Dynamic culture substrate that captures a specific extracellular matrix protein in response to light.
The development of methods for the off-on switching of immobilization or presentation of cell-adhesive peptides and proteins during cell culture is important because such surfaces are useful for the analysis of the dynamic processes of cell adhesion and migration. This paper describes a chemically functionalized gold substrate that captures a genetically tagged extracellular matrix protein in response to light. The substrate was composed of mixed self-assembled monolayers (SAMs) of three disulfide compounds containing (i) a photocleavable poly(ethylene glycol) (PEG), (ii) nitrilotriacetic acid (NTA) and (iii) hepta(ethylene glycol) (EG7). Although the NTA group has an intrinsic high affinity for oligohistidine tag (His-tag) sequences in its Ni2+-ion complex, the interaction was suppressed by the steric hindrance of coexisting PEG on the substrate surface. Upon photoirradiation of the substrate to release the PEG chain from the surface, this interaction became possible and hence the protein was captured at the irradiated regions, while keeping the non-specific adsorption of non-His-tagged proteins blocked by the EG7 underbrush. In this way, we selectively immobilized a His-tagged fibronectin fragment (FNIII7-10) to the irradiated regions. In contrast, when bovine serum albumin-a major serum protein-was added as a non-His-tagged protein, the surface did not permit its capture, with or without irradiation. In agreement with these results, cells were selectively attached to the irradiated patterns only when a His-tagged FNIII7-10 was added to the medium. These results indicate that the present method is useful for studying the cellular behavior on the specific extracellular matrix protein in cell-culturing environments.


Brain atrophy and intellectual impairment in heavy drinkers--a clinical, psychometric and computerized tomography study.
The frequency of cerebral and cerebellar atrophy was assessed by computerized tomography (CT) in 26 heavy drinkers. Findings were correlated with clinical deficits, with the results of psychometric testing using the Wechsler Adult Intelligence Scale (WAIS), and with the alcoholic history and nutritional status. Cerebral atrophy was present in 19 cases (73%), 16 of whom also had cerebellar atrophy. There was a good correlation between the degree of cerebral hemisphere atrophy and age and length of drinking history, but a poor correlation between neurological deficits and atrophy. Impairment of visuo-spatial and visuo-motor functions with sparing of other non-dominant hemisphere functions and of verbal skills was the characteristic pattern found with the WAIS in 18 subjects; four showed a more global depression of intellectual function. The degree of cerebral hemisphere atrophy correlated significantly with the impairment of non-dominant hemisphere functions but not with total IQ.


Developmental effects of maternal smoking during pregnancy on the human frontal cortex transcriptome.
Cigarette smoking during pregnancy is a major public health concern. While there are well-described consequences in early child development, there is very little known about the effects of maternal smoking on human cortical biology during prenatal life. We therefore performed a genome-wide differential gene expression analysis using RNA sequencing (RNA-seq) on prenatal (N = 33; 16 smoking-exposed) as well as adult (N = 207; 57 active smokers) human postmortem prefrontal cortices. Smoking exposure during the prenatal period was directly associated with differential expression of 14 genes; in contrast, during adulthood, despite a much larger sample size, only two genes showed significant differential expression (FDR < 10%). Moreover, 1,315 genes showed significantly different exposure effects between maternal smoking during pregnancy and direct exposure in adulthood (FDR < 10%)-these differences were largely driven by prenatal differences that were enriched for pathways previously implicated in addiction and synaptic function. Furthermore, prenatal and age-dependent differentially expressed genes were enriched for genes implicated in non-syndromic autism spectrum disorder (ASD) and were differentially expressed as a set between patients with ASD and controls in postmortem cortical regions. These results underscore the enhanced sensitivity to the biological effect of smoking exposure in the developing brain and offer insight into how maternal smoking during pregnancy affects gene expression in the prenatal human cortex. They also begin to address the relationship between in utero exposure to smoking and the heightened risks for the subsequent development of neuropsychiatric disorders.


Case report: Emergence of autochthonous cutaneous leishmaniasis in northeastern Texas and southeastern Oklahoma.
Autochthonous human cases of leishmaniasis in the United States are uncommon. We report three new cases of cutaneous leishmaniasis and details of a previously reported case, all outside the known endemic range in Texas. Surveys for enzootic rodent reservoirs and sand fly vectors were conducted around the residences of three of the case-patients during the summer of 2006; female Lutzomyia anthophora sand flies were collected at a north Texas and southeast Oklahoma residence of a case-patient, indicating proximity of a suitable vector. Urban sprawl, climatologic variability, or natural expansion of Leishmania mexicana are possible explanations for the apparent spread to the north and east. Enhanced awareness among healthcare providers in the south central region of the United States is important to ensure clinical suspicion of leishmaniasis, diagnosis, and appropriate patient management.


Evolution of the immune system in humans from infancy to old age.
This article reviews the development of the immune response through neonatal, infant and adult life, including pregnancy, ending with the decline in old age. A picture emerges of a child born with an immature, innate and adaptive immune system, which matures and acquires memory as he or she grows. It then goes into decline in old age. These changes are considered alongside the risks of different types of infection, autoimmune disease and malignancy.


Astrophysics and cosmology closing in on neutrino masses.
Massive neutrinos are expected in most grand unified theories that attempt to unify the strong and electroweak interactions. So far, heroic laboratory experiments have yielded only upper bounds on the masses of the elusive neutrinos. These bounds, however, are not very restrictive and cannot even exclude the possibility that the dark matter in the universe consists of neutrinos. The astrophysical and cosmological bounds on the masses of the muon and tau neutrinos, mv(vmicro) and mv(vtau), which already are much more restrictive than the laboratory bounds, and the laboratory bound on the mass of the electron neutrino, mv(vc), can be improved significantly by future astrophysical and cosmological observations that perhaps will pin down the neutrino masses. Indeed, the recent results from the solar neutrino experiments combined with the seesaw mechanism for generating neutrino masses suggest that mv(vc) approximately 10(-8) electron volts, mv(vmicro) approximately 10(-3) electron volts, and mv(vtau) approximately 10 electron volts, which can be tested in the near future by solar neutrino and accelerator experiments.


Simulation studies on bacteriorhodopsin alpha-helices.
Bacteriorhodopsin (BR) is a membrane protein which pumps protons through the plasma membrane. Transmembrane BR helical segments are subjected to simulation studies in order to investigate the effect of bilayer environment in various simulation conditions. A bilayer potential is introduced to the system to mimic the lipid membrane. The structures from the simulations are compared with the experimentally determined structures in terms of geometrical properties. Electrostatic contribution to the helix packing is also investigated. The simulation results show that the packing geometry of the transmembrane helices is highly affected by the bilayer potential. The results obtained from the simulations may be used for further simulation studies and analysis in investigating transmembrane helix packing.


Impairment of gradual muscle adjustment during wrist circumduction in Parkinson's disease.
Purposeful movements are attained by gradually adjusted activity of opposite muscles, or synergists. This requires a motor system that adequately modulates initiation and inhibition of movement and selectively activates the appropriate muscles. In patients with Parkinson's disease (PD) initiation and inhibition of movements are impaired which may manifest itself in e.g. difficulty to start and stop walking. At single-joint level, impaired movement initiation is further accompanied by insufficient inhibition of antagonist muscle activity. As the motor symptoms in PD primarily result from cerebral dysfunction, quantitative investigation of gradually adjusted muscle activity during execution of purposeful movement is a first step to gain more insight in the link between impaired modulation of initiation and inhibition at the levels of (i) cerebrally coded task performance and (ii) final execution by the musculoskeletal system. To that end, the present study investigated changes in gradual adjustment of muscle synergists using a manipulandum that enabled standardized smooth movement by continuous wrist circumduction. Differences between PD patients (N = 15, off-medication) and healthy subjects (N = 16) concerning the relation between muscle activity and movement performance in these groups were assessed using kinematic and electromyographic (EMG) recordings. The variability in the extent to which a particular muscle was active during wrist circumduction--defined as muscle activity differentiation--was quantified by EMG. We demonstrated that more differentiated muscle activity indeed correlated positively with improved movement performance, i.e. higher movement speed and increased smoothness of movement. Additionally, patients employed a less differentiated muscle activity pattern than healthy subjects. These specific changes during wrist circumduction imply that patients have a decreased ability to gradually adjust muscles causing a decline in movement performance. We propose that less differentiated muscle use in PD patients reflects impaired control of modulated initiation and inhibition due to decreased ability to selectively and jointly activate muscles.


Neurosurgical approach.
Glioblastoma multiforme is a highly infiltrative tumor that typically has a central region of necrosis surrounded by contrast-enhancing proliferative tumor cells surrounded by diffuse isolated tumor cells that migrate into the brain. The goal of surgery is often directed toward the central necrotic region and the imaging-defined enhancing margin. To limit morbidity from removing functional brain tissue, the infiltrative tumor cells found in surrounding brain are generally not considered part of the surgical target. This is also the site where tumors recur after treatment. It is well accepted by most surgeons and neuro-oncologists that, when possible, aggressive resection of malignant gliomas is the preferred initial step in management. Although there are limited randomized prospective studies that address extent of resection and survival, the benefit of aggressive surgical resection will not be debated in this report. Tumor resection to the maximum extent that is safely possible can decrease tumor burden and thereby enhance the effects of adjuvant therapies, improve symptoms from mass effect, reduce the frequency of seizures, and provide tissue for pathological and genomic studies to better identify and test novel therapy.Surgery for glioblastoma is highly dependent on imaging. Magnetic resonance imaging can provide an anatomic definition of the lesion and functional capacity of critical cortical regions and allow for precise localization within the brain. The common use of stereotactic guidance, intraoperative imaging, functional magnetic resonance imaging, and physiologic monitoring have enhanced the surgeon's ability to achieve aggressive tumor removal while protecting the patient from neurologic impairment. This review will address the use of these techniques as an important first step in managing patients with glioblastoma.


