Enhancement of blood coagulation by soluble fibrin complexes.
We have detected a species of soluble fibrin complexes with significant biological properties. Agarose gel chromatography of normal plasma or purified fibrinogen previously incubated with small amounts of thrombin revealed the presence of a species of high molecular weight soluble fibrin complexes, which contained only small quantities of fibrinogen by immunological assays but which exhibited enhanced sensitivity to thrombin. In addition, these complexes substantially shortened the thrombin-clotting time of normal plasma and enhanced the resistance of normal plasma to heparin action. Similar thrombin-sensitive soluble fibrin complexes were demonstrated in vivo in rabbits for up to 10 min after the infusion of 50 U of thrombin. Thrombin-sensitive soluble fibrin complexes were also demonstrated in 3 of 12 patients with documented thromboembolic disease and in 2 of 20 patients after major surgery. High molecular weight soluble fibrin complexes, which exhibit enhanced thrombin sensitivity and which are capable of increasing the rate of normal fibrinogen-to-fibrin conversion by thrombin, may appear consequent to clinical thrombosis and situations involving trauma (e.g., major surgery). Such soluble complexes, although they have no proven role in the primary pathogenesis of intravascular thrombosis, may contribute to a temporary "hypercoagulable state" and may accelerate the build-up and extension of already existing thrombotic deposits.


Plasma Cytokine Levels in Chronic Asymptomatic HIV-1 Subtype C Infection as an Indicator of Disease Progression in Botswana: A Retrospective Case Control Study.
HIV infects cells of the immune system causing immune activation and proliferation of immune cells, leading to alteration of production and activity of a number of cytokines. These changes in cytokine levels can affect the immune function, and have the potential to directly impact the course of HIV disease. We characterized plasma cytokine concentration profiles in HIV-1 subtype C chronically infected, antiretroviral therapy (ART)-naive participants to establish their influence on disease progression and viremia. Plasma levels of interleukin (IL)-1α, IL-7, IL-12p40, granulocyte macrophage-colony-stimulating factor (GM-CSF), and interferon (IFN)-γ were quantified in samples from 60 treatment-naive participants in the placebo arm of the completed Micronutrient-HIV disease progressions study, "Dikotlana" (2004-2009) in Gaborone, Botswana. Participants were stratified into progressors (P) and nonprogressors (NP) based on their rates of CD4(+) T cell depletion during the study period. Nonprogressors were those who had <1% CD4(+) T cell depletion at 24 months postenrollment. Progressors were defined as those with CD4(+) T cell depletion of >15% at 24 months postenrollment. Cytokine levels were compared between P and NP using the Mann-Whitney U-test. Logistic regression analysis was used to determine if cytokines predicted disease progression. Correlations of cytokines with CD4(+) T cell counts and viral loads were determined by the Spearman rank test. Median baseline CD4(+) T cell counts were 453 (Q1, Q3; 401, 592) and 479 (Q1, Q3; 401-592) for nonprogressors and progressors, respectively. Nonprogressors had a higher viral set point than progressors. IL-12p40 levels were significantly higher in the P than in NP at enrollment and 24 months (p < 0.05). Levels of IL-1α, IL-7, IFN-γ, and GM-CSF did not differ significantly between the two groups. Except for IL-12p40, which displayed an inverse correlation with CD4(+) T cell counts and a direct correlation with viral load, all other cytokines showed no correlations. IL-12p40 was found to be the most significant predictor of progression and its production was most likely driven by HIV replication products as evidenced by its direct correlation with viral load. In chronic HIV-1 subtype C infection, CD4(+) T cell counts and plasma cytokine levels may not necessarily evolve in parallel, suggesting the involvement of other factors in determining the rates of CD4(+) T cell depletion.


A silicon strip detector dose magnifying glass for IMRT dosimetry.
Intensity modulated radiation therapy (IMRT) allows the delivery of escalated radiation dose to tumor while sparing adjacent critical organs. In doing so, IMRT plans tend to incorporate steep dose gradients at interfaces between the target and the organs at risk. Current quality assurance (QA) verification tools such as 2D diode arrays, are limited by their spatial resolution and conventional films are nonreal time. In this article, the authors describe a novel silicon strip detector (CMRP DMG) of high spatial resolution (200 microm) suitable for measuring the high dose gradients in an IMRT delivery. A full characterization of the detector was performed, including dose per pulse effect, percent depth dose comparison with Farmer ion chamber measurements, stem effect, dose linearity, uniformity, energy response, angular response, and penumbra measurements. They also present the application of the CMRP DMG in the dosimetric verification of a clinical IMRT plan. The detector response changed by 23% for a 390-fold change in the dose per pulse. A correction function is derived to correct for this effect. The strip detector depth dose curve agrees with the Farmer ion chamber within 0.8%. The stem effect was negligible (0.2%). The dose linearity was excellent for the dose range of 3-300 cGy. A uniformity correction method is described to correct for variations in the individual detector pixel responses. The detector showed an over-response relative to tissue dose at lower photon energies with the maximum dose response at 75 kVp nominal photon energy. Penumbra studies using a Varian Clinac 21EX at 1.5 and 10.0 cm depths were measured to be 2.77 and 3.94 mm for the secondary collimators, 3.52 and 5.60 mm for the multileaf collimator rounded leaf ends, respectively. Point doses measured with the strip detector were compared to doses measured with EBT film and doses predicted by the Philips Pinnacle treatment planning system. The differences were 1.1% +/- 1.8% and 1.0% +/- 1.6%, respectively. They demonstrated the high temporal resolution capability of the detector readout system, which will allow one to investigate the temporal dose pattern of IMRT and volumetric modulated are therapy (VMAT) deliveries. The CMRP silicon strip detector dose magnifying glass interfaced to a TERA ASIC DAQ system has high spatial and temporal resolution. It is a novel and valuable tool for QA in IMRT dose delivery and for VMAT dose delivery.


A study of seroprevalence and rates of asymptomatic viremia of severe fever with thrombocytopenia syndrome virus among Chinese blood donors.
Severe fever with thrombocytopenia syndrome virus (SFTSV), an emerging tick-borne pathogen that can cause fatal severe fever with thrombocytopenia syndrome, was first identified in China in 2009. Limited evidence suggests that SFTSV can be transmitted between humans via blood contact, raising concerns over transfusion safety. A study of donor samples from three Chinese blood centers was conducted to investigate the seroprevalence and rate of SFTSV viremia among Chinese blood donors. From April 16 to October 31, 2012, a total of 17,208 plasma samples were collected from donors at Xinyang (located in an SFTSV-endemic area), Mianyang, and Luoyang Blood Centers. Assessment of anti-SFTSV total antibody was performed on all samples using enzyme-linked immunosorbent assay. Repeat-reactive samples were tested for SFTSV RNA using reverse transcription (RT)-real-time polymerase chain reaction (PCR) assay with Taqman probes. In addition, 9960 of the Xinyang samples were tested in pools of 4 by the same PCR method and each of the samples in a reactive pool was tested individually. Donor seroreactivity rates were as follows: Xinyang, 0.54% (80/14,752); Mianyang, 0.27% (3/1130); and Luoyang, 0.28% (3/1326). All seroreactive samples were negative on RT-PCR single-sample testing. Two RT-PCR-reactive donor samples were identified, both with estimated viral load of less than 20 plaque-forming units/mL. The RNA prevalence rate for SFTSV among donors in Xinyang was 0.02%. This was the first multiregion study of SFTSV sero- and viral prevalence among Chinese blood donors. Viral prevalence was low and no seroreactive sample was viremic, suggesting a limited impact of SFTSV on blood safety in China.


Setting priorities for research in blood donation and transfusion: outcome of the James Lind Alliance priority-setting partnership.
How do we decide which topics should be prioritized for research? The need for a robust process for prioritization by key stakeholders, and not just the researchers themselves, was recognized by the James Lind Alliance. A methodology has been established to enable clinicians, patients, and caregivers to identify and prioritize important uncertainties for research in different health areas. This methodology was applied to transfusion medicine to help focus the research agenda in this field. A steering group was formed in 2015 comprising four donor/patient/caregiver representatives and six clinicians and was supported by an information scientist and James Lind Alliance representatives. The scope of the priority-setting partnership included uncertainties from blood donation through transfusion but excluded laboratory aspects of transfusion and specialist blood products. Three methods were used to identify the top 10 research priorities: two widely disseminated online surveys, a search of existing literature, and a final prioritization workshop. There were 408 respondents to the first survey contributing 817 questions, which were refined into 54 indicative questions that had not already been answered by previous research. Respondents to a second survey were asked to select the three questions they believed to be the most important. The 30 most popular research questions were then brought to a workshop of donors, patients, and caregivers to produce the "top 10." This prioritized list should be of considerable value to both researchers and funding bodies when considering what research should be conducted in transfusion medicine.


5-Fluorouracil chemotherapy of gastric cancer generates residual cells with properties of cancer stem cells.
5-Fluorouracil (5Fu) chemotherapy is the first treatment of choice for advanced gastric cancer (GC), but its effectiveness is limited by drug resistance. Emerging evidence suggests that the existence of cancer stem cells (CSCs) contributes to chemoresistance. The aim of the present study was to determine whether 5Fu chemotherapy generates residual cells with CSC-like properties in GC. Human GC cell lines, SGC7901 and AGS, were exposed to increasing 5Fu concentrations. The residual cells were assessed for both chemosensitivity and CSC-like properties. B lymphoma Mo-MLV insertion region 1 (BMI1), a putative CSC protein, was analyzed by immunohistochemical staining and subjected to pairwise comparison in GC tissues treated with or without neoadjuvant 5Fu-based chemotherapy. The correlation between BMI1 expression and recurrence-free survival in GC patients who received 5Fu-based neoadjuvant chemotherapy was then examined. The residual cells exhibited 5Fu chemoresistance. These 5Fu-resistant cells displayed some CSC features, such as a high percentage of quiescent cells, increased self-renewal ability and tumorigenicity. The 5Fu-resistant cells were also enriched with cells expressing cluster of differentiation (CD)133+, CD326+ and CD44+CD24-. Moreover, the BMI1 gene was overexpressed in 5Fu-resistant cells, and BMI1 knockdown effectively reversed chemoresistance. The BMI1 protein was highly expressed consistently in the remaining GC tissues after 5Fu-based neoadjuvant chemotherapy, and BMI1 levels were correlated positively with recurrence-free survival in GC patients who received 5Fu-based neoadjuvant chemotherapy. Our data provided molecular evidence illustrating that 5Fu chemotherapy in GC resulted in acquisition of CSC-like properties. Moreover, enhanced BMI1 expression contributed to 5Fu resistance and may serve as a potential therapeutic target to reverse chemoresistance in GC patients.


