Processing of serum proteins underlies the mass spectral fingerprinting of myocardial infarction.
The MALDI-TOF spectra of peptides from the sera of normal and myocardial infarction patients produced patterns that provided an accurate diagnostic of MI. In myocardial infarction, the spectral pattern originated from the cleavage of complement C3 alpha chain to release the C3f peptide and cleavage of fibrinogen to release peptide A. The fibrinogen peptide A and complement C3f peptide were in turn progressively truncated by aminopeptidases to produce two families of fragments that formed the characteristic spectral pattern of MI. Time course and inhibitor studies demonstrated that the peptide patterns in the serum reflect the balance of disease-specific-protease and aminopeptidase activity ex vivo.


Cancer survival in Australia, Canada, Denmark, Norway, Sweden, and the UK, 1995-2007 (the International Cancer Benchmarking Partnership): an analysis of population-based cancer registry data.
Cancer survival is a key measure of the effectiveness of health-care systems. Persistent regional and international differences in survival represent many avoidable deaths. Differences in survival have prompted or guided cancer control strategies. This is the first study in a programme to investigate international survival disparities, with the aim of informing health policy to raise standards and reduce inequalities in survival. Data from population-based cancer registries in 12 jurisdictions in six countries were provided for 2·4 million adults diagnosed with primary colorectal, lung, breast (women), or ovarian cancer during 1995-2007, with follow-up to Dec 31, 2007. Data quality control and analyses were done centrally with a common protocol, overseen by external experts. We estimated 1-year and 5-year relative survival, constructing 252 complete life tables to control for background mortality by age, sex, and calendar year. We report age-specific and age-standardised relative survival at 1 and 5 years, and 5-year survival conditional on survival to the first anniversary of diagnosis. We also examined incidence and mortality trends during 1985-2005. Relative survival improved during 1995-2007 for all four cancers in all jurisdictions. Survival was persistently higher in Australia, Canada, and Sweden, intermediate in Norway, and lower in Denmark, England, Northern Ireland, and Wales, particularly in the first year after diagnosis and for patients aged 65 years and older. International differences narrowed at all ages for breast cancer, from about 9% to 5% at 1 year and from about 14% to 8% at 5 years, but less or not at all for the other cancers. For colorectal cancer, the international range narrowed only for patients aged 65 years and older, by 2-6% at 1 year and by 2-3% at 5 years. Up-to-date survival trends show increases but persistent differences between countries. Trends in cancer incidence and mortality are broadly consistent with these trends in survival. Data quality and changes in classification are not likely explanations. The patterns are consistent with later diagnosis or differences in treatment, particularly in Denmark and the UK, and in patients aged 65 years and older. Department of Health, England; and Cancer Research UK.


Tobacco industry success in Costa Rica: the importance of FCTC article 5.3.
To analyze how the tobacco industry influenced tobacco control policymaking in Costa Rica. Review of tobacco industry documents, tobacco control legislation, newspaper articles, and interviewing of key informants. During the mid-to-late 1980s, Health Ministry issued several advanced (for their time) smoking restriction decrees causing British American Tobacco (BAT) and Philip Morris International (PMI) to strengthen their political presence there, resulting in passage of a weak 1995 law, which, as of August 2011, remained in effect. Since 1995 the industry has used Costa Rica as a pilot site for Latin American programs and has dominated policymaking by influencing the Health Ministry, including direct private negotiations with the tobacco industry which violate Article 5.3's implementing guidelines of the World Health Organization Framework Convention on Tobacco Control (WHO FCTC). The Costa Rica experience demonstrates the importance of vigorous implementation of FCTC Article 5.3 which insulates public health policymaking from industry interference.


The role of biomineralization in disorders of skeletal development and tooth formation.
The major mineralized tissues are bone and teeth, which share several mechanisms governing their development and mineralization. This crossover includes the hormones that regulate circulating calcium and phosphate concentrations, and the genes that regulate the differentiation and transdifferentiation of cells. In developing endochondral bone and in developing teeth, parathyroid hormone-related protein (PTHrP) acts in chondrocytes to delay terminal differentiation, thereby increasing the pool of precursor cells. Chondrocytes and (in specific circumstances) pre-odontoblasts can also transdifferentiate into osteoblasts. Moreover, bone and teeth share outcomes when affected by systemic disorders of mineral homeostasis or of the extracellular matrix, and by adverse effects of treatments such as bisphosphonates and fluoride. Unlike bone, teeth have more permanent effects from systemic disorders because they are not remodelled after they are formed. This Review discusses the normal processes of bone and tooth development, followed by disorders that have effects on both bone and teeth, versus disorders that have effects in one without affecting the other. The takeaway message is that bone specialists should know when to screen for dental disorders, just as dental specialists should recognize when a tooth disorder should raise suspicions about a possible underlying bone disorder.


