Camera Array for Multi-Spectral Imaging.
Recently, many new applications arose for multispectral and hyper-spectral imaging. Besides modern biometric systems for identity verification, also agricultural and medical applications came up, which measure the health condition of plants and humans. Despite the growing demand, the acquisition of multi-spectral data is up to the present complicated. Often, expensive, inflexible, or low resolution acquisition setups are only obtainable for specific professional applications. To overcome these limitations, a novel camera array for multi-spectral imaging is presented in this article for generating consistent multispectral videos. As differing spectral images are acquired at various viewpoints, a geometrically constrained multi-camera sensor layout is introduced, which enables the formulation of novel registration and reconstruction algorithms to globally set up robust models. On average, the novel acquisition approach achieves a gain of 2.5 dB PSNR compared to recently published multi-spectral filter array imaging systems. At the same time, the proposed acquisition system ensures not only a superior spatial, but also a high spectral, and temporal resolution, while filters are flexibly exchangeable by the user depending on the application. Moreover, depth information is generated, so that 3D imaging applications, e.g., for augmented or virtual reality, become possible. The proposed camera array for multi-spectral imaging can be set up using off-the-shelf hardware, which allows for a compact design and employment in, e.g., mobile devices or drones, while being cost-effective.


Home as a place of caring and wellbeing? A qualitative study of informal carers and caring networks lived experiences of providing in-home end-of-life care.
Although the burden of caring is well described, the value of home as a potential place of wellbeing and support for informal caring networks when providing end-of-life care is not well recognised. Interviews and focus groups with 127 primary carers and members of informal care networks revealed their collaborative stories about caring for a dying person at home. Four themes emerged from the data: home as a place of comfort and belonging; places of social connection and collaborative caring; places of connection to nature and the non-human; places of achievement and triumph. When support is available, nurturing carer wellbeing may be best achieved at home.


Laparoscopic sleeve gastrectomy: outcomes at a military training center.
Laparoscopic sleeve gastrectomy (LSG) has gained support as a single-staged and stand-alone bariatric procedure. Reports of excess weight loss of 35 to 83 per cent, reduction in comorbidities, and decreased operative morbidity have garnered support for LSG. This study represents an initial outcome analysis of LSG performed solely at a military treatment center. This study is a retrospective analysis of all patients receiving LSG at Dwight D. Eisenhower Army Medical Center from September 2007 to December 2009. The patients were planned for a stand-alone procedure. One hundred and fifteen patients received LSG over this time period with a mean body mass index of 45.5 +/- 6.2 (range 35.1-58.3). The average age was 47.4 +/- 12.5 years. Diabetes mellitus was seen in 47 per cent and 68 per cent of patients had hypertension. The mean and median length of operation was 124 +/- 48 and 115.5 minutes. The mean percentage of excess weight loss was 16.6 +/- 6.40 per cent at 1 month, 31.5 +/- 7.6 per cent at 3 months, 41.2 +/- 13.9 per cent at 6 months, and 53.7 +/- 12.5 per cent at 1 year from surgery. One or more of patient's preoperative diabetic or hypertensive medications were improved postoperatively in 18.7 per cent and 16.3 per cent, respectively. Incidence of major complications occurred in 4.35 per cent of patients in this study to include four leaks (3.4%), one death (0.87%), and 10 readmissions. Midterm analysis of outcomes related to LSG as a single-stage bariatric procedure is promising as long-term outcome data is collected; the efficacy of this procedure as a sole bariatric procedure will continue to be borne out.


Metagenomic analysis reveals adaptations to a cold-adapted lifestyle in a low-temperature acid mine drainage stream.
An acid mine drainage (pH 2.5-2.7) stream biofilm situated 250 m below ground in the low-temperature (6-10°C) Kristineberg mine, northern Sweden, contained a microbial community equipped for growth at low temperature and acidic pH. Metagenomic sequencing of the biofilm and planktonic fractions identified the most abundant microorganism to be similar to the psychrotolerant acidophile, Acidithiobacillus ferrivorans. In addition, metagenome contigs were most similar to other Acidithiobacillus species, an Acidobacteria-like species, and a Gallionellaceae-like species. Analyses of the metagenomes indicated functional characteristics previously characterized as related to growth at low temperature including cold-shock proteins, several pathways for the production of compatible solutes and an anti-freeze protein. In addition, genes were predicted to encode functions related to pH homeostasis and metal resistance related to growth in the acidic metal-containing mine water. Metagenome analyses identified microorganisms capable of nitrogen fixation and exhibiting a primarily autotrophic lifestyle driven by the oxidation of the ferrous iron and inorganic sulfur compounds contained in the sulfidic mine waters. The study identified a low diversity of abundant microorganisms adapted to a low-temperature acidic environment as well as identifying some of the strategies the microorganisms employ to grow in this extreme environment.


Calibration-free imaging through a multicore fiber using speckle scanning microscopy.
The images produced by multicore endoscopes are pixelated, and their resolution is limited by the core-to-core spacing. Lenses can be used to improve the resolution, but this reduces the field of view proportionally. Lensless endoscopy through multicore fibers can be achieved by using wavefront shaping techniques. This requires a calibration step, and the conformation of the fiber must remain constant over time. Here we demonstrate that, without a calibration step and in the presence of core-to-core coupling, we can obtain fluorescence images with a resolution better than the core-to-core spacing. This is accomplished by taking advantage of the memory effect present in these kinds of fibers.


Integrative genome-wide chromatin signature analysis using finite mixture models.
Regulation of gene expression has been shown to involve not only the binding of transcription factor at target gene promoters but also the characterization of histone around which DNA is wrapped around. Some histone modification, for example di-methylated histone H3 at lysine 4 (H3K4me2), has been shown to bind to promoters and activate target genes. However, no clear pattern has been shown to predict human promoters. This paper proposed a novel quantitative approach to characterize patterns of promoter regions and predict novel and alternative promoters. We utilized high-throughput data generated using chromatin immunoprecipitation methods followed by massively parallel sequencing (ChIP-seq) technology on RNA Polymerase II (Pol-II) and H3K4me2. Common patterns of promoter regions are modeled using a mixture model involving double-exponential and uniform distributions. The fitted model obtained were then used to search for regions displaying similar patterns over the entire genome to find novel and alternative promoters. Regions with high correlations with the common patterns are identified as putative novel promoters. We used this proposed algorithm, RNA-seq data and several transcripts databases to find alternative promoters in MCF7 (normal breast cancer) cell line. We found 7,235 high-confidence regions that display the identified promoter patterns. Of these, 4,167 regions (58%) can be mapped to RefSeq regions. 2,444 regions are in a gene body or overlap with transcripts (non-coding RNAs, ESTs, and transcripts that are predicted by RNA-seq data). Some of these maybe potential alternative promoters. We also found 193 regions that map to enhancer regions (represented by androgen and estrogen receptor binding sites) and other regulatory regions such as CTCF (CCCTC binding factor) and CpG island. Around 5% (431 regions) of these correlated regions do not overlap with any transcripts or regulatory regions suggesting that these might be potential new promoters or markers for other annotation which are currently undiscovered.


Divergent effects of resistance and endurance exercise on plasma bile acids, FGF19, and FGF21 in humans.
Exercise has profound pleiotropic health benefits, yet the underlying mechanisms remain incompletely understood. Endocrine FGF21, bile acids (BAs), and BA-induced FGF19 have emerged as metabolic signaling molecules. Here, we investigated if dissimilar modes of exercise, resistance exercise (RE) and endurance exercise (EE), regulate plasma BAs, FGF19, and FGF21 in humans. Ten healthy, moderately trained males were enrolled in a randomized crossover study of 1 hour of bicycling at 70% of VO2peak (EE) and 1 hour of high-volume RE. Hormones and metabolites were measured in venous blood and sampled before and after exercise and at 15, 30, 60, 90, 120, and 180 minutes after exercise. We observed exercise mode-specific changes in plasma concentrations of FGF19 and FGF21. Whereas FGF19 decreased following RE (P < 0.001), FGF21 increased in response to EE (P < 0.001). Total plasma BAs decreased exclusively following RE (P < 0.05), but the composition of BAs changed in response to both types of exercise. Notably, circulating levels of the potent TGR5 receptor agonist, lithocholic acid, increased with both types of exercise (P < 0.001). This study reveals divergent effects of EE and RE on circulating concentrations of the BA species, FGF19, and FGF21. We identify temporal relationships between decreased BA and FGF19 following RE and a sharp disparity in FGF21 concentrations, with EE eliciting a clear increase parallel to that of glucagon. The Novo Nordisk Foundation (NNF17OC0026114) and the Lundbeck Foundation (R238-2016-2859).


Melatonin is a potential inhibitor of ovarian cancer: molecular aspects.
Ovarian cancer is one of the most common causes of morbidity related to gynecologic malignancies. Possible risk factors are including hereditary ovarian cancer, obesity, diabetes mellitus, alcohol consumption, aging, and smoking. Various molecular signaling pathways including inflammation, oxidative stress, apoptosis and angiogenesis are involved in this progression of ovarian cancer. Standard treatments for recently diagnosed patients are Surgery and chemotherapy such as co-treatment with other drugs such that the exploitation of neoadjuvant chemotherapy is expanding. Melatonin (N-acetyl-5-methoxy-tryptamine), an endogenous agent secreted from the pineal gland, has anti-carcinogenic features, such as regulation of estradiol production, cell cycle modulation, stimulation of apoptosis as well as anti-angiogenetic properties, anti-inflammatory activities, significant antioxidant effects and modulation of various immune system cells and cytokines. Multiple studies have shown the significant beneficial roles of melatonin in various types of cancers including ovarian cancer. This paper aims to shed light on the roles of melatonin in ovarian cancer treatment from the standpoint of the molecular aspects.