Proenergetic effects of resveratrol in the murine neuronal cell line Neuro2a.
Energy deficit is a common characteristic of neurodegenerative disorders, including Alzheimer's disease. Adenosine monophosphate activated protein kinase (AMPK) is a key enzyme maintaining energy balance by regulating the cellular uptake of glucose, β-oxidation of fatty acids, and expression of glucose transporter 4. Since resveratrol has been shown to increase the activity of AMPK, we hypothesized that it might influence energy metabolism in a model neuron-like cell line, murine Neuro2a cells. Resveratrol caused an elevation of adenosine triphosphate (ATP) and guanosine triphosphate (GTP) in a dose-dependent manner. The highest ATP and GTP levels achieved by treatment with resveratrol were 70.3 ± 8.2 nmol/mg protein (1.9-fold of control) and 27.2 ± 4.0 nmol/mg protein (1.7-fold of control), respectively, when cells were treated with 100 μM resveratrol for 6 h. Interestingly, increases in the total sum of all adenine nucleotides were found upon addition of resveratrol. Despite these increases in ATP, GTP, and the total adenine nucleotide pool, resveratrol treatment led to a decrease in glucose consumption and lactate release, suggesting that resveratrol does not increase energy production (e.g. via AMPK kinase activation) but rather inhibits energy-consuming processes. Resveratrol increases the levels of ATP and GTP, but without creating an additional glucose demand.


Effectiveness and reliability of cannulated fenestrated screws augmented with polymethylmethacrylate cement in the surgical treatment of osteoporotic vertebral fractures.
Implants' stability, especially in osteoporosis patients, is a challenging matter. Nowadays, the adoption of cannulated fenestrated screws augmented with polymethylmethacrylate cement (PMMA CSF) is described by some authors. This single-center, retrospective observational study aims to evaluate the long-term effectiveness, reliability, and mechanical performances of this type of screws in osteoporotic fractures. All the patients surgically treated from January 2009 to December 2019 with PMMA CSF were evaluated and submitted to the inclusion and exclusion criteria. Clinical and radiological evaluations were performed at pre- and post-surgery time and at the follow-up (FU). Loss of correction in the sagittal plane (bisegmental Cobb angle), kyphosis angle of the fracture (fractured vertebral angle), loosening of pedicle screws (screw's apex vertebral body's anterior cortex mean gap called SAAC gap and screw's apex vertebral body's superior endplate mean gap called SASE gap), visual analog scale, and Oswestry disability index scores were evaluated. One hundred and sixty-three patients (58 males and 105 females) aged over 65 years affected by vertebral osteoporotic fractures were included in the study. At FU, we do not found significant differences in radiological items in respect to the postoperative period. Only one case of loosening and 18 cases of cement leaking (without neurological impairments) were found. Clinical scores improvement was significant in the interval between preoperative and FU. PMMA CSF seems to can guarantee good efficacy and effectiveness in the surgical treatment of vertebral fractures in osteoporosis.


Linkage genome scan for loci predisposing to panic disorder or agoraphobia.
We conducted a 10 cM linkage genome scan in a set of 20 American pedigrees (153 subjects), ascertained through probands with panic disorder (PD). Several anxiety disorders segregate in these families; they were diagnosed on the basis of Schedule for Affective Disorders and Schizophrenia interview. In this article, we describe results for panic disorder and agoraphobia, which are closely related, common, heritable anxiety disorders. This is the first complete linkage genome scan for agoraphobia and the third for PD. A total of 407 markers (389 autosomal, 18 X chromosome) were genotyped. Multipoint LOD score and NPL analysis were completed using GENEHUNTER2. For PD, two genomic regions meet criteria for suggestive linkage. One of these regions is on chromosome 1 (LOD score = 2.04). This region coincides with a region that generated a LOD score of 1.1 in a previous genome scan by Crowe et al. [2001: Am J Med Genet (Neuropsychiatr Genet) 105:105-109]. The other (LOD score = 2.01) is located on chromosome 11p and occurs at marker CCKBR, one of eight candidate genes examined. For agoraphobia, the most promising potential linkage was on chromosome 3 (NPL score = 2.75; P = 0.005). This was accounted for primarily by a single family that by itself generated an NPL score of 10.01 (P = 0.0039) and a LOD score of 2.10. These results provide initial evidence for a genetic locus on chromosome 3 that contributes to risk for agoraphobia. They also support suggestive linkage to two risk loci for panic disorder. Additional potential loci were identified with lesser statistical support; several of these were consistent with previously reported panic disorder linkage results. Overall, the results presented here suggest that PD and agoraphobia are complex traits that share some, but not all, of their susceptibility loci. Published 2001 Wiley-Liss, Inc.


Panspecies small-molecule disruptors of heterochromatin-mediated transcriptional gene silencing.
Heterochromatin underpins gene repression, genome integrity, and chromosome segregation. In the fission yeast Schizosaccharomyces pombe, conserved protein complexes effect heterochromatin formation via RNA interference-mediated recruitment of a histone H3 lysine 9 methyltransferase to cognate chromatin regions. To identify small molecules that inhibit heterochromatin formation, we performed an in vivo screen for loss of silencing of a dominant selectable kanMX reporter gene embedded within fission yeast centromeric heterochromatin. Two structurally unrelated compounds, HMS-I1 and HMS-I2, alleviated kanMX silencing and decreased repressive H3K9 methylation levels at the transgene. The decrease in methylation caused by HMS-I1 and HMS-I2 was observed at all loci regulated by histone methylation, including centromeric repeats, telomeric regions, and the mating-type locus, consistent with inhibition of the histone deacetylases (HDACs) Clr3 and/or Sir2. Chemical-genetic epistasis and expression profiles revealed that both compounds affect the activity of the Clr3-containing Snf2/HDAC repressor complex (SHREC). In vitro HDAC assays revealed that HMS-I1 and HMS-I2 inhibit Clr3 HDAC activity. HMS-I1 also alleviated transgene reporter silencing by heterochromatin in Arabidopsis and a mouse cell line, suggesting a conserved mechanism of action. HMS-I1 and HMS-I2 bear no resemblance to known inhibitors of chromatin-based activities and thus represent novel chemical probes for heterochromatin formation and function.


A conceptual framework: redefining forest soil's critical acid loads under a changing climate.
Federal agencies of several nations have or are currently developing guidelines for critical forest soil acid loads. These guidelines are used to establish regulations designed to maintain atmospheric acid inputs below levels shown to damage forests and streams. Traditionally, when the critical soil acid load exceeds the amount of acid that the ecosystem can absorb, it is believed to potentially impair forest health. The excess over the critical soil acid load is termed the exceedance, and the larger the exceedance, the greater the risk of ecosystem damage. This definition of critical soil acid load applies to exposure of the soil to a single, long-term pollutant (i.e., acidic deposition). However, ecosystems can be simultaneously under multiple ecosystem stresses and a single critical soil acid load level may not accurately reflect ecosystem health risk when subjected to multiple, episodic environmental stress. For example, the Appalachian Mountains of western North Carolina receive some of the highest rates of acidic deposition in the eastern United States, but these levels are considered to be below the critical acid load (CAL) that would cause forest damage. However, the area experienced a moderate three-year drought from 1999 to 2002, and in 2001 red spruce (Picea rubens Sarg.) trees in the area began to die in large numbers. The initial survey indicated that the affected trees were killed by the southern pine beetle (Dendroctonus frontalis Zimm.). This insect is not normally successful at colonizing these tree species because the trees produce large amounts of oleoresin that exclude the boring beetles. Subsequent investigations revealed that long-term acid deposition may have altered red spruce forest structure and function. There is some evidence that elevated acid deposition (particularly nitrogen) reduced tree water uptake potential, oleoresin production, and caused the trees to become more susceptible to insect colonization during the drought period. While the ecosystem was not in exceedance of the CAL, long-term nitrogen deposition pre-disposed the forest to other ecological stress. In combination, insects, drought, and nitrogen ultimately combined to cause the observed forest mortality. If any one of these factors were not present, the trees would likely not have died. This paper presents a conceptual framework of the ecosystem consequences of these interactions as well as limited plot level data to support this concept. Future assessments of the use of CAL studies need to account for multiple stress impacts to better understand ecosystem response.


Molecular modeling, dynamics, and an insight into the structural inhibition of cofactor independent phosphoglycerate mutase isoform 1 from Wuchereria bancrofti using cheminformatics and mutational studies.
Phosphoglycerate mutase catalyzes the interconversion between 2-phosphoglycerate and 3-phosphoglycerate in the glycolytic and gluconeogenic pathways. They exist in two unrelated forms, that is either cofactor (2,3-diphosphoglycerate) dependent or cofactor-independent. These two enzymes have no similarity in amino acid sequence, tertiary structure, and in catalytic mechanism. Wuchereria bancrofti (WB) contains the cofactor-independent form, whereas other organisms can possess the dependent form or both. Since, independent phosphoglycerate mutase (iPGM) is an essential gene for the survival of nematodes, and it has no sequence or structural similarity to the cofactor-dependent phosphoglycerate mutase found in mammals, it represents an attractive drug target for the filarial nematodes. In this current study, a putative cofactor-iPGM gene was identified in the protein sequence of the WB. In the absence of crystal structure, a three-dimensional structure was determined using the homology modeling approximation, and the most stable protein conformation was identified through the molecular dynamics simulation studies, using GROMACS 4.5. Further, the functional or characteristic residues were identified through the sequence analysis, potential inhibitors were short-listed and validated, and potential inhibitors were ranked using the cheminformatics and molecular dynamics simulations studies, Prime MM-GBSA approach, respectively.