d-limonene ameliorates diabetes and its complications in streptozotocin-induced diabetic rats.
It is known that diabetes causes some complications including alterations in lipid profile, hepatic enzyme levels but also it causes oxidative stress. Limonene, a major component of Citrus oils, has important health beneficial effects in lowering the level of oxidative stress due to its antioxidant activity. The aim of this study was to investigate the effects of D-limonene on streptozotocin (STZ)-induced diabetes in Wistar albino rats. For this purpose, DNA damage was evaluated by alkaline comet assay. Changes in the activities of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR) and glutathione peroxidase (GSHPx) and the levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG), total glutathione (GSH), malondialdehyde (MDA), insulin, total bilirubin and BCA protein, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyl transferase (GGT), high density lipoprotein (HDL), low density lipoprotein (LDL), total cholesterol and triglyceride were also evaluated. D-limonene treatment was found to significantly decrease DNA damage, GR enzyme activities and MDA levels and significantly increase GSH levels and CAT, SOD and GSH-Px enzyme activities and altered lipid and liver enzyme parameters in diabetic rats. According to our results, it seems that D-limonene might have a role in the prevention of the complication of diabetes in rats.


Exposure to Genocide as a Risk Factor for Homicide Perpetration in Rwanda: A Population-Based Case-Control Study.
A population-based case-control study was conducted to assess the relationship between genocide exposure and homicide perpetration in Rwanda. A sample of 150 homicide perpetrators who were charged with and confessed to having committed homicide between 1 May 2011 and 31 May 2013 and 450 controls were enrolled. Cases were matched to controls by neighborhood, age and sex. Socio-demographic, background and genocide-related information was collected from study subjects' next of kin. Four characteristics of genocide exposure were: genocide survivor, genocide perpetrator, having lost a first-degree relative to genocide and having a first-degree relative convicted of genocide. We assessed the impact of each genocide-exposure variable using conditional logistic regression. Of the 150 cases, 124 (82.7%) were male and 26 (17.3%) were female. The mean age of the alleged homicide perpetrators was 33 years, with a peak in the age group 20-29 years (39.3%). After adjusting for socio-demographic characteristics and past common criminal records, having a first-degree relative who had been convicted of genocide crimes was a significant predictor for homicide perpetration (odds ratio [OR] = 14.4, 95% confidence interval [CI] = 1.6-129.4). Being a genocide perpetrator, a genocide survivor and having lost a first-degree family member to genocide were not identified as risk factors for homicide perpetration. In Rwanda, young people who experienced early exposure to trauma by witnessing their first-degree relatives' active participation in the genocide, are more likely to commit homicide. Socio-economic and psychotherapeutic programs targeting this population group are needed to rehabilitate these young people for violent behavior change.


Measuring the role of seagrasses in regulating sediment surface elevation.
Seagrass meadows provide numerous ecosystem services and their rapid global loss may reduce human welfare as well as ecological integrity. In common with the other 'blue carbon' habitats (mangroves and tidal marshes) seagrasses are thought to provide coastal defence and encourage sediment stabilisation and surface elevation. A sophisticated understanding of sediment elevation dynamics in mangroves and tidal marshes has been gained by monitoring a wide range of different sites, located in varying hydrogeomorphological conditions over long periods. In contrast, similar evidence for seagrasses is sparse; the present study is a contribution towards filling this gap. Surface elevation change pins were deployed in four locations, Scotland, Kenya, Tanzania and Saudi Arabia, in both seagrass and unvegetated control plots in the low intertidal and shallow subtidal zone. The presence of seagrass had a highly significant, positive impact on surface elevation at all sites. Combined data from the current work and the literature show an average difference of 31 mm per year in elevation rates between vegetated and unvegetated areas, which emphasizes the important contribution of seagrass in facilitating sediment surface elevation and reducing erosion. This paper presents the first multi-site study for sediment surface elevation in seagrasses in different settings and species.


Molecular Characterization of a Long-Term Survivor Double Metastatic Non-Small Cell Lung Cancer and Pancreatic Ductal Adenocarcinoma Treated with Gefitinib in Combination with Gemcitabine Plus Nab-Paclitaxel and mFOLFOX6 as First and Second Line Therapy.
The management of multiple primary cancers, an event not so infrequent in oncology practice, is a critical issue due to the lack of literature. In this study, we reported the case of a patient with non-small cell metastatic lung cancer (NSCLC) and pancreatic ductal adenocarcinoma (PDAC) who received gefitinib in combination with gemcitabine plus nab-paclitaxel and with mFOLFOX6 in first and second line, respectively. It achieved a progression-free survival and a28-months overall survival (OS) for NSCLC and PFS-1 and OS of 20 and 13 months, respectively for PDAC. Moreover, the combination of gefitinib and chemotherapy treatmentsshowed a good safety profile. Given the insignificant frequency of this case, we performed a molecular characterization of both neoplasms with the aim to investigate the existence of particular activated pathways and/or similar immunological mutations. It is interesting to note that two neoplasms shared a common mutation ofthe B7-H3 gene, with the consecutive impairment of its expressed protein. In both PDAC and NSCLC, the expression of this protein was associated with a worse survival rate. Since B7-H3 is an anti-apoptotic protein, the reduction of its expression or function should justify a pro-apoptotic activity with a leading justification of the long survival of the patient considered in this report.


