Results from Australia's 2014 Report Card on Physical Activity for Children and Youth.
Like many other countries, Australia is facing an inactivity epidemic. The purpose of the Australian 2014 Physical Activity Report Card initiative was to assess the behaviors, settings, and sources of influences and strategies and investments associated with the physical activity levels of Australian children and youth. A Research Working Group (RWG) drawn from experts around Australia collaborated to determine key indicators, assess available datasets, and the metrics which should be used to inform grades for each indicator and factors to consider when weighting the data. The RWG then met to evaluate the synthesized data to assign a grade to each indicator. Overall Physical Activity Levels were assigned a grade of D-. Other physical activity behaviors were also graded as less than average (D to D-), while Organized Sport and Physical Activity Participation was assigned a grade of B-. The nation performed better for settings and sources of influence and Government Strategies and Investments (A- to a C). Four incompletes were assigned due to a lack of representative quality data. Evidence suggests that physical activity levels of Australian children remain very low, despite moderately supportive social, environmental and regulatory environments. There are clear gaps in the research which need to be filled and consistent data collection methods need to be put into place.


Study of integrated heterogeneous data reveals prognostic power of gene expression for breast cancer survival.
Studies show that thousands of genes are associated with prognosis of breast cancer. Towards utilizing available genetic data, efforts have been made to predict outcomes using gene expression data, and a number of commercial products have been developed. These products have the following shortcomings: 1) They use the Cox model for prediction. However, the RSF model has been shown to significantly outperform the Cox model. 2) Testing was not done to see if a complete set of clinical predictors could predict as well as the gene expression signatures. We address these shortcomings. The METABRIC data set concerns 1981 breast cancer tumors. Features include 21 clinical features, expression levels for 16,384 genes, and survival. We compare the survival prediction performance of the Cox model and the RSF model using the clinical data and the gene expression data to their performance using only the clinical data. We obtain significantly better results when we used both clinical data and gene expression data for 5 year, 10 year, and 15 year survival prediction. When we replace the gene expression data by PAM50 subtype, our results are significant only for 5 year and 15 year prediction. We obtain significantly better results using the RSF model over the Cox model. Finally, our results indicate that gene expression data alone may predict long-term survival. Our results indicate that we can obtain improved survival prediction using clinical data and gene expression data compared to prediction using only clinical data. We further conclude that we can obtain improved survival prediction using the RSF model instead of the Cox model. These results are significant because by incorporating more gene expression data with clinical features and using the RSF model, we could develop decision support systems that better utilize heterogeneous information to improve outcome prediction and decision making.


Alpha1-Antitrypsin Deficiency: Transition of Care for the Child With AAT Deficiency into Adulthood.
Alpha1-antitrypsin (AAT) deficiency is a common, but an underdiagnosed genetic condition, affecting 1 in 1500 individuals. It can present insidiously with liver disease in children. Although clinical practice guidelines exist for the management of AAT deficiency, especially with regards to pulmonary involvement, there are no published recommendations that specifically relate to the management of the liver disease and monitoring for lung disease associated with this condition, particularly in children. To review the literature on the management of AAT deficiency-associated liver disease in adults and children. A systematic search for articles indexed in PubMed and published was undertaken. Some earlier selected landmark references were included in the review. Search terms included: "alpha1-antitrypsin deficiency"; "liver disease"; "end-stage liver disease"; "liver transplantation" and "preventative management". Recommendations for the management of children with suspected or confirmed AAT deficiency were made according to the Strength of Recommendation Taxonomy scale. Liver complications arising from AAT deficiency result from the accumulation of mutated AAT protein within hepatocytes. Liver disease occurs in 10% of children, manifested by cholestasis, pruritus, poor feeding, hepatomegaly, and splenomegaly, but the presentation is highly variable. A diagnostic test for AAT deficiency is recommended for these children. Baseline liver function tests should be obtained to assess for liver involvement; however, the only curative treatment for AAT deficiency-associated liver disease is organ transplantation. Conclusion and Relevance: There should be a greater vigilance for AAT deficiency testing among pediatricians. Diagnosis should prompt assessment of liver involvement. Children with AATdeficiency- associated liver disease should be referred to a liver specialist and monitored throughout their lifetimes for the symptoms of AAT-deficiency-related pulmonary involvement.


Early Visual Word Processing Is Flexible: Evidence from Spatiotemporal Brain Dynamics.
Visual word recognition is often described as automatic, but the functional locus of top-down effects is still a matter of debate. Do task demands modulate how information is retrieved, or only how it is used? We used EEG/MEG recordings to assess whether, when, and how task contexts modify early retrieval of specific psycholinguistic information in occipitotemporal cortex, an area likely to contribute to early stages of visual word processing. Using a parametric approach, we analyzed the spatiotemporal response patterns of occipitotemporal cortex for orthographic, lexical, and semantic variables in three psycholinguistic tasks: silent reading, lexical decision, and semantic decision. Task modulation of word frequency and imageability effects occurred simultaneously in ventral occipitotemporal regions-in the vicinity of the putative visual word form area-around 160 msec, following task effects on orthographic typicality around 100 msec. Frequency and typicality also produced task-independent effects in anterior temporal lobe regions after 200 msec. The early task modulation for several specific psycholinguistic variables indicates that occipitotemporal areas integrate perceptual input with prior knowledge in a task-dependent manner. Still, later task-independent effects in anterior temporal lobes suggest that word recognition eventually leads to retrieval of semantic information irrespective of task demands. We conclude that even a highly overlearned visual task like word recognition should be described as flexible rather than automatic.


Attractant-induced changes and oscillations of the extracellular Ca++ concentration in suspensions of differentiating Dictyostelium cells.
We used a Ca++-sensitive electrode to measure changes in extracellular Ca++ concentration in cell suspensions of Dictyostelium discoideum during differentiation and attractant stimulation. The cells maintained an external level of 3-8 microM Ca++ until the beginning of aggregation and then started to take up Ca++. The attractants, folic acid, cyclic AMP, and cyclic GMP, induced a transient uptake of Ca++ by the cells. The response was detectable within 6 s and peaked at 30 s. Half-maximal uptake occurred at 5 nM cyclic AMP or 0.2 microM folic acid, respectively. The apparent rate of uptake amounted to 2 X 10(7) Ca++ per cell per min. Following uptake, Ca++ was released by the cells with a rate of 5 X 10(6) ions per cell per min. Specificity studies indicated that the induced uptake of Ca++ was mediated by cell surface receptors. The amount of accumulated Ca++ remained constant as long as a constant stimulus was provided. No apparent adaptation occurred. The cyclic AMP-induced uptake of Ca++ increased during differentiation and was dependent on the external Ca++ concentration. Saturation was found above 10 microM external Ca++. The time course and magnitude of the attractant-induced uptake of external Ca++ agree with a role of Ca++ during contraction. During development the extracellular Ca++ level oscillated with a period of 6-11 min. The change of the extracellular Ca++ concentration during one cycle would correspond to a 30-fold change of the cellular free Ca++ concentration.


Control of the radiative properties via photon-plasmon interaction in Er3+ -Tm3+ -codoped tellurite glasses in the near infrared region.
The novelty of this paper is that it reports on the tuning of the spectral properties of Er3+ -Tm3+ ions in tellurite glasses in the near-infrared region through the incorporation of silver or gold nanoparticles. These noble metal nanoparticles can improve the emission intensity and expand the bandwidth of the luminescence spectrum centered at 1535 nm, covering practically all the optical telecommunication bands (S, C + L and U), and extended up to 2010 nm wavelength under excitation by a 976 nm laser diode. Both effects are obtained by the combined emission of Er3+ and Tm3+ ions due to efficient energy transfer processes promoted by the presence of silver or gold nanoparticles for the (Er3+)4I(11/2)→(Tm3+)3H5, (Er3+)4I(13/2)→(Tm3+)3H4 and (Er3+)4I(13/2)→(Tm3+)3F4 transitions. The interactions between the electronic transitions of Er3+ and Tm3+ ions that provide a tunable emission are associated with the dynamic coupling mechanism described by the variations generated by the Hamiltonian H DC in either the oscillator strength or the local crystal field, i.e. the line shape changes in the near-infrared emission band. The Hamiltonian is expressed as eigenmodes associated with the density of the conduction electron generated by the different nanoparticles through its collective free oscillations at each resonance frequency of the nanoparticle and their geometric dependence. A complete description of photon-plasmon interactions of noble metal nanoparticles with the Er3+ and Tm3+ ions is provided.


Assessing quality of health services with the SERVQUAL model in Iran. A systematic review and meta-analysis.
The five-dimension service quality (SERVQUAL) scale is one of the most common tools for evaluating gaps between clients' perceptions and expectations. This study aimed to assess the quality of health services in Iran through a meta-analysis of all Iranian studies which used the SERVQUAL tool. A systematic literature review has been performed in Web of Science, PubMed, Scopus, Google Scholar, Iran Medex, Magiran and Scientific Information Database. All relevant English or Persian studies published between January 2009 and April 2016 were have been selected. Papers were considered if they regarded all five dimensions of the SERVQUAL tool for assessing the quality of health care services. Two reviewer independently extracted mean and standard deviation of five dimensions and characteristics of studies. The quality of studies included in meta-analysis using STROBE checklist. Of 315 studies initially identified, 12 were included in our meta-analysis. All analyses were performed in Stata MP v. 14. Patients' perceptions were lower than their expectations (gap = -1.64). Responsibility (-1.22) and reliability (-1.15) had the lowest gaps, and tangibility and empathy (-1.03) had the largest gaps. Except gender, other variables had no significant associations with gaps. Patients in the cities of Arak (-3.47) and Shiraz (-3.02) had the largest gaps. All dimensions of service quality were negative, which implies that the quality of health services in Iran has not been satisfying to patients and needs to be improved.