Legislation on the preparation of medicinal products in European pharmacies and the Council of Europe Resolution.
The rights of patients should be sufficiently protected even when an appropriate authorised medicine does not exist or is unavailable on the market. The Resolution, which was adopted by the Committee of Ministers of the Council of Europe in 2011, aims at harmonising quality and safety standards for pharmacy preparation of medicinal products in Europe. The system of regulation of medicinal products is built upon two pillars: the marketing authorisation of the medicinal product and the licence for manufacturing and wholesale. This article provides insight into the recent interpretation of the European Court of Justice concerning the scope of European Union (EU) regulation of medicinal products and the circumstances in which the EU regulation does not apply: pharmacy preparations, specialties and the compassionate use of medicines, including manufacturing licence. Pharmacy preparations are allowed under certain strict conditions according to EU regulations. However, pharmacies specialised in preparation and distributing medicinal products to local pharmacies do not fulfil these strict conditions in EU regulation. Apart from the legal context, relevant standards for safety and quality assurance are needed in Europe in order to protect patients' rights and to avoid risks from pharmacy preparations. The Council of Europe Resolution provides a means of establishing standards for safety and quality assurance for pharmacy preparations through Good Manufacturing Practice Guidelines. The Resolution is available to authorities and pharmacists in order to prevent incidents with medicines prepared in pharmacies which may threaten patients' safety. The authors conclude that pharmacy practices have changed over time in Europe and this may imply a reason for a reform of EU regulation on medicinal products.


Fine discrimination of faces can be performed rapidly.
Here we measure the smallest change in a face that can be discriminated. A morphing algorithm mixed two faces in variable proportions to create a series of synthetic faces that each differed by a tiny amount. By selecting from this series, a test face could be chosen so as to reach a just noticeable difference from a sample face. Face-discrimination thresholds were about 7% of the average difference between two faces, as quantified by coefficients of a principal components decom position. This threshold remained constant as the duration of the test face was reduced from 1,000 to 100 msec, and rose quickly for shorter stimulus durations. The behavioral evidence presented here indicates that complex visual processing can be completed within the first 100 msec of the signal, suggesting involvement of feedforward neural mechanisms, and placing constraints on possible computational algorithms employed within the ventral visual pathways.


Potential for the Invasive Species Aedes Albopictus and Arboviral Transmission through the Chabahar Port in Iran.
Dengue, chikungunya, and Zika viruses are emerging infectious disease threats wherever suitable vectors, hosts, and habitat are present. The aim of the present study was to use the bioagent transport and environmental modeling system (BioTEMS) to identify the potential for arbovirus-infected Aedes species to invade the Chabahar area in southeastern Iran. ArcGIS geospatial analysis software, Statistica software, and BioTEMS were used to analyze geographic information and conduct data analysis. BioTEMS utilizes up to several hundred abiotic and biotic factors to produce risk and vulnerability assessments for biological agents and infectious diseases. The output of BioTEMS was validated using published predictive models, and most importantly published collection data of Aedes species in Iran. There appears to have been two separate invasion events by Ae. albopictus into the southern region of Iran, first preceding 2009 and then again in 2013. BioTEMS identified two probable areas of introduction during the 2009 time frame, either through one or both the Chabahar ports or the Iranshahr airport with subsequent spread through vehicular transport. BioTEMS identified the port as an introduction zone for ZIKAV with high-risk zones and identifies gap zones during the 2013 time frame. Recommended surveillance sites are provided. The air and maritime ports of Iran serve international customers, and are therefore vulnerable to import and invasion of mosquito vectors and arboviruses. Based on comparisons with other published low-resolution models, BioTEMS provides information for medical and public health professionals conducting integrated mosquito management, preventive medicine, and epidemiological surveillance.


[E-health and "Cancer outside the hospital walls", Big Data and artificial intelligence].
To heal otherwise in oncology has become an imperative of Public Health and an economic imperative in France. Patients can therefore receive live most of their care outside of hospital with more ambulatory care. This ambulatory shift will benefit from the digital revolution and the development of digital health or e-health. Cancer research will also benefit with Big Data and artificial intelligence, which gather and analyze a huge amount of data. In this synthesis, we describe the different e-health tools and their potential impacts in oncology, at the levels of education and information of patients and caregivers, prevention, screening and diagnosis, treatment, follow-up, and research. A few randomized studies have already demonstrated clinical benefits. Large Big Data projects such as ConSoRe and Health Data Hub have been launched in France. We also discuss the issues and limitations of "cancer outside the hospital walls and e-health" from the point of view of patients, health care professionals, health facilities and government. This new organization will have to provide remote support "outside the walls" with care and follow-up of quality, continuous and prolonged in total safety and equity. Ongoing and future randomized clinical trials will need to definitively demonstrate areas of interest, advantages and drawbacks not only for patients, but also for caregivers, health facilities and governments.


A new species of Ituglanis representing the southernmost record of the genus, with comments on phylogenetic relationships (Teleostei: Siluriformes: Trichomycteridae).
Ituglanis australis new species, is described from tributaries of the Laguna dos Patos and Río Uruguay, in Brazil and Uruguay. This represents the southernmost record of the genus and the first occurrence of a species of Ituglanis in those systems. It is distinguished from all its congeners, except Ituglanis parahybae and Ituglanis cahyensis, by its body pigmentation with three well-defined dark brown stripes running along each flank. Ituglanis australis differs from I. parahybae and I. cahyensis in the pectoral- and pelvic-fin ray counts, the pattern of the cephalic laterosensory system and the number of dorsal-fin basal radials. The new species, as well as several other examined congeners, has the levator internus IV muscle attached to the dorsal face of the posttemporo-supracleithrum; a condition that corroborates the inclusion of Ituglanis into a large trichomycterine clade that also includes Bullockia, Hatcheria, Scleronema and several species of Trichomycterus. Previous proposals of the affinities within Ituglanis are reviewed and, despite some advances, the phylogenetic relationships among species of the genus remain largely unknown.


Effectiveness of poly(ethylene terephthalate) and high-density polyethylene in protection of milk flavor.
The development of certain off-flavors in whole milk (3.25% milk fat) as related to packaging material [glass, high-density polyethylene (HDPE), amber poly(ethylene terephthalate) (PETE), clear PETE, and clear PETE-UV] were evaluated after exposure to fluorescent light (1100 to 1300 lx) for 18 d at 4 degrees C. Control samples packaged and stored under identical conditions were wrapped in foil to prevent light exposure. Selected flavor compounds in milk were measured analytically on d 0, 7, 14, and 18 of storage, while intensities of "oxidation," "acetaldehyde," and "lacks freshness" off-flavors were determined by sensory analysis at the same intervals. In light-exposed samples, oxidation off-flavor was significantly lower when packaged in amber PETE versus other containers. Milk packaged in HDPE containers showed a significantly higher level of oxidation off-flavor than milk packaged in PETE-UV containers but not higher than clear PETE or glass containers. No significant difference in acetaldehyde off-flavor was found between package material treatments (exposed or protected). Acetaldehyde concentration never exceeded flavor threshold levels, regardless of packaging material. Amber and PETE-UV materials proved to be a competitive packaging choice for milk in preserving fresh milk flavor.


Cost-effectiveness of antimicrobial prophylaxis for children in the RIVUR trial.
We evaluated the cost-effectiveness of antimicrobial prophylaxis to prevent recurrent UTIs in children with vesicoureteral reflux based on the RIVUR trial. A decision tree model compared strategies of antimicrobial prophylaxis vs. placebo in children with reflux using results from the RIVUR trial. Risk reduction was 50% based on intention to treat analysis. Costs were based on Medicare reimbursement and data in literature. The model incorporated costs of medications, imaging and complications such as pyelonephritis, likelihood of surgery and loss of work. One- and two-way sensitivity analyses were performed evaluating the effect of changing variables on the cost-effectiveness of antibiotic prophylaxis in preventing recurrent UTIs. Mean costs were higher for treatment vs. placebo at $3092 and $2932, respectively, with 12.7 fewer infections per 100 children. One-way sensitivity analyses showed that antibiotics would be cost equivalent if the yearly medical cost was $386, rate of recurrent UTI increased to 32%, antibiotic risk reduction was 63%, or rate of pyelonephritis in the placebo group was 48%. Two-way analyses modifying antibiotic cost, risk reduction of antibiotics and probability of infection showed areas where antibiotics could be more cost-effective than placebo. Antibiotic prophylaxis is associated with marginally higher costs compared with placebo, but significantly fewer infections. A slight decrease in antibiotic cost or increase in recurrent UTIs on placebo may result in prophylaxis being more cost-effective. We recognize that the marginal overall cost in antibiotics may have a substantial impact on the quality of life for the individual patient and family.


Genome-Scale Model Reveals Metabolic Basis of Biomass Partitioning in a Model Diatom.
Diatoms are eukaryotic microalgae that contain genes from various sources, including bacteria and the secondary endosymbiotic host. Due to this unique combination of genes, diatoms are taxonomically and functionally distinct from other algae and vascular plants and confer novel metabolic capabilities. Based on the genome annotation, we performed a genome-scale metabolic network reconstruction for the marine diatom Phaeodactylum tricornutum. Due to their endosymbiotic origin, diatoms possess a complex chloroplast structure which complicates the prediction of subcellular protein localization. Based on previous work we implemented a pipeline that exploits a series of bioinformatics tools to predict protein localization. The manually curated reconstructed metabolic network iLB1027_lipid accounts for 1,027 genes associated with 4,456 reactions and 2,172 metabolites distributed across six compartments. To constrain the genome-scale model, we determined the organism specific biomass composition in terms of lipids, carbohydrates, and proteins using Fourier transform infrared spectrometry. Our simulations indicate the presence of a yet unknown glutamine-ornithine shunt that could be used to transfer reducing equivalents generated by photosynthesis to the mitochondria. The model reflects the known biochemical composition of P. tricornutum in defined culture conditions and enables metabolic engineering strategies to improve the use of P. tricornutum for biotechnological applications.