Continuous wavelet transform of laser-Doppler signals from facial microcirculation reveals vasomotion asymmetry.
Facial laser Doppler flux (FLDF) consists of a mean value and complex oscillatory components called vasomotion. Vasomotion can be expressed as spectral amplitudes (SA) after the application of continuous wavelet transform. Vasomotion is influenced by cardiac activity (frequency interval I), respiration (interval II), intrinsic myogenic activity (interval III), neural mechanisms (interval IV) and endothelial mechanisms (intervals V and VI). Asymmetry of FLDF mean value was previously documented and hereby we present homogeneity assessment for FLDF vasomotion. Mean FLDF (p<0.001), total SA (p<0.001) and SA within frequency intervals I-VI were significantly higher in both cheeks compared to forehead. Total SA (p=0.009) and SA within frequency intervals I (p=0.041), II (p=0.005), III (p=0.009), IV (p=0.001) and V (p=0.036) were significantly higher in right than in left forehead. Human face is a heterogeneous microvascular region. Angiographic characteristics of deep horizontal sub-dermal plexus, endothelial and vascular smooth muscle cell heterogeneity, and plasticity of the microvasculature, autonomic asymmetry and facial neuropsychological asymmetry are possible causes of microvascular asymmetry. The origin and significance of microvascular imbalance need to be elucidated further.


Effectiveness of oils rich in linoleic and linolenic acids to enhance conjugated linoleic acid in milk from dairy cows.
Forty Holstein dairy cows were used to determine the effectiveness of linoleic or linolenic-rich oils to enhance C18:2 cis-9, trans-11 conjugated linoleic acid (CLA) and C18:1 trans-11 (vaccenic acid; VA) in milk. The experimental design was a complete randomized design for 9 wk with measurements made during the last 6 wk. Cows were fed a basal diet containing 59% forage (control) or a basal diet supplemented with either 4% soybean oil (SO), 4% flaxseed oil (FO), or 2% soybean oil plus 2% flaxseed oil (SFO) on a dry matter basis. Total fatty acids in the diet were 3.27, 7.47, 7.61, and 7.50 g/100 g in control, SO, FO, and SFO diets, respectively. Feed intake, energy-corrected milk (ECM) yield, and ECM produced/kg of feed intake were similar among treatments. The proportions of VA were increased by 318, 105, and 206% in milk fat from cows in the SO, FO, and SFO groups compared with cows in the control group. Similar increases in C18:2 cis-9, trans-11 CLA were 273, 150, and 183% in SO, FO, and SFO treatments, respectively. Under similar feeding conditions, oils rich in linoleic acid (soybean oil) were more effective in enhancing VA and C18:2 cis-9, trans-11 CLA in milk fat than oils containing linolenic acid (flaxseed oil) in dairy cows fed high-forage diets (59% forage). The effects of mixing linoleic and linolenic acids (50:50) on enhancing VA and C18:2 cis-9, trans-11 CLA were additive, but not greater than when fed separately. Increasing the proportion of healthy fatty acids (VA and CLA) by feeding soybean or flaxseed oil would result in milk with higher nutritive and therapeutic value.


TARGET-Seq: A Protocol for High-Sensitivity Single-Cell Mutational Analysis and Parallel RNA Sequencing.
Single-cell RNA-sequencing technologies are ideally placed to resolve intratumoral heterogeneity. However, the lack of coverage across key mutation hotspots has precluded the correlation of genetic and transcriptional readouts from the same single cell. To overcome this, we developed TARGET-seq, a protocol for TARGETed high-sensitivity single-cell mutational analysis with extremely low allelic dropout rates, parallel RNA SEQuencing, and cell-surface proteomics. Here, we present a detailed step-by-step protocol for TARGET-seq, including troubleshooting tips, approaches for automation, and methods for high-throughput multiplexing of libraries. For complete details on the use and execution of this protocol, please refer to Rodriguez-Meira et al. (2019).


Beyond Auditory Sensory Processing Deficits: Lexical Tone Perception Deficits in Chinese Children With Developmental Dyslexia.
Increasing evidence suggests that children with developmental dyslexia exhibit a deficit not only at the segmental level of phonological processing but also, by extension, at the suprasegmental level. However, it remains unclear whether such a suprasegmental phonological processing deficit is due to a difficulty in processing acoustic cues of speech rhythm, such as rise time and intensity. This study set out to investigate to what extent suprasegmental phonological processing (i.e., Cantonese lexical tone perception) and rise time sensitivity could distinguish Chinese children with dyslexia from typically developing children. Sixteen children with dyslexia and 44 age-matched controls were administered a Cantonese lexical tone perception task, psychoacoustic tasks, a nonverbal reasoning ability task, and word reading and dictation tasks. Children with dyslexia performed worse than controls on Cantonese lexical tone perception, rise time, and intensity. Furthermore, Cantonese lexical tone perception appeared to be a stable indicator that distinguishes children with dyslexia from controls, even after controlling for basic auditory processing skills. These findings suggest that suprasegmental phonological processing (i.e., lexical tone perception) is a potential factor that accounts for reading difficulty in Chinese.