A Randomized, Controlled Trial of Fusion Surgery for Lumbar Spinal Stenosis.
The efficacy of fusion surgery in addition to decompression surgery in patients who have lumbar spinal stenosis, with or without degenerative spondylolisthesis, has not been substantiated in controlled trials. We randomly assigned 247 patients between 50 and 80 years of age who had lumbar spinal stenosis at one or two adjacent vertebral levels to undergo either decompression surgery plus fusion surgery (fusion group) or decompression surgery alone (decompression-alone group). Randomization was stratified according to the presence of preoperative degenerative spondylolisthesis (in 135 patients) or its absence. Outcomes were assessed with the use of patient-reported outcome measures, a 6-minute walk test, and a health economic evaluation. The primary outcome was the score on the Oswestry Disability Index (ODI; which ranges from 0 to 100, with higher scores indicating more severe disability) 2 years after surgery. The primary analysis, which was a per-protocol analysis, did not include the 14 patients who did not receive the assigned treatment and the 5 who were lost to follow-up. There was no significant difference between the groups in the mean score on the ODI at 2 years (27 in the fusion group and 24 in the decompression-alone group, P=0.24) or in the results of the 6-minute walk test (397 m in the fusion group and 405 m in the decompression-alone group, P=0.72). Results were similar between patients with and those without spondylolisthesis. Among the patients who had 5 years of follow-up and were eligible for inclusion in the 5-year analysis, there were no significant differences between the groups in clinical outcomes at 5 years. The mean length of hospitalization was 7.4 days in the fusion group and 4.1 days in the decompression-alone group (P<0.001). Operating time was longer, the amount of bleeding was greater, and surgical costs were higher in the fusion group than in the decompression-alone group. During a mean follow-up of 6.5 years, additional lumbar spine surgery was performed in 22% of the patients in the fusion group and in 21% of those in the decompression-alone group. Among patients with lumbar spinal stenosis, with or without degenerative spondylolisthesis, decompression surgery plus fusion surgery did not result in better clinical outcomes at 2 years and 5 years than did decompression surgery alone. (Funded by an Uppsala institutional Avtal om Läkarutbildning och Forskning [Agreement concerning Cooperation on Medical Education and Research] and others; Swedish Spinal Stenosis Study ClinicalTrials.gov number, NCT01994512.).


3D velocity field and flow profile reconstruction from arbitrarily sampled Doppler ultrasound data.
With the need for adequate analysis of blood flow dynamics, different maging modalities have been developed to measure varying blood velocities over time. Due to its numerous advantages, Doppler ultrasound sonography remains one of the most widely used techniques in clinical routine, but requires additional preprocessing to recover 3D velocity information. Despite great progress in the last years, recent approaches do not jointly consider spatial and temporal variation in blood flow. In this work, we present a novel gating- and compounding-free method to simultaneously reconstruct a 3D velocity field and a temporal flow profile from arbitrarily sampled Doppler ultrasound measurements obtained from multiple directions. Based on a laminar flow assumption, a patch-wise B-spline formulation of blood velocity is coupled for the first time with a global waveform model acting as temporal regularization. We evaluated our method on three virtual phantom datasets, demonstrating robustness in terms of noise, angle between measurements and data sparsity, and applied it successfully to five real case datasets of carotid artery examination.


Increasing inequalities in health: is it an artefact caused by the selective movement of people?
Changes in health socio-economic inequalities are currently measured by comparing the mortality gradient across aggregates of small administrative areas at two points in time. However, this methodology may be flawed as it ignores population movement, which previous research has shown to be selective, with a net loss of the more affluent (and possibly healthier) residents from deprived to more affluent areas. This paper investigates whether selective migration contributed sufficiently to the observed socio-economic gradients in mortality in England and Wales throughout the 1990s so as to invalidate the current method of monitoring health inequalities. The ONS Longitudinal Study for England and Wales was used to calculate directly standardised mortality rates (DSR) by decile of deprivation in 1991 and 2001. The DSRs for 2001 were calculated twice, once according to decile of residence in 2001, and also according to the original decile in 1991. Selective migration was found to make an important contribution in explaining increases in inequalities between areas, accounting for about 50% of the increase for those aged less than 75. At the older age groups, however, selective migration was responsible for a narrowing of mortality differentials over time. These results indicate that caution should be exercised when using repeated ecological studies in assessing the extent of changes in inequalities over time.


Sorting Linear Genomes with Rearrangements and Indels.
Rearrangements are mutations that can change the organization of a genome, but not its content. Examples are inversions of DNA segments, translocations of chromosome ends, fusions and fissions of chromosomes. All mentioned rearrangements can be represented by the generic Double Cut and Join (DCJ) operation. However, the DCJ operation also allows circular chromosomes to be created at intermediate steps, even if the compared genomes are linear. In this case it is more plausible to consider a restriction in which the reincorporation of a circular chromosome has to be done immediately after its creation. We call these two consecutive operations an ER composition. It has been shown that an ER composition mimics either an internal block interchange (when two segments in the same chromosome exchange their positions), or an internal transposition (the special case of a block interchange when the two segments are adjacent). The DCJ distance of two genomes is the same, regardless of this restriction, and can be computed in linear time. For comparing two genomes with unequal contents, in addition to rearrangements we have to allow insertions and deletions of DNA segments-named indels. It is already known that the distance in the model combining DCJ and indel operations can be exactly computed. Again, for linear genomes it would be more plausible to adopt a restricted version with ER compositions. This model was studied recently by da Silva et al. (BMC Bioinformatics 13, Suppl. 19, S14, 2012), but only an upper bound for the restricted DCJ-indel distance was provided. Here we first solve an open problem posed in that paper and present a very simple proof showing that the distance, which can be computed in linear time, is the same for both the unrestricted and the restricted DCJ-indel models. We then give a simpler algorithm for computing an optimal restricted DCJ-indel sorting scenario in O(n log n) time. We also relate the DCJ-indel distance to the restricted DCJ-substitution distance, which instead of indels considers a more powerful operation that allows the substitution of a DNA segment by another DNA segment. We show that the DCJ-indel distance is a 2-approximation for the restricted DCJ-substitution distance.


Contribution of phenazine antibiotic biosynthesis to the ecological competence of fluorescent pseudomonads in soil habitats.
Phenazine antibiotics produced by Pseudomonas fluorescens 2-79 and Pseudomonas aureofaciens 30-84, previously shown to be the principal factors enabling these bacteria to suppress take-all of wheat caused by Gaeumannomyces graminis var. tritici, also contribute to the ecological competence of these strains in soil and in the rhizosphere of wheat. Strains 2-79 and 30-84, their Tn5 mutants defective in phenazine production (Phz-), or the mutant strains genetically restored for phenazine production (Phz+) were introduced into Thatuna silt loam (TSL) or TSL amended with G. graminis var. tritici. Soils were planted with three or five successive 20-day plant-harvest cycles of wheat. Population sizes of Phz- derivatives declined more rapidly than did population sizes of the corresponding parental or restored Phz+ strains. Antibiotic biosynthesis was particularly critical to survival of these strains during the fourth and fifth cycles of wheat in the presence of G. graminis var. tritici and during all five cycles of wheat in the absence of take-all. In pasteurized TSL, a Phz- derivative of strain 30-84 colonized the rhizosphere of wheat to the same extent that the parental strain did. The results indicate that production of phenazine antibiotics by strains 2-79 and 30-84 can contribute to the ecological competence of these strains and that the reduced survival of the Phz- strains is due to a diminished ability to compete with the resident microflora.


Workflow for highly porous resorbable custom 3D printed scaffolds using medical grade polymer for large volume alveolar bone regeneration.
This study investigates the design, workflow, and manufacture of highly porous, resorbable additively manufactured, 3-dimensional (3D) custom scaffolds for the regeneration of large volume alveolar bone defects. Computed tomography (CT) scans of 5 posterior mandibular vertical bone defects were obtained. Surface masks (3D surface contours) of the recipient site were first isolated using a contrast threshold, transformed into 3D objects, and used to guide the formation of custom implant template models. To determine model accuracy and fit, the gap and overlap between the patient geometry models and the idealized template 3D models were quantified. Models were 3D printed from medical grade polycaprolactone (PCL) into porous scaffolds. For scaffold dimensional quantification, scaffolds were scanned using a micro-computed tomography (µCT) scanner. The design and printing processes each achieved dimensional errors of <200 µm on average. The average gap between the template implant model and the scanned scaffold model was found to be 74 ± 14 µm. The printed scaffold was confirmed as having a porosity of 83.91%, a mean polymer or filament thickness of 200 ± 46 µm, and a mean pore size of 590 ± 243 µm. The approach described in this study is straightforward, adaptable to a range of patient geometries, and results in the formation of reproducible, dimensionally accurate custom implants. These highly porous 3D structures manufactured from resorbable medical grade material represent a potentially transformative technology toward the clinical implementation of scaffold-guided bone regeneration procedures.


Surgical amputation of a limb 31,000 years ago in Borneo.
The prevailing view regarding the evolution of medicine is that the emergence of settled agricultural societies around 10,000 years ago (the Neolithic Revolution) gave rise to a host of health problems that had previously been unknown among non-sedentary foraging populations, stimulating the first major innovations in prehistoric medical practices1,2. Such changes included the development of more advanced surgical procedures, with the oldest known indication of an 'operation' formerly thought to have consisted of the skeletal remains of a European Neolithic farmer (found in Buthiers-Boulancourt, France) whose left forearm had been surgically removed and then partially healed3. Dating to around 7,000 years ago, this accepted case of amputation would have required comprehensive knowledge of human anatomy and considerable technical skill, and has thus been viewed as the earliest evidence of a complex medical act3. Here, however, we report the discovery of skeletal remains of a young individual from Borneo who had the distal third of their left lower leg surgically amputated, probably as a child, at least 31,000 years ago. The individual survived the procedure and lived for another 6-9 years, before their remains were intentionally buried in Liang Tebo cave, which is located in East Kalimantan, Indonesian Borneo, in a limestone karst area that contains some of the world's earliest dated rock art4. This unexpectedly early evidence of a successful limb amputation suggests that at least some modern human foraging groups in tropical Asia had developed sophisticated medical knowledge and skills long before the Neolithic farming transition.


Regulation of arachidonic acid metabolism in macrophages by immune and nonimmune interferons.
Mouse resident peritoneal M phi release AAA and metabolize it into cyclooxygenase- and lipoxygenase-derived eicosanoids, when triggered in vitro with different stimuli. Pretreatment of M phi with nonimmune IFN-alpha and IFN-beta dramatically decreased AA liberation from M phi phospholipids and eicosanoid formation after stimulation of M phi with Zy, A23187, or PMA. M phi exposed to immune IFN-gamma also showed a substantial impairment of both AA liberation and eicosanoid production upon exposure to Zy. However, AA and eicosanoid release was increased by IFN-gamma, rather than depressed, in PMA-triggered M phi. In addition, IFN-gamma showed differential effects on M phi stimulated with A23187. In fact, it inhibited AA release as well as formation of lipoxygenase-derived LTC4, but it highly increased the release of the cyclooxygenase products PGE2 and 6-keto PGF1 alpha. The ability of IFN-gamma to differentially modulate AA metabolism of M phi, depending on the nature of the triggering agent, sets forth the high specificity of the regulatory capacity of this molecule. This is at variance with the down regulation of AA metabolism that is generally observed with nonimmune IFN.