The N-terminal region of alpha-dystroglycan is an autonomous globular domain.
The structure of the N-terminal region of mouse alpha-dystroglycan (DGN) was investigated by expression of two protein fragments (residues 30-180 and 30-438) in Escherichia coli cells. Trypsin susceptibility experiments show the presence of a stable alpha-dystroglycan N-terminal region (approximately from residue 30 to 315). In addition, guanidinium hydrochloride (Gdn/HCl) denaturation of DGN-(30-438)-peptide, monitored by means of tryptophan fluorescence, produces a cooperative transition typical of folded protein structures. These results strongly suggest that the alpha-dystroglycan N-terminal is an autonomous folding unit preluding a flexible mucin-like region and that its folding is not influenced by the absence of glycosylation. In order to obtain more information on the structural features of the N-terminal domain we have also used circular dichroism, analytical sedimentation and electron microscopy analysis. Circular dichroic spectra show the absence of typical secondary structure (e.g. alpha-helix or beta-sheet) and closely resemble those recorded for loop-containing proteins. This is consistent with a sequence similarity of the alpha-dystroglycan domain with the loop-containing protein elastase. Analytical ultracentrifugation and electron microscopy analysis reveal that the N-terminal domain has a globular structure. DGN-(30-438)-peptide does not bind in the nanomolar range to an iodinated agrin fragment which binds with high affinity to tissue purified alpha-dystroglycan. No binding was detected also to laminin. This result suggests that the alpha-dystroglycan N-terminal domain does not contain the binding site to its extracellular matrix binding partners. It is less likely than the lack of glycosylation reduces its binding affinity, because the N-terminal globular domain only contains two glycosylation sites.


Characteristics and phylogeny of Bacillus cereus strains isolated from Maari, a traditional West African food condiment.
Maari is a spontaneously fermented food condiment made from baobab tree seeds in West African countries. This type of product is considered to be safe, being consumed by millions of people on a daily basis. However, due to the spontaneous nature of the fermentation the human pathogen Bacillus cereus occasionally occurs in Maari. This study characterizes succession patterns and pathogenic potential of B. cereus isolated from the raw materials (ash, water from a drilled well (DW) and potash), seed mash throughout fermentation (0-96h), after steam cooking and sun drying (final product) from two production sites of Maari. Aerobic mesophilic bacterial (AMB) counts in raw materials were of 10(5)cfu/ml in DW, and ranged between 6.5×10(3) and 1.2×10(4)cfu/g in potash, 10(9)-10(10)cfu/g in seed mash during fermentation and 10(7) - 10(9) after sun drying. Fifty three out of total 290 AMB isolates were identified as B. cereus sensu lato by use of ITS-PCR and grouped into 3 groups using PCR fingerprinting based on Escherichia coli phage-M13 primer (M13-PCR). As determined by panC gene sequencing, the isolates of B. cereus belonged to PanC types III and IV with potential for high cytotoxicity. Phylogenetic analysis of concatenated sequences of glpF, gmk, ilvD, pta, pur, pycA and tpi revealed that the M13-PCR group 1 isolates were related to B. cereus biovar anthracis CI, while the M13-PCR group 2 isolates were identical to cereulide (emetic toxin) producing B. cereus strains. The M13-PCR group 1 isolates harboured poly-γ-D-glutamic acid capsule biosynthesis genes capA, capB and capC showing 99-100% identity with the environmental B. cereus isolate 03BB108. Presence of cesB of the cereulide synthetase gene cluster was confirmed by PCR in M13-PCR group 2 isolates. The B. cereus harbouring the cap genes were found in potash, DW, cooking water and at 8h fermentation. The "emetic" type B. cereus were present in DW, the seed mash at 48-72h of fermentation and in the final product, while the remaining isolates (PanC type IV) were detected in ash, at 48-72h fermentation and in the final product. This work sheds light on the succession and pathogenic potential of B. cereus species in traditional West African food condiment and clarifies their phylogenetic relatedness to B. cereus biovar anthracis. Future implementation of GMP and HACCP and development of starter cultures for controlled Maari fermentations will help to ensure a safe product.


Glycemic Control among Pregnant Diabetic Women on Insulin Who Fasted During Ramadan.
Ramadan fasting for pregnant women with diabetes remains controversial and underreported. The objective of this study was to determine the glycemic control in pregnant diabetic women on insulin who fasted during Ramadan. This was a retrospective study carried out over a period of three years including pregnant diabetic women, who were on short-acting, intermediate-acting, or a combination of them, and opted to carry out Ramadan fasting. Glycemic control was assessed before, middle and after Ramadan fasting. Thirty seven women opted to fast with 24 (64.9%) of them had type 2 diabetes mellitus and 83.8% of them required combined insulin (short- acting, intermediate-acting) therapy. The age of the participants was 32.13±4.68 years, and the age of their pregnancies was 25.60±7.12 weeks when the study was performed. The median number of days fasted was 25 days, and most of the women were able to fast for more than 15 days. There was no difference between glycemic control of type 2 diabetes mellitus and gestational diabetes mellitus women prior to fasting. In the middle of Ramadan, serum fructosamine decreased in both groups. However, only serum HbA1c reduced in gestational diabetes mellitus after Ramadan. the findings indicate that pregnant diabetic women on insulin were able to fast during Ramadan and that their glycemic control was improved during fasting period. They may also suggest that instead of absolute ban on fasting for pregnant diabetic women more practical approach and close consultation with health care providers might be more helpful.


Factors that Impact Self-reported Wellness Scores in Elite Australian Footballers.
This study aimed to 1) identify the impact of external load variables on changes in wellness and 2) identify the impact of age, training/playing history, strength levels, and preseason loads on changes in wellness in elite Australian footballers. Data were collected from one team (45 athletes) during the 2017 season. Self-reported wellness was collected daily (4, best score possible; 28, worst score possible). External load/session availability variables were calculated using global positioning systems and session availability data from every training session and match. Additional variables included demographic data, preseason external loads, and strength/power measures. Linear mixed models were built and compared using root mean square error (RMSE) to determine the impact of variables on wellness. The external load variables explained wellness to a large degree (RMSE = 1.55, 95% confidence intervals = 1.52 to 1.57). Modeling athlete ID as a random effect appeared to have the largest impact on wellness, improving the RMSE by 1.06 points. Aside from athlete ID, the variable that had the largest (albeit negligible) impact on wellness was sprint distance covered across preseason. Every additional 2.1 km covered across preseason worsened athletes' in-season wellness scores by 1.2 points (95% confidence intervals = 0.0-2.3). The isolated impact of the individual variables on wellness was negligible. However, after accounting for the individual athlete variability, the external load variables examined collectively were able to explain wellness to a large extent. These results validate the sensitivity of wellness to monitor individual athletes' responses to the external loads imposed on them.


Isolating influential regions of electrostatic focusing in protein and DNA structure.
Electrostatic focusing is a general phenomenon that occurs in cavities and grooves on the molecular surface of biomolecules. Narrow surface features can partially shield charged atoms from the high-dielectric solvent, enhancing electrostatic potentials inside the cavity and projecting electric field lines outward into the solvent. This effect has been observed in many instances and is widely considered in the human examination of molecular structure, but it is rarely integrated into the digital representations used in protein structure comparison software. To create a computational representation of electrostatic focusing, that is compatible with structure comparison algorithms, this paper presents an approach that generates three-dimensional solids that approximate regions where focusing occurs. We verify the accuracy of this representation against instances of focusing in proteins and DNA. Noting that this representation also identifies thin focusing regions on the molecular surface that are unlikely to affect binding, we describe a second algorithm that conservatively isolates larger focusing regions. The resulting 3D solids can be compared with Boolean set operations, permitting a new range of analyses on the regions where electrostatic focusing occurs. They also represent a novel integration of molecular shape and electrostatic focusing into the same structure comparison framework.


Targeted assessment of fracture risk in women at midlife.
This study establishes a profile for women at midlife, referred for a dual energy X-ray absorptiometry (DXA), most likely to have osteoporosis, and from this, a pre-DXA screening tool has been developed. These findings inform much needed evidence-based guidelines for targeted and effective screening for osteoporosis and osteoporotic fracture prevention in women at midlife. There is no consensus as to whether women at midlife should undergo screening dual energy X-ray absorptiometry (DXA) to identify osteoporosis (T-score < -2.5). We investigated the prevalence of osteoporosis in women, aged 40-65 years, referred to 42 community-based Australian radiology centres, and identified the characteristics that best predict osteoporosis in women having a screening DXA. One thousand four hundred and two women completed the study questionnaire and had DXA reports available. After excluding women with an established indication for a DXA (58 %), users of bone-specific medication (10.5 %) and cancer (7.6 %), 466 women were classified as having a screening DXA. Forty of these women had osteoporosis at the lumbar spine (n = 32, 6.9 %) or femoral neck (n = 17, 3.6 %). Three predictors of osteoporosis (postmenopausal, nonuse of hormonal therapy and body mass index) were identified and incorporated into the Monash Osteoporosis Risk Score for women at midlife (MORS). In the screened study population, the MORS had a sensitivity of 70 % and specificity of 66 %, with a positive predictive value of 16.2 % and negative predictive value of 95.9 % for osteoporosis. Very few women referred for a screening DXA scan will be found to have osteoporosis. The MORS, a simple decision tool, would have identified 70 % of the women in our screening DXA study population and would have eliminated over 60 % of the screening DXA studies. Hence, use of the MORS may reduce unnecessary DXA scans and facilitate identification of the majority of cases of osteoporosis in women aged 40 to 65 years.


Salvage therapy with thalidomide in multiple myeloma patients relapsing after autologous peripheral blood stem cell transplantation.
The introduction of high-dose therapy with stem cell support has significantly improved the outcome of patients with multiple myeloma (MM) in terms of increased complete remission (CR) rate and extended survival, both disease-free and overall. Few options, however, are presently available for patients who relapse after single or double autologous stem cell transplantation (SCT). Thalidomide, a glutamic acid derivative with anti-angiogenetic properties, has been recently proposed as salvage treatment for such patients. The present study was aimed at evaluating thalidomide as single agent therapy for patients who had previously received autologous peripheral blood stem cell transplantation. From October 1999 to August 2000, 11 patients (7 males/4 females) who had relapsed after single (n = 4) or double (n = 7) autologous peripheral blood SCT were enrolled in the trial. Thalidomide, always employed as a single agent, was initially administered at a dose of 100 mg/day; if well tolerated, the dose was increased serially by 200 mg every other week to a maximum of 800 mg/day. The median administered dose was 600 mg/day. WHO grade > II toxic effects were constipation, lethargy, and leukopenia. Four patients (36%) showed > 50% reduction in serum M protein concentration and 4 showed > 25% reduction, for a total response rate averaging 72%. After a median follow-up of 5 months, 3 out of 8 responding patients are alive and progression-free and 5 patients have relapsed. INTERPRETATION AND CONCLUSIONS; These data confirm that thalidomide is active in poor-prognosis MM patients such as those relapsing after autologous SCT, and could thus deserve further testing in combination therapy.