Role of ghrelin in the pathophysiology of eating disorders: implications for pharmacotherapy.
Ghrelin is the only known circulating orexigenic hormone. It increases food intake by interacting with hypothalamic and brainstem circuits involved in energy balance, as well as reward-related brain areas. A heightened gut-brain ghrelin axis is an emerging feature of certain eating disorders such as anorexia nervosa and Prader-Willi syndrome. In common obesity, ghrelin levels are lowered, whereas post-meal ghrelin levels remain higher than in lean individuals. Agents that interfere with ghrelin signalling have therapeutic potential for eating disorders, including obesity. However, most of these drugs are only in the preclinical phase of development. Data obtained so far suggest that ghrelin agonists may have potential in the treatment of anorexia nervosa, while ghrelin antagonists seem promising for other eating disorders such as obesity and Prader-Willi syndrome. However, large clinical trials are needed to evaluate the efficacy and safety of these drugs.


Chewing cycles in 4- and 5-year-old normal children: an index of eating efficacy.
Eating movements in the preschool child undergo change between the ages of 2 and 5 years. There is a lack of objective clinical data from normal children against which eating movements of feeding-impaired children can be compared. In this study, chewing movements were measured to complement tongue movements described in an earlier study. The movements were monitored in 40 children: 20 were four years old and 20 were five years old. Each group had ten boys and ten girls. Chewing movements were measured by time (sec), number of cycles, and a time/cycle ratio. A chewing cycle was defined as an upward and downward movement of the chin. Total time from the moment food was placed in the mouth until the final swallow occurred was divided by the number of cycles counted for the same period. Age and sex did not affect time, cycles, or the time/cycle ratio. However, the measures were strongly affected by the type of food eaten. These findings suggest that the texture of food strongly influences both the number of chewing cycles performed and the time used for chewing. The occupational therapist administering eating evaluations to children should carefully choose the foods offered for initial evaluations and use the same food consistently during re-evaluations.


Plasma and urine dimercaptopropanesulfonate concentrations after dermal application of transdermal DMPS (TD-DMPS).
2,3-Dimercaptopropane-1-sulfonate (DMPS) is a metal chelator approved in Europe for oral or intravenous use for heavy metal poisoning. Transdermally applied DMPS (TD-DMPS) is used by some alternative practitioners to treat autism, despite the absence of evidence for its efficacy. We found no literature evaluating the pharmacokinetics of the transdermal route of delivery or the ability of TD-DMPS to enhance urinary mercury elimination. We hypothesized that TD-DMPS is not absorbed. Eight adult volunteers underwent application of 1.5-3 drops/kg of TD-DMPS. Subjects provided 12-h urine collections the day before and day of application. Subjects underwent blood draws at 0, 30, 60,90, 120, and 240 min after TD-DMPS application. Plasma and urine were assayed for the presence of DMPS. Urine was assayed for any change in urinary mercury excretion after DMPS. One control subject ingested 250 mg of oral DMPS and underwent the same urine and blood collections and analyses. No subject had detectable urine DMPS or increased urine mercury excretion after TD-DMPS. One subject had detectable levels of DMPS in the 30-min plasma sample, suspected to be contamination. All other samples for that subject and the other seven subjects showed no detectable plasma DMPS. The control subject had detectable urine and plasma DMPS levels and increased urine mercury excretion. These results indicate that TD-DMPS is not absorbed. There was no increase in urine mercury excretion after TD-DMPS. Our results argue that TD-DMPS is an ineffective metal chelator.


Assessment for self-blame and trauma symptoms during the medical evaluation of suspected sexual abuse.
The purpose of this study was to describe behavioural and emotional symptoms and to examine the effect of abuse-related factors, family responses to disclosure, and child self-blame on these symptoms in children presenting for medical evaluations after disclosure of sexual abuse. A retrospective review was conducted of 501 children ages 8-17. Trauma symptoms were determined by two sets of qualitative measures. Abstracted data included gender, ethnicity, and age; severity of abuse and abuser relationship to child; child responses regarding difficulty with sleep, school, appetite/weight, sadness, or self-harm, parent belief in abuse disclosure, and abuse-specific self-blame; responses to the Trauma Symptom Checklist in Children-Alternate; and the parent's degree of belief in the child's sexual abuse disclosure. Overall, 83% of the children had at least one trauma symptom; 60% had difficulty sleeping and one-third had thoughts of self-harm. Child age and abuse severity were associated with 3 of 12 trauma symptoms, and abuse-specific self-blame was associated with 10 trauma symptoms, after controlling for other variables. The children of parents who did not completely believe the initial disclosure of abuse were twice as likely to endorse self-blame as children of parents who completely believed the initial disclosure. Screening for behavioural and emotional problems during the medical assessment of suspected sexual abuse should include assessment of self-blame and family responses to the child's disclosures. In addition, parents should be informed of the importance of believing their child during the initial disclosure of abuse and of the impact this has on the child's emotional response to the abuse.


An Intervention to Improve Physical Function and Caregiver Perceptions in Family Caregivers of Persons With Heart Failure.
Objective: This randomized controlled trial was conducted to determine whether a 12-week home-based aerobic and resistance exercise program would improve physical function and caregiving perceptions among family caregivers (FCGs) of persons with heart failure. Method: Overall, 127 FCGs were randomized to one of three groups: usual care attention control (UCAC), psychoeducation only (PE), and psychoeducation plus exercise (PE + EX). Physical function measures (6-min walk test, handgrip, and upper and lower strength) and caregiving perceptions (Bakas Caregiving Outcomes Scale) were obtained at baseline and at 6 months. Results: FCGs in the PE + EX showed significant improvement in 6-min walk distance (p = .012), handgrip, and lower extremity strength compared with the PE and UCAC groups. The combined group had the greatest improvement in caregiver perceptions (p < .001). Conclusion: FCGs in the PE + EX group improved the most in physical function and caregiver perception outcomes. Directions for future research are provided.