Creating a model program for influenza surveillance in California: results from the 2005-2006 influenza season.
Influenza surveillance is valuable for monitoring trends in influenza-related morbidity and mortality. Using the 2005-2006 influenza season as an example, this paper describes a comprehensive influenza surveillance program used by the California Department of Public Health (CDPH). Data collected from patients evaluated for acute respiratory illness in a given week were reported and summarized the following week, including (1) electronic hospital pneumonia and influenza admission and antiviral usage records from Kaiser Permanente, (2) sentinel provider influenza-like illness (ILI) reports, (3) severe pediatric influenza case reports (e.g., children either hospitalized in intensive care or expired), (4) school clinic ILI evaluations, and (5) positive influenza test results from a network of academic, hospital, commercial, and public health laboratories and the state CDPH Viral and Rickettsial Disease Laboratory. Influenza activity in California in the 2005-2006 season was moderate in severity; all clinical and laboratory markers rose and fell consistently. Extensive laboratory characterization identified the predominant circulating virus strain as A/California/7/2004(H3N2), which was a component of the 2005-2006 influenza vaccine; 96% of samples tested showed adamantane resistance. By using multiple, complementary surveillance methods coupled with a strong laboratory component, the CDPH has developed a simple, flexible, stable, and widely accepted influenza surveillance system that can monitor trends in statewide influenza activity, ascertain the correlation between circulating strains with vaccine strains, and assist with detection of new strain variants. The methods described can serve as a model for influenza surveillance in other states.


Primary Sjogren's syndrome 1976-2005 and associated interstitial lung disease: a population-based study of incidence and mortality.
A very few studies describe the epidemiology of primary Sjögren's syndrome (pSS). The reported frequency of pulmonary involvement in pSS varies widely depending on the detection method employed, and consists mainly of various forms of airways disease. We aimed to evaluate the incidence and mortality of pSS and of lung disease in pSS, focusing on interstitial lung disease (ILD). A population-based incidence cohort of patients diagnosed with pSS in 1976-2005 was assembled. Diagnosis was based on the 2002 American-European Consensus Group criteria for pSS. Cumulative incidence adjusted for the competing risk of death was estimated. A Cox model with a time-dependent covariate was used to determine the incidence and the standardised mortality HR of pSS. 85 patients with pSS were identified (mean age 59.9 years; 91% women). The annual incidence of pSS was 4.2, 95% CI (3.3 to 5.1)/100 000 population and it increased with higher age at pSS diagnosis (18-44 years: 2.1/100 000 vs ≥75 years: 12.3/100 000). Standardised mortality ratio in pSS compared with the general population was 0.92, 95% CI (0.57 to 1.41). A total of 105 patients with pSS and ILD were identified (mean age 58.1 years; 91% women). Among patients with pSS without prior ILD, the cumulative incidence of ILD in patients with pSS was 10% (±3%) at 1 year after diagnosis of pSS and increased to 20% (±4%) by 5 years after pSS. The development of lung disease in pSS was associated with poor survival (HR 2.16; 95% CI 0.99 to 4.74). pSS incidence seems to be almost the same as was reported in a previous study conducted among Olmsted County Minnesota population. Survival among patients with pSS and general population does not differ substantially. However, patients with pSS who have ILD likely have increased premature mortality.


Two histone deacetylases, FfHda1 and FfHda2, are important for Fusarium fujikuroi secondary metabolism and virulence.
Histone modifications are crucial for the regulation of secondary metabolism in various filamentous fungi. Here we studied the involvement of histone deacetylases (HDACs) in secondary metabolism in the phytopathogenic fungus Fusarium fujikuroi, a known producer of several secondary metabolites, including phytohormones, pigments, and mycotoxins. Deletion of three Zn(2+)-dependent HDAC-encoding genes, ffhda1, ffhda2, and ffhda4, indicated that FfHda1 and FfHda2 regulate secondary metabolism, whereas FfHda4 is involved in developmental processes but is dispensable for secondary-metabolite production in F. fujikuroi. Single deletions of ffhda1 and ffhda2 resulted not only in an increase or decrease but also in derepression of metabolite biosynthesis under normally repressing conditions. Moreover, double deletion of both the ffhda1 and ffhda2 genes showed additive but also distinct phenotypes with regard to secondary-metabolite biosynthesis, and both genes are required for gibberellic acid (GA)-induced bakanae disease on the preferred host plant rice, as Δffhda1 Δffhda2 mutants resemble the uninfected control plant. Microarray analysis with a Δffhda1 mutant that has lost the major HDAC revealed differential expression of secondary-metabolite gene clusters, which was subsequently verified by a combination of chemical and biological approaches. These results indicate that HDACs are involved not only in gene silencing but also in the activation of some genes. Chromatin immunoprecipitation with the Δffhda1 mutant revealed significant alterations in the acetylation state of secondary-metabolite gene clusters compared to the wild type, thereby providing insights into the regulatory mechanism at the chromatin level. Altogether, manipulation of HDAC-encoding genes constitutes a powerful tool to control secondary metabolism in filamentous fungi.


Trends from 1996 to 2007 in incidence and mortality outcomes of heart failure after acute myocardial infarction: a population-based study of 20,812 patients with first acute myocardial infarction in Western Australia.
Advances in treatment for acute myocardial infarction (AMI) are likely to have had a beneficial impact on the incidence of and deaths attributable to heart failure (HF) complicating AMI, although limited data are available to support this contention. Western Australian linked administrative health data were used to identify 20 812 consecutive patients, aged 40 to 84 years, without prior HF hospitalized with an index (first) AMI between 1996 and 2007. We assessed the temporal incidence of and adjusted odds ratio/hazard ratio for death associated with HF concurrent with AMI admission and within 1 year after discharge. Concurrent HF comprised 75% of incident HF cases. Between the periods 1996-1998 and 2005-2007, the prevalence of HF after AMI declined from 28.1% to 16.5%, with an adjusted odds ratio of 0.50 (95% CI, 0.44 to 0.55). The crude 28-day case-fatality rate for patients with concurrent HF declined marginally from 20.5% to 15.9% (P < 0.05) compared with those without concurrent HF, in whom the case-fatality rate declined from 11.0% to 4.8% (P < 0.001). Concurrent HF was associated with a multivariate-adjusted odds ratio of 2.2 for 28-day mortality and a hazard ratio of 2.2 for 1-year mortality in 28-day survivors. Occurrence of HF within 90 days of the index AMI was associated with an adjusted hazard ratio of 2.7 for 1-year mortality in 90-day survivors. Despite encouraging declines in the incidence of HF complicating AMI, it remains a common problem with high mortality. Increased attention to these high-risk patients is needed given the lack of improvement in their long-term prognosis.


Preventing Healthcare-Associated Legionellosis: Results after 3 Years of Continuous Disinfection of Hot Water with Monochloramine and an Effective Water Safety Plan.
The purpose of this study is to report the experience of the implementation and application of a 3-year Water Safety Plan (WSP) together with the secondary disinfection of water by monochloramine to control and prevent healthcare-associated legionellosis in an Italian hospital strongly colonized by Legionella. Risk assessment was carried out by the WSP team. The main critical control points focused on in developing the WSP for the control of Legionella was the water distribution system. A sampling plan for the detection of Legionella was implemented. A widespread contamination of the hot water distribution system by L. pneumophila sg5 was found. Results after 3 years of the continuous disinfection of hot water with monochloramine indicate the eradication of Legionella. The implementation and application of a WSP in a hospital, together with the disinfection of the water distribution system with monochloramine, can be effective in controlling the growth of Legionella and in preventing nosocomial legionellosis.


eNOS plays a major role in adiponectin synthesis in adipocytes.
Nitric oxide (NO) stimulates mitochondrial biogenesis. We recently reported that adiponectin synthesis is regulated by mitochondrial function in adipocytes. This study was undertaken to test the hypothesis that endothelial NO synthase (eNOS) plays an important role in adiponectin synthesis by producing NO and enhancing mitochondrial function in adipocytes. We examined the effects of eNOS knockdown on adiponectin synthesis in 3T3-L1 adipocytes and also examined plasma adiponectin levels and the mitochondria in adipose tissue of eNOS knockout (eNOS(-/-)) mice with and without chronic administration of a NO donor. In cultured 3T3-L1 adipocytes, eNOS siRNA decreased rosiglitazone-induced adiponectin secretion, which was associated with decreases in mitochondrial proteins and biogenesis factors. Plasma adiponectin concentrations were reduced in adult eNOS(-/-) mice compared with age-matched wild-type mice. Mitochondrial contents in adipose tissue were reduced in eNOS(-/-) mice, and this was associated with decreased expression of mitochondrial biogenesis factors, increased levels of 8-hydroxyguanosine, a biomarker of oxidative stress, and morphological abnormalities in mitochondria. Rosiglitazone-induced increases in adiponectin expression and mitochondrial content were also reduced significantly in eNOS(-/-) mice. Chronic administration of a NO donor reversed mitochondrial abnormalities and increased adiponectin expression in adipose tissue of eNOS(-/-) mice. eNOS plays an important role in adiponectin synthesis in adipocytes by increasing mitochondrial biogenesis and enhancing mitochondrial function.