Responsible Data Governance of Neuroscience Big Data.
Current discussions of the ethical aspects of big data are shaped by concerns regarding the social consequences of both the widespread adoption of machine learning and the ways in which biases in data can be replicated and perpetuated. We instead focus here on the ethical issues arising from the use of big data in international neuroscience collaborations. Neuroscience innovation relies upon neuroinformatics, large-scale data collection and analysis enabled by novel and emergent technologies. Each step of this work involves aspects of ethics, ranging from concerns for adherence to informed consent or animal protection principles and issues of data re-use at the stage of data collection, to data protection and privacy during data processing and analysis, and issues of attribution and intellectual property at the data-sharing and publication stages. Significant dilemmas and challenges with far-reaching implications are also inherent, including reconciling the ethical imperative for openness and validation with data protection compliance and considering future innovation trajectories or the potential for misuse of research results. Furthermore, these issues are subject to local interpretations within different ethical cultures applying diverse legal systems emphasising different aspects. Neuroscience big data require a concerted approach to research across boundaries, wherein ethical aspects are integrated within a transparent, dialogical data governance process. We address this by developing the concept of "responsible data governance," applying the principles of Responsible Research and Innovation (RRI) to the challenges presented by the governance of neuroscience big data in the Human Brain Project (HBP).


Long-acting oral vs injectable antipsychotic drugs in schizophrenics: a one-year double-blind comparison in multiple episode schizophrenics.
Sixty patients meeting the criteria established for schizophrenia who attained a clinical plateau following hospital discharge were randomized to receive for one year either penfluridol, 20 to 160 mg orally once each week, or fluphenazine decanoate, 0.5 to 4 ml every two weeks. The relapse rate for both treatments was low and equal. The rate of recurrence of psychosis for patients receiving penfluridol was 7% and for those receiving fluphenazine decanoate 10%. A retrospective comparison of the penfluridol group was made to a similar group of patients assigned to placebo in an earlier study. Placebo-treated patients had a relapse rate of 68%. Penfluridol patients had statistically fewer psychotic relapses. Questions about the possible carcinogenicity of penfluridol in animals will have to be resolved before it can be widely used. This study demonstrates the feasibility of using an oral, long-acting antipsychotic agent. It would be a useful psychopharmacologic addition in the treatment of outpatient schizophrenics.


Persistence of oral antidiabetic treatment for type 2 diabetes characterized by drug class, patient characteristics and severity of renal impairment: A Japanese database analysis.
To evaluate the persistence with oral antidiabetic drug (OAD) treatment characterized by drug class, patient characteristics and severity of renal impairment (RI) in patients with type 2 diabetes (T2DM) in Japan. This retrospective, observational study extracted data from a large-scale hospital database (April 2008 to September 2016). Patients with T2DM aged ≥40 years on the day of their first prescription (index date) of any OAD (biguanides [BGs], thiazolidinediones [TZDs], sulphonylureas [SUs], glinides, dipeptidyl peptidase-4 [DPP-4] inhibitors, or α-glucosidase inhibitors [α-GIs]) available between January 1, 2014 and September 30, 2016 were identified. Sodium-glucose co-transporter-2 inhibitors were not available at study initiation. Treatment persistence was assessed by Kaplan-Meier survival curves. Patients were also categorized by RI status using estimated glomerular filtration rate: ≥90 mL/min/1.73 m2 (G1); 60 to <90 mL/min/1.73 m2 (G2); 30 to <60 mL/min/1.73 m2 (G3); and <30 mL/min/1.73 m2 (G4+). We identified 206 406 index dates from 162 116 eligible patients. The largest number of index dates (91634) was observed for DPP-4 inhibitors, followed by BGs, SUs, α-GIs, glinides and TZDs. Treatment persistence was longest for DPP-4 inhibitors (median 17.0 months, 95% confidence interval [CI] 16.4-17.5) and BGs (median 17.3 months, 95% CI 16.6-18.2), and shortest for α-GIs (median 5.6 months, 95% CI 5.4-5.9) and SUs (median 4.3 months, 95% CI 4.2-4.6). Persistence was longest with DPP-4 inhibitors at all RI stages (G1-G4+), followed by BGs at stages G1/G2. The longest OAD persistence was observed for BGs and DPP-4 inhibitors at RI stages G1/G2, and for DPP-4 inhibitors at RI stages G3/G4+.


System theory and physiological processes. A physiologist looks at engineering.
Over the past centuries physiology has weathered a series of revolutions generated by the introduction of powerful tools and concepts from other disciplines, notably physics and chemistry. On each occasion such infusion of new ideas has enormously strengthened and enriched physiology. Today, the introduction of tools and concepts from engineering promises another revolution with another strengthening and enrichment. Among the things engineering has to offer are some that are close to the heart of physiology. These must be adapted and thoroughly integrated into the body of physiological concepts, attitudes, and operations. This goal presents a challenge which physiologists cannot afford to ignore. It calls for an "operation bootstrap" on the part of those of us long past our student days. It will entail a changing pattern of undergraduate preparation and graduate education for those who will become the physiologists of the coming generation. But a welcoming of the challenge and an encouragement of efforts to meet it will transform this opportunity, as others have been transformed, into another revolutionary advance in physiology.


Morphometric abnormalities in spleen and kidney of the progeny of mice fed American cranberry extract (Vaccinium macrocarpon) during pregnancy and lactation.
Cranberries and cranberry-derived diet supplements are often recommended for the treatment of urinary tract infections, also during pregnancy. These products contain strongly anti-angiogenic chemical compounds which could not be indifferent to the developing fetus. In the present work we evaluated the effect of feeding pregnant and lactating mice American cranberry extract (daily dose 0.88 mg) on the morphology and some parameters of spleen and kidney function of their adult progeny. Six weeks after delivery the morphometry of spleen and kidney, cytometric analysis of spleen lymphocytes, evaluation of humoral response to SRBC (Sheep Red Blood Cells), and examination of serum creatinine/urea concentration, were performed in the offspring. Spleens of progeny from experimental (E) group differed from the spleens of progeny of control mice in the lower number of lymphatic nodules and their larger diameter. Cytometry of spleen cells from progeny of E mothers revealed more CD19+ and CD8+ lymphocytes than in the control group. No difference was seen in the response to immunization by red blood cells of sheep (SRBC) between control and E offspring. An increase in the diameter of glomeruli was observed in the kidneys of the experimental group in comparison with the control group. No abnormalities in creatinine and urea serum level were observed. A higher concentration of VEGF and bFGF in E offspring sera in comparison to the controls was seen. Although the observed differences between the control and experimental group were not large, caution is recommended in using cranberries and their extracts during pregnancy until more research will be done on this topic.


Common variants at 7p21 are associated with frontotemporal lobar degeneration with TDP-43 inclusions.
Frontotemporal lobar degeneration (FTLD) is the second most common cause of presenile dementia. The predominant neuropathology is FTLD with TAR DNA-binding protein (TDP-43) inclusions (FTLD-TDP). FTLD-TDP is frequently familial, resulting from mutations in GRN (which encodes progranulin). We assembled an international collaboration to identify susceptibility loci for FTLD-TDP through a genome-wide association study of 515 individuals with FTLD-TDP. We found that FTLD-TDP associates with multiple SNPs mapping to a single linkage disequilibrium block on 7p21 that contains TMEM106B. Three SNPs retained genome-wide significance following Bonferroni correction (top SNP rs1990622, P = 1.08 x 10(-11); odds ratio, minor allele (C) 0.61, 95% CI 0.53-0.71). The association replicated in 89 FTLD-TDP cases (rs1990622; P = 2 x 10(-4)). TMEM106B variants may confer risk of FTLD-TDP by increasing TMEM106B expression. TMEM106B variants also contribute to genetic risk for FTLD-TDP in individuals with mutations in GRN. Our data implicate variants in TMEM106B as a strong risk factor for FTLD-TDP, suggesting an underlying pathogenic mechanism.


Direct interaction of natural and synthetic catechins with signal transducer activator of transcription 1 affects both its phosphorylation and activity.
Our previous studies showed that (-)-epigallocatechin-3-gallate (EGCG) inhibits signal transducer activator of transcription 1 (STAT1) activation. Since EGCG may be a promising lead compound for new anti-STAT1 drug design, 15 synthetic catechins, characterized by the (-)-gallocatechin-3-gallate stereochemistry, were studied in the human mammary MDA-MB-231 cell line to identify the minimal structural features that preserve the anti-STAT1 activity. We demonstrate that the presence of three hydroxyl groups of B ring and one hydroxyl group in D ring is essential to preserve their inhibitory action. Moreover, a possible molecular target of these compounds in the STAT1 pathway was investigated. Our results demonstrate a direct interaction between STAT1 protein and catechins displaying anti-STAT1 activity. In particular, surface plasmon resonance (SPR) analysis and molecular modeling indicate the presence of two putative binding sites (a and b) with different affinity. Based on docking data, site-directed mutagenesis was performed, and interaction of the most active catechins with STAT1 was studied with SPR to test whether Gln518 on site a and His568 on site b could be important for the catechin-STAT1 interaction. Data indicate that site b has higher affinity for catechins than site a as the highest affinity constant disappears in the H568A-STAT1 mutant. Furthermore, Janus kinase 2 (JAK2) kinase assay data suggest that the contemporary presence in vitro of STAT1 and catechins inhibits JAK2-elicited STAT1 phosphorylation. The very tight catechin-STAT1 interaction prevents STAT1 phosphorylation and represents a novel, specific and efficient molecular mechanism for the inhibition of STAT1 activation.


Source sector and fuel contributions to ambient PM2.5 and attributable mortality across multiple spatial scales.
Ambient fine particulate matter (PM2.5) is the world's leading environmental health risk factor. Reducing the PM2.5 disease burden requires specific strategies that target dominant sources across multiple spatial scales. We provide a contemporary and comprehensive evaluation of sector- and fuel-specific contributions to this disease burden across 21 regions, 204 countries, and 200 sub-national areas by integrating 24 global atmospheric chemistry-transport model sensitivity simulations, high-resolution satellite-derived PM2.5 exposure estimates, and disease-specific concentration response relationships. Globally, 1.05 (95% Confidence Interval: 0.74-1.36) million deaths were avoidable in 2017 by eliminating fossil-fuel combustion (27.3% of the total PM2.5 burden), with coal contributing to over half. Other dominant global sources included residential (0.74 [0.52-0.95] million deaths; 19.2%), industrial (0.45 [0.32-0.58] million deaths; 11.7%), and energy (0.39 [0.28-0.51] million deaths; 10.2%) sectors. Our results show that regions with large anthropogenic contributions generally had the highest attributable deaths, suggesting substantial health benefits from replacing traditional energy sources.