Protection of the Ovine Fetal Gut against Ureaplasma-Induced Chorioamnionitis: A Potential Role for Plant Sterols.
Chorioamnionitis, clinically most frequently associated with Ureaplasma, is linked to intestinal inflammation and subsequent gut injury. No treatment is available to prevent chorioamnionitis-driven adverse intestinal outcomes. Evidence is increasing that plant sterols possess immune-modulatory properties. Therefore, we investigated the potential therapeutic effects of plant sterols in lambs intra-amniotically (IA) exposed to Ureaplasma. Fetal lambs were IA exposed to Ureaplasma parvum (U. parvum, UP) for six days from 127 d-133 d of gestational age (GA). The plant sterols β-sitosterol and campesterol, dissolved with β-cyclodextrin (carrier), were given IA every two days from 122 d-131 d GA. Fetal circulatory cytokine levels, gut inflammation, intestinal injury, enterocyte maturation, and mucosal phospholipid and bile acid profiles were measured at 133 d GA (term 150 d). IA plant sterol administration blocked a fetal inflammatory response syndrome. Plant sterols reduced intestinal accumulation of proinflammatory phospholipids and tended to prevent mucosal myeloperoxidase-positive (MPO) cell influx, indicating an inhibition of gut inflammation. IA administration of plant sterols and carrier diminished intestinal mucosal damage, stimulated maturation of the immature epithelium, and partially prevented U. parvum-driven reduction of mucosal bile acids. In conclusion, we show that β-sitosterol and campesterol administration protected the fetus against adverse gut outcomes following UP-driven chorioamnionitis by preventing intestinal and systemic inflammation.


Foxa2 regulates multiple pathways of insulin secretion.
The regulation of insulin secretion by pancreatic beta cells is perturbed in several diseases, including adult-onset (type 2) diabetes and persistent hyperinsulinemic hypoglycemia of infancy (PHHI). The first mouse model for PHHI has a conditional deletion of the gene encoding the winged-helix transcription factor Foxa2 (Forkhead box a2, formerly Hepatocyte nuclear factor 3beta) in pancreatic beta cells. Using isolated islets, we found that Foxa2 deficiency resulted in excessive insulin release in response to amino acids and complete loss of glucose-stimulated insulin secretion. Most PHHI cases are associated with mutations in SUR1 (Sulfonylurea receptor 1) or KIR6.2 (Inward rectifier K(+) channel member 6.2), which encode the subunits of the ATP-sensitive K(+) channel, and RNA in situ hybridization of mutant mouse islets revealed that expression of both genes is Foxa2 dependent. We utilized expression profiling to identify additional targets of Foxa2. Strikingly, one of these genes, Hadhsc, encodes short-chain L-3-hydroxyacyl-coenzyme A dehydrogenase, deficiency of which has been shown to cause PHHI in humans. Hadhsc is a direct target of Foxa2, as demonstrated by cotransfection as well as in vivo chromatin immunoprecipitation experiments using isolated islets. Thus, we have established Foxa2 as an essential activator of genes that function in multiple pathways governing insulin secretion.


Serum ghrelin levels but not GH, IGF-1 and IGFBP-3 levels are altered in patients with fibromyalgia syndrome.
Both hypothalamo-pituitary-insulin-like growth factor-1 (IGF-1) axis and ghrelin levels may be altered in fibromyalgia syndrome (FMS) due to increased somatostatin tone. The aim of this study is to compare hypothalamo-pituitary-IGF-1 axis, ghrelin concentrations and their relations in premenopausal women with FMS and premenopausal healthy controls. Seventy-five women (47 FMS and 28 healthy women) were enrolled in the study. Fasting plasma glucose, serum growth hormone (GH), insulin, C-peptide, IGF-1, insulin-like growth factor binding protein-3 (IGFBP-3) and ghrelin levels were measured. Depressive symptoms were assessed using beck depression inventory. Pain intensity and sleep disturbance were recorded on a visual analog scale. The activity of daily living was assessed by fibromyalgia impact questionnaire. There were no significant differences in GH, IGF-1, IGFBP-3, glucose, insulin, and C-peptide levels between patients and controls (p>0.05), whereas ghrelin levels were significantly lower in patients than controls (p<0.05). Ghrelin levels were not correlated with GH, IGF-1, IGFBP-3, glucose, insulin, and C-peptide levels while they were positively correlated with tender point score and sleep disturbance score and negatively correlated with pain intensity score. Our results suggest that low levels of ghrelin in FMS are not related to the changes in hypothalamo-pituitary-IGF-1 axis but may be related to some symptoms of FMS. Our results need to be clarified by further studies.


The impact of the school-based Psychosocial Structured Activities (PSSA) program on conflict-affected children in Northern Uganda.
Children in northern Uganda have undergone significant psychosocial stress during the region's lengthy conflict. A Psychosocial Structured Activities (PSSA) program was implemented in 21 schools identified as amongst those most severely affected by conflict-induced displacement across Gulu and Amuru Districts. The PSSA intervention comprised a series of 15 class sessions designed to progressively increase children's resilience through structured activities involving drama, movement, music and art (with additional components addressing parental support and community involvement). Eight schools were selected by random quota sampling from those schools receiving the PSSA intervention. Two hundred and three children were identified in these schools as being scheduled to receive intervention, and were followed up 12 months later following engagement with PSSA activities. A comparison group comprised 200 children selected from schools that had met inclusion criteria for receipt of intervention, but were not scheduled for intervention coverage until later. Preliminary research used participatory focus group methodology to determine local indicators of child well-being as viewed by parents, teachers, and children respectively. Pre- and post- assessments focused on ratings for each child - by parents, teachers and children - with respect to these indicators. Significant increases in ratings of child well-being were observed in both intervention and comparison groups over a 12-month period. However, the well-being of children who had received the PSSA intervention increased significantly more than for children in the comparison group, as judged by child and parent (but not teacher) report. This effect was evident despite considerable loss-to-follow-up at post-testing as a result of return of many households to communities of origin. General improvement in child well-being over a 12-month period suggests that recovery and reconstruction efforts in Northern Uganda following the onset of peace had a substantive impact on the lives of children. However, exposure to the PSSA program had an additional positive impact on child well-being, suggesting its value in post-conflict recovery contexts.


Mother's curse: the effect of mtDNA on individual fitness and population viability.
The mitochondrial genome is considered generally to be an innocent bystander in adaptive evolution; however, there is increasing evidence that mitochondrial DNA (mtDNA) is an important contributor to viability and fecundity. Some of this evidence is now well documented, with mtDNA mutations having been shown to play a causal role in degenerative diseases, ageing, and cancer. However, most research on mtDNA has ignored the possibility that other instances exist where mtDNA mutations could have profound fitness consequences. Recent work in humans and other species now indicates that mtDNA mutations play an important role in sperm function, male fertility, and male fitness. Ironically, deleterious mtDNA mutations that affect only males, such as those that impair sperm function, will not be subject to natural selection because mitochondria are generally maternally inherited and could reach high frequencies in populations if the mutations are not disadvantageous in females. Here, we review how such mtDNA mutations might affect the viability of natural populations. We consider factors that increase or decrease the strength of the effect of mtDNA mutations on population viability and discuss what mechanisms exist to mitigate deleterious mtDNA effects.


Cortical firing and sleep homeostasis.
The need to sleep grows with the duration of wakefulness and dissipates with time spent asleep, a process called sleep homeostasis. What are the consequences of staying awake on brain cells, and why is sleep needed? Surprisingly, we do not know whether the firing of cortical neurons is affected by how long an animal has been awake or asleep. Here, we found that after sustained wakefulness cortical neurons fire at higher frequencies in all behavioral states. During early NREM sleep after sustained wakefulness, periods of population activity (ON) are short, frequent, and associated with synchronous firing, while periods of neuronal silence are long and frequent. After sustained sleep, firing rates and synchrony decrease, while the duration of ON periods increases. Changes in firing patterns in NREM sleep correlate with changes in slow-wave activity, a marker of sleep homeostasis. Thus, the systematic increase of firing during wakefulness is counterbalanced by staying asleep.


A Worldwide Survey on the Practices and Perceptions of Submaximal Fitness Tests in Team Sports.
To survey team-sport practitioners on current practices and perceptions of submaximal fitness tests (SMFTs). A convenience sample of team-sport practitioners completed an online survey (September to November 2021). Descriptive statistics were used to obtain information of frequencies. A mixed-model quantile (median) regression was employed to assess the differences between the perceived influence of extraneous factors. A total of 66 practitioners (74 discrete protocols) from 24 countries completed the survey. Time-efficient and nonexhaustive nature were considered the most important features of implementation. Practitioners prescribed a range of SMFTs, administered mostly on a monthly or weekly basis, but scheduling strategies appeared to differ across SMFT categories. Cardiorespiratory/metabolic outcome measures were collected in most protocols (n = 61; 82%), with the majority monitoring heart-rate-derived indices. Subjective outcome measures (n = 33; 45%) were monitored exclusively using ratings of perceived exertion. Mechanical outcome measures (n = 19; 26%) included either a combination of locomotor outputs (eg, distance covered) or variables derived from microelectrical mechanical systems. The perceived influence of extraneous factors on measurement accuracy varied according to outcome measure, and there was a lack of consensus among practitioners. Our survey showcases the methodological frameworks, practices, and challenges of SMFTs in team sports. The most important features for implementation perhaps support the use of SMFTs as a feasible and sustainable tool for monitoring in team sports. The wide variety of protocols, scheduling strategies, and outcome measures, along with their associated collection and analytical techniques, may reflect the absence of robust evidence regarding the application of SMFTs in team sports.


Enhancing the collectivist critique: accounts of the human enhancement debate.
Individualist ethical analyses in the enhancement debate have often prioritised or only considered the interests and concerns of parents and the future child. The collectivist critique of the human enhancement debate argues that rather than pure individualism, a focus on collectivist, or group-level ethical considerations is needed for balanced ethical analysis of specific enhancement interventions. Here, I defend this argument for the insufficiency of pure individualism. However, existing collectivist analyses tend to take a negative approach that hinders them from adequately contributing to balanced ethical analysis, and often leads to a prohibitive stance. I argue this is due to two common problems with collectivist analyses: inappropriate acceptance of individualist assumptions, and failure to appropriately weigh individual vs collective ethical considerations. To further develop the collectivist critique in the enhancement debate, I suggest we may look to collectivism in public health ethics, which avoids these problems.


mStruct: inference of population structure in light of both genetic admixing and allele mutations.
Traditional methods for analyzing population structure, such as the Structure program, ignore the influence of the effect of allele mutations between the ancestral and current alleles of genetic markers, which can dramatically influence the accuracy of the structural estimation of current populations. Studying these effects can also reveal additional information about population evolution such as the divergence time and migration history of admixed populations. We propose mStruct, an admixture of population-specific mixtures of inheritance models that addresses the task of structure inference and mutation estimation jointly through a hierarchical Bayesian framework, and a variational algorithm for inference. We validated our method on synthetic data and used it to analyze the Human Genome Diversity Project-Centre d'Etude du Polymorphisme Humain (HGDP-CEPH) cell line panel of microsatellites and HGDP single-nucleotide polymorphism (SNP) data. A comparison of the structural maps of world populations estimated by mStruct and Structure is presented, and we also report potentially interesting mutation patterns in world populations estimated by mStruct.


