor. We assessed the role of Ets2 in endothelial cells during atherosclerotic lesion progression toward plaque instability. In 91 patients treated for carotid artery disease, Ets2 levels showed modest correlations with capillary growth, thrombogenicity, and rising levels of tumor necrosis factor-α (TNFα), monocyte chemoattractant protein 1, and interleukin-6 in the atherosclerotic lesions. Experiments in ApoE(-/-) mice, using a vulnerable plaque model, showed that Ets2 expression was increased under atherogenic conditions and was augmented specifically in the vulnerable versus stable lesions. In endothelial cell cultures, Ets2 expression and activation was responsive to the atherogenic cytokine TNFα. In the murine vulnerable plaque model, overexpression of Ets2 promoted lesion growth with neovessel formation, hemorrhaging, and plaque destabilization. In contrast, Ets2 silencing, using a lentiviral shRNA construct, promoted lesion stabilization. In vitro studies showed that Ets2 was crucial for TNFα-induced expression of monocyte chemoattractant protein 1, interleukin-6, and vascular cell adhesion molecule 1 in endothelial cells. In addition, Ets2 promoted tube formation and amplified TNFα-induced loss of vascular endothelial integrity. Evaluation in a murine retina model further validated the role of Ets2 in regulating vessel inflammation and endothelial leakage. We provide the first evidence for the plaque-destabilizing role of Ets2 in atherosclerosis development by induction of an intraplaque proinflammatory phenotype in endothelial cells.


Ancient DNA analysis of dental calculus.
Dental calculus (calcified tartar or plaque) is today widespread on modern human teeth around the world. A combination of soft starchy foods, changing acidity of the oral environment, genetic pre-disposition, and the absence of dental hygiene all lead to the build-up of microorganisms and food debris on the tooth crown, which eventually calcifies through a complex process of mineralisation. Millions of oral microbes are trapped and preserved within this mineralised matrix, including pathogens associated with the oral cavity and airways, masticated food debris, and other types of extraneous particles that enter the mouth. As a result, archaeologists and anthropologists are increasingly using ancient human dental calculus to explore broad aspects of past human diet and health. Most recently, high-throughput DNA sequencing of ancient dental calculus has provided valuable insights into the evolution of the oral microbiome and shed new light on the impacts of some of the major biocultural transitions on human health throughout history and prehistory. Here, we provide a brief historical overview of archaeological dental calculus research, and discuss the current approaches to ancient DNA sampling and sequencing. Novel applications of ancient DNA from dental calculus are discussed, highlighting the considerable scope of this new research field for evolutionary biology and modern medicine.


Ursodeoxycholic Acid Treatment Preferentially Improves Overall Survival Among African Americans With Primary Biliary Cholangitis.
We used data from the Fibrotic Liver Disease Consortium to evaluate the impact of ursodeoxycholic acid (UDCA) treatment across race/ethnicity, gender, and clinical status among patients with primary biliary cholangitis. Data were collected from "index date" (baseline) through December 31, 2016. Inverse Probability of Treatment Weighting was used to adjust for UDCA treatment selection bias. Cox regression, focusing on UDCA-by-risk factor interactions, was used to assess the association between treatment and mortality and liver transplant/death. Among 4,238 patients with primary biliary cholangitis (13% men; 8% African American, 7% Asian American/American Indian/Pacific Island [ASINPI]; 21% Hispanic), 78% had ever received UDCA. The final multivariable model for mortality retained age, household income, comorbidity score, total bilirubin, albumin, alkaline phosphatase, and interactions of UDCA with race, gender, and aspartate aminotransferase/alanine aminotransferase ≥1.1. Among untreated patients, African Americans and ASINPIs had higher mortality than whites (adjusted hazard ratio [aHR] = 1.34, 95% confidence interval [CI] 1.08-1.67 and aHR = 1.40, 95% CI 1.11-1.76, respectively). Among treated patients, this relationship was reversed (aHR = 0.67, 95% CI 0.51-0.86 and aHR = 0.88, 95% CI 0.67-1.16). Patterns were similar for liver transplant/death. UDCA reduced the risk of liver transplant/death in all patient groups and mortality across all groups except white women with aspartate aminotransferase/alanine aminotransferase ≥1.1. As compared to patients with low-normal bilirubin at baseline (≤0.4 mg/dL), those with high-normal (1.0 > 0.7) and mid-normal bilirubin (0.7 > 0.4) had significantly higher liver transplant/death and all-cause mortality. African American and ASINPI patients who did not receive UDCA had significantly higher mortality than white patients. Among African Americans, treatment was associated with significantly lower mortality. Regardless of UDCA treatment, higher baseline bilirubin, even within the normal range, was associated with increased mortality and liver transplant/death compared with low-normal levels.