What is the role of calcium scoring in the age of coronary computed tomographic angiography?
Non-contrast-enhanced CT for coronary artery calcification (CAC) as a marker of coronary atherosclerosis has been studied extensively in the primary prevention setting. With rapidly evolving multidetector CT technology, contrast-enhanced coronary CT angiography (CCTA) has emerged as the non-invasive method of choice for detailed imaging of the coronary tree. In this review, we systematically evaluate the role of CAC testing in the age of CCTA in both asymptomatic and symptomatic patients, across varying levels of risk. Although the role of CAC testing is well established in asymptomatic subjects, its use in evaluating those with stable symptoms that represent possible obstructive coronary artery disease is controversial. Nevertheless, available data suggest that in low-to-intermediate risk symptomatic patients, CAC scanning may serve as an appropriate gatekeeper to further testing with either CCTA (if no or only mild CAC present) versus functional imaging or invasive coronary angiography (when moderate or severe CAC present). Given the strong short-term prognostic value of CAC = 0, studies are needed to further evaluate the role of CAC scanning in low-risk patients with acute chest pain presenting to the emergency room.


Quantitative imaging of magnetization transfer using multiple selective pulses.
New spectroscopic and imaging methods have been developed for quantitatively measuring magnetization transfer (MT). These methods use trains of radiofrequency (rf) pulses with pulse separations much longer than 1/k(mf) and pulse durations much shorter than 1/k(mf), where k(mf) is the rate of MT from the immobile (macromolecular) protons to the mobile (free water) protons. Signal sensitivity to MT occurs when these pulses affect the mobile and immobile proton pools to different degrees. The signal from water may be quantitatively related to the macromolecular content of the sample using theory. The method has been used to make quantitative measurements of macromolecular content in cross-linked bovine serum albumin and employed in conjunction with echoplanar imaging to produce maps of the spatial distribution of the macromolecular content.


Fatty acid binding protein facilitates sarcolemmal fatty acid transport but not mitochondrial oxidation in rat and human skeletal muscle.
The transport of long-chain fatty acids (LCFAs) across mitochondrial membranes is regulated by carnitine palmitoyltransferase I (CPTI) activity. However, it appears that additional fatty acid transport proteins, such as fatty acid translocase (FAT)/CD36, influence not only LCFA transport across the plasma membrane, but also LCFA transport into mitochondria. Plasma membrane-associated fatty acid binding protein (FABPpm) is also known to be involved in sacrolemmal LCFA transport, and it is also present on the mitochondria. At this location, it has been identified as mitochondrial aspartate amino transferase (mAspAT), despite being structurally identical to FABPpm. Whether this protein is also involved in mitochondrial LCFA transport and oxidation remains unknown. Therefore, we have examined the ability of FABPpm/mAspAT to alter mitochondrial fatty acid oxidation. Muscle contraction increased (P < 0.05) the mitochondrial FAT/CD36 content in rat (+22%) and human skeletal muscle (+33%). By contrast, muscle contraction did not alter the content of mitochondrial FABPpm/mAspAT protein in either rat or human muscles. Electrotransfecting rat soleus muscles, in vivo, with FABPpm cDNA increased FABPpm protein in whole muscle (+150%; P < 0.05), at the plasma membrane (+117%; P < 0.05) and in mitochondria (+80%; P < 0.05). In these FABPpm-transfected muscles, palmitate transport into giant vesicles was increased by +73% (P < 0.05), and fatty acid oxidation in intact muscle was increased by +18% (P < 0.05). By contrast, despite the marked increase in mitochondrial FABPpm/mAspAT protein content (+80%), the rate of mitochondrial palmitate oxidation was not altered (P > 0.05). However, electrotransfection increased mAspAT activity by +70% (P < 0.05), and the mitochondrial FABPpm/mAspAT protein content was significantly correlated with mAspAT activity (r = 0.75). It is concluded that FABPpm has two distinct functions depending on its subcellular location: (a) it contributes to increasing sarcolemmal LCFA transport while not contributing directly to LCFA transport into mitochondria; and (b) its primary role at the mitochondria level is to transport reducing equivalents into the matrix.


RBC transfusion independence among lower risk MDS patients receiving hypomethylating agents: a population-level analysis.
Most patients with lower risk myelodysplastic syndromes (LR-MDS) become red blood cell (RBC) transfusion dependent at some time during their disease course. Hypomethylating agents (HMAs) are frequently used in this setting; however, reported rates of in RBC transfusion independence (TI) achieved with HMA therapy vary significantly between studies. Here we study the real-life clinical effectiveness of HMA in inducing RBC TI in anemic LR-MDS patients using the Surveillance, Epidemiology and End Results (SEER)-Medicare database. We find that approximately 40% of LR-MDS patients who were receiving RBC transfusions and 33% who were dependent on RBC transfusions at HMA initiation ultimately achieved TI. The receipt of ≥3 transfusions in the 8-week period before HMA initiation was significantly associated with lower odds of achieving TI. Our study provides important population level estimates of clinical effectiveness of HMAs in LR-MDS.