[Investigation of Chlamydophila pneumoniae seropositivity and risk factors in patients with atherosclerotic vascular disease in Sivas, Turkey].
The role of various microorganisms including Chlamydophila (formerly Chlamydia) pneumoniae, have been frequently investigated in the pathogenesis of atherosclerosis. In our study, the relationship between C.pneumoniae seropositivity and risk factors for atherosclerosis have been evaluated. A total of 90 atherosclerotic patients (71 of them were male; age range: 45-87 years; mean age: 65.3 ± 8.7 years) and 90 control subjects without coronary diseases (41 of them were male; age range: 42-84 years; mean age: 61.6 ± 9.6 years) were included in the study. Both groups were also evaluated for the presence of risk factors such as age, gender, smoking, hypertension, diabetes, obesity, dyslipidemia, familial history and the high levels of ferritin, cholesterol (total, HDL and LDL) and HS (high sensitive)-CRP. The presence of C.pneumoniae IgG, IgM and IgA antibodies were investigated by micro-immunofluorescence (MIF) and ELISA methods using commercial kits (Euroimmun, Germany). The total antibody seropositivity rate was found 100% (90/90) in patient group by both MIF and ELISA methods, while this rate in control group was 94% (85/90) by MIF and 92% (83/90) by ELISA. When MIF test results were taken into consideration (since it is accepted as the reference method for C.pneumoniae serology), IgG, IgM and IgA positivity rates in patient and control groups were found as 100% (90/90) and 89% (80/90); 70% (63/90) and 59% (53/90); 3% (3/90) and 2% (2/90), respectively. Statistically significant difference between patient and control groups was detected only for IgG positivity (p< 0.05) and for total antibody positivities (100% and 94%, respectively) (p< 0.05). The evaluation of the risk factors revealed that age, hypertension, dyslipidemia and HS-CRP levels exhibited statistically significant differences between patient and control groups (p< 0.05 for each parameter tested). Statistically significant relation was detected only between high HS-CRP levels and C.pneumoniae seropositivity (p< 0.05). It was concluded that in areas with high C.pneumoniae infection prevalence, early diagnosis and specific treatment of C.pneumoniae infections, may prevent establishment of chronic infection and eliminate a risk factor for the development of atherosclerosis.


Cell-Free Hemoglobin Is Associated With Increased Vascular Resistance and Reduced Peripheral Perfusion in Severe Malaria.
In severe falciparum malaria, unlike sepsis, hypotension on admission is uncommon. We hypothesized that low nitric oxide bioavailability due to the presence of cell-free hemoglobin (CFH) increases vascular tone in severe malaria. Patients with severe malaria (n = 119), uncomplicated malaria (n = 91), or suspected bacterial sepsis (n = 56), as well as healthy participants (n = 50), were recruited. The systemic vascular resistance index (SVRI) was estimated from the echocardiographic cardiac index and the mean arterial pressure. SVRI and hematocrit levels were lower and plasma CFH and asymmetric dimethylarginine levels were higher in patients with malaria, compared with healthy participants. In multivariate linear regression models for mean arterial pressure or SVRI in patients with severe malaria, hematocrit and CFH but not asymmetric dimethylarginine were significant predictors. The SVRI was lower in patients with suspected bacterial sepsis than in those with severe malaria, after adjustment for hematocrit and age. Plasma CFH levels correlated positively with the core-peripheral temperature gradient and plasma lactate levels and inversely with the perfusion index. Impaired peripheral perfusion, as reflected by a low perfusion index or a high core-peripheral temperature gradient, predicted mortality in patients with severe malaria. CFH is associated with mean arterial pressure, SVRI, and peripheral perfusion in patients with severe malaria. This may be mediated through the nitric oxide scavenging potency of CFH, increasing basal vascular tone and impairing tissue perfusion.


Communication between Bacteria and Their Hosts.
It is clear that a dialogue is occurring between microbes and their hosts and that chemical signals are the language of this interkingdom communication. Microbial endocrinology shows that, through their long coexistence with animals and plants, microorganisms have evolved sensors for detecting eukaryotic hormones, which the microbe uses to determine that they are within proximity of a suitable host and to optimally time the expression of genes needed for host colonisation. It has also been shown that some prokaryotic chemical communication signals are recognized by eukaryotes. Deciphering what is being said during the cross-talk between microbe and host is therefore important, as it could lead to new strategies for preventing or treating bacterial infections.


Glycosome turnover in Leishmania major is mediated by autophagy.
Autophagy is a central process behind the cellular remodeling that occurs during differentiation of Leishmania, yet the cargo of the protozoan parasite's autophagosome is unknown. We have identified glycosomes, peroxisome-like organelles that uniquely compartmentalize glycolytic and other metabolic enzymes in Leishmania and other kinetoplastid parasitic protozoa, as autophagosome cargo. It has been proposed that the number of glycosomes and their content change during the Leishmania life cycle as a key adaptation to the different environments encountered. Quantification of RFP-SQL-labeled glycosomes showed that promastigotes of L. major possess ~20 glycosomes per cell, whereas amastigotes contain ~10. Glycosome numbers were significantly greater in promastigotes and amastigotes of autophagy-defective L. major Δatg5 mutants, implicating autophagy in glycosome homeostasis and providing a partial explanation for the previously observed growth and virulence defects of these mutants. Use of GFP-ATG8 to label autophagosomes showed glycosomes to be cargo in ~15% of them; glycosome-containing autophagosomes were trafficked to the lysosome for degradation. The number of autophagosomes increased 10-fold during differentiation, yet the percentage of glycosome-containing autophagosomes remained constant. This indicates that increased turnover of glycosomes was due to an overall increase in autophagy, rather than an upregulation of autophagosomes containing this cargo. Mitophagy of the single mitochondrion was not observed in L. major during normal growth or differentiation; however, mitochondrial remnants resulting from stress-induced fragmentation colocalized with autophagosomes and lysosomes, indicating that autophagy is used to recycle these damaged organelles. These data show that autophagy in Leishmania has a central role not only in maintaining cellular homeostasis and recycling damaged organelles but crucially in the adaptation to environmental change through the turnover of glycosomes.


The N terminus of pro-endothelial monocyte-activating polypeptide II (EMAP II) regulates its binding with the C terminus, arginyl-tRNA synthetase, and neurofilament light protein.
Pro-endothelial monocyte-activating polypeptide II (EMAP II), one component of the multi-aminoacyl tRNA synthetase complex, plays multiple roles in physiological and pathological processes of protein translation, signal transduction, immunity, lung development, and tumor growth. Recent studies have determined that pro-EMAP II has an essential role in maintaining axon integrity in central and peripheral neural systems where deletion of the C terminus of pro-EMAP II has been reported in a consanguineous Israeli Bedouin kindred suffering from Pelizaeus-Merzbacher-like disease. We hypothesized that the N terminus of pro-EMAP II has an important role in the regulation of protein-protein interactions. Using a GFP reporter system, we defined a putative leucine zipper in the N terminus of human pro-EMAP II protein (amino acid residues 1-70) that can form specific strip-like punctate structures. Through GFP punctum analysis, we uncovered that the pro-EMAP II C terminus (amino acids 147-312) can repress GFP punctum formation. Pulldown assays confirmed that the binding between the pro-EMAP II N terminus and its C terminus is mediated by a putative leucine zipper. Furthermore, the pro-EMAP II 1-70 amino acid region was identified as the binding partner of arginyl-tRNA synthetase, a polypeptide of the multi-aminoacyl tRNA synthetase complex. We also determined that the punctate GFP pro-EMAP II 1-70 amino acid aggregate colocalizes and binds to the neurofilament light subunit protein that is associated with pathologic neurofilament network disorganization and degeneration of motor neurons. These findings indicate the structure and binding interaction of pro-EMAP II protein and suggest a role of this protein in pathological neurodegenerative diseases.


N tau-methylhistidine release: contributions of rat skeletal muscle, GI tract, and skin.
The relative contributions of skeletal muscle, gastrointestinal tract, and skin to urinary N tau-methylhistidine (MH) excretion were estimated during in vitro studies using the rat hemicorpus preparation. After 0.5 h of perfusion, MH release into the perfusate was linear for 3 h and averaged 29.8 nmol . h-1 . 100 g hemicorpus-1. In vivo, 24-h urinary MH excretion averaged 37.3 nmol . h-1 . 100 g body wt-1. The ratio of soft tissue to skin weight is equal (3.2:1) in the whole rat and in the hemicorpus. The gastrointestinal tract released 16.0 nmol . h-1 . 100 g body wt-1 or approximately 41% of the total urinary MH excretion. Preparations perfused with or without skin showed modest differences in the rate of MH release that were not statistically significant. Skeletal muscle contains 89.8% of total body MH content, whereas gastrointestinal tract and skin contain 3.8 and 6.4%, respectively. Gastrointestinal tract actomyosin turns over rapidly with a fractional catabolic rate of 24%/day versus 1.4%/day for skeletal muscle actomyosin.