Estimating a preference-based single index from the Overactive Bladder Questionnaire.
The aim of the study is to estimate a preference-based single index for calculating quality-adjusted life years for patients with overactive bladder (OAB), based on a survey of the UK general population using the 5-dimensional health classification system OAB-5D, derived from the validated Overactive Bladder Questionnaire (OAB-q). An interview valuation survey of members of public in the South Yorkshire, UK was undertaken using the time-trade-off method. Each respondent was randomly allocated to one of 14 blocks, and valued seven states each plus the "pits" state, so that a total of 99 states were valued. A number of multivariate regression models were estimated for predicting a total of 3125 health state values defined by the classification. Models were compared and selected using a set of criteria, including overall diagnosis by adjusted R-squared, the sign and significance of individual parameter estimates, the relative size of coefficients within a given dimension and predictive ability. The mean model was recommended for use in economic evaluation. This will permit the cost-effectiveness of new interventions to be assessed in patients with OAB using existing and future OAB-q data sets.


Detection of a specific bacterial antigen in urine of rats with Bacteroides fragilis infection.
Human intra-abdominal infections frequently yield Bacteroides fragilis and require specific antimicrobial and surgical therapy. Noninvasive immunologic assessment of this organism might allow more optimum therapy. Therefore we raised antisera in goats to Bacteroides fragilis ATCC 23745 and allowed it to react with a solid-phase capsular polysaccharide-protein antigen extracted from the same organism. Preliminary work disclosed that 10 ng/ml antigen could be detected in competition assays in both saline and dialyzed rat urine. Results were manifest by diminution of bound antiglobulin alkaline phosphatase conjugate in an antigen-mediated antibody-inhibition enzyme-linked immunosorbent assay. Rats were then infected intra-abdominally with (1) B. fragilis ATCC 23745; (2) one of eight recent clinical isolates of B. fragilis; or (3) one of nine isolates representative of Enterobacteriaceae. Seventy-two rat urine samples obtained prior to infection disclosed essentially no assay inhibition: 98.3% +/- 10.3 (1 S.D.). Mean values of reagent antibody activity after incubation with urine aliquots from 24 hr samples collected between 24 and 72 hr were (1) strain 23745 (n = 35) 70.9% +/- 2.6 (S.E.); (2) eight isolates of B. fragilis (n = 49) 86.8% +/- 1.9; (3) nine isolates of Enterobacteriaceae (n = 47) 100.9% +/- 1.0; and (4) shams (n = 29) 95.5% +/- 1.55. Ascribing values less than or equal to 77.7% (2 S.D.) as positive, seven of the eight clinical B. fragilis isolates causing infection were detected in at least one 24 hr urine sample (sensitivity = 87% by organism); 12 of 17 infected rats were correctly identified as positive by at least one urine (sensitivity = 70.6% by rat). Specificity, as assessed in the Enterobacteriaceae group, was 89% (by organism) and 94.5% (by rat). Collectively, these results suggest the presence of a potentially specific, soluble antigen excreted in the urine of rats with B. fragilis infection.


Applying generalizability theory methods to assess continuity and change on the Adolescent Quality of Life-Mental Health Scale (AQOL-MHS).
The Adolescent Quality of Life-Mental Health Scale (AQOL-MHS) is designed to measure quality of life in clinical samples of Latino adolescents aged 12-18 years. Initial findings support its reliability, validity and conceptual model for use of its three domains (emotional regulation, self-concept and social context). Our current study tests the usefulness of the AQOL-MHS for tracking changes in HRQOL during the course of service use. Three waves of data were collected from 59 participants who were recruited from mental health clinics, where waves were spaced 4 months apart. Participants were receiving services at baseline assessment and were tracked for follow-up appointments regardless of treatment status. We analyze conventional reliability statistics for individual differences (e.g., Cronbach's alpha and test-retest correlations), and to estimate the reliability of change, we carried out a variance decomposition analysis. The psychometric analyses from Chavez et al. (Qual Life Res 23(4):1327-1335, 2014) were replicated with comparable results. A generalizability theory (GT) analysis revealed that the AQOL-MHS domains can measure change reliably. The reliability estimates varied from .65 to .78. Although there was reliable change at the individual level, on the average the AQOL-MHS means improved only slightly over time. The reliability of change for all three scales in the AQOL-MHS was acceptable, but consistently lower than the Cronbach's alpha values for each wave. Future work will use this approach to adapt our current measure to provide a better reliability of within-person change and thus broaden its applicability for prospective use.


CD44 enhances invasion of basal-like breast cancer cells by upregulating serine protease and collagen-degrading enzymatic expression and activity.
Basal-like breast cancers (BL-BCa) have the worst prognosis of all subgroups of this disease. Hyaluronan (HA) and the HA receptor CD44 have a long-standing association with cell invasion and metastasis of breast cancer. The purpose of this study was to establish the relation of CD44 to BL-BCa and to characterize how HA/CD44 signaling promotes a protease-dependent invasion of breast cancer (BrCa) cells. CD44 expression was determined with immunohistochemistry (IHC) analysis of a breast cancer tissue microarray (TMA). In vitro experiments were performed on a panel of invasive BL-BCa cell lines, by using quantitative polymerase chain reaction (PCR), immunoblotting, protease activity assays, and invasion assays to characterize the basis of HA-induced, CD44-mediated invasion. Expression of the hyaluronan (HA) receptor CD44 associated with the basal-like subgroup in a cohort of 141 breast tumor specimens (P = 0.018). Highly invasive cells of the representative BL-BCa cell line, MDA-MB-231 (MDA-MB-231Hi) exhibited increased invasion through a basement membrane matrix (Matrigel) and collagen. In further experiments, HA-induced promotion of CD44 signaling potentiated expression of urokinase plasminogen activator (uPA) and its receptor uPAR, and underpinned an increased cell-associated activity of this serine protease in MDA-MB-231Hi and a further BL-BCa cell line, Hs578T cells. Knockdown of CD44 attenuated both basal and HA-stimulated uPA and uPAR gene expression and uPA activity. Inhibition of uPA activity by using (a) a gene-targeted RNAi or (b) a small-molecule inhibitor of uPA attenuated HA-induced invasion of MDA-MB-231Hi cells through Matrigel. HA/CD44 signaling also was shown to increase invasion of MDA-MB-231 cells through collagen and to potentiate the collagen-degrading activity of MDA-MB-231Hi cells. CD44 signaling was subsequently shown to upregulate expression of two potent collagen-degrading enzymes, the cysteine protease cathepsin K and the matrix metalloprotease MT1-MMP. RNAi- or shRNA-mediated depletion of CD44 in MDA-MB-231Hi cells decreased basal and HA-induced cathepsin K and MT1-MMP expression, reduced the collagen-degrading activity of the cell, and attenuated cell invasion through collagen. Pharmacologic inhibition of cathepsin K or RNAi-mediated depletion of MT1-MMP also attenuated MDA-MB-231Hi cell invasion through collagen. HA-induced CD44 signaling increases a diverse spectrum of protease activity to facilitate the invasion associated with BL-BCa cells, providing new insights into the molecular basis of CD44-promoted invasion.


Multiple Signaling Pathways Involved in Human Dendritic Cell Maturation Are Affected by the Fungal Quorum-Sensing Molecule Farnesol.
The quorum-sensing molecule farnesol is produced by the opportunistic human fungal pathogen Candida albicans Aside from its primary function of blocking the transition from yeast to hyphal morphotype, it has an immunomodulatory role on human dendritic cells (DC) through the alteration of surface markers, cytokine secretion, and their ability to activate T cells. Nonetheless, the molecular mechanisms by which farnesol modulates DC differentiation and maturation remained unknown. In this study, we demonstrate through transcriptional and functional assays that farnesol influences several signaling pathways during DC differentiation and in response to TLR agonists. In particular, farnesol increases the expression of the Ag-presenting glycoprotein CD1d through the nuclear receptors PPARγ and RARα, as well as p38 MAPK. However, the higher expression of CD1d did not confer these DC with an enhanced capacity to activate CD1d-restricted invariant NKT cells. In the presence of farnesol, there is reduced secretion of the Th1-inducing cytokine, IL-12, and increased release of proinflammatory cytokines, as well as the anti-inflammatory cytokine IL-10. These changes are partially independent of nuclear receptor activity but, in the case of TNF-α and IL-10, dependent on NF-κB and MAPK pathways. Interestingly, renewal of the IL-12/IL-10 milieu restores the ability of farnesol-differentiated DC to activate invariant NKT, Th1, and FOXP3+ regulatory T cells. Our results show that farnesol modulates nuclear receptors, NF-κB, and MAPK-signaling pathways, thereby impairing the capacity of DC to activate several T cells subsets and potentially conferring C. albicans, an advantage in overcoming DC-mediated immunity.


System combining the best features of heterodyne and direct detection receivers.
A system is presented and analyzed that combines the best features of heterodyne and direct detection systems. It provides the noise suppression characteristics of a heterodyne system; at the same time, it has the advantages of a direct detection system in being able to process equally efficiently received fields that are spatially incoherent or coherent, and in being able to use wide field-of-view receiver optics. Moreover, in the presence of strong background noise and detector thermal noise its performance can be even better than that of a direct detection system for which the only receiver noise present is the signal shot noise. When receiving a spatially incoherent field, its performance could be better than a heterodyne system having the same collector area but requiring a spatially coherent received field. The system uses a cascade combination of a wide-band and narrow-band quantum amplifier in the receiver. In contrast to a previously presented system of this type, which used amplitude modulation, the system now described and analyzed permits almost any type of modulation.