Dual-functional peptide conjugated gold nanorods for the detection and photothermal ablation of pathogenic bacteria.
It is urgent to find an effective and convenient method to detect and kill microbial strains due to the threat of pathogenic bacterial infection and the low effectiveness of antibiotics. In the present work, a dual-functional peptide coupling gold nanoconjugate (Au@P937 NRs) was facilely fabricated by conjugating an adhesion peptide (WGLHTSATNLYLHGGGC, P937) with Au nanorods (Au NRs), of which the peptide P937 was designed by the phage display biopanning technology and showed specific binding affinity to bacteria. After the modification of P937, the longitudinal surface plasmon resonance (LSPR) absorbance peak of the Au@P937 NRs was found to be very sensitive to the variation of bacterial concentration. The Au@P937 NRs exhibited enhanced NIR-based photothermal conversion efficiency. Moreover, the Au@P937 NRs exhibited good biocompatibility. Hence, the Au@P937 NRs could be used as a dual-functional nanoconjugate for the detection and photothermal ablation of bacteria. For the bacterial detection by the Au@P937 NRs, the detection limit could reach 46 cfu mL-1 for E. coli and 89 cfu mL-1 for S. aureus, respectively. Both E. coli and S. aureus could be killed completely in 10 min by the Au@P937 NRs under 808 nm laser irradiation. This work provided a facile, convenient and effective strategy for detecting and sterilizing bacteria.


Exploiting gas diffusion for non-invasive sampling in flow analysis: determination of ethanol in alcoholic beverages.
A tubular gas diffusion PTFE membrane is exploited for non-invasive sampling in flow analysis, aiming to develop an improved spectrophotometric determination of ethanol in alcoholic beverages. The probe is immersed into the sample, allowing ethanol to diffuse through the membrane. It is collected into the acceptor stream (acidic dichromate solution), leading to formation of Cr(III), monitored at 600 nm. The analytical curve is linear up to 50% (v/v) ethanol, baseline drift is < 0.005 absorbance during four working-hours, and sample throughput is 30 h(-1), meaning 0.6 mmol K2Cr2O7 per determination. Results are precise (r.s.d.< 2%) and in agreement with an official procedure.


Musculoskeletal Complaints in Transverse Upper Limb Reduction Deficiency and Amputation in The Netherlands: Prevalence, Predictors, and Effect on Health.
(1) To determine the prevalence of musculoskeletal complaints (MSCs) in individuals with upper limb absence in The Netherlands, (2) to assess the health status of individuals with upper limb absence in general and in relation to the presence of MSCs, and (3) to explore the predictors of development of MSCs and MSC-related disability in this population. Cross-sectional study: national survey. Twelve rehabilitation centers and orthopedic workshops. Individuals (n=263; mean age, 50.7±16.7y; 60% men) ≥18 years old, with transverse upper limb reduction deficiency (42%) or amputation (58%) at or proximal to the carpal level (response, 45%) and 108 individuals without upper limb reduction deficiency or amputation (n=108; mean age, 50.6±15.7y; 65% men) (N=371). Not applicable. Point and year prevalence of MSCs, MSC-related disability (Pain Disability Index), and general health perception and mental health (RAND-36 subscales). Point and year prevalence of MSCs were almost twice as high in individuals with upper limb absence (57% and 65%, respectively) compared with individuals without upper limb absence (27% and 34%, respectively) and were most often located in the nonaffected limb and upper back/neck. MSCs were associated with decreased general health perception and mental health and higher perceived upper extremity work demands. Prosthesis use was not related to presence of MSCs. Clinically relevant predictors of MSCs were middle age, being divorced/widowed, and lower mental health. Individuals with upper limb absence experienced more MSC-related disability than individuals without upper limb absence. Higher age, more pain, lower general and mental health, and not using a prosthesis were related to higher disability. Presence of MSCs is a frequent problem in individuals with upper limb absence and is associated with decreased general and mental health. Mental health and physical work demands should be taken into account when assessing such a patient. Clinicians should note that MSC-related disability increases with age.


Mice deficient in L-12/15 lipoxygenase show increased vulnerability to 3-nitropropionic acid neurotoxicity.
Considerable evidence supports a contributory role for leukocyte-type 12/15 Lipoxygenase (L-12/15 LO) in mediating hippocampal and cortical neuronal injury in models of Alzheimer's disease and stroke. Whether L-12/15 LO contributes to neuronal injury in a model of Huntington's disease (HD) has yet to be determined. HD is characterized by marked striatal neuronal loss, which can be mimicked in humans and animals by inhibition of mitochondrial complex II using 3-Nitropropionic acid (3-NP). Herein, we compared histological and behavioral outcomes between mice that were wild-type or null for L-12/15 LO following systemic injection of 3NP. We found that mice deficient in L-12/15 LO had a higher incidence of striatal lesions coincident with an increase in morbidity as compared to their wild-type littermate controls. This could not be explained by differential metabolism of 3-NP as striatal succinate dehydrogenase activity was inhibited to the same extent in both genotypes. The present results show that deleting L-12/15 LO is detrimental to the striatum in the setting of chronic, systemic 3-NP exposure and are consistent with the overall conclusion that region-specific effects may determine the ultimate outcome of L-12/15 LO activation in the setting of brain injury.


The role of diabetes mellitus in the higher prevalence of tuberculosis among Hispanics.
This research studied the relative contribution of diabetes mellitus to the increased prevalence of tuberculosis in Hispanics. A case-control study was conducted involving all 5290 discharges from civilian hospitals in California during 1991 who had a diagnosis of tuberculosis, and 37,366 control subjects who had a primary discharge diagnosis of deep venous thrombosis, pulmonary embolism, or acute appendicitis. Risk of tuberculosis was estimated as the odds ratio (OR) across race/ethnicity, with adjustment for other factors. Diabetes mellitus was found to be an independent risk factor for tuberculosis. The association of diabetes and tuberculosis was higher among Hispanics (adjusted OR [ORadj] = 2.95: 95% confidence interval [CI] = 2.61, 3.33) than among non-Hispanic Whites (ORadj = 1.31: 95% CI = 1.19. 1.45): among non-Hispanic Blacks, diabetes was not found to be associated with tuberculosis (ORadj = 0.93: 95% CI = 0.78, 1.09). Among Hispanics aged 25 to 54, the estimated risk of tuberculosis attributable to diabetes (25.2%) was equivalent to that attributable to HIV infection (25.5%). Diabetes mellitus remains a significant risk factor for tuberculosis in the United States. The association is especially notable in middle-aged Hispanics.


Banning front-of-package food labels: first Amendment constraints on public health policy.
In recent months, the FDA has begun a crackdown on misleading nutrition and health claims on the front of food packages by issuing warning letters to manufacturers and promising to develop stricter regulatory standards. Leading nutrition policy experts Marion Nestle and David Ludwig have called for an even tougher approach: a ban on all nutrition and health claims on the front of food packages. Nestle and Ludwig argue that most of these claims are scientifically unsound and misleading to consumers and that eliminating them would 'aid educational efforts to encourage the public to eat whole or minimally processed foods and to read the ingredients list on processed foods'. Nestle and Ludwig are right to raise concerns about consumer protection and public health when it comes to front-of-package food labels, but an outright ban on front-of-package nutrition and health claims would violate the First Amendment. As nutrition policy experts develop efforts to regulate front-of-package nutrition and health claims, they should be mindful of First Amendment constraints on government regulation of commercial speech.


Bioaerosol sampling optimization for community exposure assessment in cities with poor sanitation: A one health cross-sectional study.
Evidence of exposure to enteric pathogens through the air and associated risk of infection is scarce in the literature outside of animal- or human-waste handling settings. Cities with poor sanitation are important locations to investigate this aerial exposure pathway as their rapid growth will pose unprecedented challenges in waste management. To address this issue, simple surveillance methods are needed. Therefore, the objectives of this study were to optimize a community exposure bioaerosol surveillance strategy for urban outdoor locations with poor sanitation, and to determine which bioaerosols could contribute to exposure. Passive and active bioaerosol sampling methods were used to characterize the fate and transport of sanitation-related bioaerosols during the rainy and dry seasons in La Paz, Bolivia. Median coliform bacteria fluxes were 71 CFU/(m2 × h) during the rainy season and 64 CFU/(m2 × h) during the dry season, with 38% of the dry season samples testing positive for E. coli. Wind speed, relative humidity and UVB irradiance were identified as significant covariates to consider in bioaerosol transport models in La Paz. Active sampling yielded one positive sample (10%) for human adenovirus (HadV) and one sample (10%) for influenza A virus during the rainy season. HadV was detected at the site with the highest bacterial flux. Four samples (8%) were positive for influenza A virus in the dry season. These findings suggest that aerosols can contribute to community exposure to potentially pathogenic microorganisms in cities with poor sanitation. The use of passive sampling, despite its limitations, can provide quantitative data on microorganisms' viability within realistic timeframes of personal exposure.


Reexamining the vocabulary spurt.
The authors asked whether there is evidence to support the existence of the vocabulary spurt, an increase in the rate of word learning that is thought to occur during the 2nd year of life. Using longitudinal data from 38 children, they modeled the rate of word learning with two functions, one with an inflection point (logistic), which would indicate a spurt, and one without an inflection point (quadratic). Comparing the fits of these two functions using likelihood ratios, they found that just 5 children had a better logistic fit, which indicated that these children underwent a spurt. The implications for theories of cognitive and language development are considered.


Genetic control of the immune response to staphylococcal nuclease. III. Time-course and correlation between the response to native nuclease and the response to its polypeptide fragments.
The progression of the Ir gene-controlled antibody response to staphylococcal nuclease in mice with repeated immunizations has been examined. H-2-linked control of the response to a single immunization with 100 mug of nuclease in complete Freund's adjuvant was confirmed. However, among strains of the high responder H-2a haplotype, the response of the A/J mice was about 10-fold higher than that of the B10.A, indicating additional non-H-2-linked control. In addition, the low responder C57BL/10 (H-2b) strain produced antibody levels as high as or higher than those of the congenic high responder B10.A (H-2a) strain when both strains were repeatedly immunized, indicating complexity even in the H-2-linked control of the response to this small monomeric protein. Polypeptide fragments of nuclease were also studied as immunogens. The antibody response to one fragment (residues 99-149) was found to follow the same pattern among five strains tested as that to whole nuclease. However, in this case the C57BL/10 was found to be a nonresponder rather than a low responder, failing to develop a response despite repeated immunizations. In contrast, the C57BL/10 showed a low but significant response to another fragment (residues 1-126) of nuclease. These results suggest that the apparent H-2-linked control of the response to whole nuclease is a reflection of the ability to recognize a determinant(s) in the region from residues 99 to 149, and that the eventual response of the C57BL/10 strain after hyperimmunization reflects the recognition of other determinants. If these observations reflect the common recognition of a determinant on native nuclease and on a random-conformation fragment, they have implications about the conformational specificity of the receptors, or the flexibility of the determinants, involved in H-2-linked Ir-gene control. In addition, evidence is presented for a possible second H-2-linked gene (or genes) controlling the response to other determinants of nuclease expressed on the polypeptide fragments.