Bacterial dynamics associated with algal antibacterial substances during post harvest desiccation process of Sargassum stolonifolium Phang et Yoshida.
Brown algae of genus Sargassum are known to produce relatively higher amount of alginic acid. Optimal extraction of this algalcolloid for local consumption requires in-depth studies on post-harvest treatment of the algal fronds. Present investigation endeavors to establish the dynamics and inter-relationship of moisture content and bacteria found on the surface of the alga and alginic acid content during post-harvest desiccation of Sargassum stolonifolium Phang et Yoshida. Harvested fronds were subjected to desiccation for 31 days and bacterial dynamics were monitored with relation to moisture content and water activity index (a(w)). There was 85% decrease in moisture content, however, a(w) showed a more gradual decrease. Total bacterial count increased during the first week and attained maximal value on day 7. Thereafter, a drastic decrease was seen until day 14, followed by a gradual decline. Six species of bacteria were isolated and identified, i.e. Azomonas punctata, Azomonas sp., Escherichia coli, Micrococcus sp., Proteus vulgaris and Vibrio alginolyticus. Calculated ratios for increase in alginic acid content and decrease in moisture content were almost the same throughout the desiccation process, implying that extracellular alginase-producing bacteria did not use the alginic acid produced by the algae as its carbon source. It became apparent that drastic decrease in bacterial count after day 7 could not be attributed to salinity, moisture content, a(w) or lack of carbon source for the bacteria. The possible exposure of these bacteria to algal cell sap which is formed due to the rupture of algal cells was seen as the most likely reason for the drop in bacterial population. Scanning electron microscope (SEM) micrograph taken on day 10 of desiccation showed the presence of cracks and localities where bacteria were exposed to algal cell sap. In vitro antibacterial tests were carried out to verify the effect of algal extracts. Separation and purification of crude algal extracts via bioassay guided separation methodology revealed the identity of active compounds (i.e. gylcolipids and free fatty acids) involved in this inherently available antibacterial defense mechanism during algal desiccation.


Single-detector polarization-sensitive optical frequency domain imaging using high-speed intra A-line polarization modulation.
We demonstrate a novel high-speed polarization-sensitive optical frequency domain imaging system employing high-speed polarization modulation. Rapid and continuous polarization modulation of light prior to illumination of the sample is accomplished by shifting the frequency of one polarization eigenstate by an amount equal to one quarter of the digitization sampling frequency. This approach enables polarization-sensitive imaging with a single detection channel and overcomes artifacts that may arise from temporal variations of the birefringence in fiber-optic imaging probes and spatial variation of birefringence in the sample.


Thermal strategies vary with life history stage.
With both global surface temperatures and the incidence and intensity of extreme temperature events projected to increase, the assessment of species' sensitivity to chronic and acute changes in temperature has become crucial. Sensitivity predictions are based predominantly on adult responses, despite the fact that early life stages may be more vulnerable to thermal challenge. Here, we compared the sensitivity of different life history stages of the intertidal gastropod Littorina obtusata using thermal death time curves, which incorporate the intensity and duration of heat stress, and used these to calculate upper critical thermal limits (CTmax) and sensitivity to temperature change (z). Early (larval) life stages had both a lower CTmax and a lower z than adults, suggesting they are less good at withstanding short-term extreme thermal challenges but better able to survive moderate temperatures in the long term. This result supports the predicted trade-off between acute and chronic tolerance to thermal stress, and is consistent with the different thermal challenges that these stages encounter in the intertidal zone. We conclude that different life history stages employ different thermal strategies that may be adaptive. Our findings caution against the use of predictions of the impact of global warming that are based on only adult responses and, hence, which may underestimate vulnerability.


Arsenic in drinking water and lung cancer: a systematic review.
Exposure to inorganic arsenic via drinking water is a growing public health concern. We conducted a systematic review of the literature examining the association between arsenic in drinking water and the risk of lung cancer in humans. Towards this aim, we searched electronic databases for articles published through April 2006. Nine ecological studies, two case-control studies, and six cohort studies were identified. The majority of the studies were conducted in areas of high arsenic exposure (100 microg/L) such as southwestern Taiwan, the Niigata Prefecture, Japan, and Northern Chile. Most of the studies reported markedly higher risks of lung cancer mortality or incidence in high arsenic areas compared to the general population or a low arsenic exposed reference group. The quality assessment showed that, among the studies identified, only four assessed arsenic exposure at the individual level. Further, only one of the ecological studies presented results adjusted for potential confounders other than age; of the cohort and case-control studies, only one-half adjusted for cigarette smoking status in the analysis. Despite these methodologic limitations, the consistent observation of strong, statistically significant associations from different study designs carried out in different regions provide support for a causal association between ingesting drinking water with high concentrations of arsenic and lung cancer. The lung cancer risk at lower exposure concentrations remains uncertain.


Long-term behavioral and NMDA receptor effects of young-adult corticosterone treatment in BDNF heterozygous mice.
Psychiatric illnesses, such as schizophrenia, are most likely caused by an interaction between genetic predisposition and environmental factors, including stress during development. The neurotrophin, brain-derived neurotrophic factor (BDNF) has been implicated in this illness as BDNF levels are decreased in the brain of patients with schizophrenia. The aim of the present study was to assess the combined effect of reduced BDNF levels and postnatal stress, simulated by chronic young-adult treatment with the stress hormone, corticosterone. From 6 weeks of age, female and male BDNF heterozygous mice and their wild-type controls were chronically treated with corticosterone in their drinking water for 3 weeks. At 11 weeks of age, male, but not female BDNF heterozygous mice treated with corticosterone exhibited a profound memory deficit in the Y-maze. There were no differences between the groups in baseline prepulse inhibition (PPI), a measure of sensorimotor gating, or its disruption by treatment with MK-801. However, an increase in startle caused by MK-801 treatment was absent in male, but not female BDNF heterozygous mice, irrespective of corticosterone treatment. Analysis of protein levels of the NMDA receptor subunits NR1, NR2A, NR2B and NR2C, showed a marked increase of NR2B levels in the dorsal hippocampus of male BDNF heterozygous mice treated with corticosterone. In the ventral hippocampus, significantly reduced levels of NR2A, NR2B and NR2C were observed in male BDNF heterozygous mice. The NMDA receptor effects in hippocampal sub-regions could be related to the spatial memory deficits and the loss of the effect of MK-801 on startle in these mice, respectively. No significant changes in NMDA receptor subunit levels were observed in any of the female groups. Similarly, no significant changes in levels of BDNF or its receptor, TrkB, were found other than the expected reduced levels of BDNF in heterozygous mice. In conclusion, the data show differential interactive effects of reduced levels of BDNF expression and corticosterone treatment on spatial memory and startle in male and female mice, accompanied by significant, but region-specific changes in NMDA receptor subunit levels in the dorsal and ventral hippocampus. These results could be important for our understanding of the interaction of neurodevelopmental stress and BDNF deficiency in cognitive and anxiety-related symptoms of psychiatric illnesses, such as schizophrenia.


Identical skin toxins by convergent molecular adaptation in frogs.
The Tree of Life is rife with adaptive convergences at all scales and biological levels of complexity. However, natural selection is not likely to result in the independent evolution of identical gene products. Here we report such a striking example of evolutionary convergence in the toxic skin secretions of two distantly related frog lineages. Caeruleins are important decapeptides in pharmacological and clinical research [1] and are commonly believed to represent a single evolutionary class of peptides [2-4]. Instead, our phylogenetic analyses combining transcriptome and genome data reveal that independently evolved precursor genes encode identical caeruleins in Xenopus and Litoria frogs. The former arose by duplication from the cholecystokinin (cck) gene, whereas the latter was derived from the gastrin gene. These hormone genes that are involved in many physiological processes diverged early in vertebrate evolution, after a segmental duplication during the Cambrian period. Besides implicating convergent mutations of the peptide-encoding sequence, recurrent caerulein origins entail parallel shifts of expression from the gut-brain axis to skin secretory glands. These results highlight extreme structural convergence in anciently diverged genes as an evolutionary mechanism through which recurrent adaptation is attained across large phylogenetic distances.


Subconjunctival bevacizumab to augment trabeculectomy with mitomycin C in the management of failed glaucoma surgery.
To provide a feasible solution to the problem of failed glaucoma surgery. The aim was to evaluate the efficacy and safety of the additional effects of a combined surgical approach. This approach augments the application of trabeculectomy with mitomycin C (MMC) by adding subconjunctival bevacizumab injection. The results were compared with those of trabeculectomy with only adjunctive MMC. A randomized controlled prospective clinical trial included 28 eyes diagnosed with failed scarred bleb of a previous trabeculectomy. The eyes were divided into two equal groups: combined group A, "trabeculectomy with adjunctive MMC and subconjunctival bevacizumab," and control group B, "trabeculectomy with adjunctive MMC only." The main outcome results included the cumulative probability of surgical success, intraocular pressure (IOP) values, and number of IOP-lowering medications needed to achieve the target IOP. Group A achieved a cumulative probability of complete success of 0.769 and of qualified success of 0.231 at the end of the 24 month study period; group B achieved cumulative probabilities of 0.538 and 0.308, respectively. Group A achieved a lower mean IOP value than group B, with fewer antiglaucoma drugs at all postoperative visits, but this lower value did not reach a statistically significant level (P>0.05). There was no statistically significant difference between both groups regarding best corrected visual acuity, visual field parameters, operative and/or postoperative complications, and additional interventions. No significant adverse effects were caused by this combined approach. Bevacizumab was not found to add much to the favorable long-term outcome of conventional trabeculectomy with MMC as a solution to the problem of scarred failed bleb.


Child mental health and maternal depression history in Pakistan.
We address the significant gaps in knowledge of prevalence and correlates of child mental health (CMH) problems outside of high income countries. We describe the prevalence of CMH problems and their correlates with a focus on the association with maternal depression in a sample of seven-year-old children in rural Pakistan. This study was nested in a long-term follow-up of a perinatal depression intervention together with a reference group of non-depressed women, yielding a population representative sample. The Total Difficulties (TD) and component scores of the Strength and Difficulties Questionnaire (SDQ) were used to measure emotional and behavioral difficulties. The mean SDQ TD score was 10.6 (standard deviation = 8.3), with 12.5 % of children categorized as "abnormal" using standard cutoffs. Boys had a roughly 1 point higher (worse) SDQ TD score than girls (p value = 0.04). Children of mothers who were depressed prenatally as well as currently had SDQ TD scores 2.87 points higher than children whose mothers were not depressed at either time point (p value < 0.01). This association was stronger for boys. There was no evidence of elevated SDQ TD score among children whose mothers were depressed only prenatally or only currently. Some deviations from this pattern were observed with specific components of the SDQ. In this low resource, South Asian setting, we found evidence of elevated levels of emotional and behavioral problems, highlighting the need for effective interventions. Given the strong association of CMH with maternal depression, any intervention efforts should give strong consideration to maternal mental health.