Vasodilatory action of endogenous atrial natriuretic factor in a rat model of chronic heart failure as determined by monoclonal ANF antibody.
Elucidation of the role of (elevated) endogenous atrial natriuretic factor (ANF) in chronic heart failure has been hampered by a lack of specific inhibitors. We used a newly developed monoclonal antibody that has been shown to specifically block both exogenously and endogenously released ANF in vivo. For assessment of the vasodilatory action of ANF in chronic heart failure, either this antibody against ANF or ascites (control serum) was injected in rats with myocardial infarction and failure and in sham animals. Ascites did not alter central hemodynamics in either the sham or infarcted group. Antibody significantly increased right atrial pressure, left ventricular end-diastolic pressure, and systemic vascular resistance (SVR) in the infarction group but did not affect these variables in the sham group. Because renal blood flow, as measured by radioactive microspheres, decreased significantly in all four groups, probably due to nonspecific renal vasoconstrictor effects of the ascites, a separate group of infarcted animals was treated with purified ANF antibody (devoid of nonspecific effects) or mouse IgG as a control injection. In these animals, right atrial pressure increased from 1.1 +/- 0.7 to 2.6 +/- 0.7 mm Hg (p less than 0.001). Although SVR, renal blood flow velocity (measured by Doppler probe), and renal vascular resistance did not change in the infarcted animals after administration of purified ANF antibody, a significant correlation was found between baseline plasma ANF values and the change in SVR exerted by purified ANF antibody (r = 0.758, p less than 0.02, n = 9); that is, SVR increased in rats with high baseline plasma ANF (greater than 350 pg/ml), but decreased in animals with plasma ANF less than 200 pg/ml. These results suggest that moderately elevated endogenous plasma ANF levels in chronic heart failure do affect central hemodynamics, primarily by reducing venous pressure (e.g., by decreasing intravascular volume or by venous dilation). Arterial vasodilation, however, appears to emerge when plasma ANF is greatly increased.


Urine Steroid Metabolomics as a Novel Tool for Detection of Recurrent Adrenocortical Carcinoma.
Urine steroid metabolomics, combining mass spectrometry-based steroid profiling and machine learning, has been described as a novel diagnostic tool for detection of adrenocortical carcinoma (ACC). This proof-of-concept study evaluated the performance of urine steroid metabolomics as a tool for postoperative recurrence detection after microscopically complete (R0) resection of ACC. 135 patients from 14 clinical centers provided postoperative urine samples, which were analyzed by gas chromatography-mass spectrometry. We assessed the utility of these urine steroid profiles in detecting ACC recurrence, either when interpreted by expert clinicians or when analyzed by random forest, a machine learning-based classifier. Radiological recurrence detection served as the reference standard. Imaging detected recurrent disease in 42 of 135 patients; 32 had provided pre- and post-recurrence urine samples. 39 patients remained disease-free for ≥3 years. The urine "steroid fingerprint" at recurrence resembled that observed before R0 resection in the majority of cases. Review of longitudinally collected urine steroid profiles by 3 blinded experts detected recurrence by the time of radiological diagnosis in 50% to 72% of cases, improving to 69% to 92%, if a preoperative urine steroid result was available. Recurrence detection by steroid profiling preceded detection by imaging by more than 2 months in 22% to 39% of patients. Specificities varied considerably, ranging from 61% to 97%. The computational classifier detected ACC recurrence with superior accuracy (sensitivity = specificity = 81%). Urine steroid metabolomics is a promising tool for postoperative recurrence detection in ACC; availability of a preoperative urine considerably improves the ability to detect ACC recurrence.


High-throughput structural characterisation of therapeutic protein targets.
The overarching scientific objective for high-throughput structural biology projects targeting human proteins or relevant proteins of human pathogens is to generate a detailed structural insight of biologically relevant structural features of soluble or membrane protein families of therapeutic relevance. These data will better enable structure-guided drug discovery in pharmaceutical and biotechechnology companies. In particular, detailed structural descriptions for entire protein families will be highly valuable for the design of drugs and other bioactive compounds that are selective for individual members or groups of proteins within protein families. In addition to determined structural information, targeted high-throughput structural biology will also provide reagents, methodologies, technologies and small molecules that bind selectively to targets that are suitable as intervention points in the treatment of disease.


Meta-analysis of soy intake and breast cancer risk.
High intake of soy foods has been proposed to contribute to the low breast cancer risk in Asian countries. However, results of epidemiologic studies of this association are highly variable, and experimental data suggest that soy constituents can be estrogenic and potentially risk enhancing. Thus, rigorous evaluation of available epidemiologic data is necessary before appropriate recommendations can be made, especially for women at high risk of breast cancer or those who have survived the disease. We performed a meta-analysis of 18 epidemiologic studies (12 case-control and six cohort or nested case-control) published from 1978 through 2004 that examined soy exposure and breast cancer risk. Pooled relative risk estimates were based on either the original soy exposure measure defined in each study or on an estimate of daily soy protein intake. Risk estimates, levels and measures of soy exposure, and control for confounding factors varied considerably across studies. In a pooled analysis, among all women, high soy intake was modestly associated with reduced breast cancer risk (odds ratio [OR] = 0.86, 95% confidence interval [CI] = 0.75 to 0.99); the association was not statistically significant among women in Asian countries (OR = 0.89, 95% CI = 0.71 to 1.12). Among the 10 studies that stratified by menopausal status the inverse association between soy exposure and breast cancer risk was somewhat stronger in premenopausal women (OR = 0.70, 95% CI = 0.58 to 0.85) than in postmenopausal women (OR = 0.77, 95% CI = 0.60 to 0.98); however, eight studies did not provide menopause-specific results, six of which did not support an association. When exposure was analyzed by soy protein intake in grams per day, a statistically significant association with breast cancer risk was seen only among premenopausal women (OR = 0.94, 95% CI = 0.92 to 0.97). Soy intake may be associated with a small reduction in breast cancer risk. However, this result should be interpreted with caution due to potential exposure misclassification, confounding, and lack of a dose response. Given these caveats and results of some experimental studies that suggest adverse effects from soy constituents, recommendations for high-dose isoflavone supplementation to prevent breast cancer or prevent its recurrence are premature.


Disentangling the effects of spatial inconsistency of targets and distractors when searching in realistic scenes.
Previous research has suggested that correctly placed objects facilitate eye guidance, but also that objects violating spatial associations within scenes may be prioritized for selection and subsequent inspection. We analyzed the respective eye guidance of spatial expectations and target template (precise picture or verbal label) in visual search, while taking into account any impact of object spatial inconsistency on extrafoveal or foveal processing. Moreover, we isolated search disruption due to misleading spatial expectations about the target from the influence of spatial inconsistency within the scene upon search behavior. Reliable spatial expectations and precise target template improved oculomotor efficiency across all search phases. Spatial inconsistency resulted in preferential saccadic selection when guidance by template was insufficient to ensure effective search from the outset and the misplaced object was bigger than the objects consistently placed in the same scene region. This prioritization emerged principally during early inspection of the region, but the inconsistent object also tended to be preferentially fixated overall across region viewing. These results suggest that objects are first selected covertly on the basis of their relative size and that subsequent overt selection is made considering object-context associations processed in extrafoveal vision. Once the object was fixated, inconsistency resulted in longer first fixation duration and longer total dwell time. As a whole, our findings indicate that observed impairment of oculomotor behavior when searching for an implausibly placed target is the combined product of disruption due to unreliable spatial expectations and prioritization of inconsistent objects before and during object fixation.


Enhanced Growth and Activities of the Dominant Functional Microbiota of Chicken Manure Composts in the Presence of Maize Straw.
As a consequence of intensive feeding, the bulk deposition of livestock manure causes severe environmental problems. Composting is a promising method for waste disposal, and the fermentation process is driven by microbial communities. However, chicken manure contains diverse gut microbes, mainly species derived from Proteobacteria, which may include pathogens that threaten human health. To evaluate composting as a harmless treatment of livestock manure, the dynamics of the microbiota in two chicken manure composts were studied, and the influences of adding maize straw on the compost microbiota were compared. The results revealed that microbes from Firmicutes including Bacillus and Lentibacillus are the most dominant degraders with a strong amino acid metabolism, and they secrete a diverse array of proteases as revealed in metaproteomics data. The addition of maize straw to the chicken manure compost accelerated species succession at the initial stage, and stimulated carbohydrate metabolism in the dominant microbiota. Besides, under the resulting high temperature (>70°C) conditions, the relative abundance of Proteobacteria was reduced by 78% in composts containing maize straw by day 4, which was faster than in compost without added maize straw, in which the abundance was reduced by 66%. Adding maize straw to chicken manure composts can therefore increase the fermentation temperature and inhibit the growth of Proteobacteria. In general, these findings provide increased insight into the dynamic changes among the dominant functional microbiota in chicken manure composts, and may contribute to the optimization of livestock manure composting on an industrial scale.


Predictors of Persistent Milk Allergy in Children: A Retrospective Cohort Study.
Cow's milk (CM) allergy is the second most common food allergy developed during infancy in Japan. To identify predictors of persistent CM allergy, we investigated the tolerance acquisition rate based on an oral food challenge in children under 6 years of age, diagnosed with immediate-type CM allergy. This retrospective cohort study included 131 children born in 2005 with a history of immediate allergic reaction to CM, of whom 39 were excluded because of ongoing oral immunotherapy (n = 18) or a lack of follow-up data (n = 21). The 92 remaining participants were followed for 6 years. Tolerance was defined as no adverse reaction to 200 mL of CM and regular intake of milk at home. Subjects were divided into 3 groups based on age at tolerance acquisition: group I (<3 years; n = 31), group II (3-6 years; n = 42), and group III (persistent allergic group; n = 19). Tolerance acquisition rates by 3, 5, and 6 years of age were 32.6% (30/92), 64.1% (59/92), and 84.8% (70/92), respectively. Age at first hospital visit was significantly higher in groups II and III than in group I (p < 0.001). The incidence of anaphylaxis to other foods was also higher in group III than in group I (p = 0.04), as was CM-induced anaphylaxis (p = 0.03). Furthermore, milk and casein-specific immunoglobulin E (IgE) levels were significantly higher in group III than in group II after birth and remained high thereafter (p < 0.05). The history of anaphylaxis and high milk-specific IgE levels were associated with persistent CM allergy.