The secretin/secretin receptor axis modulates liver fibrosis through changes in transforming growth factor-β1 biliary secretion in mice.
The secretin/secretin receptor (SR) axis is up-regulated by proliferating cholangiocytes during cholestasis. Secretin stimulates biliary proliferation by down-regulation of let-7a and subsequent up-regulation of the growth-promoting factor, nerve growth factor (NGF). It is not known whether the secretin/SR axis plays a role in subepithelial fibrosis observed during cholestasis. Our aim was to determine the role of the secretin/SR axis in activation of biliary fibrosis in animal models and human primary sclerosing cholangitis (PSC). Studies were performed in wild-type (WT) mice with bile duct ligation (BDL), BDL SR(-/-) mice, or Mdr2(-/-) mouse models of cholestatic liver injury. In selected studies, the SR antagonist (Sec 5-27) was used to block the secretin/SR axis. Biliary proliferation and fibrosis were evaluated as well as secretion of secretin (by cholangiocytes and S cells), expression of markers of fibrosis, transforming growth factor-β1 (TGF-β1), transforming growth factor-β1 receptor (TGF-β1R), let-7a, and downstream expression of NGF. Correlative studies were performed in human control and PSC liver tissue biopsies, serum, and bile. SR antagonist reduced biliary proliferation and hepatic fibrosis in BDL WT and Mdr2(-/-) mice. There was decreased expression of let-7a in BDL and Mdr2(-/-) cholangiocytes that was associated with increased NGF expression. Inhibition of let-7a accelerated liver fibrosis was attributed to cholestasis. There was increased expression of TGF-β1 and TGF-β1R. Significantly higher expression of secretin, SR, and TGF-β1 was observed in PSC patient liver samples compared to healthy controls. In addition, there was higher expression of fibrosis genes and remarkably decreased expression of let-7a and increased expression of NGF compared to the control. The secretin/SR axis plays a key role in regulating the biliary contribution to cholestasis-induced hepatic fibrosis. (Hepatology 2016;64:865-879).


The 4-Dimensional Plant: Effects of Wind-Induced Canopy Movement on Light Fluctuations and Photosynthesis.
Physical perturbation of a plant canopy brought about by wind is a ubiquitous phenomenon and yet its biological importance has often been overlooked. This is partly due to the complexity of the issue at hand: wind-induced movement (or mechanical excitation) is a stochastic process which is difficult to measure and quantify; plant motion is dependent upon canopy architectural features which, until recently, were difficult to accurately represent and model in 3-dimensions; light patterning throughout a canopy is difficult to compute at high-resolutions, especially when confounded by other environmental variables. Recent studies have reinforced the expectation that canopy architecture is a strong determinant of productivity and yield; however, links between the architectural properties of the plant and its mechanical properties, particularly its response to wind, are relatively unknown. As a result, biologically relevant data relating canopy architecture, light- dynamics, and short-scale photosynthetic responses in the canopy setting are scarce. Here, we hypothesize that wind-induced movement will have large consequences for the photosynthetic productivity of our crops due to its influence on light patterning. To address this issue, in this study we combined high resolution 3D reconstructions of a plant canopy with a simple representation of canopy perturbation as a result of wind using solid body rotation in order to explore the potential effects on light patterning, interception, and photosynthetic productivity. We looked at two different scenarios: firstly a constant distortion where a rice canopy was subject to a permanent distortion throughout the whole day; and secondly, a dynamic distortion, where the canopy was distorted in incremental steps between two extremes at set time points in the day. We find that mechanical canopy excitation substantially alters light dynamics; light distribution and modeled canopy carbon gain. We then discuss methods required for accurate modeling of mechanical canopy excitation (here coined the 4-dimensional plant) and some associated biological and applied implications of such techniques. We hypothesize that biomechanical plant properties are a specific adaptation to achieve wind-induced photosynthetic enhancement and we outline how traits facilitating canopy excitation could be used as a route for improving crop yield.


Enzyme-inhibitor interactions at the plant-pathogen interface.
The plant apoplast during plant-pathogen interactions is an ancient battleground that holds an intriguing range of attacking enzymes and counteracting inhibitors. Examples are pathogen xylanases and polygalacturonases that are inhibited by plant proteins like TAXI, XIP, and PGIP; and plant glucanases and proteases, which are targeted by pathogen proteins such as GIP1, EPI1, EPIC2B, and AVR2. These seven well-characterized inhibitors have different modes of action and many probably evolved from inactive enzymes themselves. Detailed studies of the structures, sequence variation, and mutated proteins uncovered molecular struggles between these enzymes and their inhibitors, resulting in positive selection for variant residues at the contact surface, where single residues determine the outcome of the interaction.


Investigating the effect of chitosan/ polycaprolactone blends in differentiation of corneal endothelial cells and extracellular matrix compositions.
This study aimed to investigate the underlying mechanisms of corneal endothelial cells (CECs) differentiation and identify the extracellular matrix (ECM) compositions using chitosan/polycaprolactone (PCL) blended membrane, hence exploring the potential use of chitosan/PCL blends in tissue engineering of CECs. We utilized the chitosan/PCL blends named as PCL25 consisting of PCL at 25% by weight. The surface characteristics of PCL25 were confirmed by using Fourier Transform Infrared Spectroscopy (FTIR) and Atomic Force Microscope (AFM). Bovine CECs were cultured on the blends, compared with TCPS and pure chitosan membrane. Cell behaviors in terms of cell attachment, proliferation, differentiation phenotype and expression of differentiation proteins were examined. Furthermore, ECM protein productions were also analyzed. From the experiments, we found the topography (roughness) of PCL25 membrane examined by AFM was greater than pure chitosan membrane. FTIR results confirmed the functional groups of C=O bond of PCL. The CECs displayed hexagonal morphology and similar proliferation rate on both PCL25 membrane and TCPS. In addition, the immunofluorescence evidence showed well-localized ZO-1 and Na+/K+ ATPase expression of membrane proteins. ECM protein productions of CECs on PCL were no inferior to TCPS. Moreover, western blot results verified the higher amount of collagen type IV, and reduced TGF-β2 expression on PCL25 membrane compared to TCPS substrate. In conclusions, chitosan/PCL blends membrane provided a favorable environment for CECs in terms of ECM compositions, therefore enhancing the growth and differentiation. Accordingly, for CEC tissue engineering applications, PCL 25 might be a suitable alternative for cadaveric cornea transplantation in the near future.


Inhibition of thrombomodulin surface expression and protein C activation by the thrombogenic agent homocysteine.
Elevated levels of plasma homocysteine are associated with both venous and arterial thrombosis. Homocysteine inhibits the function of thrombomodulin, an anticoagulant glycoprotein on the endothelial surface that serves as a cofactor for the activation of protein C by thrombin. The effects of homocysteine on thrombomodulin expression and protein C activation were investigated in cultured human umbilical vein endothelial cells and CV-1(18A) cells that express recombinant human thrombomodulin. Addition of 5 mM homocysteine to endothelial cells produced slight increases in thrombomodulin mRNA and thrombomodulin synthesis without affecting cell viability. In both cell types, thrombomodulin synthesized in the presence of homocysteine remained sensitive to digestion with endoglycosidase H and failed to appear on the cell surface, suggesting impaired transit along the secretory pathway. In a cell-free protein C activation assay, homocysteine irreversibly inactivated both thrombomodulin and protein C in a process that required free thiol groups and was inhibited by the oxidizing agents diamide or N-ethylmaleimide. By inhibiting both thrombomodulin surface expression and protein C activation, homocysteine may contribute to the development of thrombosis in patients with cystathionine beta-synthase deficiency.


Resting-state fMRI in the Human Connectome Project.
Resting-state functional magnetic resonance imaging (rfMRI) allows one to study functional connectivity in the brain by acquiring fMRI data while subjects lie inactive in the MRI scanner, and taking advantage of the fact that functionally related brain regions spontaneously co-activate. rfMRI is one of the two primary data modalities being acquired for the Human Connectome Project (the other being diffusion MRI). A key objective is to generate a detailed in vivo mapping of functional connectivity in a large cohort of healthy adults (over 1000 subjects), and to make these datasets freely available for use by the neuroimaging community. In each subject we acquire a total of 1h of whole-brain rfMRI data at 3 T, with a spatial resolution of 2×2×2 mm and a temporal resolution of 0.7s, capitalizing on recent developments in slice-accelerated echo-planar imaging. We will also scan a subset of the cohort at higher field strength and resolution. In this paper we outline the work behind, and rationale for, decisions taken regarding the rfMRI data acquisition protocol and pre-processing pipelines, and present some initial results showing data quality and example functional connectivity analyses.


ACE inhibitor use is associated with hospitalization for severe hypoglycemia in patients with diabetes. DARTS/MEMO Collaboration. Diabetes Audit and Research in Tayside, Scotland. Medicines Monitoring Unit.
To evaluate the association between the use of ACE inhibitors and hospital admission for severe hypoglycemia and to explore the effects of potential confounding variables on this relationship. The association between the use of ACE inhibitors and the incidence of hypoglycemia is controversial. A recent study reported that 14% of all hospital admissions for hypoglycemia might be attributable to ACE inhibitors. We performed a nested case-control study, using a cohort of 6,649 diabetic patients taking insulin or oral antidiabetic drugs, on the Diabetes Audit and Research in Tayside, Scotland (DARTS) database. From 1 January 1993 to 30 April 1994, we identified 64 patients who had been admitted to Tayside hospitals with hypoglycemia and selected 440 control patients from the same cohort. Hypoglycemia was associated with the use of ACE inhibitors (odds ratio [OR] 3.2, 95% CI 1.2-8.3, P = 0.023), whereas use of beta-blockers and calcium antagonists was not associated with an increased risk of hospitalization for hypoglycemia with ORs of 0.9 (95% CI 0.3-3.3) and 1.7 (95% CI 0.2-2.1), respectively. There were significant differences between case and control patients in type of diabetes treatment, diabetes duration, place of routine diabetes care, and congestive cardiac failure. These differences did not confound the relationship between ACE inhibitors and hypoglycemia (adjusted OR 4.3, 95% CI 1.2-16.0). The results show that the association between ACE inhibitor therapy and hospital admission for severe hypoglycemia is not explained by these confounding factors. Although ACE inhibitors have distinct advantages over other antihypertensive drugs in diabetes, the risk of hypoglycemia should be considered.


Saddle pressure distributions of three saddles used for Icelandic horses and their effects on ground reaction forces, limb movements and rider positions at walk and tölt.
Icelandic horse riding practices aim to place the rider further caudally on the horse's back than in English riding, claiming that a weight shift toward the hindquarters improves the quality of the tölt (e.g. giving the shoulder more freedom to move). This study compared saddle pressure patterns and the effects on limb kinetics and kinematics of three saddles: an Icelandic saddle (S(Icel), lowest point of seat in the hind part of the saddle), a treeless saddle cushion (S(Cush)) and a dressage-style saddle (S(Dres)). Twelve Icelandic horses were ridden with S(Icel), S(Cush) and S(Dres) on an instrumented treadmill at walk and tölt. Saddle pressure, limb forces and kinematics were recorded simultaneously. With S(Cush), pressure was highest under the front part of the saddle, whereas the saddles with trees had more pressure under the hind area. The saddles had no influence on the motion patterns of the limbs. The slight weight shift to the rear with S(Cush) and S(Icel) may be explained by the more caudal position of the rider relative to the horse's back.