Calcitriol modulates receptor for advanced glycation end products (RAGE) in diabetic hearts.
Receptor for advanced glycation end products (RAGE) signaling pathway plays a vital role in diabetic cardiovascular complications. Calcitriol has been shown to exert various beneficial cardiovascular effects. The purpose of this study is to determine whether calcitriol can modulate RAGE expression, and study the potential mechanisms in diabetic hearts. Streptozotocin (65 mg/kg, intraperitoneal injection once) induced diabetic rats were treated with or without subcutaneous injections of calcitriol at a dose of 150 ng/kg/day for 4 weeks. Western blot was used to evaluate protein expressions of myocardial RAGE, TNF-α, p65 subunit of NF-κB (p65), α subunit of inhibitor of κB (IκBα), subunits of NADPH oxidase (NOX4 and p22(phox)), angiotensin II type 1 receptor (AT1R), TGF-β1, TGF-β receptor I, total and phosphorylated SMAD2/3 and ERK, matrix metalloproteinases 2 (MMP2), tissue inhibitors of metalloproteinases 2 (TIMP2) and procollagen I. As compared to control, diabetic rats had increased expressions of cardiac RAGE, TNF-α, p22(phox), AT1R, and TGF-β1, which were significantly attenuated in the diabetic rats treated with calcitriol. Calcitriol-treated diabetic hearts also had lesser expressions of p-SMAD2/3 and p-ERK signaling than those of diabetic hearts. Moreover, diabetic hearts had increased expressions of MMP2 and procollagen I and decreased TIMP2. However, calcitriol reverted the diabetic effects in procollagen I but not in MMP2 or TIMP2. Calcitriol decreased diabetic effects on RAGE and fibrosis, which may be caused by its modulation on AT1R and the anti-inflammatory and antioxidative potentials. Therefore, calcitriol may attenuate diabetic cardiomyopathy.


Differential reports of suicidal ideation and attempts of questioning adults compared to heterosexual, lesbian, gay, and bisexual individuals.
Many sexual minority individuals attempt suicide each year, but little is known about the suicidality of individuals who are questioning their sexual orientation. This study assessed suicidal ideation and attempts of questioning individuals compared to lesbian/gay, bisexual, and heterosexual individuals. This cross-sectional study enrolled participants (N = 2,841) from a community health center. Questioning (OR = 4.286, 95% CI [2.119-8.671]), lesbian/gay (OR = 3.024, 95% CI [2.351-3.890]), and bisexual (OR = 4.389, 95% CI [2.942-6.575]) individuals had significantly greater odds of considering suicide compared to heterosexuals. However, questioning individuals had non-significant odds of attempting suicide compared to heterosexuals. We discuss possible explanations for these findings.


Systemic Measures and Legislative and Organizational Frameworks Aimed at Preventing or Mitigating Drug Shortages in 28 European and Western Asian Countries.
Drug shortages have been identified as a public health problem in an increasing number of countries. This can negatively impact on the quality and efficiency of patient care, as well as contribute to increases in the cost of treatment and the workload of health care providers. Shortages also raise ethical and political issues. The scientific evidence on drug shortages is still scarce, but many lessons can be drawn from cross-country analyses. The objective of this study was to characterize, compare, and evaluate the current systemic measures and legislative and organizational frameworks aimed at preventing or mitigating drug shortages within health care systems across a range of European and Western Asian countries. The study design was retrospective, cross-sectional, descriptive, and observational. Information was gathered through a survey distributed among senior personnel from ministries of health, state medicines agencies, local health authorities, other health or pharmaceutical pricing and reimbursement authorities, health insurance companies and academic institutions, with knowledge of the pharmaceutical markets in the 28 countries studied. Our study found that formal definitions of drug shortages currently exist in only a few countries. The characteristics of drug shortages, including their assortment, duration, frequency, and dynamics, were found to be variable and sometimes difficult to assess. Numerous information hubs were identified. Providing public access to information on drug shortages to the maximum possible extent is a prerequisite for performing more advanced studies on the problem and identifying solutions. Imposing public service obligations, providing the formal possibility to prescribe unlicensed medicines, and temporary bans on parallel exports are widespread measures. A positive finding of our study was the identification of numerous bottom-up initiatives and organizational frameworks aimed at preventing or mitigating drug shortages. The experiences and lessons drawn from these initiatives should be carefully evaluated, monitored, and presented to a wider international audience for careful appraisal. To be able to find solutions to the problem of drug shortages, there is an urgent need to develop a set of agreed definitions for drug shortages, as well as methodologies for their evaluation and monitoring. This is being progressed.


Malignant melanoma metastatic to the thyroid gland: a case report and review of the literature.
The thyroid gland is a relatively uncommon site for a secondary malignancy; even less common is a case of malignant melanoma metastatic to the thyroid. We describe the case of a 68-year-old man who presented with a neck mass in the posterior triangle. Fine-needle aspiration biopsy (FNAB) identified the mass as a malignant melanoma. The patient had had no known primary skin melanoma. He underwent a left modified radical neck dissection, and the mass was discovered to be a positive lymph node. Postoperatively, he declined to undergo radio- and chemotherapy. Eighteen months later, he returned with a diffusely enlarged thyroid. FNAB again attributed the enlargement to malignant melanoma. Soon thereafter, the patient began experiencing seizures, and on magnetic resonance imaging, he was found to have metastatic disease to the brain. He developed ventilator-dependent respiratory failure and required a subtotal thyroidectomy for the placement of a tracheostomy tube. Patients who present with a thyroid nodule and who have a history of malignancy present a diagnostic dilemma: Is the nodule benign, a new primary, or a distant metastasis? The findings of this case and a review of the literature strengthen the argument that any patient with a thyroid mass and a history of malignancy should be considered to have a metastasis until proven otherwise.


Association of non-alcoholic fatty liver disease with chronic kidney disease: a systematic review and meta-analysis.
Chronic kidney disease (CKD) is a frequent, under-recognized condition and a risk factor for renal failure and cardiovascular disease. Increasing evidence connects non-alcoholic fatty liver disease (NAFLD) to CKD. We conducted a meta-analysis to determine whether the presence and severity of NAFLD are associated with the presence and severity of CKD. English and non-English articles from international online databases from 1980 through January 31, 2014 were searched. Observational studies assessing NAFLD by histology, imaging, or biochemistry and defining CKD as either estimated glomerular filtration rate (eGFR) <60 ml/min/1.73 m2 or proteinuria were included. Two reviewers extracted studies independently and in duplicate. Individual participant data (IPD) were solicited from all selected studies. Studies providing IPD were combined with studies providing only aggregate data with the two-stage method. Main outcomes were pooled using random-effects models. Sensitivity and subgroup analyses were used to explore sources of heterogeneity and the effect of potential confounders. The influences of age, whole-body/abdominal obesity, homeostasis model of insulin resistance (HOMA-IR), and duration of follow-up on effect estimates were assessed by meta-regression. Thirty-three studies (63,902 participants, 16 population-based and 17 hospital-based, 20 cross-sectional, and 13 longitudinal) were included. For 20 studies (61% of included studies, 11 cross-sectional and nine longitudinal, 29,282 participants), we obtained IPD. NAFLD was associated with an increased risk of prevalent (odds ratio [OR] 2.12, 95% CI 1.69-2.66) and incident (hazard ratio [HR] 1.79, 95% CI 1.65-1.95) CKD. Non-alcoholic steatohepatitis (NASH) was associated with a higher prevalence (OR 2.53, 95% CI 1.58-4.05) and incidence (HR 2.12, 95% CI 1.42-3.17) of CKD than simple steatosis. Advanced fibrosis was associated with a higher prevalence (OR 5.20, 95% CI 3.14-8.61) and incidence (HR 3.29, 95% CI 2.30-4.71) of CKD than non-advanced fibrosis. In all analyses, the magnitude and direction of effects remained unaffected by diabetes status, after adjustment for other risk factors, and in other subgroup and meta-regression analyses. In cross-sectional and longitudinal studies, the severity of NAFLD was positively associated with CKD stages. Limitations of analysis are the relatively small size of studies utilizing liver histology and the suboptimal sensitivity of ultrasound and biochemistry for NAFLD detection in population-based studies. The presence and severity of NAFLD are associated with an increased risk and severity of CKD. Please see later in the article for the Editors' Summary.


Selecting the aspect ratio of a scatter plot based on its delaunay triangulation.
Scatter plots are diagrams that visualize two-dimensional data as sets of points in the plane. They allow users to detect correlations and clusters in the data. Whether or not a user can accomplish these tasks highly depends on the aspect ratio selected for the plot, i.e., the ratio between the horizontal and the vertical extent of the diagram. We argue that an aspect ratio is good if the Delaunay triangulation of the scatter plot at this aspect ratio has some nice geometric property, e.g., a large minimum angle or a small total edge length. More precisely, we consider the following optimization problem. Given a set Q of points in the plane, find a scale factor s such that scaling the x-coordinates of the points in Q by s and the y-coordinates by 1=s yields a point set P(s) that optimizes a property of the Delaunay triangulation of P(s), over all choices of s. We present an algorithm that solves this problem efficiently and demonstrate its usefulness on real-world instances. Moreover, we discuss an empirical test in which we asked 64 participants to choose the aspect ratios of 18 scatter plots. We tested six different quality measures that our algorithm can optimize. In conclusion, minimizing the total edge length and minimizing what we call the 'uncompactness' of the triangles of the Delaunay triangulation yielded the aspect ratios that were most similar to those chosen by the participants in the test.