Clinical Outcomes of Monofocal Toric IOLs Using Digital Tracking and Intraoperative Aberrometry.
To evaluate clinical outcomes of a toric IOL using digital tracking (DT) and intraoperative aberrometry (IA). This was a retrospective, single surgeon study examining 151 eyes of 106 patients. Inclusion criteria were subjects who presented with visually significant cataracts (or as a candidate for clear lens extraction) and were implanted with a toric intraocular lens. Spherical equivalent prediction errors for IA and preoperative planning were calculated and compared. Preoperative and postoperative refractive data and monocular uncorrected distance visual acuity (UDVA) and corrected distance visual acuity (CDVA) were also collected at 3 months postoperatively. Postoperative actual residual refractive astigmatism with IA was 0.50 D or less in 140 eyes (92.8%) and was 0.50 D or less in 88 eyes (58.3%) with back-calculations based on preoperative planning. The absolute spherical equivalent prediction error was 0.50 D or less in 135 eyes (89.4%) for IA compared to 123 eyes (85.4%) for preoperative planning. Postoperative monocular UDVA was 0.10 logMAR or better in 124 eyes (82.1%) and 0.00 logMAR or better in 90 eyes (59.6%). Postoperative CDVA was 0.10 logMAR or better in 147 eyes (97.4%) and 134 eyes (88.7%) were 0.00 logMAR or better. The results demonstrate that toric implantation with DT and IA can provide excellent refractive and visual outcomes.


Heart rate variability is an independent predictor of morbidity and mortality in hemodynamically stable trauma patients.
Reduced heart rate variability (HRV) reflects autonomic dysfunction and can triage patients better than routine trauma criteria or vital signs. However, there is questionable specificity and no consensus measurement technique. The purpose of this study was to analyze whether factors that alter autonomic function affect the specificity of HRV for assessing traumatic injury. We evaluated 216 hemodynamically stable adults (3:1 M:F; 97:3 blunt:penetrating; age 49 years ± 1 year, mean ± standard error) undergoing computed axial tomography (CT) scan to rule out traumatic brain injury (TBI). All were prospectively instrumented with a Mars Holter system (GE Healthcare, Milwaukee, WI). HRV was determined offline using time domain (standard deviation of normal-normal intervals, root-mean-square successive difference) and frequency domain (very low frequency [VLF], LF, wideband frequency, high frequency [HF], low to HF index ratio) calculations from 15-minute electrocardiogram and correlated with routine vital signs, mortality, TBI, morbidity, length of stay (LOS), and comorbidities. Significance (p ≤ 0.05) was determined using nonparametric analysis, Student's t test, analysis of variance, or multiple logistic regression. VLF alone predicted survival, severity of TBI, intensive care unit LOS, and hospital LOS (all p < 0.05). Beta-blockers or diabetes had no effect, whereas age, sedation, mechanical ventilation, spinal cord injury, and intoxication influenced one or more of the variables with age being the most powerful confounder (all p < 0.05). Except for the Glasgow Coma Scale, no other routine trauma or hemodynamic criteria correlated with any of these outcomes. Decreased VLF is an independent predictor of mortality and morbidity in hemodynamically stable trauma patients. Other time and other frequency domain variables correlated with some, but not all, outcomes. All were heavily influenced by factors that alter autonomic function, especially patient age.


Addressing gaps in surgical skills training by means of low-cost simulation at Muhimbili University in Tanzania.
Providing basic surgical and emergency care in rural settings is essential, particularly in Tanzania, where the mortality burden addressable by emergency and surgical interventions has been estimated at 40%. However, the shortages of teaching faculty and insufficient learning resources have hampered the traditionally intensive surgical training apprenticeships. The Muhimbili University of Health and Allied Sciences consequently has experienced suboptimal preparation for graduates practising surgery in the field and a drop in medical graduates willing to become surgeons. To address the decline in circumstances, the first step was to enhance technical skills in general surgery and emergency procedures for senior medical students by designing and implementing a surgical skills practicum using locally developed simulation models. A two-day training course in nine different emergency procedures and surgical skills based on the Canadian Network for International Surgery curriculum was developed. Simulation models for the surgical skills were created with locally available materials. The curriculum was pilot-tested with a cohort of 60 senior medical students who had completed their surgery rotation at Muhimbili University. Two measures were used to evaluate surgical skill performance: Objective Structured Clinical Examinations and surveys of self-perceived performance administered pre- and post-training. Thirty-six students participated in the study. Prior to the training, no student was able to correctly perform a surgical hand tie, only one student was able to correctly perform adult intubation and three students were able to correctly scrub, gown and glove. Performance improved after training, demonstrated by Objective Structured Clinical Examination scores that rose from 6/30 to 15/30. Students perceived great benefit from practical skills training. The cost of the training using low-tech simulation was four United States dollars per student. Simulation is valued to gain experience in practising surgical skills prior to working with patients. In the context of resource-limited settings, an additional benefit is that of learning skills not otherwise obtainable. Further testing of this approach will determine its applicability to other resource-limited settings seeking to develop skill-based surgical and emergency procedure apprenticeships. Additionally, skill sustainability and readiness for actual surgical and emergency experiences need to be assessed.


GATA3 expression is decreased in psoriasis and during epidermal regeneration; induction by narrow-band UVB and IL-4.
Psoriasis is characterized by hyperproliferation of keratinocytes and by infiltration of activated Th1 and Th17 cells in the (epi)dermis. By expression microarray, we previously found the GATA3 transcription factor significantly downregulated in lesional psoriatic skin. Since GATA3 serves as a key switch in both epidermal and T helper cell differentiation, we investigated its function in psoriasis. Because psoriatic skin inflammation shares many characteristics of epidermal regeneration during wound healing, we also studied GATA3 expression under such conditions.Psoriatic lesional skin showed decreased GATA3 mRNA and protein expression compared to non-lesional skin. GATA3 expression was also markedly decreased in inflamed skin of mice with a psoriasiform dermatitis induced with imiquimod. Tape-stripping of non-lesional skin of patients with psoriasis, a standardized psoriasis-triggering and skin regeneration-inducing technique, reduced the expression of GATA3. In wounded skin of mice, low GATA3 mRNA and protein expression was detected. Taken together, GATA3 expression is downregulated under regenerative and inflammatory hyperproliferative skin conditions. GATA3 expression could be re-induced by successful narrow-band UVB treatment of both human psoriasis and imiquimod-induced psoriasiform dermatitis in mice. The prototypic Th2 cytokine IL-4 was the only cytokine capable of inducing GATA3 in skin explants from healthy donors. Based on these findings we argue that GATA3 serves as a key regulator in psoriatic inflammation, keratinocyte hyperproliferation and skin barrier dysfunction.


Site-specific factors associated with clinical trial recruitment efficiency in general practice settings: a comparative descriptive analysis.
Recruitment of participants is crucial to the success of randomised control trials (RCTs) but can be challenging and expensive. Current research on trial efficiency is often focused at the patient-level with an emphasis on effective recruitment strategies. Less is known about selection of study sites to optimise recruitment. We examine site-level factors that are associated with patient recruitment and cost efficiency using data from an RCT conducted across 25 general practices (GP) in Victoria, Australia. Data on number of participants screened, excluded, eligible, recruited, and randomised from each study site were extracted from a clinical trial. Details regarding site characteristics, recruitment practices, and staff time commitment were collected using a three-part survey. The key outcomes assessed were recruitment efficiency (ratio of screened to randomised), average time, and cost for each participant recruited and randomised. To identify practice-level factors associated with efficient recruitment and lower cost, outcomes were dichotomised (25th percentile vs others) and each practice-level factor assessed against the outcomes to determine its association. Across 25 GP study sites, 1968 participants were screened of which 299 (15.2%) were recruited and randomised. The mean recruitment efficiency was 7.2, varying from 1.4 to 19.8 across sites. The strongest factor associated with efficiency was assigning clinical staff to identify potential participants (57.14% vs. 22.2%). The more efficient sites were smaller practices and were more likely to be rural locations and in areas of lower socioeconomic status. The average time used for recruitment was 3.7 h (SD2.4) per patient randomised. The mean cost per patient randomised was $277 (SD161), and this varied from $74 to $797 across sites. The sites identified with the 25% lowest recruitment cost (n = 7) were more experienced in research participation and had high levels of nurse and/or administrative support. Despite the small sample size, this study quantified the time and cost used to recruit patients and provides helpful indications of site-level characteristics that can help improve feasibility and efficiency of conducting RCT in GP settings. Characteristics indicative of high levels of support for research and rural practices, which often tends to be overlooked, were observed to be more efficient in recruiting.


Gonadal hormones and frontocortical expression of vascular endothelial growth factor in male stroke-prone, spontaneously hypertensive rats, a model for attention-deficit/hyperactivity disorder.
Attention-deficit/hyperactivity disorder (AD/HD) is a common pediatric behavioral disorder associated, in part, with male preponderance and reduced regional cerebral blood flow (rCBF). However, mechanism(s) underlying male preponderance and reduced rCBF in AD/HD are unclear. The present study profiles the expression of angiogenic and hormonal factors likely to underlie these symptoms using a recently characterized AD/HD animal model, juvenile male stroke-prone spontaneously hypertensive rats (SHRSP). Because vascular endothelial growth factor (VEGF) signaling cascade and gonadal steroids are key regulators of angiogenesis and gender-based behavior, respectively, we profiled their patterns of expression in the frontal cortex of SHRSP to elucidate their roles in the genesis of AD/HD male preponderance and rCBF. Interestingly, levels of VEGF, VEGF receptors (KDR, Flt-1), endothelial nitric oxide synthase, phosphorylated Akt (pAkt), estrogen receptor-alpha, aromatase, and capillary density in sham-operated SHRSP were remarkably down-regulated, whereas androgen receptor levels were up-regulated, compared with age-matched genetic control, Wistar-Kyoto rats. Castration, estrogen, and androgen receptor antagonist (flutamide) counteracted these effects. Dihydrotestosterone, but not testosterone, reversed the beneficiary effects of castration. Estrogen receptor-beta levels remained unchanged in all groups examined. We postulate that changes in androgen metabolism that tend to up-regulate local dihydrotestosterone concentration and diminish estrogen synthesis, in the frontal cortex of juvenile male SHRSP, may lower levels and/or activity of VEGF and its signaling cascade and, subsequently, reduce rCBF. These findings could, in part, help explain the pathogenesis of reduced rCBF and male preponderance in AD/HD.