Reactivation of DNA viruses in association with histone deacetylase inhibitor therapy: a case series report.
Histone deacetylase inhibitors are a class of anti-neoplastic agents that induce growth arrest, differentiation, and/or apoptotic cell death of transformed cells in vitro and in vivo. A phase II study exploring the efficacy of romidepsin, an histone deacetylase inhibitor, in patients with cutaneous or peripheral T-cell lymphomas was initiated at the National Cancer Institute. To date, over 120 patients with T-cell lymphoma have been treated on a multi-institutional phase II trial of romidepsin. Reactivation of latent DNA viruses including EBV, HBV, and VZV is well described as a consequence of the immune suppression associated with systemic chemotherapy. The incidence of viral reactivation in patients treated with histone deacetylase inhibitors is not yet known. We report the observation of EBV-associated illnesses in 2 patients and the reactivation of HBV in an additional patient treated with romidepsin. These cases may represent reactivation of DNA viruses due to histone deacetylase inhibitor induced immunosuppression, or direct promotion of viral replication via histone deacetylase inhibitor induced chromatin remodeling, or, alternatively, may be related to the underlying disease process. These observations suggest that vigilance for DNA virus reactivation is needed to quantify the risk in patients treated with histone deacetylase inhibitors.


Utility of commercially available reagent test strips for estimation of blood urea nitrogen concentration and detection of azotemia in pet rabbits (Oryctolagus cuniculus) and ferrets (Mustela putorius furo).
To evaluate the utility of commercially available reagent test strips for estimation of BUN concentration and detection of azotemia in pet rabbits (Oryctolagus cuniculus) and ferrets (Mustela putorius furo). 65 blood samples from 53 rabbits and 71 blood samples from 50 ferrets of various health statuses. BUN concentrations were measured with a clinical laboratory biochemical analyzer and estimated with a reagent test strip. Results obtained with both methods were assigned to a BUN category (range, 1 to 4; higher categories corresponded to higher BUN concentrations). Samples with a biochemical analyzer BUN concentration ≥ 27 mg/dL (rabbits) or ≥ 41 mg/dL (ferrets) were considered azotemic. A test strip BUN category of 3 or 4 (rabbits) or 4 (ferrets) was considered positive for azotemia. Test strip and biochemical analyzer BUN categories were concordant for 46 of 65 (71%) rabbit blood samples and 58 of 71 (82%) ferret blood samples. Sensitivity, specificity, and accuracy of the test strips for detection of azotemia were 92%, 79%, and 82%, respectively, for rabbit blood samples and 80%, 100%, and 96%, respectively, for ferret blood samples. Test strips provided reasonable estimates of BUN concentration but, for rabbits, were more appropriate for ruling out than for ruling in azotemia because of false-positive test strip results. False-negative test strip results for azotemia were more of a concern for ferrets than rabbits. Testing with a biochemical analyzer remains the gold standard for measurement of BUN concentration and detection of azotemia in rabbits and ferrets.


Neural substrates of social status inference: roles of medial prefrontal cortex and superior temporal sulcus.
The negotiation of social order is intimately connected to the capacity to infer and track status relationships. Despite the foundational role of status in social cognition, we know little about how the brain constructs status from social interactions that display it. Although emerging cognitive neuroscience reveals that status judgments depend on the intraparietal sulcus, a brain region that supports the comparison of targets along a quantitative continuum, we present evidence that status judgments do not necessarily reduce to ranking targets along a quantitative continuum. The process of judging status also fits a social interdependence analysis. Consistent with third-party perceivers judging status by inferring whose goals are dictating the terms of the interaction and who is subordinating their desires to whom, status judgments were associated with increased recruitment of medial pFC and STS, brain regions implicated in mental state inference.


Routine childhood immunisation during the COVID-19 pandemic in Africa: a benefit-risk analysis of health benefits versus excess risk of SARS-CoV-2 infection.
National immunisation programmes globally are at risk of suspension due to the severe health system constraints and physical distancing measures in place to mitigate the ongoing COVID-19 pandemic. We aimed to compare the health benefits of sustaining routine childhood immunisation in Africa with the risk of acquiring severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection through visiting routine vaccination service delivery points. We considered a high-impact scenario and a low-impact scenario to approximate the child deaths that could be caused by immunisation coverage reductions during COVID-19 outbreaks. In the high-impact scenario, we used previously reported country-specific child mortality impact estimates of childhood immunisation for diphtheria, tetanus, pertussis, hepatitis B, Haemophilus influenzae type b, Streptococcus pneumoniae, rotavirus, measles, meningitis A, rubella, and yellow fever to approximate the future deaths averted before 5 years of age by routine childhood vaccination during a 6-month COVID-19 risk period without catch-up campaigns. In the low-impact scenario, we approximated the health benefits of sustaining routine childhood immunisation on only the child deaths averted from measles outbreaks during the COVID-19 risk period. We assumed that contact-reducing interventions flattened the outbreak curve during the COVID-19 risk period, that 60% of the population will have been infected by the end of that period, that children can be infected by either vaccinators or during transport, and that upon child infection the whole household will be infected. Country-specific household age structure estimates and age-dependent infection-fatality rates were applied to calculate the number of deaths attributable to the vaccination clinic visits. We present benefit-risk ratios for routine childhood immunisation, with 95% uncertainty intervals (UIs) from a probabilistic sensitivity analysis. In the high-impact scenario, for every one excess COVID-19 death attributable to SARS-CoV-2 infections acquired during routine vaccination clinic visits, 84 (95% UI 14-267) deaths in children could be prevented by sustaining routine childhood immunisation in Africa. The benefit-risk ratio for the vaccinated children is 85 000 (4900-546 000), for their siblings (<20 years) is 75 000 (4400-483 000), for their parents or adult carers (aged 20-60 years) is 769 (148-2700), and for older adults (>60 years) is 96 (14-307). In the low-impact scenario that approximates the health benefits to only the child deaths averted from measles outbreaks, the benefit-risk ratio to the households of vaccinated children is 3 (0-10); if the risk to only the vaccinated children is considered, the benefit-risk ratio is 3000 (182-21 000). The deaths prevented by sustaining routine childhood immunisation in Africa outweigh the excess risk of COVID-19 deaths associated with vaccination clinic visits, especially for the vaccinated children. Routine childhood immunisation should be sustained in Africa as much as possible, while considering other factors such as logistical constraints, staff shortages, and reallocation of resources during the COVID-19 pandemic. Gavi, the Vaccine Alliance; Bill & Melinda Gates Foundation.


Demand for contraception to delay first pregnancy among young married women in India.
In India, evidence is sparse regarding the demand for contraception to delay first pregnancy among married young people. Using data drawn from a study conducted in six Indian states among 9,572 women aged 15-24 who were married for five or fewer years, we explore the scope of this demand, the extent to which it has been satisfied, and, using logistic regression analyses, the factors correlated with contraceptive use to delay first pregnancy among those reporting demand. Findings confirm considerable demand for contraception to postpone first pregnancy (51 percent). Of those with demand, only 10 percent had practiced contraception. Contraception was more likely to have been practiced by educated women, those aware of family planning methods before they were married, those exposed to quality sexuality education, and those who participated in marriage-related decision making. Women who reported feeling pressure to prove their fertility were less likely to have practiced contraception.


Impact of population structure on genetic diversity of a potential vaccine target in the canine hookworm (Ancylostoma caninum).
Ancylostoma caninum is a globally distributed canine parasitic nematode. To test whether positive selection, population structure, or both affect genetic variation at the candidate vaccine target Ancylostoma secreted protein 1 (asp-1), we have quantified the genetic variation in A. caninum at asp-1 and a mitochondrial gene, cytochrome oxidase subunit 1 (cox-1), using the statistical population analysis tools found in the SNAP Workbench. The mitochondrial gene cox-1 exhibits moderate diversity within 2 North American samples, comparable to the level of variation observed in other parasitic nematodes. The protein coding portion for the C-terminal half of asp-1 shows similar levels of genetic variation in a Wake County, North Carolina, sample as cox-1. Standard tests of neutrality provide little formal evidence for selection acting on this locus, but haplotype networks for 2 of the exon regions have significantly different topologies, consistent with different evolutionary forces shaping variation at either end of a 1.3-kilobase stretch of sequence. Evidence for gene flow among geographically distinct samples suggests that the mobility of hosts of A. caninum is an important contributing factor to the population structure of the parasite.


The role of xylem class III peroxidases in lignification.
Lignification is a cell wall fortifying process which occurs in xylem tissue in a scheduled manner during tissue differentiation. In this review, enzymes and the genes responsible for lignin biosynthesis have been studied with an emphasis on lignin polymerizing class III secretable plant peroxidases. Our aim is to understand the cell and molecular biology of the polymerization of lignin especially in tracheids and vessels of woody species but much of the experimental evidence comes from herbaceous plants. Class III peroxidases pose many problems for empirical work as their encoding genes are variable, their substrate specificities are wide and the half-life of many of the isozymes is very long. However, there is some evidence for the role of specific peroxidases in lignin polymerization through antisense mutants in tobacco and poplar and from tissue and cell culture lines of Picea abies and Zinnia elegans. Peroxidase enzyme action has been shown by substrate specificity studies and, for example, RT-PCR results have pointed out that many peroxidases have tissue-specific expression patterns. Tissue-level location of gene expression of some peroxidases has been studied by in situ hybridization and their cellular localization with antibodies and using EGFP-fusion genes. From these, it can be concluded that, although many of the xylem class III peroxidases have the potential for functioning in the synthesis of the lignin polymer, the combined information of catalytic properties, expression, and localization can reveal differences in the significance of different peroxidases in the lignification process.