Lightweight Adaptation of Classifiers to Users and Contexts: Trends of the Emerging Domain.
Intelligent computer applications need to adapt their behaviour to contexts and users, but conventional classifier adaptation methods require long data collection and/or training times. Therefore classifier adaptation is often performed as follows: at design time application developers define typical usage contexts and provide reasoning models for each of these contexts, and then at runtime an appropriate model is selected from available ones. Typically, definition of usage contexts and reasoning models heavily relies on domain knowledge. However, in practice many applications are used in so diverse situations that no developer can predict them all and collect for each situation adequate training and test databases. Such applications have to adapt to a new user or unknown context at runtime just from interaction with the user, preferably in fairly lightweight ways, that is, requiring limited user effort to collect training data and limited time of performing the adaptation. This paper analyses adaptation trends in several emerging domains and outlines promising ideas, proposed for making multimodal classifiers user-specific and context-specific without significant user efforts, detailed domain knowledge, and/or complete retraining of the classifiers. Based on this analysis, this paper identifies important application characteristics and presents guidelines to consider these characteristics in adaptation design.


The influence of pH on iron speciation in podzol extracts: iron complexes with natural organic matter, and iron mineral nanoparticles.
The quantities of natural organic matter (NOM) and associated iron (Fe) in soil extracts are known to increase with increasing extractant pH. However, it was unclear how the extraction pH affects Fe speciation for particles below 30 nm. We used flow field-flow fractionation (FlowFFF) and transmission electron microscopy (TEM) to investigate the association of Fe and trace elements with NOM and nanoparticulate iron (oxy)hydroxides in podzol extracts. For extracts prepared at the native soil pH (~4), and within a 1-30 nm size range, Fe was associated with NOM. In extracts with a pH≥7 from the E and B soil horizons, Fe was associated with NOM as well as with iron (oxy)hydroxide nanoparticles with a size of approximately 10 nm. The iron (oxy)hydroxide nanoparticles may have either formed within the soil extracts in response to the increase in pH, or they were mobilized from the soil. Additionally, pH shift experiments showed that iron (oxy)hydroxides formed when the native soil pH (~4) was increased to 9 following the extraction. The iron (oxy)hydroxide nanoparticles aggregated if the pH was decreased from 9 to 4. The speciation of Fe also influenced trace element speciation: lead was partly associated with the iron (oxy)hydroxides (when present), while copper binding to NOM remained unaffected by the presence of iron (oxy)hydroxide nanoparticles. The results of this study are important for interpreting the representativeness of soil extracts prepared at a pH other than the native soil pH, and for understanding the changes in Fe speciation that occur along a pH gradient.


Rabies Outbreak in Captive Big Brown Bats (Eptesicus fuscus) Used in a White-Nose Syndrome Vaccine Trial.
An outbreak of rabies occurred in a captive colony of wild-caught big brown bats (Eptesicus fuscus). Five of 27 bats exhibited signs of rabies virus infection 22-51 d after capture or 18-22 d after contact with the index case. Rabid bats showed weight loss, aggression, increased vocalization, hypersalivation, and refusal of food. Antigenic typing and virus sequencing confirmed that all five bats were infected with an identical rabies virus variant that circulates in E. fuscus in the US. Two bats with no signs of rabies virus infection were seropositive for rabies virus-neutralizing antibodies; the brains of these bats had no detectable viral proteins by the direct fluorescence antibody test. We suspect bat-to-bat transmission of rabies virus occurred among our bats because all rabies-infected bats were confined to the cage housing the index case and were infected with viruses having identical sequences of the entire rabies nucleoprotein gene. This outbreak illustrates the risk of rabies virus infection in captive bats and highlights the need for researchers using bats to assume that all wild bats could be infected with rabies virus.


Common genetic variants regulating ADD3 gene expression alter biliary atresia risk.
Biliary atresia (BA) is a rare and most severe cholestatic disease in neonates, but the pathogenic mechanisms are unknown. Through a previous genome wide association study (GWAS) on Han Chinese, we discovered association of the 10q24.2 region encompassing ADD3 and XPNPEP1 genes, which was replicated in Chinese and Thai populations. This study aims to fully characterize the genetic architecture at 10q24.2 and to reveal the link between the genetic variants and BA. We genotyped 107 single nucleotide polymorphisms (SNPs) in 10q24.2 in 339 Han Chinese patients and 401 matched controls using Sequenom. Exhaustive follow-up studies of the association signals were performed. The combined BA-association p-value of the GWAS SNP (rs17095355) achieved 6.06×10(-10). Further, we revealed the common risk haplotype encompassing 5 tagging-SNPs, capturing the risk-predisposing alleles in 10q24.2 [p=5.32×10(-11); odds ratio, OR: 2.38; confidence interval, CI: (2.14-2.62)]. Through Sanger sequencing, no deleterious rare variants (RVs) residing in the risk haplotype were found, dismissing the theory of "synthetic" association. Moreover, in bioinformatics and in vivo genotype-expression investigations, the BA-associated potentially regulatory SNPs correlated with ADD3 gene expression (n=36; p=0.0030). Remarkably, the risk haplotype frequency coincides with BA incidences in the population, and, positive selection (favoring the derived alleles that arose from mutations) was evident at the ADD3 locus, suggesting a possible role for the BA-associated common variants in shaping the general population diversity. Common genetic variants in 10q24.2 can alter BA risk by regulating ADD3 expression levels in the liver, and may exert an effect on disease epidemiology and on the general population.


Proteasome and thiol involvement in quality control of glycosylphosphatidylinositol anchor addition.
Improperly processed secretory proteins are degraded by a hydrolytic system that is associated with the endoplasmic reticulum (ER) and appears to involve re-export of lumenal proteins into the cytoplasm for ultimate degradation by the proteasome. The chimaeric protein hGHDAF28, which contains a crippled glycosylphosphatidylinositol (GPI) C-terminal signal peptide, is degraded by a pathway highly similar to that for other ER-retained proteins and is characterized by formation of disulphide-linked aggregates, failure to reach the Golgi complex and intracellular degradation with a half life of approximately 2 h. Here we show that N-acetyl-leucinal-leucinal-norleucinal, MG-132 and lactacystin, all inhibitors of the proteasome, protect hGHDAF28; hGHDAF28 is still proteolytically cleaved in the presence of lactacystin or MG-132, by the removal of approximately 2 kDa, but the truncated fragment is not processed further. We demonstrate that the ubiquitination system accelerates ER-degradation of hGHDAF28, but is not essential to the process. Overall, these findings indicate that GPI quality control is mediated by the cytoplasmic proteasome. We also show that the presence of a cysteine residue in the GPI signal of hGHDAF28 is required for retention and degradation, as mutation of this residue to serine results in secretion of the fusion protein, implicating thiol-mediated retention as a mechanism for quality control of some GPI signals. Removal of the cysteine also prevents inclusion of hGHDAF28 in disulphide-linked aggregates, indicating that aggregate formation is an additional retention mechanism for this class of protein. Therefore our data suggest that an unpaired terminal cysteine is the retention motif of the hGHDAF28 GPI-processing signal and that additional information may be required for efficient engagement of ER quality control systems by the majority of GPI signals which lack cysteine residues.


CIPRO 2.5: Ciona intestinalis protein database, a unique integrated repository of large-scale omics data, bioinformatic analyses and curated annotation, with user rating and reviewing functionality.
The Ciona intestinalis protein database (CIPRO) is an integrated protein database for the tunicate species C. intestinalis. The database is unique in two respects: first, because of its phylogenetic position, Ciona is suitable model for understanding vertebrate evolution; and second, the database includes original large-scale transcriptomic and proteomic data. Ciona intestinalis has also been a favorite of developmental biologists. Therefore, large amounts of data exist on its development and morphology, along with a recent genome sequence and gene expression data. The CIPRO database is aimed at collecting those published data as well as providing unique information from unpublished experimental data, such as 3D expression profiling, 2D-PAGE and mass spectrometry-based large-scale analyses at various developmental stages, curated annotation data and various bioinformatic data, to facilitate research in diverse areas, including developmental, comparative and evolutionary biology. For medical and evolutionary research, homologs in humans and major model organisms are intentionally included. The current database is based on a recently developed KH model containing 36,034 unique sequences, but for higher usability it covers 89,683 all known and predicted proteins from all gene models for this species. Of these sequences, more than 10,000 proteins have been manually annotated. Furthermore, to establish a community-supported protein database, these annotations are open to evaluation by users through the CIPRO website. CIPRO 2.5 is freely accessible at http://cipro.ibio.jp/2.5.


Functional hyperemia in the brain: hypothesis for astrocyte-derived vasodilator metabolites.
Cerebral blood flow is tightly coupled to neuronal metabolic activity, a phenomenon referred to as functional hyperemia. The mechanisms underlying functional hyperemia in the brain have been extensively studied, but the link between neuronal activation and nutritive blood flow has yet to be defined. Recent investigations by our laboratory and others have identified a potential role for astrocytes as an intermediary cell type in this process. This short review will develop the hypothesis that cytochrome P450 epoxygenase activity in astrocytes catalyzes formation of epoxyeicosatrienoic acids (EETs), which act as potent dilators of cerebral vessels and are released in response to glutamate receptor activation within astrocytes. Neuronal activity stimulates release of arachidonic acid from the phospholipid pool of astrocytic membranes. We provide evidence that the arachidonic acid released on stimulation of glutamate receptors within astrocytes is metabolized by cytochrome P450 2C11 cDNA enzymes into EETs. The EETs thus formed will be released and activate K+ channels, increase outward K+ current, and hyperpolarize the plasma membrane. The resulting membrane hyperpolarization inhibits voltage-gated Ca2+ channels and leads to arteriolar dilation, thereby increasing regional nutritive blood flow in response to neuronal activity.


Acute and chronic sympathetic reflex activation and antihypertensive response to nifedipine.
The importance of counterregulatory mechanisms triggered by arterial vasodilation for the antihypertensive response to the calcium entry blocking agent nifedipine was investigated in 13 men with mild to moderate essential hypertension. Blood pressure and systemic vascular resistance were significantly reduced 30 minutes after sublingual administration of 10 mg of nifedipine while heart rate, cardiac index and plasma norepinephrine concentrations increased (all p less than 0.01). Also, changes in mean blood pressure correlated inversely with arterial baroreflex sensitivity (r = -0.74, p less than 0.01), suggesting that arterial baroreflex mechanisms by means of sympathetic activation tend to limit the acute antihypertensive response. Blood pressure, but not systemic vascular resistance, decreased further (p less than 0.01) after 6 weeks of therapy with nifedipine 20 mg three times daily, while average heart rate, cardiac index and plasma norepinephrine concentrations had returned toward pretreatment values. Thus, a reduction of acutely increased sympathetic activity toward pretreatment values during long-term nifedipine therapy was associated with further decreases in blood pressure. The importance of sympathetic activity was also stressed by the finding of an inverse relation between chronic changes in heart rate and blood pressure (percent of control, r = -0.76, p less than 0.01). Blood volume did not change during long-term nifedipine therapy. The results suggest that the degree of sympathetic reflex activation in part determines the antihypertensive response to nifedipine monotherapy.