Low-grade astrocytoma: treatment with conventionally fractionated stereotactic radiation therapy.
To assess the efficacy of fractionated stereotactic radiation therapy for low-grade astrocytoma in terms of improvements to therapeutic ratio, patient survival, and quality of life. Since 1987, 32 patients with inoperable grade II astrocytoma were irradiated. Stereotactic radiation therapy was given only to patients with progressive symptoms. The mean total dose applied was 59.8 Gy (range, 54.0-65.0 Gy). Four years after therapy, the overall survival rate calculated with Kaplan-Meier analysis was 76%. Median progression-free survival was 48 months. On the basis of clinical symptoms, 91% (29 of 32) of patients showed improvement or stable disease after stereotactic radiation therapy. Eleven local failures were observed, 10 of which were within the planning target volume; one patient had a continuously enlarging mass. Acute toxic effects of radiation did not exceed grade II of the World Health Organization classification. Two patients developed reversible contrast enhancement without clinical symptoms on MR images within 1 year after stereotactic radiation therapy. Stereotactic radiation therapy for grade II astrocytoma appears to improve patients' quality of life or stabilize disease and is not correlated with marginal misses.


Contemporary Characterization of Injury Patterns, Initial Management, and Disparities in Treatment of Facial Fractures Using the National Trauma Data Bank.
Contemporary description of facial fracture patterns and factors associated with early operative intervention at trauma centers is lacking. The purpose of this study was to characterize injuries and initial treatment patterns of patients suffering from facial fractures. Using the National Trauma Databank from 2007 to 2015, patients suffering from facial fractures were included. Demographics, injury characteristics, and outcomes including operative interventions were assessed. Over 9 years, 626,270 patients were included: 74.5% were male, 39.0% suffered traumatic brain injury (TBI), and 23.3% severe noncraniofacial injuries (chest, abdomen, and/or extremity Abbreviated Injury Score ≥3). A total of 537,594 (85.8%) patients were admitted and 184,206 (34.3%) underwent operations for their facial injuries during the index hospitalization. The frequency and patterns of fractures varied by age, sex, race, and mechanism of injury. Operative intervention rates were highest for mandible (63.2%) and lowest for orbit fractures (1.0%). Multiple regression revealed that multiple factors were independently associated with the odds of early fracture repair including: female versus male (odds ratio [95% confidence interval]: 0.96 [0.94-0.98]), age >65 versus <18 years (0.62 [0.59-0.64]), non-white race (0.95 [0.94-0.97]), uninsured versus Medicaid (0.88 [0.86-0.90]), hospital bed size (>600 vs ≤200 beds, 1.67 [1.61-1.73]), TBI (0.70 [0.69-0.71]), and C-spine injury (0.93 [0.90-0.96]). Facial fractures are common among many demographic cohorts, and multiple patient and injury-specific factors influenced fracture patterns and management. Early operative intervention was highest for mandible fractures and lowest for orbit fractures. Multiple factors including age, sex, insurance status, hospital characteristics, and race/ethnicity were independently associated with early operative intervention, highlighting disparities in care.


Aerosolization of recombinant SLPI to augment antineutrophil elastase protection of pulmonary epithelium.
In a variety of lung diseases the respiratory epithelial surface must contend with an increased burden of neutrophil elastase (NE). One candidate for augmenting epithelial anti-NE protection is the secretory leukoprotease inhibitor (SLPI). In vitro evaluation demonstrated that 96 +/- 1% of the recombinant SLPI (rSLPI) molecules were capable of inhibiting NE, with an association rate constant of 7.1 +/- 0.1 X 10(6) M-1.s-1. Evaluation of rSLPI after in vitro and in vivo aerosolization showed that aerosolization did not alter rSLPI. Aerosolization of a single dose of 50 mg rSLPI to sheep resulted in a fourfold increase of the anti-NE capacity in epithelial lining fluid (ELF) at 3 h, with a half-life in ELF of 12 h. After aerosolization some rSLPI appeared in lung lymph. Simultaneous aerosolization of rSLPI and recombinant alpha 1-antitrypsin (rAAT) demonstrated a molar ratio of the concentration in lymph to the concentration in ELF 3 h after the aerosol eightfold higher for rAAT than for rSLPI. Overall, these observations demonstrate that it is feasible to use aerosolized rSLPI to directly augment the anti-NE capacity of the lung, particularly on the pulmonary epithelial surface.