Plant perceptions of plant growth-promoting Pseudomonas.
Plant-associated Pseudomonas live as saprophytes and parasites on plant surfaces and inside plant tissues. Many plant-associated Pseudomonas promote plant growth by suppressing pathogenic micro-organisms, synthesizing growth-stimulating plant hormones and promoting increased plant disease resistance. Others inhibit plant growth and cause disease symptoms ranging from rot and necrosis through to developmental dystrophies such as galls. It is not easy to draw a clear distinction between pathogenic and plant growth-promoting Pseudomonas. They colonize the same ecological niches and possess similar mechanisms for plant colonization. Pathogenic, saprophytic and plant growth-promoting strains are often found within the same species, and the incidence and severity of Pseudomonas diseases are affected by environmental factors and host-specific interactions. Plants are faced with the challenge of how to recognize and exclude pathogens that pose a genuine threat, while tolerating more benign organisms. This review examines Pseudomonas from a plant perspective, focusing in particular on the question of how plants perceive and are affected by saprophytic and plant growth-promoting Pseudomonas (PGPP), in contrast to their interactions with plant pathogenic Pseudomonas. A better understanding of the molecular basis of plant-PGPP interactions and of the key differences between pathogens and PGPP will enable researchers to make more informed decisions in designing integrated disease-control strategies and in selecting, modifying and using PGPP for plant growth promotion, bioremediation and biocontrol.


Carbonaceous Materials Coated Carbon Fibre Reinforced Polymer Matrix Composites.
Carbon fibre reinforced polymer composites have high mechanical properties that make them exemplary engineered materials to carry loads and stresses. Coupling fibre and matrix together require good understanding of not only fibre morphology but also matrix rheology. One way of having a strongly coupled fibre and matrix interface is to size the reinforcing fibres by means of micro- or nanocarbon materials coating on the fibre surface. Common coating materials used are carbon nanotubes and nanofibres and graphene, and more recently carbon black (colloidal particles of virtually pure elemental carbon) and graphite. There are several chemical, thermal, and electrochemical processes that are used for coating the carbonous materials onto a carbon fibre surface. Sizing of fibres provides higher interfacial adhesion between fibre and matrix and allows better fibre wetting by the surrounded matrix material. This review paper goes over numerous techniques that are used for engineering the interface between both fibre and matrix systems, which is eventually the key to better mechanical properties of the composite systems.


Racial-Ethnic Protective Factors and Mechanisms in Psychosocial Prevention and Intervention Programs for Black Youth.
Extending previous reviews related to cultural responsiveness in the treatment of ethnic minority youth, the current review provides a critical assessment and synthesis of both basic and applied research on the integration of three racial-ethnic protective factors (racial identity, racial socialization, Africentric worldview) in psychosocial prevention and intervention programs for Black children and adolescents. Seventeen programs meeting inclusion and exclusion criteria were evaluated for the extent to which racial-ethnic protective factors and related mechanisms were integrated, applied, and tested in such programs. A systematic assessment of these programs revealed that several prevention and intervention programs drew upon the three factors, particularly Africentric worldview. In addition, a number of studies hypothesized and assessed mechanisms, both those previously identified in conceptual literature and those that emerged from the interventions themselves. A set of recommendations encouraging the implementation of these factors into future prevention and intervention programs, examples of how clinicians can infuse these factors into psychotherapy, and areas for future research are discussed.


Epidemiological differences between back pain of sudden and gradual onset.
To explore possible differences in risk factors for low back pain according to its speed of onset. We analyzed longitudinal data from 1366 hospital nurses in England who initially had been free from low back pain for at least one month. Risk factors were ascertained from a self-administered baseline questionnaire, and outcomes from serial followup questionnaires. Hazard ratios (HR) for developing a first new episode of low back pain during followup were derived by Cox regression. Low back pain with gradual onset was significantly associated with psychological symptoms measured at baseline [HR 1.7 (95% CI 1.2, 2.4) and higher], but no such association was seen for sudden pain. Low back pain with sudden onset while at work was associated with exposure to specific patient-handling tasks [HR 1.8 (95% CI 1.1, 3.0) to 2.8 (95% CI 1.4, 5.5)]. However, symptoms that came on suddenly elsewhere were not related to occupational activity, and low back pain of gradual onset showed little relation to patient-handling. These findings suggest that a useful distinction can be made according to the speed and circumstances of onset of low back pain. If confirmed, they have important implications for the evaluation of ergonomic interventions aimed at reducing back pain.


How prior preferences determine decision-making frames and biases in the human brain.
Understanding how option values are compared when making a choice is a key objective for decision neuroscience. In natural situations, agents may have a priori on their preferences that create default policies and shape the neural comparison process. We asked participants to make choices between items belonging to different categories (e.g., jazz vs. rock music). Behavioral data confirmed that the items taken from the preferred category were chosen more often and more rapidly, which qualified them as default options. FMRI data showed that baseline activity in classical brain valuation regions, such as the ventromedial Prefrontal Cortex (vmPFC), reflected the strength of prior preferences. In addition, evoked activity in the same regions scaled with the default option value, irrespective of the eventual choice. We therefore suggest that in the brain valuation system, choices are framed as comparisons between default and alternative options, which might save some resource but induce a decision bias.


Structural brain imaging studies offer clues about the effects of the shared genetic etiology among neuropsychiatric disorders.
Genomewide association studies have found significant genetic correlations among many neuropsychiatric disorders. In contrast, we know much less about the degree to which structural brain alterations are similar among disorders and, if so, the degree to which such similarities have a genetic etiology. From the Enhancing Neuroimaging Genetics through Meta-Analysis (ENIGMA) consortium, we acquired standardized mean differences (SMDs) in regional brain volume and cortical thickness between cases and controls. We had data on 41 brain regions for: attention-deficit/hyperactivity disorder (ADHD), autism spectrum disorder (ASD), bipolar disorder (BD), epilepsy, major depressive disorder (MDD), obsessive compulsive disorder (OCD), and schizophrenia (SCZ). These data had been derived from 24,360 patients and 37,425 controls. The SMDs were significantly correlated between SCZ and BD, OCD, MDD, and ASD. MDD was positively correlated with BD and OCD. BD was positively correlated with OCD and negatively correlated with ADHD. These pairwise correlations among disorders were correlated with the corresponding pairwise correlations among disorders derived from genomewide association studies (r = 0.494). Our results show substantial similarities in sMRI phenotypes among neuropsychiatric disorders and suggest that these similarities are accounted for, in part, by corresponding similarities in common genetic variant architectures.


Therapeutic Taping for Scapular Stabilization in Children With Brachial Plexus Birth Palsy.
In this study, we aimed to assess whether therapeutic taping for scapular stabilization affected scapulothoracic, glenohumeral, and humerothoracic joint function in children with brachial plexus birth palsy and scapular winging. Motion capture data were collected with and without therapeutic taping to assist the middle and lower trapezius in seven positions for 26 children. Data were compared with one-way multivariate analyses of variance. With therapeutic taping, scapular winging decreased considerably in all positions except abduction. Additionally, there were increased glenohumeral cross-body adduction and internal rotation angles in four positions. The only change in humerothoracic function was an increase of 3° of external rotation in the external rotation position. Therapeutic taping for scapular stabilization resulted in a small but statistically significant decrease in scapular winging. Overall performance of positions was largely unchanged. The increased glenohumeral joint angles with therapeutic taping may be beneficial for joint development; however, the long-term impact remains unknown.


Myeloperoxidase/nitrite-mediated lipid peroxidation of low-density lipoprotein as modulated by flavonoids.
In the presence of a H(2)O(2)-generating system, myeloperoxidase (MPO) caused conjugated diene formation in low-density lipoprotein (LDL), indicating lipid peroxidation which was dependent on nitrite but not on chloride. The oxidation of LDL was inhibited by micromolar concentrations of flavonoids such as (-)-epicatechin, quercetin, rutin, taxifolin and luteolin, presumably via scavenging of the MPO-derived NO(2) radical. The flavonoids served as substrates of MPO leading to products with distinct absorbance spectra. The MPO-catalyzed oxidation of flavonoids was accelerated in the presence of nitrite.


Rhizosphere microbiome metagenomics of gray mangroves (Avicennia marina) in the Red Sea.
Mangroves are unique, and endangered, coastal ecosystems that play a vital role in the tropical and subtropical environments. A comprehensive description of the microbial communities in these ecosystems is currently lacking, and additional studies are required to have a complete understanding of the functioning and resilience of mangroves worldwide. In this work, we carried out a metagenomic study by comparing the microbial community of mangrove sediment with the rhizosphere microbiome of Avicennia marina, in northern Red Sea mangroves, along the coast of Saudi Arabia. Our results revealed that rhizosphere samples presented similar profiles at the taxonomic and functional levels and differentiated from the microbiome of bulk soil controls. Overall, samples showed predominance by Proteobacteria, Bacteroidetes and Firmicutes, with high abundance of sulfate reducers and methanogens, although specific groups were selectively enriched in the rhizosphere. Functional analysis showed significant enrichment in 'metabolism of aromatic compounds', 'mobile genetic elements', 'potassium metabolism' and 'pathways that utilize osmolytes' in the rhizosphere microbiomes. To our knowledge, this is the first metagenomic study on the microbiome of mangroves in the Red Sea, and the first application of unbiased 454-pyrosequencing to study the rhizosphere microbiome associated with A. marina. Our results provide the first insights into the range of functions and microbial diversity in the rhizosphere and soil sediments of gray mangrove (A. marina) in the Red Sea.


The K1 capsule is the critical determinant in the development of Escherichia coli meningitis in the rat.
Although Escherichia coli strains possessing the K1 capsule are predominant among isolates from neonatal E. coli meningitis and most of these K1 isolates are associated with a limited number of 0 lipopolysaccharide (LPS) types, the basis of this association of K1 and certain 0 antigens with neonatal E. coli meningitis is not clear. The present study examined in experimental E. coli bacteremia and meningitis in newborn and adult rats whether or not the K1 capsule and/or O-LPS antigen are critical determinants in the development of meningitis. Rats received subcutaneously at K1 E. coli strain (018+K1+) or mutants lacking either the K1 capsule (018+K1-) or 0 side-chain (018-K1+). 12-24 h later, blood and cerebrospinal fluid (CSF) specimens were obtained for quantitative cultures. The isolation of E. coli from CSF was observed in both newborn and adult rats infected with K1+ strains regardless of LPS phenotype (018+ or 18-) who also developed a high degree of bacteremia (e.g., greater than 10(4) CFU/ml of blood). In contrast, none of the newborn and adult rats infected with 018+K1- and developing bacteremia of greater than 10(4) were found to have positive CSF cultures. These findings indicate that the presence of the K1 capsule and a high degree of bacteremia are key determinants in the development of E. coli meningitis, suggesting that there may be specific binding sites present in the brain which have an affinity for the K1 capsule and thus may be responsible for the entry of K1-encapsulated E. coli into the meninges.