Addition of glucagon to lipid-free total parenteral nutrition reduces production of prostaglandin E2 by stimulated splenic macrophages.
Sepsis is a major complication of total parenteral nutrition (TPN). Impaired immunity has been suggested as being responsible for TPN-related sepsis, but it is unknown how the immune system is affected by TPN. We recently found that administration of lipid-free TPN resulted in an increase in prostaglandin E2 (PGE2) release by stimulated splenic macrophages. This observation suggested that TPN may impair immunity through the prominent immunosuppressive effects of PGE2. In the present study, we tested the hypothesis that addition of glucagon to TPN solution may protect against the immunosuppressive effect of TPN by modifying PGE2 secretion. Adult, male Sprague-Dawley rats (n = 18) underwent jugular vein cannulation: group 1 (n = 7) received intravenous saline and chow ad libitum; group 2 (n = 6) received TPN (80 mL/24 h); and group 3 (n = 5) received TPN (80 mL/24 h) plus glucagon (100 micrograms/24 h). After 10 days, spleens were removed and splenic macrophages were isolated and cultured for 24 h in plain M199 medium (nonstimulated) or in medium containing Escherichia coli lipopolysaccharide (5 micrograms/mL) (stimulated). PGE2 release was determined by enzyme-linked immunosorbent assay. There were no differences in PGE2 release between the groups of nonstimulated cells, but when stimulated with lipopolysaccharide, the macrophages from the TPN rats (group 2) released more PGE2 (81.68 +/- 25.99 ng/2.5 x 10(6) cells) than the control group (16.04 +/- 3.26 ng/2.5 x 10(6) cells). The release of PGE2 was normalized in the TPN animals treated with glucagon (15.71 +/- 3.33 ng/2.5 x 10(6) cells). This difference was significant, with p < .05 by Tukey's test after analysis of variance.(ABSTRACT TRUNCATED AT 250 WORDS)


Arsenic speciation and reactivity in poultry litter.
Recent U.S. government action to lower the maximum concentration levels (MCL) of total arsenic (As) (10 ppb) in drinking water has raised serious concerns about the agricultural use of As-containing biosolids such as poultry litter (PL). In this study, solid-state chemical speciation, desorbability, and total levels of As in PL and long-term amended soils were investigated using novel synchrotron-based probing techniques (microfocused (micro) synchrotron X-ray fluorescence (SXRF) and micro-X-ray absorption near-edge structure (XANES) spectroscopies) coupled with chemical digestion and batch experiments. The total As levels in the PL were as high as approximately 50 mg kg(-1), and As(II/III and V) was always concentrated in abundant needle-shaped microscopic particles (approximately 20 microm x 850 microm) associated with Ca, Cu, and Fe and to a lesser extent with S, Cl, and Zn. Post-edge XANES features of litter particles are dissimilar to those of the organo-As(V) compound in poultry feed (i.e., roxarsone), suggesting possible degradation/transformation of roxarsone in the litter and/or in poultry digestive tracts. The extent of As desorption from the litter increased with increasing time and pH from 4.5 to 7, but at most 15% of the total As was released after 5 d at pH 7, indicating the presence of insoluble phases and/or strongly retained soluble compounds. No significant As accumulation (< 15 mg kg(-1)) was found in long-term PL-amended agricultural surface soils. This suggests that As in the PL may have undergone surface and subsurface transport processes. Our research results raise concerns about long-term PL amendment effects on As contamination in surrounding soil-water environments.


Insights into the growth mechanism of REF3 (RE = La-Lu, Y) nanocrystals: hexagonal and/or orthorhombic.
The synthesis of REF3 (RE = La-Lu, Y) nanocrystals with controlled phase structures has so far remained a challenge. Herein we have developed a one-for-all synthetic procedure that allows the successful synthesis of REF3 nanocrystals in a controlled manner. Experimental results showed that the radius of RE ions determines the phase structure: pure hexagonal REF3 (RE = La-Eu), a mixture of hexagonal and orthorhombic REF3 (RE = Gd), and pure orthorhombic REF3 (RE = Tb-Lu, Y) nanocrystals are obtained along with the decrease of the ionic radius. As Gd is positioned exactly in the middle of the lanthanides row, GdF3 nanocrystals were used as a model to further investigate how the molar ratio of F- : Gd3+, the doping of RE ions with different ionic radii, and the doping concentration of certain RE ions affects the crystal structure of the final product.


Short communication: Genetic evaluation of milking speed for Brown Swiss dairy cattle in the United States.
Genetic parameters and relative breeding values were estimated for milking speed of US Brown Swiss dairy cattle. Owner-recorded milking-speed scores on a scale of 1 (slow) to 8 (fast) were collected by the Brown Swiss Association as part of its linear type appraisal program starting in 2004. Data were 7,366 records for 6,666 cows in 393 herds. The pedigree file included information for 21,458 animals born in 1985 or later. Six unknown-parent groups that each included 4 birth years were defined. The model included fixed effects for herd appraisal date and parity-lactation stage and random effects for permanent environment, animal, and error. Within parity (1, 2, and > or =3), 6 groups were defined: unknown calving date, four 90-d lactation stages, and lactations with >400 d in milk. Heritability of 0.22 and repeatability of 0.42 were estimated by average-information REML; residual variance was 1.13. Little trend in estimated breeding value was found for cows born from 1999 through 2002. Although solutions increased with lactation stage for first-parity cows by 0.37, no clear trend was found for later parities. Genetic evaluations for milking speed were expressed as relative breeding values with a mean of 100 and a standard deviation of 5. The 121 bulls with > or =10 daughters had milking speed evaluations that ranged from 83 to 112 and had correlations of 0.56 with productive life evaluations and -0.40 with somatic cell score evaluations. The association of faster milking speed with lower somatic cell score was not expected. The moderate heritability found for milking speed indicates that the evaluations (first released in May 2006) should be useful in detecting bulls with slow-milking daughters.


The relationship of pretreatment family functioning to drinking behavior during follow-up by alcoholic patients.
The relationship of alcoholics' perceptions of the pretreatment functioning of their families to drinking outcomes during an 18-month follow-up was examined. Family functioning was hypothesized to be predictive of drinking behavior, particularly in subjects with low assertion of autonomy scores. These individuals report greater dependency and attachment, and therefore might be more affected by the state of important relationships. The results indicated that with low autonomy male alcoholics, greater family dysfunction predicted significantly fewer days abstinent during Months 1-6 and 13-18 and more severe drinking episodes during the first year of the follow-up. In the high autonomy males, family dysfunction was unrelated to subsequent drinking behavior. In women, on the other hand, greater family dysfunction predicted more days abstinent in those high in autonomy and was unrelated to the drinking behavior of those low in autonomy. Implications for patient-treatment matching, differences between male and female alcoholics, and the need for additional studies of family functioning and drinking behavior in women are discussed.


Humor in systemic lupus erythematosus.
Humor has neurophysiological effects influencing the release of cortisol, which may have a direct impact on the immune system. Laughter is associated with a decreased production of inflammatory cytokines both in the general population and in rheumatoid arthritis (RA). Our objective was to explore the effects of humor on serum cytokines [particularly interleukin-6 (IL-6)] and cortisol levels in systemic lupus erythematosus (SLE), after a standard intervention (120 min of visual comedy). We enrolled 58 females with SLE from consecutive patients assessed in the Montreal General Hospital lupus clinic. The subjects who consented to participate were randomized in a 1:1 ratio to the intervention (watching 120 min of comedy) or control group (watching a 120 min documentary). Measurements of cytokine and serum cortisol levels as well as 24-h urine cortisol were taken before, during, and after the interventions. We compared serum cytokine levels and serum and 24-h urine cortisol levels in the humor and control groups and performed regression analyses of these outcomes, adjusting for demographics and the current use of prednisone. There were no significant differences between the control and humor groups in demographics or clinical variables. Baseline serum levels of IL-6, IL-10, tumor necrosis factor-alpha, and B-cell activating factor were also similar in both groups. There was no evidence of a humor effect in terms of decreasing cytokine levels, although there was some suggestion of lowered cortisol secretion in the humor group based the 24-h urinary cortisol levels in a subgroup. In contrast to what has been published for RA, we saw no clear effects of humor in altering cytokine levels in SLE, although interesting trends were seen for lower cortisol levels after humor intervention compared with the control group.


Gunshot injuries to the liver: the role of selective nonoperative management.
Selective nonoperative management of blunt liver injuries has become standard practice in most trauma centers. We evaluated the role of selective nonoperative management of gunshot wounds to the liver. This was a retrospective review of gunshot wounds to the liver treated in a level I trauma center. Patients with peritoneal signs or hemodynamic instability were operated on without delay. Patients with a soft, nontender abdomen and no signs of heavy bleeding were selected for nonoperative management. Liver injury was diagnosed by CT scan. If peritonitis or signs of substantial internal bleeding developed, an operation was performed; otherwise the patient was discharged within a few days of admission. Analysis was restricted to the group of patients with isolated liver injuries. During a 42-month period, 928 patients were admitted with abdominal gunshot injuries, 152 of whom (16%) had a liver injury. In 52 patients (34% of all liver injuries), the liver was the only injured intraabdominal organ (4 patients had associated kidney or splenic injuries that did not require surgical repair). Thirty-six of the patients (69%) with isolated liver injuries had an emergent operation because of signs of peritonitis or hemodynamic instability. The remaining 16 patients (31%) were selected for nonoperative management (3 patients had associated right kidney injury). Five patients in the observed group required delayed operation because of development of signs of peritonitis (4 patients) or abdominal compartment syndrome (1 patient). The remaining 11 patients (7% of all liver injuries or 21% of isolated liver injuries) were managed successfully without operation. One patient with delayed operation developed multiple complications from abdominal compartment syndrome, and 1 patient in the nonoperative group had a biloma, which was treated with percutaneous drainage. Selected patients with isolated grades I and II gunshot wounds to the liver can be managed nonoperatively.