n-3 PUFA esterified to glycerol or as ethyl esters reduce non-fasting plasma triacylglycerol in subjects with hypertriglyceridemia: a randomized trial.
To date, treatment of hypertriglyceridemia with long-chain n-3 polyunsaturated fatty acids (n-3 PUFA) has been investigated solely in fasting and postprandial subjects. However, non-fasting triacylglycerols are more strongly associated with risk of cardiovascular disease. The objective of this study was to investigate the effect of long-chain n-3 PUFA on non-fasting triacylglycerol levels and to compare the effects of n-3 PUFA formulated as acylglycerol (AG-PUFA) or ethyl esters (EE-PUFA). The study was a double-blinded randomized placebo-controlled interventional trial, and included 120 subjects with non-fasting plasma triacylglycerol levels of 1.7-5.65 mmol/L (150-500 mg/dL). The participants received approximately 3 g/day of AG-PUFA, EE-PUFA, or placebo for a period of eight weeks. The levels of non-fasting plasma triacylglycerols decreased 28% in the AG-PUFA group and 22% in the EE-PUFA group (P < 0.001 vs. placebo), with no significant difference between the two groups. The triacylglycerol lowering effect was evident after four weeks, and was inversely correlated with the omega-3 index (EPA + DHA content in erythrocyte membranes). The omega-3 index increased 63.2% in the AG-PUFA group and 58.5% in the EE-PUFA group (P < 0.001). Overall, the heart rate in the AG-PUFA group decreased by three beats per minute (P = 0.045). High-density lipoprotein (HDL) cholesterol increased in the AG-PUFA group (P < 0.001). Neither total nor non-HDL cholesterol changed in any group. Lipoprotein-associated phospholipase A2 (LpPLA2) decreased in the EE-PUFA group (P = 0.001). No serious adverse events were observed. Supplementation with long-chain n-3 PUFA lowered non-fasting triacylglycerol levels, suggestive of a reduction in cardiovascular risk. Regardless of the different effects on heart rate, HDL, and LpPLA2 that were observed, compared to placebo, AG-PUFA, and EE-PUFA are equally effective in reducing non-fasting triacylglycerol levels.


Effects of electronic cigarette smoking on human health.
Electronic cigarette smoking is gaining dramatic popularity and is steadily spreading among the adolescents, high income, urban population around the world. The aim of this study is to highlight the hazards of e-cigarette smoking on human health. In this study, we identified 38 published studies through a systematic database searches including ISI-web of science and pub-med. We searched the related literature by using the key words including Electronic cigarette, E-cigarette, E-vapers, incidence, hazards. Studies in which electronic cigarette smoking hazards was investigated were included in the study. No limitations on publication status, study design of publication were implemented. Finally we included 28 publications and remaining 10 were excluded. E-smoking can cause, nausea, vomiting, headache, dizziness, choking, burn injuries, upper respiratory tract irritation, dry cough, dryness of the eyes and mucous membrane, release of cytokines and pro-inflammatory mediators, allergic airway inflammation, decreased exhaled nitric oxide (FeNO) synthesis in the lungs, change in bronchial gene expression and risk of lung cancer. Electronic cigarettes are swiftly promoted as an alternative to conventional cigarette smoking, although its use is highly controversial. Electronic cigarettes are not a smoking cessation product. Non-scientific claims about e-cigarettes are creating confusion in public perception about e-cigarette and people believe that e-cigarettes are safe and less addictive, but its use is unsafe and hazardous to human health. E-cigarette smoking should be regulated in the same way as traditional cigarettes and must be prohibited to children and adolescents.


The [NiFe]-hydrogenase of the cyanobacterium Synechocystis sp. PCC 6803 works bidirectionally with a bias to H2 production.
Protein film electrochemistry (PFE) was utilized to characterize the catalytic activity and oxidative inactivation of a bidirectional [NiFe]-hydrogenase (HoxEFUYH) from the cyanobacterium Synechocystis sp. PCC 6803. PFE provides precise control of the redox potential of the adsorbed enzyme so that its activity can be monitored under changing experimental conditions as current. The properties of HoxEFUYH are different from those of both the standard uptake and the "oxygen-tolerant" [NiFe]-hydrogenases. First, HoxEFUYH is biased toward proton reduction as opposed to hydrogen oxidation. Second, despite being expressed under aerobic conditions in vivo, HoxEFUYH is clearly not oxygen-tolerant. Aerobic inactivation of catalytic hydrogen oxidation by HoxEFUYH is total and nearly instantaneous, producing two inactive states. However, unlike the Ni-A and Ni-B inactive states of standard [NiFe]-hydrogenases, both of these states are quickly (<90 s) reactivated by removal of oxygen and exposure to reducing conditions. Third, proton reduction continues at 25-50% of the maximal rate in the presence of 1% oxygen. Whereas most previously characterized [NiFe]-hydrogenases seem to be preferential hydrogen oxidizing catalysts, the cyanobacterial enzyme works effectively in both directions. This unusual catalytic bias as well as the ability to be quickly reactivated may be essential to fulfilling the physiological role in cyanobacteria, organisms expected to experience swings in cellular reduction potential as they switch between aerobic conditions in the light and dark anaerobic conditions. Our results suggest that the uptake [NiFe]-hydrogenases alone are not representative of the catalytic diversity of [NiFe]-hydrogenases, and the bidirectional heteromultimeric enzymes may serve as valuable models to understand the diverse mechanisms of tuning the reactivity of the hydrogen activating site.