GP views on strategies to cope with increasing workload: a qualitative interview study.
The existence of a crisis in primary care in the UK is in little doubt. GP morale and job satisfaction are low, and workload is increasing. In this challenging context, finding ways for GPs to manage that workload is imperative. To explore what existing or potential strategies are described by GPs for dealing with their workload, and their views on the relative merits of each. Semi-structured, qualitative interviews with GPs working within NHS England. All GPs working within NHS England were eligible. Of those who responded to advertisements, a maximum-variation sample was selected and interviewed until data saturation was reached. Data were analysed thematically. Responses were received from 171 GPs, and, from these, 34 were included in the study. Four main themes emerged for workload management: patient-level, GP-level, practice-level, and systems-level strategies. A need for patients to take greater responsibility for self-management was clear, but many felt that GPs should not be responsible for this education. Increased delegation of tasks was felt to be key to managing workload, with innovative use of allied healthcare professionals and extended roles for non-clinical staff suggested. Telephone triage was a commonly used tool for managing workload, although not all participants found this helpful. This in-depth qualitative study demonstrates an encouraging resilience among GPs. They are proactively trying to manage workload, often using innovative local strategies. GPs do not feel that they can do this alone, however, and called repeatedly for increased recruitment and more investment in primary care.


Lynch syndrome screening should be considered for all patients with newly diagnosed endometrial cancer.
Lynch syndrome (LS) is an autosomal dominant inherited disorder caused by germline mutations in DNA mismatch repair (MMR) genes. Mutation carriers are at substantially increased risk of developing cancers of the colorectum and endometrium, among others. Given recent recommendations for universal, cost-effective screening of all patients with newly diagnosed colorectal cancer using MMR protein immunohistochemistry, we evaluated MMR protein expression in a series of endometrial cancers in the general population. A total of 605 consecutive cases of primary endometrial cancer at a single institution (1997 to 2013) were evaluated regardless of age, family history, or histologic features. Evaluation methods consisted of immunohistochemistry for the MMR proteins MLH1, MSH2, MSH6, and PMS2, followed by DNA methylation analysis for cases with MLH1/PMS2 deficiency. Germline mutation testing was performed on a subset of cases. Forty MMR-deficient, nonmethylated endometrial cancers were identified: 3 MLH1/PMS2 and 37 MSH6/MSH2 protein deficiencies. Only 25% occurred in women below 50 years of age (range, 39 to 88 y), 1 of which was in a risk-reducing hysterectomy specimen. Only 15% of patients had a prior history of carcinoma, including only 2 patients with prior colorectal carcinoma. Most (80%) of the endometrial cancers were purely endometrioid; there were 2 mixed endometrioid/mucinous, 1 mucinous, 1 serous, 2 clear cell, and 2 carcinosarcoma cases. When grading was applicable, 40% of the endometrial malignancies were FIGO grade 1, 34% grade 2, and 26% grade 3. Thirteen percent arose in the lower uterine segment, and 23% had tumor-infiltrating lymphocytes. Of the tumors with known germline testing, 41% with a LS-associated germline mutation were not associated with any of the traditional indicators that have been recommended for LS screening (ie, age 50 y or younger, personal/family cancer pedigree that meets Bethesda guideline criteria, presence of MMR-associated tumor morphology, or location in the lower uterine segment). These data suggest that a significant number of LS-associated endometrial carcinomas are missed using clinical, histologic, and locational screening parameters and provide support for universal screening of all newly diagnosed endometrial cancers.


Nasal balloon autoinflation for glue ear in primary care: a qualitative interview study.
Nasal balloon autoinflation is an effective, non-surgical treatment for symptomatic children with glue ear, although uptake is variable and evidence about acceptability and feasibility is limited. To explore parent and healthcare professional views and experiences of nasal balloon autoinflation for children with glue ear in primary care. Qualitative study using semi-structured interviews with a maximum-variety sample of parents, GPs, and practice nurses. The study took place between February 2013 and September 2014. Semi-structured face-to-face and telephone interviews were audiorecorded, transcribed verbatim, and analysed using inductive thematic analysis. In all, 14 parents, 31 GPs, and 19 nurses were included in the study. Parents described the nasal balloon as a natural, holistic treatment that was both acceptable and appealing to children. GPs and nurses perceived the method to be a low-cost, low-risk strategy, applicable to the primary care setting. Good instruction and demonstration ensured children mastered the technique and engaged with the treatment, but uncertainties were raised about training provision and potential impact on the GP consultation. Making nasal balloon autoinflation part of a child's daily routine enhances compliance, but difficulties can arise if children are unwell or refuse to cooperate. Nasal balloon autoinflation is an acceptable, low-cost treatment option for children with glue ear in primary care. Provision of educational materials and demonstration of the method are likely to promote uptake and compliance. Wider use of the nasal balloon has the potential to enhance early management, and may help to fill the management gap arising from forthcoming changes to care pathways.


Cognitive performance and serotonergic function in users of ecstasy.
(+/-) 3,4-Methylenedioxymethamphetamine (MDMA or "ecstasy") has been shown to cause long term damage to serotonergic cerebral neurons in animals. The neurotoxic effects in humans are less clear and little is known about the functional consequences, although some studies suggest memory impairment. Given the widespread use of MDMA, our lack of knowledge raises concerns. We investigated, in humans, the relation between past use of ecstasy and cognitive performance as well as serotonergic function. Two groups of 21 males with moderate and heavy recreational use of MDMA, respectively, and a control group of 20 males without use of MDMA were compared. All were from the same subculture. Reaction time, direct recall, and recognition were assessed. Serotonergic function was measured by the neuro-endocrine response to a placebo-controlled, crossover challenge with dexfenfluramine. Ecstasy users showed a broad pattern of statistically significant, but clinically small, impairment of memory and prolonged reaction times. Heavy users were affected stronger than moderate users. Release of cortisol but not of prolactin after dexfenfluramine administration was significantly reduced in both groups of ecstasy users compared with the controls. Analyses of covariance showed that likely confounding variables including recent exposure to ecstasy, psychosocial profiles and use of other drugs did not explain the differences found between the groups. These results provide further evidence that use of ecstasy may be associated with impairment of memory and of serotonergic function. These findings are compatible with neurotoxicity of ecstasy as shown in animals.


Glycemic load, carbohydrate intake, and risk of colorectal cancer in women: a prospective cohort study.
Mounting evidence suggests that high circulating levels of insulin might be associated with increased colorectal cancer risk. The glycemic effects of diets high in refined starch may increase colorectal cancer risk by affecting insulin and/or insulin-like growth factor-I levels. We examined the association between dietary intake and colorectal cancer risk in a cohort of 49 124 women participating in a randomized, controlled trial of screening for breast cancer in Canada. Linkages to Canadian mortality and cancer databases yielded data on mortality and cancer incidence up to December 31, 2000. During an average 16.5 years of follow-up, we observed 616 incident cases of colorectal cancer (436 colon cancers, 180 rectal cancers). Rate ratios for colorectal cancer for the highest versus the lowest quintile level were 1.05 (95% confidence interval [CI] = 0.73 to 1.53; P(trend) =.94) for glycemic load, 1.01 (95% CI = 0.68 to 1.51; P(trend) =.66) for total carbohydrates, and 1.03 (95% CI = 0.73 to 1.44; P(trend) =.71) for total sugar. Our data do not support the hypothesis that diets high in glycemic load, carbohydrates, or sugar increase colorectal cancer risk.


The role of asymmetric interactions on the effect of habitat destruction in mutualistic networks.
Plant-pollinator mutualistic networks are asymmetric in their interactions: specialist plants are pollinated by generalist animals, while generalist plants are pollinated by a broad range involving specialists and generalists. It has been suggested that this asymmetric--or disassortative--assemblage could play an important role in determining the observed equal susceptibility of specialist and generalist plants under habitat destruction. At the core of the analysis of the phenomenon lies the observation that specialist plants, otherwise candidates to extinction, could cope with the disruption thanks to their interaction with a few generalist pollinators. We present a theoretical framework that supports this thesis. We analyze a dynamical model of a system of mutualistic plants and pollinators, subject to the destruction of their habitat. We analyze and compare two families of interaction topologies, ranging from highly assortative to highly disassortative ones, as well as real pollination networks. We found that several features observed in natural systems are predicted by the mathematical model. First, there is a tendency to increase the asymmetry of the network as a result of the extinctions. Second, an entropy measure of the differential susceptibility to extinction of specialist and generalist species show that they tend to balance when the network is disassortative. Finally, the disappearance of links in the network, as a result of extinctions, shows that specialist plants preserve more connections than the corresponding plants in an assortative system, enabling them to resist the disruption.


Effect of Moisture and Temperature on Disease of Green Foxtail Caused by Drechslera gigantea and Pyricularia setariae.
Leaf wetness duration, temperature, intermittent leaf wetness, and delayed leaf wetness were investigated for their influence on disease of green foxtail caused by Drechslera gigantea and Pyricularia setariae to determine the potential of these two fungi as bioherbicide agents in the Canadian prairies. For both fungi, disease severity increased with increasing leaf wetness duration at 15, 20, 25, 30, and 32°C. At 10°C, conidia of both fungi showed minimal germination, regardless of leaf wetness duration; however, an increase in conidial germination, appressoria formation, and disease occurred at 15°C. Conidia of both species showed 80% or greater germination at all temperatures above 15°C, whereas the optimum temperatures for appressoria formation by D. gigantea and P. setariae were 23 and 25°C, respectively. Maximum disease occurred after 48 h of leaf wetness at 32°C for D. gigantea and at 25°C for P. setariae. Disease caused by both fungi decreased when 4 h of continuous leaf wetness was followed by a 20-h dry period, and after an 8-h delay in leaf wetness following inoculation. Both fungi required immediate and prolonged periods of leaf wetness at temperatures of 15°C and above to cause severe disease on green foxtail. The moisture requirements of these fungi may limit their effectiveness as bioherbicide agents in the semi-arid Canadian prairies.