A Fast Approximate Algorithm for Mapping Long Reads to Large Reference Databases.
Emerging single-molecule sequencing technologies from Pacific Biosciences and Oxford Nanopore have revived interest in long-read mapping algorithms. Alignment-based seed-and-extend methods demonstrate good accuracy, but face limited scalability, while faster alignment-free methods typically trade decreased precision for efficiency. In this article, we combine a fast approximate read mapping algorithm based on minimizers with a novel MinHash identity estimation technique to achieve both scalability and precision. In contrast to prior methods, we develop a mathematical framework that defines the types of mapping targets we uncover, establish probabilistic estimates of p-value and sensitivity, and demonstrate tolerance for alignment error rates up to 20%. With this framework, our algorithm automatically adapts to different minimum length and identity requirements and provides both positional and identity estimates for each mapping reported. For mapping human PacBio reads to the hg38 reference, our method is 290 × faster than Burrows-Wheeler Aligner-MEM with a lower memory footprint and recall rate of 96%. We further demonstrate the scalability of our method by mapping noisy PacBio reads (each ≥5 kbp in length) to the complete NCBI RefSeq database containing 838 Gbp of sequence and >60,000 genomes.


Horseshoe Kidney Protection with Histidine-Tryptophan-Ketoglutarate Solution during Surgical Abdominal Aortic Aneurysm Repair.
We report an unusual case of a 65-year-old male patient with horseshoe kidney, who underwent a successful open repair for an abdominal aortic aneurysm (AAA). The accessory renal arteries were perfused with histidine-tryptophan-ketoglutarate (HTK) solution (Custodiol®; Dr. Franz-Kohler Chemie GmbH, Bensheim, Germany) during the vascular reconstruction. There were no creatinine and estimated glomerular filtration rate (eGFR) modifications in the postoperative time. In the literature, only two cases of Custodiol solution for kidney protection during aortic surgery are reported. To the best of our knowledge, this is the first case of Custodiol perfusion for horseshoe kidney protection. The concomitant presence of horseshoe kidney and an AAA requires a specific preoperative planning. This is necessary to define the appropriate surgical procedure and strategy. The onset of acute kidney injury is an aspect that must always be taken into consideration during aortic surgery, even more in the case we are reporting. Indeed, despite the complexity of the kidney anatomy, the use of Custodiol solution allowed a proper maintenance of the perioperative renal function, as shown by the postoperative levels of creatinine and eGFR. Preoperative planning and organ preservation are crucial in AAA open repair, especially in the presence of congenital anomalies such as horseshoe kidney.


[Ethnic differences in forensic psychiatry: an exploratory study at a Dutch forensic psychiatric centre].
Compared to the percentage of ethnic minorities in the general population, ethnic minorities are overrepresented in forensic psychiatry. If these minorities are to be treated successfully, we need to know more about this group. So far, however, little is known about the differences between mental disorders and types of offences associated with patients of non-Dutch descent and those associated with patients of Dutch descent.
 AIM: To take the first steps to obtain the information we need in order to provide customised care for patients of non-Dutch descent.
 METHOD: It proved possible to identify differences between patients of Dutch and non-Dutch descent with regard to treatment, diagnosis and offences committed within a group of patients who were admitted to the forensic psychiatric centre Oostvaarderskliniek during the period 2001 - 2014.
 RESULTS: The treatment of patients of non-Dutch descent lasted longer than the treatment of patients of Dutch descent (8.5 year versus 6.6 year). Furthermore, patients from ethnic minority groups were diagnosed more often with schizophrenia (49.1% versus 21.4%), but less often with pervasive developmental disorders or sexual disorders. Patients of non-Dutch descent were more often convicted for sexual crimes where the victim was aged 16 years or older, whereas patients of Dutch descent were convicted of sexual crimes where the victim was under 16.
 CONCLUSION: There are differences between patients of Dutch and non-Dutch descent with regard to treatment duration, diagnosis and offences they commit. Future research needs to investigate whether these results are representative for the entire field of forensic psychiatry and to discover the reasons for these differences.


Antiplatelet effect of sanguinarine is correlated to calcium mobilization, thromboxane and cAMP production.
Sanguinarine is a plant alkaloid present in the root of Sanguinaria canadensis and Poppy fumaria species. Sanguinarine has been used as an antiseptic mouth rinse and a toothpaste additive to reduce dental plaque and gingival inflammation. In this study, we investigated the antiplatelet effects of sanguinarine, aiming to extend its potential pharmacological applications. Sanguinarine inhibited platelet aggregation induced by arachidonic acid (AA), collagen, U46619 and sub-threshold concentration of thrombin (0.05 U/ml) with IC(50) concentrations of 8.3, 7.7, 8.6 and 4.4 microM, respectively. Sanguinarine (5-10 microM) inhibited 10-31% of platelet TXB(2) production, but not platelet aggregation induced by higher concentration of thrombin (0.1 U/ml). SQ29548, a thromboxane receptor antagonist, inhibited the AA-induced platelet aggregation but not TXB(2) production. Sanguinarine suppressed cyclooxygenase-1 (COX-1) activity (IC(50)=28 microM), whereas its effect on COX-2 activity was minimal. Sanguinarine (8, 10 microM) further inhibited the AA-induced Ca(2+) mobilization by 27-62%. In addition, SQ22536, an adenylate cyclase inhibitor, attenuated the inhibitory effect of sanguinarine toward AA-induced platelet Ca(2+) mobilization and aggregation. These results suggest that sanguinarine is a potent antiplatelet agent, which activates adenylate cyclase, inhibits platelet Ca(2+) mobilization, TXB(2) production as well as suppresses COX-1 enzyme activity. Sanguinarine may have therapeutic potential for treatment of cardiovascular diseases related to platelet aggregation.


Biofeedback and the behavioral treatment of disorders of disregulation.
This paper reviews biofeedback research from the perspective of cybernetic/feedback theory and applies the theory to the behavioral treatment of psychosomatic disorders. The concept of disregulation is used to elucidate how environmental factors can modulate the central nervous system and effect homeostatic, self-regulatory control of peripheral organs. When feedback from peripheral organs is disrupted, it is hypothesized that disregulation occurs, leading to physiological instability and functional disease. Within this framework, biofeedback provides a new feedback loop that can help individuals regain physiological self-control. Basic research using biofeedback to enhance self-regulation of cardiovascular responses is reviewed. The use of biofeedback in the behavioral treatment of disorders such as tension and migraine headache, hypertension, and epilepsy are selectively reviewed and critically evaluated. The need to consider feedback mechanisms in behavioral and biomedical approaches to treatment is highlighted. Predictions regarding the potential inadvertent perpetuation of disregulation and disease through inappropriate biomedical intervention is also considered.


Staphylococcus aureus autoinducer-2 quorum sensing decreases biofilm formation in an icaR-dependent manner.
Staphylococcus aureus is an important pathogen that causes biofilm-associated infection in humans. Autoinducer 2 (AI-2), a quorum-sensing (QS) signal for interspecies communication, has a wide range of regulatory functions in both Gram-positive and Gram-negative bacteria, but its exact role in biofilm formation in S. aureus remains unclear. Here we demonstrate that mutation of the AI-2 synthase gene luxS in S. aureus RN6390B results in increased biofilm formation compared with the wild-type (WT) strain under static, flowing and anaerobic conditions and in a mouse model. Addition of the chemically synthesized AI-2 precursor in the luxS mutation strain (ΔluxS) restored the WT phenotype. Real-time RT-PCR analysis showed that AI-2 activated the transcription of icaR, a repressor of the ica operon, and subsequently a decreased level of icaA transcription, which was presumably the main reason why luxS mutation influences biofilm formation. Furthermore, we compared the roles of the agr-mediated QS system and the LuxS/AI-2 QS system in the regulation of biofilm formation using the ΔluxS strain, RN6911 and the Δagr ΔluxS strain. Our data indicate a cumulative effect of the two QS systems on the regulation of biofilm formation in S. aureus. These findings demonstrate that AI-2 can decrease biofilm formation in S. aureus via an icaR-activation pathway. This study may provide clues for therapy in S. aureus biofilm-associated infection.


Posttranscriptional regulation of p21WAF1/CIP1 expression in human breast carcinoma cells.
p21WAF1/CIP1 plays a major role in the induction of G1 arrest following DNA damage. Although p21WAF1/CIP1 expression is regulated by the tumor suppressor p53, induction of p21WAF1/CIP1 expression through p53-independent pathways has been described in numerous cell types. In this report, we describe the mechanism by which the retinoid 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalene carboxylic acid (CD437) induces p21WAF1/CIP1 in breast carcinoma cells possessing either a wild-type (MCF-7 cells) or mutated (MDA-MB-468 cells) p53. Exposure of MDA-MB-468 cells to this retinoid results in an approximately 10-fold increase in p21WAF1/CIP1 mRNA levels, whereas less than a 2-fold increase in p21WAF1/CIP1 gene transcription was observed as indicated by transient transfection experiments utilizing a p21WAF1/CIP1 promoter firefly luciferase reporter gene construct and nuclear run-off studies. We found similar results in the MCF-7 cells (Z-M. Shao et al., Oncogene, 11: 493-504, 1995). We have now found that while enhancing p21WAF1/CIP1 gene transcription minimally, this retinoid increases p21WAF1/CIP1 mRNA stability by 3-fold in both cell types. We also demonstrate that approximately 1.5 kb of the 3' untranslated region causes enhanced instability of p21WAF1/CIP1 mRNA. The retinoid-dependent increase in p21WAF1/CIP1 mRNA stability is accompanied by an increase in p21WAF1/CIP1 protein expression, as indicated by Western blot experiments utilizing anti-p21WAF1/CIP1 monoclonal antibody. This increase in p21WAF1/CIP1 is subsequently followed by the onset of programmed cell death in both cell types. Thus, CD437 is a novel retinoid which enhances p21WAF1/CIP1 mRNA levels through stabilization of the message regardless of the p53 status of the cell.