Morningness-eveningness correlates with sleep time, quality, and hygiene in secondary school students: a multilevel analysis.
At the onset of puberty, students shift their sleep to later hours, but school starts early. It is suggested that evening orientation and early school start times do not go well together. Therefore, the aim of this study was to investigate sleep problems in adolescence, and it was expected that the adolescents' eveningness orientation is associated with many sleep-related problems. Students of secondary education (n = 3201; mean = 13.8 ± 1.8 years) filled out a self-report questionnaire containing measures of morningness-eveningness, sleep time (midpoint of sleep, social "jetlag"), sleep length (on schooldays and on weekends), sleep quality, and sleep hygiene as well as questions on electronic screen media use. The impact of circadian preference on sleep time (midpoint of sleep and social jetlag), sleep length (on schooldays and on weekends), sleep quality, and sleep hygiene of adolescents was tested via multilevel analyses while controlling for covariates on the student level (age, sex, screen media use, and time leaving home) and on the class level (school type, grade level, and school start time). Morningness-eveningness was a significant predictor of all dependent variables and associations were highest (β > 0.40) for midpoint of sleep, social jetlag, problems with going to bed, problems with falling asleep, and problems with returning to wakefulness. Providing guidance for parents on sleep hygiene behavior routines for their child, an educational program in sleep hygiene, and later school start times could help to synchronize adolescents' circadian rhythms to daily educational and social demands.


Exploration of a hypothesized independent association of a common 9p21.3 gene variant and ischemic stroke in patients with and without angiographic coronary artery disease.
Single-nucleotide polymorphisms (SNPs) at the chromosome 9p21.3 locus are associated with coronary artery disease (CAD). An association of this genomic region with ischemic stroke independent of its effect on CAD could suggest an additional, stroke-specific pathophysiological relationship. Medical record review was used to identify 548 patients without a history of cerebrovascular disease and 232 who had a verified ischemic stroke or transient ischemic attack (TIA) from the Duke CATHGEN biorepository of patients who had a cardiac catheterization. ANCOVA and multivariable logistic regression modeling were performed to determine independent genetic associations between the key chromosome 9p21.3 SNP, rs10757278, and ischemic stroke by comparing allele frequencies between 229 patients with stroke or TIA and an equal number of matched nonstroke controls, adjusting for other risk factors. In a secondary analysis, controls were further divided based on the presence (n = 353) or absence (n = 195) of angiographic CAD. Allele frequencies were similar between patients with and without a history of ischemic stroke in both additive (p = 0.83) and dominant (p = 0.92) models of genetic risk. There was no association between rs10757278 allele frequency and stroke status based on the presence or absence of angiographically demonstrated CAD in nonstroke controls (ANCOVA, p = 0.99). These results provide no evidence of a stroke-specific association of the 9p21.3 locus regardless of the presence or absence of angiographic CAD and highlight the need for larger studies to further evaluate this hypothesized relationship.


Trans-Cyclooctene-Functionalized PeptoBrushes with Improved Reaction Kinetics of the Tetrazine Ligation for Pretargeted Nuclear Imaging.
Tumor targeting using agents with slow pharmacokinetics represents a major challenge in nuclear imaging and targeted radionuclide therapy as they most often result in low imaging contrast and high radiation dose to healthy tissue. To address this challenge, we developed a polymer-based targeting agent that can be used for pretargeted imaging and thus separates tumor accumulation from the imaging step in time. The developed targeting agent is based on polypeptide-graft-polypeptoid polymers (PeptoBrushes) functionalized with trans-cyclooctene (TCO). The complementary 111In-labeled imaging agent is a 1,2,4,5-tetrazine derivative, which can react with aforementioned TCO-modified PeptoBrushes in a rapid bioorthogonal ligation. A high degree of TCO loading (up to 30%) was achieved, without altering the physicochemical properties of the polymeric nanoparticle. The highest degree of TCO loading resulted in significantly increased reaction rates (77-fold enhancement) compared to those with small molecule TCO moieties when using lipophilic tetrazines. Based on computer simulations, we hypothesize that this increase is a result of hydrophobic effects and significant rearrangements within the polymer framework, in which hydrophobic patches of TCO moieties are formed. These patches attract lipophilic tetrazines, leading to increased reaction rates in the bioorthogonal ligation. The most reactive system was evaluated as a targeting agent for pretargeted imaging in tumor-bearing mice. After the setup was optimized, sufficient tumor-to-background ratios were achieved as early as 2 h after administration of the tetrazine imaging agent, which further improved at 22 h, enabling clear visualization of CT-26 tumors. These findings show the potential of PeptoBrushes to be used as a pretargeting agent when an optimized dose of polymer is used.


Use of statins in patients with liver disease.
Statins are generally well tolerated and shown to have a good safety profile. In clinical trials a similar proportion of patients randomized to statins and placebo treated developed abnormal liver tests. However, idiosyncratic drug-induced liver injury (DILI) is a rare adverse reaction and clinical trials are underpowered to detect uncommon side effects. Although probably very rare, idiosyncratic DILI due to statins has been reported among other drugs in all major prospective and retrospective series on DILI. A summary of 40 cases of statin hepatoxicity has been published and, a series of 76 cases suspected statin induced liver injury has recently been published. In the last mentioned series, three patients died and/or underwent liver transplantation and three cases had a rechallenge with the same statin which produced a similar pattern of liver injury. Statin therapy appears to be safe in the treatment of non-alcoholic fatty liver disease, in compensated patients with chronic hepatitis B or C. Statins have also been shown to have an inhibitory effect on hepatitis C virus replication in vitro. Retrospective data as well as data from a recent randomized controlled trial suggest that statin therapy might be a useful adjunct to standard combination antiviral therapy in patients with chronic hepatitis C. Statin therapy seems to be related to a reduced risk for hepatocellular carcinoma in patients with chronic liver disease, but randomized data are lacking.


Resolving biodegradation patterns of persistent saturated hydrocarbons in weathered oil samples from the Deepwater Horizon disaster.
Biodegradation plays a major role in the natural attenuation of oil spills. However, limited information is available about biodegradation of different saturated hydrocarbon classes in surface environments, despite that oils are composed mostly of saturates, due to the limited ability of conventional gas chromatography (GC) to resolve this compound group. We studied eight weathered oil samples collected from four Gulf of Mexico beaches 12-19 months after the Deepwater Horizon disaster. Using comprehensive two-dimensional gas chromatography (GC × GC), we successfully separated, identified, and quantified several distinct saturates classes in these samples. We find that saturated hydrocarbons eluting after n-C22 dominate the GC-amenable fraction of these weathered samples. This compound group represented 8-10%, or 38-68 thousand metric tons, of the oil originally released from Macondo well. Saturates in the n-C22 to n-C29 elution range were found to be partly biodegraded, but to different relative extents, with ease of biodegradation decreasing in the following order: n-alkanes > methylalkanes and alkylcyclopentanes+alkylcyclohexanes > cyclic and acyclic isoprenoids. We developed a new quantitative index designed to characterize biodegradation of >n-C22 saturates. These results shed new light onto the environmental fate of these persistent, hydrophobic, and mostly overlooked compounds in the unresolved complex mixtures (UCM) of weathered oils.


Heterogeneous Integration of Carbon-Nanotube-Graphene for High-Performance, Flexible, and Transparent Photodetectors.
Low-dimensional carbon materials, such as semiconducting carbon nanotubes (CNTs), conducting graphene, and their hybrids, are of great interest as promising candidates for flexible, foldable, and transparent electronics. However, the development of highly photoresponsive, flexible, and transparent optoelectronics still remains limited due to their low absorbance and fast recombination rate of photoexcited charges, despite the considerable potential of photodetectors for future wearable and foldable devices. This work demonstrates a heterogeneous, all-carbon photodetector composed of graphene electrodes and porphyrin-interfaced single-walled CNTs (SWNTs) channel, exhibiting high photoresponse, flexibility, and full transparency across the device. The porphyrin molecules generate and transfer photoexcited holes to the SWNTs even under weak white light, resulting in significant improvement of photoresponsivity from negligible to 1.6 × 10-2 A W-1 . Simultaneously, the photodetector exhibits high flexibility allowing stable light detection under ≈50% strain (i.e., a bending radius of ≈350 µm), and retaining a sufficient transparency of ≈80% at 550 nm. Experimental demonstrations as a wearable sunlight sensor highlight the utility of the photodetector that can be conformally mounted on human skin and other curved surfaces without any mechanical and optical constraints. The heterogeneous integration of porphyrin-SWNT-graphene may provide a viable route to produce invisible, high-performance optoelectronic systems.


A prospective study of in-office diagnostic vitreous sampling in patients with vitreoretinal pathology.
To evaluate the safety and efficacy of in-office vitreous sampling (50-100 microL). A prospective, consecutive case series of 578 diagnostic vitreous sample attempts performed between February 23, 2007 and September 31, 2008. A standardized technique of vitreous sampling followed by intravitreal injection of therapeutic medication was performed. All subjects were followed for adverse events after sampling/injection. Out of a total of 578 vitreous sampling attempts, a total of 550 positive diagnostic vitreous samples were collected from 157 eyes of 144 patients. There were a total of 28 "dry" samples in 19 subjects (13 females and 6 males). The rate of dry sample was 4.8% (28 of 578). Complications included transient vitreous hemorrhage (n = 3), transient intraocular inflammation (n = 3), localized, chronic retinal detachment (n = 1). None of the complications resulted in permanent visual sequelae. These complications were no different than those expected from intravitreal injection alone. This prospective study has shown that in-office vitreous sampling of 50 microL to 100 microL is a safe and reproducible procedure. Larger randomized studies will be helpful to further assess the safety and long-term effects of in-office diagnostic vitreous sampling.


Revisiting absorption and electronic circular dichroism spectra of cholesterol in solution: a joint experimental and theoretical study.
Cholesterol is doubtless one of the most studied bio-molecules, which unfortunately features low emitting properties, precluding its in vivo study by fluorescence experiments. The design of fluorescent analogues of cholesterol is thus an appealing challenge in biochemistry, which simultaneously requires minor changes in its chemical structure (to retain main biological properties) and considerable enhancement of light emission. To this aim, the photochemical behaviour of the native molecule has to be deeply understood. In this work, we focused our attention on the electronic absorption of cholesterol in several common organic solutions, combining experimental (through ultraviolet-visible and electronic circular dichroism spectroscopy) and theoretical approaches (at the time-dependent density functional theory level) in order to solve the important discrepancies previously reported in the literature on the maximum absorption wavelengths and on the nature (Rydberg and/or π → π*) of the associated electronic transition.