Recent advances in the design, synthesis, and biological evaluation of selective DYRK1A inhibitors: a new avenue for a disease modifying treatment of Alzheimer's?
With 24.3 million people affected in 2005 and an estimated rise to 42.3 million in 2020, dementia is currently a leading unmet medical need and costly burden on public health. Seventy percent of these cases have been attributed to Alzheimer's disease (AD), a neurodegenerative pathology whose most evident symptom is a progressive decline in cognitive functions. Dual specificity tyrosine phosphorylation regulated kinase-1A (DYRK1A) is important in neuronal development and plays a variety of functional roles within the adult central nervous system. The DYRK1A gene is located within the Down syndrome critical region (DSCR) on human chromosome 21 and current research suggests that overexpression of DYRK1A may be a significant factor leading to cognitive deficits in people with Alzheimer's disease (AD) and Down syndrome (DS). Currently, treatment options for cognitive deficiencies associated with Down syndrome, as well as Alzheimer's disease, are extremely limited and represent a major unmet therapeutic need. Small molecule inhibition of DYRK1A activity in the brain may provide an avenue for pharmaceutical intervention of mental impairment associated with AD and other neurodegenerative diseases. We herein review the current state of the art in the development of DYRK1A inhibitors.


Important miRs of pathways in different tumor types.
We computationally determined miRs that are significantly connected to molecular pathways by utilizing gene expression profiles in different cancer types such as glioblastomas, ovarian and breast cancers. Specifically, we assumed that the knowledge of physical interactions between miRs and genes indicated subsets of important miRs (IM) that significantly contributed to the regression of pathway-specific enrichment scores. Despite the different nature of the considered cancer types, we found strongly overlapping sets of IMs. Furthermore, IMs that were important for many pathways were enriched with literature-curated cancer and differentially expressed miRs. Such sets of IMs also coincided well with clusters of miRs that were experimentally indicated in numerous other cancer types. In particular, we focused on an overlapping set of 99 overall important miRs (OIM) that were found in glioblastomas, ovarian and breast cancers simultaneously. Notably, we observed that interactions between OIMs and leading edge genes of differentially expressed pathways were characterized by considerable changes in their expression correlations. Such gains/losses of miR and gene expression correlation indicated miR/gene pairs that may play a causal role in the underlying cancers.


Positive Selection Drives Preferred Segment Combinations during Influenza Virus Reassortment.
Influenza A virus (IAV) has a segmented genome that allows for the exchange of genome segments between different strains. This reassortment accelerates evolution by breaking linkage, helping IAV cross species barriers to potentially create highly virulent strains. Challenges associated with monitoring the process of reassortment in molecular detail have limited our understanding of its evolutionary implications. We applied a novel deep sequencing approach with quantitative analysis to assess the in vitro temporal evolution of genomic reassortment in IAV. The combination of H1N1 and H3N2 strains reproducibly generated a new H1N2 strain with the hemagglutinin and nucleoprotein segments originating from H1N1 and the remaining six segments from H3N2. By deep sequencing the entire viral genome, we monitored the evolution of reassortment, quantifying the relative abundance of all IAV genome segments from the two parent strains over time and measuring the selection coefficients of the reassorting segments. Additionally, we observed several mutations coemerging with reassortment that were not found during passaging of pure parental IAV strains. Our results demonstrate how reassortment of the segmented genome can accelerate viral evolution in IAV, potentially enabled by the emergence of a small number of individual mutations.


Distribution of genetic variation underlying adult migration timing in steelhead of the Columbia River basin.
Fish migrations are energetically costly, especially when moving between freshwater and saltwater, but are a viable strategy for Pacific salmon and trout (Oncorhynchus spp.) due to the advantageous resources available at various life stages. Anadromous steelhead (O. mykiss) migrate vast distances and exhibit variation for adult migration phenotypes that have a genetic basis at candidate genes known as greb1L and rock1. We examined the distribution of genetic variation at 13 candidate markers spanning greb1L, intergenic, and rock1 regions versus 226 neutral markers for 113 populations (n = 9,471) of steelhead from inland and coastal lineages in the Columbia River. Patterns of population structure with neutral markers reflected genetic similarity by geographic region as demonstrated in previous studies, but candidate markers clustered populations by genetic variation associated with adult migration timing. Mature alleles for late migration had the highest frequency overall in steelhead populations throughout the Columbia River, with only 9 of 113 populations that had a higher frequency of premature alleles for early migration. While a single haplotype block was evident for the coastal lineage, we identified multiple haplotype blocks for the inland lineage. The inland lineage had one haplotype block that corresponded to candidate markers within the greb1L gene and immediately upstream in the intergenic region, and the second block only contained candidate markers from the intergenic region. Haplotype frequencies had similar patterns of geographic distribution as single markers, but there were distinct differences in frequency between the two haplotype blocks for the inland lineage. This may represent multiple recombination events that differed between lineages where phenotypic differences exist between freshwater entry versus arrival timing as indicated by Micheletti et al. (2018a). Redundancy analyses were used to model environmental effects on allelic frequencies of candidate markers, and significant variables were migration distance, temperature, isothermality, and annual precipitation. This study improves our understanding of the spatial distribution of genetic variation underlying adult migration timing in steelhead as well as associated environmental factors and has direct conservation and management implications.


Head CT for nontrauma patients in the emergency department: clinical predictors of abnormal findings.
To identify predictors of clinically important abnormal findings in computed tomography (CT) images of the head among emergency department (ED) patients without a history of trauma. Approval was obtained from the institutional research ethics board, and informed consent from patients was not required. This study was a retrospective review of consecutive unenhanced head CT examinations in patients aged 18 years or older who did not have trauma or known intracranial pathologic processes in ED from January 2004 through June 2006. Multivariable logistic regression was used to identify predictors of clinically important abnormal CT findings in the derivation cohort (CT examinations from January 1, 2004, through August 15, 2005), and the reproducibility of findings in a validation cohort (all subsequent CT scans through to June 30, 2006) was assessed. The strength of association of each variable was expressed with clinically important abnormal CT findings as adjusted odds ratio (OR) and 95% confidence interval (CI). Of 29 469 consecutive head CT images performed at a single institution between January 1, 2004, and June 30, 2006, 3967 were eligible for this study. Of the CT images in these patients, 548 (13.8%) revealed clinically important abnormalities. Six independent clinical predictors of important abnormal findings on head CT were identified: age (adjusted OR per 10-year increase: 1.17; 95% CI: 1.08, 1.28), focal neurologic deficit (adjusted OR: 5.39; 95% CI: 3.90, 7.47), altered mental status (adjusted OR: 2.32; 95% CI: 1.66, 3.25), history of malignancy (adjusted OR: 4.11; 95% CI: 2.28, 7.42), nausea and/or vomiting (adjusted OR: 2.22; 95% CI: 1.14, 4.33), and derangements in coagulation profile (adjusted OR: 1.91; 95% CI: 1.07, 3.41). This study identified several potential clinical predictors of abnormal head CT findings in ED patients who did not sustain trauma. Prospective validation of a clinical prediction rule in this population is warranted.


Systematic review: the effects of autologous stem cell therapy for patients with liver disease.
As morbidity and mortality from liver disease continues to rise, new strategies are necessary. Liver transplantation is not only an expensive resource committing the patient to lifelong immunosuppression but also suitable donor organs are in short supply. Against this background, autologous stem cell therapy has emerged as a potential treatment option. To evaluate if it is possible to make a judgement on the safety, feasibility and effect of autologous stem cell therapy for patients with liver disease. MEDLINE and EMBASE were searched up until July 2013 to identify studies where autologous stem cell therapy was administered to patients with liver disease. Of 1668 studies identified, 33 were eligible for inclusion evaluating a median sample size of 10 patients for a median follow-up of 6 months. Although there was marked heterogeneity between studies with regards to type, dose and route of delivery of stem cell, the treatment was shown to be safe and feasible largely when a peripheral route of administration was used. Of the studies which also looked at biochemical outcome, statistically significant improvement in liver function tests was seen in 16 studies post-treatment. Although autologous stem cell therapy is a much needed possibility in the treatment of liver disease, further robust clinical trials and collaborative protocols are required.


Feature-based alignment of volumetric multi-modal images.
This paper proposes a method for aligning image volumes acquired from different imaging modalities (e.g. MR, CT) based on 3D scale-invariant image features. A novel method for encoding invariant feature geometry and appearance is developed, based on the assumption of locally linear intensity relationships, providing a solution to poor repeatability of feature detection in different image modalities. The encoding method is incorporated into a probabilistic feature-based model for multi-modal image alignment. The model parameters are estimated via a group-wise alignment algorithm, that iteratively alternates between estimating a feature-based model from feature data, then realigning feature data to the model, converging to a stable alignment solution with few pre-processing or pre-alignment requirements. The resulting model can be used to align multi-modal image data with the benefits of invariant feature correspondence: globally optimal solutions, high efficiency and low memory usage. The method is tested on the difficult RIRE data set of CT, T1, T2, PD and MP-RAGE brain images of subjects exhibiting significant inter-subject variability due to pathology.


Differential alterations in the activity of matrix metalloproteinases within the nervous tissue of dogs in distinct manifestations of visceral leishmaniasis.
Canine visceral leishmaniasis is an important emerging disease with a multitude of clinical symptoms, including neurological alterations. Matrix metalloproteinases (MMP) are proteases implicated with the extracellular matrix remodelling and, within the central nervous system, these enzymes are involved with blood-brain-barrier disruption and inflammation. To establish the involvement of MMP-2 and -9 within the nervous tissue of dogs with spontaneous visceral leishmaniasis, fragments of nervous tissue from oligosymptomatic (n=9), symptomatic (n=8), neurological (n=12) and normal dogs (n=8) were subjected to zymographic and to immunohistochemical analysis. Immunohistochemistry evidenced MMP-2 staining in inflammatory cells inside and outside blood vessels. MMP-9 was found in endothelial cells and in the ependyma. Zymographic evaluation revealed only the latent forms of MMP-2 and -9 within the nervous tissue. ProMMP-9 activity in the infected animals was found higher than the normal dogs, but with no difference among the infected dogs. Oligosymptomatic dogs presented the highest proMMP-2 activity, followed by the symptomatic and then, by the neurological and the normal dogs. In summary, the nervous tissue compartment seems to be preserved in dogs with VL, due to the absence of active MMPs, even though the elevated levels of proMMP-2 and -9 would indicate a pro-inflammatory state in the brain.