Smartphones for community health in rural Cambodia: A feasibility study.
Background: Village Malaria Workers (VMWs) are lay people trained to provide a valuable role in frontline testing and treatment of malaria in rural villages in Cambodia. Emergence of artemisinin-resistant malaria highlights the essential role of such VMWs in surveillance and early treatment of malaria. Smartphone technology offers huge potential to support VMWs in isolated and resource-poor settings. Methods: We investigated the feasibility of issuing established VMWs with a smartphone, bespoke Android application and solar charger to support their role. 27 VMWs in Kampong Cham and Kratie provinces participated. Results: 26/27 of the smartphones deployed were working well at study completion twelve months later. Interviews with VMWs using quantitative and qualitative methods revealed pride, ease of use and reports of faster communication with the smartphone. VMWs also expressed a strong wish to help people presenting with non-malarial fever, for which further potential supportive smartphone applications are increasingly available. Conclusions: As a result of this pilot study, two smartphone based reporting systems for malaria have been developed at the Cambodian National Malaria Center, and the programme is now being extended nationwide. The full code for the smartphone application is made available to other researchers and healthcare providers with this article. Smartphones represent a feasible platform for developing the VMW role to include other health conditions, thus maintaining the relevance of these important community health workers.


From phenotypic to molecular polymorphisms involved in naturally occurring variation of plant development.
An enormous amount of naturally occurring genetic variation affecting development is found within wild and domesticated plant species. This diversity is presumably involved in plant adaptation to different natural environments or in human preferences. In addition, such intraspecific variation provides the basis for the evolution of plant development at larger evolutionary scales. Natural phenotypic differences are now amenable to genetic dissection up to the identification of causal DNA polymorphisms. Here we describe 30 genes and their functional nucleotide polymorphisms currently found as underlying allelic variation accounting for plant intraspecific developmental diversity. These studies provide molecular and cellular mechanisms that determine natural variation for quantitative and qualitative traits such as: fruit and seed morphology, colour and composition; flowering time; seedling emergence; plant architecture and inflorescence or flower morphology. Besides, analyses of flowering time variation within several distant species allow molecular comparisons between species, which are detecting homologous genes with partly different functions and unrelated genes with analogous functions. Thus, considerable gene function differences are being revealed also among species. Inspection of a catalogue of intraspecific nucleotide functional polymorphisms shows that transcriptional regulators are the main class of genes involved. Furthermore, barely more than half of the polymorphisms described are located in coding regions and affect protein structure, while the rest are regulatory changes altering gene expression. These limited analyses of intraspecific developmental variation support Doebley and Lukens's proposition (1998) that modifications in cis -regulatory regions of transcriptional regulators represent a predominant mode for the evolution of novel forms, but await more detailed studies in wild plant species.


1H NMR study of the role of heme carboxylate side chains in modulating heme pocket structure and the mechanism of reconstitution of cytochrome b5.
1H nuclear magnetic resonance spectroscopy was used to assign the hyperfine-shifted resonances and determine the position of a side chain in the heme cavity of wild-type rat apocytochrome b5 reconstituted with a series of synthetic hemins possessing systematically perturbed carboxylate side chains. The hemins included protohemin derivatives with individually removed or pairwise shortened and lengthened carboxylate side chains, as well as (propionate)n(methyl)8-nporphine-iron(III) isomers with n = 1-3 designed to force occupation of nonnative propionate sites. The resonance assignments were effected on the basis of available empirical heme contact shift correlations and steady-state nuclear Overhauser effect measurements in the low-spin oxidized proteins. The failure to detect holoproteins with certain hemins dictates that the stable holoproteins, unlike the case of myoglobin, demand the axial iron-His bonds and cannot accommodate carboxylate side chains at interior positions in the binding pocket. Hence, the heme pocket interior in cytochrome b5 is judged much less polar and less sterically accommodating than that of myoglobin. The propionate occupational preference was greatest as the native 7-propionate site, but also possible at the nonnative crystallographic 5-methyl or 8-methyl positions. Only for a propionate at the crystallographic 8-methyl position was a significant perturbation of the native molecular/electronic structure observed, and this was attributed to an alternative propionate-protein hydrogen bond at the crystallographic 8-methyl position. The structures of the transient protein complexes detected only shortly after reconstitution reveal that the initial encounter complexes during assembly of holoprotein from apoprotein and hemin involve one of the two alternate propionate-protein links at either the 7-propionate or native 8-methyl position. In a monopropionate hemin, this leads to the characterization of a new type of heme orientational disorder involving rotation about a N-Fe-N axis.


High muscular fitness has a powerful protective cardiometabolic effect in adults: influence of weight status.
Low levels of muscular fitness (MF) are recognized as an important marker of nutritional status and a predictor of metabolic complications, cardiovascular disease and death, however, the relationship between MF, body mass index (BMI) and the subsequent cardiometabolic protective effects has been less studied among Latin American populations. This study identified an association between MF and the cardiometabolic risk score index (CMRSI) and the lipid-metabolic cardiovascular risk index (LMCRI) in a wide sample of university students grouped according to their BMI. Six thousand ninety five healthy males (29.6 ± 11.7 year-old) participated in the study. Absolute strength was measured using a T.K.K. analogue dynamometer (handgrip), and the participant's strength was then calculated relative to their body mass (MF/BM). The LMCRI was derived from the levels of triglycerides, low-density lipoprotein cholesterol (LDL-c), high-density lipoprotein cholesterol (HDL-c), and glucose levels in a blood sample. The CMRSI was calculated by summing the standardized residuals (z-score) for waist circumference, total cholesterol, LDL-c, triglycerides, HDL-c, and median blood pressure. Subjects were divided into six subgroups according to BMI (normal vs. overweight/obese) and MF/BM tertiles (unfit, average, fit). The group of participants with low and moderate levels of MF/BM showed higher CMRSI values independent of BMI (P < 0.001). The group with normal BMI and high MF/BM had the highest levels of cardiometabolic protection. All overweight/obese BMI groups had significantly higher LMCRI values independent of the level of MF/BM (P < 0.001). Participants with high MF/BM showed reduced cardiometabolic risk, which increased significantly when they were within normal parameters.


Characterization of a new strain of HHV-6 (HHV-6SF) recovered from the saliva of an HIV-infected individual.
An isolate of the human herpesvirus-6 (HHV-6SF) recovered from the saliva of an HIV-infected individual differs in its cellular host range and certain genomic properties from other HHV-6 strains described. HHV-6SF replicates in adult peripheral blood mononuclear cells (PMC) substantially better than in fetal cord blood PMC and can be grown only in the MT-4 established T cell line. It preferentially infects CD4+ lymphocytes but can replicate in CD8+ cells and peripheral blood macrophages. It also infects neuroblastoma cells and cell lines derived from the gastrointestinal tract. These latter results suggest that this herpesvirus could play a role in disorders affecting these tissues. Finally, the restriction enzyme pattern of HHV-6SF differs from that of other HHV-6 strains. The identification of this distinct HHV-6 strain could indicate an unusual biologic variation among viral isolates thus far not observed with other herpesviruses.


A 3-month-old baby with H1N1 and Guillain-Barré syndrome.
Majority of children with pandemic influenza A (H1N1)pdm09 experience mild illness with full recovery without treatment. A previously healthy two and a half month-old girl was admitted to our paediatric intensive care unit because of severe respiratory failure with A (H1N1)pdm09 infection. Despite initial clinical improvement all attempts to extubate to non-invasive ventilation were unsuccessful and 2 to 3 weeks after symptom onset she started periods of cardiovascular instability and a progressive neurological deterioration with distal symmetrical progressive motor weakness and areflexia. All investigations were normal except elevated liver enzymes and cerebrospinal fluid examination that revealed elevated protein without pleocytosis. A possible diagnosis of Guillain-Barré syndrome (GBS) was considered and electromyogram was compatible with axonal form of GBS. To our knowledge this is the youngest case of GBS acquired postnatally and the first in children associated with H1N1 virus.


The Sanger FASTQ file format for sequences with quality scores, and the Solexa/Illumina FASTQ variants.
FASTQ has emerged as a common file format for sharing sequencing read data combining both the sequence and an associated per base quality score, despite lacking any formal definition to date, and existing in at least three incompatible variants. This article defines the FASTQ format, covering the original Sanger standard, the Solexa/Illumina variants and conversion between them, based on publicly available information such as the MAQ documentation and conventions recently agreed by the Open Bioinformatics Foundation projects Biopython, BioPerl, BioRuby, BioJava and EMBOSS. Being an open access publication, it is hoped that this description, with the example files provided as Supplementary Data, will serve in future as a reference for this important file format.


Systemic accident analysis: examining the gap between research and practice.
The systems approach is arguably the dominant concept within accident analysis research. Viewing accidents as a result of uncontrolled system interactions, it forms the theoretical basis of various systemic accident analysis (SAA) models and methods. Despite the proposed benefits of SAA, such as an improved description of accident causation, evidence within the scientific literature suggests that these techniques are not being used in practice and that a research-practice gap exists. The aim of this study was to explore the issues stemming from research and practice which could hinder the awareness, adoption and usage of SAA. To achieve this, semi-structured interviews were conducted with 42 safety experts from ten countries and a variety of industries, including rail, aviation and maritime. This study suggests that the research-practice gap should be closed and efforts to bridge the gap should focus on ensuring that systemic methods meet the needs of practitioners and improving the communication of SAA research.


Meta-ethnography to understand healthcare professionals' experience of treating adults with chronic non-malignant pain.
We aimed to explore healthcare professionals' experience of treating chronic non-malignant pain by conducting a qualitative evidence synthesis. Understanding this experience from the perspective of healthcare professionals will contribute to improvements in the provision of care. Qualitative evidence synthesis using meta-ethnography. We searched five electronic bibliographic databases from inception to November 2016. We included studies that explore healthcare professionals' experience of treating adults with chronic non-malignant pain. We used the GRADE-CERQual framework to rate confidence in review findings. We screened the 954 abstracts and 184 full texts and included 77 published studies reporting the experiences of over 1551 international healthcare professionals including doctors, nurses and other health professionals. We abstracted six themes: (1) a sceptical cultural lens, (2) navigating juxtaposed models of medicine, (3) navigating the geography between patient and clinician, (4) challenge of dual advocacy, (5) personal costs and (6) the craft of pain management. We rated confidence in review findings as moderate to high. This is the first qualitative evidence synthesis of healthcare professionals' experiences of treating people with chronic non-malignant pain. We have presented a model that we developed to help healthcare professionals to understand, think about and modify their experiences of treating patients with chronic pain. Our findings highlight scepticism about chronic pain that might explain why patients feel they are not believed. Findings also indicate a dualism in the biopsychosocial model and the complexity of navigating therapeutic relationships. Our model may be transferable to other patient groups or situations.