Efficient cellular solid-state NMR of membrane proteins by targeted protein labeling.
Solid-state NMR spectroscopy (ssNMR) has made significant progress towards the study of membrane proteins in their native cellular membranes. However, reduced spectroscopic sensitivity and high background signal levels can complicate these experiments. Here, we describe a method for ssNMR to specifically label a single protein by repressing endogenous protein expression with rifampicin. Our results demonstrate that treatment of E. coli with rifampicin during induction of recombinant membrane protein expression reduces background signals for different expression levels and improves sensitivity in cellular membrane samples. Further, the method reduces the amount of time and resources needed to produce membrane protein samples, enabling new strategies for studying challenging membrane proteins by ssNMR.


I-J loop involvement in the pharmacological profile of CLC-K channels expressed in Xenopus oocytes.
CLC-K chloride channels and their subunit, barttin, are crucial for renal NaCl reabsorption and for inner ear endolymph production. Mutations in CLC-Kb and barttin cause Bartter syndrome. Here, we identified two adjacent residues, F256 and N257, that when mutated hugely alter in Xenopus oocytes CLC-Ka's biphasic response to niflumic acid, a drug belonging to the fenamate class, with F256A being potentiated 37-fold and N257A being potently blocked with a KD~1μM. These residues are localized in the same extracellular I-J loop which harbors a regulatory Ca(2+) binding site. This loop thus can represent an ideal and CLC-K specific target for extracellular ligands able to modulate channel activity. Furthermore, we demonstrated the involvement of the barttin subunit in the NFA potentiation. Indeed the F256A mutation confers onto CLC-K1 a transient potentiation induced by NFA which is found only when CLC-K1/F256A is co-expressed with barttin. Thus, in addition to the role of barttin in targeting and gating, the subunit participates in the pharmacological modulation of CLC-K channels and thus represents a further target for potential drugs.


Task-specific transfer of perceptual learning across sensory modalities.
It is now widely accepted that primary cortical areas of the brain that were once thought to be sensory-specific undergo significant functional reorganisation following sensory deprivation. For instance, loss of vision or audition leads to the brain areas normally associated with these senses being recruited by the remaining sensory modalities [1]. Despite this, little is known about the rules governing crossmodal plasticity in people who experience typical sensory development, or the potential behavioural consequences. Here, we used a novel perceptual learning paradigm to assess whether the benefits associated with training on a task in one sense transfer to another sense. Participants were randomly assigned to a spatial or temporal task that could be performed visually or aurally, which they practiced for five days; before and after training, we measured discrimination thresholds on all four conditions and calculated the extent of transfer between them. Our results show a clear transfer of learning between sensory modalities; however, generalisation was limited to particular conditions. Specifically, learned improvements on the spatial task transferred from the visual domain to the auditory domain, but not vice versa. Conversely, benefits derived from training on the temporal task transferred from the auditory domain to visual domain, but not vice versa. These results suggest a unidirectional transfer of perceptual learning from dominant to non-dominant sensory modalities and place important constraints on models of multisensory processing and plasticity.


Neopterin levels in patients with cerebrovascular disease.
The recovery period in ischemic cerebrovascular disease is sometimes long and complicated. It is thought that a systemic inflammatory response plays an important role in this process. Therefore, promising studies have reported that neopterin and other cytokines serve as indicators of the inflammatory response. The present study investigated the effects of cerebrovascular diseases on plasma neopterin. We studied 68 consecutive patients under 65 years of age with cerebrovascular disease; the mean age was 52.08±5.74 years (yrs) (ranging from 39 yrs to 64 yrs; 28 female and 40 male). We also studied 29 randomly selected age-matched healthy subjects (control); the mean age was 49.76±13.11 years (yrs) (ranging from 41 yrs to 62 yrs; 12 female and 17 male). Blood samples for assessing plasma levels of neopterin were usually taken within seven days after admission to the hospital and stored at -20°C until analysis. Serum neopterin levels in all the subjects were measured by Enzyme Immunoassay (EÝA) using the BRAHMS method (Neopterin; Diagnostic GmbH, 16761 Berlin, Germany). Differences between the groups' means were analyzed with the Mann-Whitney U test. The mean values of neopterin levels in patients (mean neopterin levels in patients, 18.51 ± 11.56 nmol/L; vs. control, 12.26 ± 3.87 nmol/L [p=.001]) were significantly different when compared with the controls. It has been suggested that the mortality and morbidity associated with cerebrovascular disease could be prevented by a reduction in the inflammatory response. We suggest that plasma neopterin levels were significantly increased. So, the levels of plasma neopterin may be useful monitoring of treatment and course of diseases.


A comparison of single molecule and amplification based sequencing of cancer transcriptomes.
The second wave of next generation sequencing technologies, referred to as single-molecule sequencing (SMS), carries the promise of profiling samples directly without employing polymerase chain reaction steps used by amplification-based sequencing (AS) methods. To examine the merits of both technologies, we examine mRNA sequencing results from single-molecule and amplification-based sequencing in a set of human cancer cell lines and tissues. We observe a characteristic coverage bias towards high abundance transcripts in amplification-based sequencing. A larger fraction of AS reads cover highly expressed genes, such as those associated with translational processes and housekeeping genes, resulting in relatively lower coverage of genes at low and mid-level abundance. In contrast, the coverage of high abundance transcripts plateaus off using SMS. Consequently, SMS is able to sequence lower- abundance transcripts more thoroughly, including some that are undetected by AS methods; however, these include many more mapping artifacts. A better understanding of the technical and analytical factors introducing platform specific biases in high throughput transcriptome sequencing applications will be critical in cross platform meta-analytic studies.


Six3 demarcates the anterior-most developing brain region in bilaterian animals.
The heads of annelids (earthworms, polychaetes, and others) and arthropods (insects, myriapods, spiders, and others) and the arthropod-related onychophorans (velvet worms) show similar brain architecture and for this reason have long been considered homologous. However, this view is challenged by the 'new phylogeny' placing arthropods and annelids into distinct superphyla, Ecdysozoa and Lophotrochozoa, together with many other phyla lacking elaborate heads or brains. To compare the organisation of annelid and arthropod heads and brains at the molecular level, we investigated head regionalisation genes in various groups. Regionalisation genes subdivide developing animals into molecular regions and can be used to align head regions between remote animal phyla. We find that in the marine annelid Platynereis dumerilii, expression of the homeobox gene six3 defines the apical region of the larval body, peripherally overlapping the equatorial otx+ expression. The six3+ and otx+ regions thus define the developing head in anterior-to-posterior sequence. In another annelid, the earthworm Pristina, as well as in the onychophoran Euperipatoides, the centipede Strigamia and the insects Tribolium and Drosophila, a six3/optix+ region likewise demarcates the tip of the developing animal, followed by a more posterior otx/otd+ region. Identification of six3+ head neuroectoderm in Drosophila reveals that this region gives rise to median neurosecretory brain parts, as is also the case in annelids. In insects, onychophorans and Platynereis, the otx+ region instead harbours the eye anlagen, which thus occupy a more posterior position. These observations indicate that the annelid, onychophoran and arthropod head develops from a conserved anterior-posterior sequence of six3+ and otx+ regions. The six3+ anterior pole of the arthropod head and brain accordingly lies in an anterior-median embryonic region and, in consequence, the optic lobes do not represent the tip of the neuraxis. These results support the hypothesis that the last common ancestor of annelids and arthropods already possessed neurosecretory centres in the most anterior region of the brain. In light of its broad evolutionary conservation in protostomes and, as previously shown, in deuterostomes, the six3-otx head patterning system may be universal to bilaterian animals.


Genetic Polymorphisms Complicate COVID-19 Therapy: Pivotal Role of HO-1 in Cytokine Storm.
Coronaviruses are very large RNA viruses that originate in animal reservoirs and include severe acute respiratory distress syndrome (SARS) and Middle East respiratory syndrome (MERS) and other inconsequential coronaviruses from human reservoirs like the common cold. SARS-CoV-2, the virus that causes COVID-19 and is believed to originate from bat, quickly spread into a global pandemic. This RNA virus has a special affinity for porphyrins. It invades the cell at the angiotensin converting enzyme-2 (ACE-2) receptor and binds to hemoproteins, resulting in a severe systemic inflammatory response, particularly in high ACE-2 organs like the lungs, heart, and kidney, resulting in systemic disease. The inflammatory response manifested by increased cytokine levels and reactive oxygen species results in inhibition of heme oxygenase (HO-1), with a subsequent loss of cytoprotection. This has been seen in other viral illness like human immunodeficiency virus (HIV), Ebola, and SARS/MERS. There are a number of medications that have been tried with some showing early clinical promise. This illness disproportionately affects patients with obesity, a chronic inflammatory disease with a baseline excess of cytokines. The majority of the medications used in the treatment of COVID-19 are metabolized by cytochrome P450 (CYP) enzymes, primarily CYP2D6. This is further complicated by genetic polymorphisms of CYP2D6, HO-1, ACE, and ACE-2. There is a potential role for HO-1 upregulation to treat/prevent cytokine storm. Current therapy must focus on antivirals and heme oxygenase upregulation. Vaccine development will be the only magic bullet.


Temporal and spatial scaling of the genetic structure of a vector-borne plant pathogen.
The ecology of plant pathogens of perennial crops is affected by the long-lived nature of their immobile hosts. In addition, changes to the genetic structure of pathogen populations may affect disease epidemiology and management practices; examples include local adaptation of more fit genotypes or introduction of novel genotypes from geographically distant areas via human movement of infected plant material or insect vectors. We studied the genetic structure of Xylella fastidiosa populations causing disease in sweet orange plants in Brazil at multiple scales using fast-evolving molecular markers (simple-sequence DNA repeats). Results show that populations of X. fastidiosa were regionally isolated, and that isolation was maintained for populations analyzed a decade apart from each other. However, despite such geographic isolation, local populations present in year 2000 were largely replaced by novel genotypes in 2009 but not as a result of migration. At a smaller spatial scale (individual trees), results suggest that isolates within plants originated from a shared common ancestor. In summary, new insights on the ecology of this economically important plant pathogen were obtained by sampling populations at different spatial scales and two different time points.