Biomimetic collagen/elastin meshes for ventral hernia repair in a rat model.
Ventral hernia repair remains a major clinical need. Herein, we formulated a type I collagen/elastin crosslinked blend (CollE) for the fabrication of biomimetic meshes for ventral hernia repair. To evaluate the effect of architecture on the performance of the implants, CollE was formulated both as flat sheets (CollE Sheets) and porous scaffolds (CollE Scaffolds). The morphology, hydrophylicity and in vitro degradation were assessed by SEM, water contact angle and differential scanning calorimetry, respectively. The stiffness of the meshes was determined using a constant stretch rate uniaxial tensile test, and compared to that of native tissue. CollE Sheets and Scaffolds were tested in vitro with human bone marrow-derived mesenchymal stem cells (h-BM-MSC), and finally implanted in a rat ventral hernia model. Neovascularization and tissue regeneration within the implants was evaluated at 6weeks, by histology, immunofluorescence, and q-PCR. It was found that CollE Sheets and Scaffolds were not only biomechanically sturdy enough to provide immediate repair of the hernia defect, but also promoted tissue restoration in only 6weeks. In fact, the presence of elastin enhanced the neovascularization in both sheets and scaffolds. Overall, CollE Scaffolds displayed mechanical properties more closely resembling those of native tissue, and induced higher gene expression of the entire marker genes tested, associated with de novo matrix deposition, angiogenesis, adipogenesis and skeletal muscles, compared to CollE Sheets. Altogether, this data suggests that the improved mechanical properties and bioactivity of CollE Sheets and Scaffolds make them valuable candidates for applications of ventral hernia repair. Due to the elevated annual number of ventral hernia repair in the US, the lack of successful grafts, the design of innovative biomimetic meshes has become a prime focus in tissue engineering, to promote the repair of the abdominal wall, avoid recurrence. Our meshes (CollE Sheets and Scaffolds) not only showed promising mechanical performance, but also allowed for an efficient neovascularization, resulting in new adipose and muscle tissue formation within the implant, in only 6weeks. In addition, our meshes allowed for the use of the same surgical procedure utilized in clinical practice, with the commercially available grafts. This study represents a significant step in the design of bioactive acellular off-the-shelf biomimetic meshes for ventral hernia repair.


Polymorphisms of matrix metalloproteinase gene and adiposity indices in European children: results of the IDEFICS study.
We investigated the relationship between matrix metalloproteinase 3 (MMP3) polymorphisms and adiposity indices in European children of the IDEFICS (Identification and Prevention of Dietary- and Lifestyle-Induced Health Effects in Children and Infants) project. A total of 16 224 Caucasian children (2-9 years) were recruited into a population-based survey from eight European countries. In all, 4540 children were randomly selected for genetic studies (T0); 3238 children were re-examined 2 years later (T1). Anthropometric measures were collected by standardized protocols at T0 and T1. Six variants of MMP3 gene were genotyped. Homozygotes for the variant A allele of rs646910 and for the H3 haplotype had higher hip circumference (P=0.002 and 0.001; age, sex and country adjusted) at T0. The association remained significant after false discovery rate (FDR) correction. At T1, subjects carrying rs646910 A/A genotype or H3/H3 diplotype showed significantly higher values of body mass index, waist and hip circumference and sum of tricipital and subscapular skinfolds, all associations remaining significant after FDR correction (P=0.020-0.048). We showed for the first time an association between the MMP3 rs646910 variant and indices of adiposity in European children, highlighting the involvement of metalloproteinase genes in adipose tissue remodeling and growth.


The Research Agenda for General Practice/Family Medicine and Primary Health Care in Europe. Part 1. Background and methodology.
At the WONCA Europe conference 2009 the recently published 'Research Agenda for General Practice/Family Medicine and Primary Health Care in Europe' was presented. The Research Agenda is a background paper and reference manual for GPs/ family doctors, researchers and policy makers, providing advocacy of general practice/family medicine GP/FM in Europe. The Research Agenda summarizes the evidence relating to the core competencies and characteristics of the WONCA Europe definition of GP/FM, and its meaning for researchers and policy makers. Evidence gaps and research needs are pointed out to provide a basis for planning research for which there is a need and for action that may influence health and research policy, i.e. applying/lobbying for research funds. WONCA Europe and its associated networks and special interest groups could consider the agenda's research priorities when planning future conferences, courses, or projects, and for funding purposes. The European Journal of General Practice will publish a series of articles based on this document. In this first article, background, objectives, methodology and relevant literature are discussed. In subsequent articles, the results will be presented.


An integrated, directed mass spectrometric approach for in-depth characterization of complex peptide mixtures.
LC-MS/MS has emerged as the method of choice for the identification and quantification of protein sample mixtures. For very complex samples such as complete proteomes, the most commonly used LC-MS/MS method, data-dependent acquisition (DDA) precursor selection, is of limited utility. The limited scan speed of current mass spectrometers along with the highly redundant selection of the most intense precursor ions generates a bias in the pool of identified proteins toward those of higher abundance. A directed LC-MS/MS approach that alleviates the limitations of DDA precursor ion selection by decoupling peak detection and sequencing of selected precursor ions is presented. In the first stage of the strategy, all detectable peptide ion signals are extracted from high resolution LC-MS feature maps or aligned sets of feature maps. The selected features or a subset thereof are subsequently sequenced in sequential, non-redundant directed LC-MS/MS experiments, and the MS/MS data are mapped back to the original LC-MS feature map in a fully automated manner. The strategy, implemented on an LTQ-FT MS platform, allowed the specific sequencing of 2,000 features per analysis and enabled the identification of more than 1,600 phosphorylation sites using a single reversed phase separation dimension without the need for time-consuming prefractionation steps. Compared with conventional DDA LC-MS/MS experiments, a substantially higher number of peptides could be identified from a sample, and this increase was more pronounced for low intensity precursor ions.


Invasive, gangrenous mucormycosis of arm: A fatal opportunistic infection in a highly immunocompromised host.
Opportunistic fungal infections occur predominantly in immunocompromised (IC) patients. Mucormycosis has the highest mortality among fungal infections. The organism is ubiquitous in its presence. The infection is commonly acquired by inhalation of fungal spores or by inoculation by direct trauma. Rhinocerebral and pulmonary mucormycosis present commonly with high mortality rates. Cutaneous mucormycosis (CM) is rarely reported and usually presents in two forms, superficial and disseminated. The superficial infection occurs commonly in immunocompetent hosts, and it can sometimes turn angioinvasive and become a fatal and very aggressive disseminated disease, especially in IC hosts. Coexistant risk factors increase the mortality rate. We report a case of posttraumatic, highly lethal, angioinvasive, and gangrenous type of CM of the arm in an IC patient. Diabetic ketoacidosis and cirrhosis due to chronic alcoholic liver disease were the risk factors.


[Excess respiratory diseases in children and elderly people in a community of Santiago with high particulate air pollution].
The relationship between air pollution and health damage has been sufficiently documented. In station "R" of the air quality monitoring system, located in a community of Metropolitan Santiago (Cerro Navia), the Chilean standard of 150 microg/m3, averaged in 24 hours, for particles with a diameter of 10 micrometers or less (PM10), has been exceeded more days than in the rest of the city stations. To investigate if the population living near that station has a higher proportion of lower respiratory infections than the Metropolitan Region (MR) as a whole. An outpatient clinic located near station "R" (Centro Albertz), was implemented as a sentinel center according to UNICEF methodology, used since 1992 by the Acute Respiratory Infections National Program. Daily information was collected between May and December 2004. Monitoring data included total number of consults by children less than 15 years old for lower respiratory tract infections, pneumonia, obstructive bronchitis syndrome in children and by adults over 64 years old for lower airway disease, chronic obstructive pulmonary disease (COPD), and pneumonia. Results were compared with those of the rest of MR. Compared with the MR, children from the sentinel clinic had a significantly higher proportion of consults for obstructive bronchial syndrome (20.1% and 26.4% respectively, in p<0.01) and pneumonia (1.3 and 2.7% respectively, p<0.01). In the elderly, the average consults for lower airway disease were 17% in the sentinel clinic and 12.2% in MR (p<0.04). Children and elderly subjects at the sentinel clinic had a significantly higher proportion of respiratory infections (pneumonia and obstructive bronchial syndrome in children and lower airway disease in the elderly) as compared to the Metropolitan Region.


Allopurinol Effects on Residual Renal Function in End-Stage Renal Disease Patients Undergoing Peritoneal Dialysis: Randomized Controlled Trial.
There is increasing evidence to show that hyperuricemia may have a pathogenic role in the progression of renal diseases. We performed a prospective, randomized, controlled trial to investigate the renal effects of allopurinol treatment in hyperuricemic patients with end-stage renal disease (ESRD) who undergo peritoneal dialysis. This was a unicenter, randomized, controlled clinical trial conducted in "Alzahra Hospital, Isfahan, Iran." Patients were randomly assigned into treatment or control group. Treatment-group patients were administered a starting allopurinol dose of 100 mg/day. The dose was adjusted according to serum uric acid level, aiming to maintain uric acid levels within the normal range. Participants were followed up for 6 months after receiving the medicine. Residual renal function (RRF) was assessed by measuring the renal component of Kt/V urea and estimating the patient's glomerular filtration rate (GFR) by calculating the mean of urea and creatinine clearance. In addition, systolic and diastolic blood pressure and serum level of creatinine were measured every 3 months during the follow-up period. Eighty patients were enrolled in the study and divided into two groups, including 40 ESRD patients receiving allopurinol and 40 ESRD did not receive allopurinol and considered as the control group. GFR measurements showed that there was not a significant difference between patients' RRF of two groups. However, allopurinol group had higher RRF than the control group during the follow-up period. Evaluating RRF by Kt/V showed the same results. Our study demonstrated significant effects of allopurinol on decreasing serum levels of uric acid in ESRD patients undergoing peritoneal dialysis. On the other hand, renal residual function of patients under treatment with allopurinol was better than the control group. We recommend that further studies should be conducted on the effects of allopurinol with greater sample size and longer time of follow-up.