Therapeutic vaccination targeting the tumour vasculature.
Therapeutic vaccination targeting self-molecules could provide a cost-efficient alternative to monoclonal antibody-based therapies for cancer and various inflammatory diseases. However, development of cancer vaccines targeting self-molecules has proven difficult. One complicating factor is that tumour cells have developed strategies to escape recognition by the immune system. Antigens specifically expressed by the tumour vasculature can therefore provide alternative targets. The present mini-review highlights potential target molecules associated with tumour angiogenesis and the approaches made to direct an immune response against them. Furthermore, the requirements on a vaccine targeting self-molecules, in contrast with those directed against virus or bacteria, are discussed.


Continual lifelong learning with neural networks: A review.
Humans and animals have the ability to continually acquire, fine-tune, and transfer knowledge and skills throughout their lifespan. This ability, referred to as lifelong learning, is mediated by a rich set of neurocognitive mechanisms that together contribute to the development and specialization of our sensorimotor skills as well as to long-term memory consolidation and retrieval. Consequently, lifelong learning capabilities are crucial for computational learning systems and autonomous agents interacting in the real world and processing continuous streams of information. However, lifelong learning remains a long-standing challenge for machine learning and neural network models since the continual acquisition of incrementally available information from non-stationary data distributions generally leads to catastrophic forgetting or interference. This limitation represents a major drawback for state-of-the-art deep neural network models that typically learn representations from stationary batches of training data, thus without accounting for situations in which information becomes incrementally available over time. In this review, we critically summarize the main challenges linked to lifelong learning for artificial learning systems and compare existing neural network approaches that alleviate, to different extents, catastrophic forgetting. Although significant advances have been made in domain-specific learning with neural networks, extensive research efforts are required for the development of robust lifelong learning on autonomous agents and robots. We discuss well-established and emerging research motivated by lifelong learning factors in biological systems such as structural plasticity, memory replay, curriculum and transfer learning, intrinsic motivation, and multisensory integration.


Blue and UV LED-induced fluorescence in cotton foreign matter.
Cotton is an important domesticated fiber used to manufacture a variety of products and industrial goods. During harvesting with cotton strippers and cotton pickers, it is contaminated with foreign matter from botanical and non-botanical sources which adversely affect the quality and consistency of cotton, and therefore reduces its market value. To improve the current grading done by the High Volume Instrument (HVI) and human inspectors, it was explored whether fluorescence imaging can be used for cotton foreign matter detection. Eight types of botanical foreign matter (bark, bract, brown leaf, green leaf, hull, seed coat, seed, stem), and four types of non-botanical foreign matter (paper, twine, plastic bale packaging, plastic bag) were subjected to a fluorescence spectroscopy analysis to determine their optimal excitation and emission wavelength range. Matrix 3D scans were performed in the excitation range from 300 nm to 500 nm, and emission range from 320 nm to 700 nm, and the results indicated the photo-excitable fluorescence in the aforementioned excitation range for all the selected foreign matter categories. Consequently, the blue and the UV LEDs were selected as the excitation sources. The blue LED light provided optimal excitation light for bark, brown leaf, bract, green leaf, hull, and stem, while the UV LED light provided optimal excitation light for paper, plastic bag, plastic packaging, seed, seed coat, and twine. UV and blue light induces fluorescence in 12 types of botanical and non-botanical cotton foreign matter. An imaging apparatus with blue and UV light excitation sources, and a consumer grade SLR camera was successfully developed to capture and characterize fluorescent images of cotton foreign matter. Based on the results, fluorescent imaging could be a promising method for cotton foreign matter detection. Future studies will focus on the classification of cotton foreign matter categories and to further refine the image processing sequence.


Dynamic reorganization of chromatin structure and selective DNA demethylation prior to stable enhancer complex formation during differentiation of primary hematopoietic cells in vitro.
In order to gain insights in the true molecular mechanisms involved in cell fate decisions, it is important to study the molecular details of gene activation where such decisions occur, which is at the level of the chromatin structure of individual genes. In the study presented here we addressed this issue and examined the dynamic development of an active chromatin structure at the chicken lysozyme locus during the differentiation of primary myeloid cells from transgenic mouse bone marrow. Using in vivo footprinting we found that stable enhancer complex assembly and high-level gene expression are late events in cell differentiation. However, even before the onset of gene expression and stable transcription factor binding, specific chromatin alterations are observed. This includes changes in DNA topology and the selective demethylation of CpG dinucleotides located in the cores of critical transcription factor binding sites, but not in flanking DNA. These results firmly support the idea that epigenetic programs guiding blood cell differentiation are engraved into the chromatin of lineage-specific genes and that such chromatin changes are implemented before cell lineage specification.


Surface runoff losses of phosphorus from Virginia soils amended with turkey manure using phytase and high available phosphorus corn diets.
Many states have passed legislation that regulates agricultural P applications based on soil P levels and crop P uptake in an attempt to protect surface waters from nonpoint P inputs. Phytase enzyme and high available phosphorus (HAP) corn supplements to poultry feed are considered potential remedies to this problem because they can reduce total P concentrations in manure. However, less is known about their water solubility of P and potential nonpoint-source P losses when land-applied. This study was conducted to determine the effects of phytase enzyme and HAP corn supplemented diets on runoff P concentrations from pasture soils receiving surface applications of turkey manure. Manure from five poultry diets consisting of various combinations of phytase enzyme, HAP corn, and normal phytic acid (NPA) corn were surface-applied at 60 kg P ha(-1) to runoff boxes containing tall fescue (Festuca arundinacea Schreb.) and placed under a rainfall simulator for runoff collection. The alternative diets caused a decrease in manure total P and water soluble phosphorus (WSP) compared with the standard diet. Runoff dissolved reactive phosphorus (DRP) concentrations were significantly higher from HAP manure-amended soils while DRP losses from other manure treatments were not significantly different from each other. The DRP concentrations in runoff were not directly related to manure WSP. Instead, because the mass of manure applied varied for each treatment causing different amounts of manure particles lost in runoff, the runoff DRP concentrations were influenced by a combination of runoff sediment concentrations and manure WSP.


Squamous cell carcinoma of the skin (non-metastatic).
Cutaneous squamous cell carcinoma is a malignant tumour of keratinocytes arising in the epidermis, with histological evidence of dermal invasion. Incidence varies by country and skin colour, and is as high as 400/100,000 in Australia. People with fair skin colour who sunburn easily without tanning, people with xeroderma pigmentosum, and people who are immunosuppressed are most susceptible to squamous cell carcinoma. We conducted a systematic review and aimed to answer the following clinical questions: Does the use of sunscreen help prevent cutaneous squamous cell carcinoma and solar keratosis? What is the optimal margin for primary excision of cutaneous squamous cell carcinoma (non-metastatic)? Does micrographically controlled surgery result in lower rates of local recurrence than standard primary excision in people with squamous cell carcinoma of the skin (non-metastatic)? Does radiotherapy after surgery affect local recurrence of cutaneous squamous cell carcinoma in people with squamous cell carcinoma of the skin (non-metastatic)? We searched: Medline, Embase, The Cochrane Library, and other important databases up to October 2009 (Clinical Evidence reviews are updated periodically, please check our website for the most up-to-date version of this review). We included harms alerts from relevant organisations such as the US Food and Drug Administration (FDA) and the UK Medicines and Healthcare products Regulatory Agency (MHRA). We found 11 systematic reviews, RCTs, or observational studies that met our inclusion criteria. We performed a GRADE evaluation of the quality of evidence for interventions. In this systematic review we present information relating to the effectiveness and safety of the following interventions: micrographically controlled surgery, primary excision, radiotherapy after surgery, and sunscreens.


Cyclin D1 gene amplification in human laryngeal squamous cell carcinomas: prognostic significance and clinical implications.
The cyclin D1 (CCND1) gene is amplified, rearranged, and overexpressed frequently in human cancer, including squamous cell carcinoma. The gene dosage of CCND1 was examined in 51 primary laryngeal squamous cell carcinomas, and amplification of the gene was found in 9 (17.6%) cases. CCND1 amplification did not correlate with age, tumor localization and extension, cervical lymph node involvement, histopathological grading, and epidermal growth factor receptor levels. In a univariate analysis, CCND1 amplification, tumor extension, lymph node involvement, poor histological differentiation, and high epidermal growth factor receptor levels were correlated significantly with shorter overall survival. In a median follow-up period of 29 months, the overall survival rate was 71.4% for patients affected with tumors displaying a normal CCND1 dosage and only 25.0% for patients affected with tumors carrying amplified CCND1 (P = 0.0288). In a multivariate analysis, only CCND1 and tumor extension retained statistically significant prognostic values (P = 0.037 and 0.041, respectively). This is the first report in which CCND1 amplification is identified as a significant independent prognostic factor in laryngeal carcinoma. Evaluation of CCND1 amplification could be applicable to the clinical management of laryngeal cancer, allowing identification of patients with poor prognoses.


Early Cellular Response to Radiation in Human Vocal Fold Fibroblasts.
Radiation therapy is a common treatment strategy for laryngeal carcinoma. However, radiation is not without adverse side effects, especially toward healthy vocal fold tissue, which can lead to long-term impairments in vocal function. The objective of this preliminary study was to investigate early responses of healthy human vocal fold fibroblasts (VFF) to radiation. VFF were exposed to a single or fractionated dose radiation scheme. Nonradiated VFF served as controls. Morphology of radiated and control VFF was subjectively examined. Quantitative polymerase chain reaction was used to evaluate the effect of radiation on extracellular matrix and inflammatory-related genes. VFF viability was investigated using a LIVE/DEAD and clonogenic assay. Single or fractioned dose radiated VFF were morphologically indistinguishable from control VFF. No significant differences in gene expression were observed following either radiation scheme and as compared to controls. Clonogenic assay revealed reduced VFF viability following the fractionated but not single dose scheme. No changes in viability were detected using the LIVE/DEAD assay. We present one of the first investigations to evaluate early responses of healthy VFF to radiation. Findings will contribute to a growing body of literature seeking to elucidate the biological mechanisms underlying voice changes following radiation therapy for laryngeal carcinoma.