Endoscopic pelvic osteotomy for the treatment of hip dysplasia.
Adolescent and adult hip dysplasia can be surgically treated by rotating the acetabulum into a better weight-supporting position; however, open pelvic osteotomies are among the most invasive of all pediatric orthopaedic procedures. Endoscopic pelvic osteotomy offers the theoretical advantages of magnified visualization of the bone cuts, minimized surgical dissection, and rapid postoperative recovery. The technique of endoscopically assisted triple innominate osteotomy requires the combination of endoscopic skills and facility with more standard surgical approaches.


Effects of static muscular contraction on impulse activity of groups III and IV afferents in cats.
Static contraction of the hindlimb muscles, induced by electrical stimulation of the ventral roots, reflexly increases arterial blood pressure and heart rate. Although stimulation of groups III and IV muscle afferents is believed to cause these reflex increases, the responses of these afferents to a level of static contraction that increases arterial pressure have not yet been determined. Therefore, in barbiturate-anesthetized cats, afferent impulses arising from endings in the gastrocnemius muscle were recorded from the L7 or S1 dorsal roots, while the cut peripheral end of the L7 ventral root was stimulated. In addition, the effects of capsaicin (100-200 micrograms) and bradykinin (25 micrograms) on the activity of the groups III and IV afferents stimulated by contraction were examined. Contraction of the gastrocnemius muscle to a level equal to or greater than that needed to cause a pressor response stimulated 12 of 19 (63%) group III afferents and 13 of 19 (68%) group IV afferents. However, the discharge patterns of the group III afferents stimulated by contraction were very different from those of the group IV fibers. No relationship was found between those fibers stimulated by contraction and those stimulated by chemicals. Our results suggest that although both groups III and IV muscle afferents contribute to the reflex cardiovascular increases evoked by static exercise, group III fibers were likely to be stimulated by the mechanical effects of muscular contraction, whereas at least some group IV fibers were likely to be stimulated by the metabolic products of muscular contraction.


Marketing little cigars and cigarillos: advertising, price, and associations with neighborhood demographics.
We have documented little cigar and cigarillo (LCC) availability, advertising, and price in the point-of-sale environment and examined associations with neighborhood demographics. We used a multimodal real-time surveillance system to survey LCCs in 750 licensed tobacco retail outlets that sold tobacco products in Washington, DC. Using multivariate models, we examined the odds of LCC availability, the number of storefront exterior advertisements, and the price per cigarillo for Black & Mild packs in relation to neighborhood demographics. The odds of LCC availability and price per cigarillo decreased significantly in nearly a dose-response manner with each quartile increase in proportion of African Americans. Prices were also lower in some young adult neighborhoods. Having a higher proportion of African American and young adult residents was associated with more exterior LCC advertising. Higher availability of LCCs in African American communities and lower prices and greater outdoor advertising in minority and young adult neighborhoods may establish environmental triggers to smoke among groups susceptible to initiation, addiction, and long-term negative health consequences.


Novel MMP-9 substrates in cancer cells revealed by a label-free quantitative proteomics approach.
Matrix metalloproteinase-9 (MMP-9) is implicated in tumor metastasis as well as a variety of inflammatory and pathological processes. Although many substrates for MMP-9, including components of the extracellular matrix, soluble mediators such as chemokines, and cell surface molecules have been identified, we undertook a more comprehensive proteomics-based approach to identify new substrates to further understand how MMP-9 might contribute to tumor metastasis. Previous proteomics approaches to identify protease substrates have depended upon differential labeling of each sample. Instead we used a label-free quantitative proteomics approach based on ultraperformance LC-ESI-high/low collision energy MS. Conditioned medium from a human metastatic prostate cancer cell line, PC-3ML, in which MMP-9 had been down-regulated by RNA interference was compared with that from the parental cells. From more than 200 proteins identified, 69 showed significant alteration in levels after depletion of the protease (>+/-2-fold), suggesting that they might be candidate substrates. Levels of six of these (amyloid-beta precursor protein, collagen VI, leukemia inhibitory factor, neuropilin-1, prostate cancer cell-derived growth factor (PCDGF), and protease nexin-1 (PN-1)) were tested in the conditioned media by immunoblotting. There was a strong correlation between results by ultraperformance LC-ESI-high/low collision energy MS and by immunoblotting giving credence to the label-free approach. Further information about MMP-9 cleavage was obtained by comparison of the peptide coverage of collagen VI in the presence and absence of MMP-9 showing increased sensitivity of the C- and N-terminal globular regions over the helical regions. Susceptibility of PN-1 and leukemia inhibitory factor to MMP-9 degradation was confirmed by in vitro incubation of the recombinant proteins with recombinant MMP-9. The MMP-9 cleavage sites in PN-1 were sequenced. This study provides a new label-free method for degradomics cell-based screening leading to the identification of a series of proteins whose levels are affected by MMP-9, some of which are clearly direct substrates for MMP-9 and become candidates for involvement in metastasis.


Gastric adenocarcinoma in Zambia: a case-control study of HIV, lifestyle risk factors, and biomarkers of pathogenesis.
Gastric cancer is a leading cause of cancer deaths worldwide but there are few data from Africa. We recently observed a trend towards diagnosis in younger patients. To test the hypothesis that HIV might have altered risk factors for acquisition of gastric cancer, in a case-control study in the University Teaching Hospital, Lusaka, Zambia. Patients (n=52) with confirmed gastric adenocarcinoma and controls (n=94) undergoing endoscopy but with no macroscopic gastric pathology. Established risk factors and HIV status were compared. HIV status did not differ significantly between cases and controls (odds ratio 1.03; 95% CI 0.2 - 4.3; p=1.00) and seroprevalence in cases was similar to that of the Zambian population. Smoking, regular alcohol intake, and gastric atrophy were all associated with cancer in univariate and multivariate analysis. Helicobacter pylori serology was positive in 84% of patients studied and cagA serology in 66%; neither serological marker was associated with cancer. Atrophy was common in cases (57%) and controls (30%) and associated with both smoking and alcohol use. Intestinal metaplasia was present in 17% of the controls, but was not associated with atrophy. HIV was not associated with gastric cancer and does not explain the apparent younger age distribution. Atrophy was common and was not essential for the development of intestinal metaplasia, suggesting that gastric carcinogenesis in Africa does not always follow the pathway from atrophy to intestinal metaplasia to gastric carcinoma (the so-called Correa pathway).


Cystic adventitial disease of the popliteal artery: an infrequent cause of intermittent claudication.
Intermittent claudication is frequently associated with atherosclerotic disease, but differential diagnosis must be sought in patients with no traditional risk factors. Cystic adventitial disease, of unknown etiology, most frequently affects the popliteal artery, and occasionally presents as intermittent claudication. We report a case of this disease and the surgical treatment, and discuss some aspects related to etiopathogenesis, diagnosis and treatment of this condition.


An ex vivo exothermal and mechanical evaluation of two-solution bone cements in vertebroplasty.
Previous ex vivo studies showed that the properties of commercial cements modified for use in vertebroplasty are not optimal and are associated with several drawbacks, including high exothermic reaction, low cement viscosity and consequent extravasation, and unpredictable wait time after cement preparation. Additionally, strength and stiffness restoration are controversial varying with the cement type, volume injected, and technique used. To investigate maximum polymerization temperatures and mechanical performance of novel two-solution bone cement (TSBC) modified by the addition of cross-linked poly(methyl methacrylate) nanospheres (η-TSBC) and microspheres (μ-TSBC) in a cadaver vertebroplasty model in comparison to a commercially available cement (KyphX). To study the viability of application of these novel cement formulations in the treatment of vertebral compression fractures. Ex vivo biomechanical and exothermal evaluation of TSBCs using cadaveric vertebral bodies (VBs). Thirty-one cadaveric vertebrae (age, 74±2 years; T score, -1.5±0.5) were disarticulated. Thirteen vertebrae were assigned into three groups and instrumented with thermocouples positioned midbody along the intersection of the midsagittal and midcoronal axes, as well as along the intersection of the midsagittal axis and posterior VB wall. After equilibration at 37°C, 5 mL of cement was injected and temperatures were recorded for 1 hour. The groups were injected with η-TSBC, μ-TSBC, or KyphX. The remaining 18 vertebrae were biomechanically tested. After randomization into three groups, each specimen was fractured in compression and stabilized with 5 mL of each cement type. Each specimen was then retested in axial compression. Temperatures in the central region of the vertebrae were significantly lower (p<.05) when injected with η-TSBC (44°C) in comparison to KyphX (75°C) and μ-TSBC (64°C). A significant difference was not detected between the pre- and postcementing strength (p>.05) of the three groups. There was no significant difference between the average values of stiffness among the cements (p>.05), however there was a significant difference between intact and treated stiffness (p<.05). The TSBC cements decreased the local temperature within the VB while providing similar mechanical strength when compared with vertebrae treated with KyphX.


14-3-3zeta is an effector of tau protein phosphorylation.
Neurofibrillary tangles associated with Alzheimer's disease are composed mainly of paired helical filaments that are formed by the aggregation of abnormally phosphorylated microtubule-associated protein tau. 14-3-3, a highly conserved protein family that exists as seven isoforms and regulates diverse cellular processes is present in neurofibrillary tangles (Layfield, R., Fergusson, J., Aitken, A., Lowe, J., Landon, M., Mayer, R. J. (1996) Neurosci. Lett. 209, 57-60). The role of 14-3-3 in Alzheimer's disease pathogenesis is not known. In this study, we found that the 14-3-3zeta isoform is associated with tau in brain extract and profoundly stimulates cAMP-dependent protein kinase catalyzed in vitro phosphorylation on Ser(262)/Ser(356) located within the microtubule-binding region of tau. 14-3-3zeta binds to both phosphorylated and nonphosphorylated tau, and the binding site is located within the microtubule-binding region of tau. From brain extract, 14-3-3zeta co-purifies with microtubules, and tubulin blocks 14-3-3zeta-tau binding. Among four 14-3-3 isoforms tested, beta and zeta but not gamma and epsilon associate with tau. Our data suggest that 14-3-3zeta is a tau protein effector and may be involved in the abnormal tau phosphorylation occurring during Alzheimer's disease ontogeny.