Development of 19F-NMR chemical shift detection of DNA B-Z equilibrium using 19F-NMR.
Various DNA conformational changes are in correlation with biological events. In particular, DNA B-Z equilibrium showed a high correlation with translation and transcription. In this study, we developed a DNA probe containing 5-trifluoromethylcytidine or 5-trifluoromethylthymidine to detect DNA B-Z equilibrium using 19F-NMR. Its probe enabled the quantitative detection of B-, Z-, and ss-DNA based on 19F-NMR chemical shift change.


The structure of NMB1585, a MarR-family regulator from Neisseria meningitidis.
The structure of the MarR-family transcription factor NMB1585 from Neisseria meningitidis has been solved using data extending to a resolution of 2.1 A. Overall, the dimeric structure resembles those of other MarR proteins, with each subunit comprising a winged helix-turn-helix (wHtH) domain connected to an alpha-helical dimerization domain. The spacing of the recognition helices of the wHtH domain indicates that NMB1585 is pre-configured for DNA binding, with a putative inducer pocket that is largely occluded by the side chains of two aromatic residues (Tyr29 and Trp53). NMB1585 was shown to bind to its own promoter region in a gel-shift assay, indicating that the protein acts as an auto-repressor.


Stress and gastrointestinal tract. II. Stress and intestinal barrier function.
The influence of stress on the clinical course of a number of intestinal diseases is increasingly being recognized, but the underlying mechanisms are largely unknown. This themes article focuses on recent findings related to the effects of stress on mucosal barrier function in the small intestine and colon. Experiments using animal models demonstrate that various types of psychological and physical stress induce dysfunction of the intestinal barrier, resulting in enhanced uptake of potentially noxious material (e.g., antigens, toxins, and other proinflammatory molecules) from the gut lumen. Evidence from several studies indicates that in this process, mucosal mast cells play an important role, possibly activated via neurons releasing corticotropin-releasing hormone and/or acetylcholine. Defining the role of specific cells and mediator molecules in stress-induced barrier dysfunction may provide clues to novel treatments for intestinal disorders.


Plasma LTE4/PGF2α Ratio and Blood Eosinophil Count Are Increased in Elderly Asthmatics With Previous Asthma Exacerbation.
The tools for asthma control assessment recommended by the current guideline are cognitive function- and effort-dependent, which is substantially impaired in the elderly. The aim of this study is to investigate objective assessment tools of asthma control status and previous asthma exacerbation (AE) in elderly subjects. Asthmatics aged >60 years who were treated with step 2 or 3 by the Global Initiative for Asthma (GINA) guideline were enrolled. During the 12-week study period, the subjects used either 400 μg of budesonide plus 10 mg of montelukast or 800 μg of inhaled budesonide. The occurrence of AE during the 4-week run-in and 12-week treatment period was monitored. After 12-week of treatment, sputum eosinophil count, peripheral eosinophil count, the plasma leukotriene E4 (LTE4), and prostaglandin F2α (PGF2α) metabolite levels were measured using the UHPLC/Q-ToF MS system. The study subjects were divided into group 1 (asthmatics who experienced AE during the study period) and group 2 (those who did not). A total of 101 patients aged 60-85 years were enrolled. Twenty-three patients (22.8%) had experienced AE. The plasma LTE4 level, LTE4/PGF2α ratio, and peripheral eosinophil count were significantly higher in group 1 than in group 2 (P=0.023, P=0.010, P=0.033, respectively). The plasma LTE4/PGF2α ratio and peripheral eosinophil count at week 12 were significantly associated with previous AE (odds ratio [OR]=1.748, P=0.013; OR=1.256, P=0.027). Receiver operating characteristic (ROC) curves to discriminate the subjects with previous AE, including these 2 parameters, showed that the area under the curve was 0.700 (P=0.004), with 73.9% sensitivity and 47.9% specificity. In conclusion, a combination of plasma LTE4/PGF2α ratio and peripheral eosinophil count can be an objective assessment tool which is significantly associated with asthma control status in elderly asthmatics.


Cold truths: how winter drives responses of terrestrial organisms to climate change.
Winter is a key driver of individual performance, community composition, and ecological interactions in terrestrial habitats. Although climate change research tends to focus on performance in the growing season, climate change is also modifying winter conditions rapidly. Changes to winter temperatures, the variability of winter conditions, and winter snow cover can interact to induce cold injury, alter energy and water balance, advance or retard phenology, and modify community interactions. Species vary in their susceptibility to these winter drivers, hampering efforts to predict biological responses to climate change. Existing frameworks for predicting the impacts of climate change do not incorporate the complexity of organismal responses to winter. Here, we synthesise organismal responses to winter climate change, and use this synthesis to build a framework to predict exposure and sensitivity to negative impacts. This framework can be used to estimate the vulnerability of species to winter climate change. We describe the importance of relationships between winter conditions and performance during the growing season in determining fitness, and demonstrate how summer and winter processes are linked. Incorporating winter into current models will require concerted effort from theoreticians and empiricists, and the expansion of current growing-season studies to incorporate winter.


Allele frequency data of 15 autosomal STR loci in four major population groups of South Africa.
Allele frequency distributions for 15 tetrameric short tandem repeat (STR) loci were determined using the AmpFlSTR® Identifiler Plus™ PCR amplification kit. There was little evidence of departures from Hardy-Weinberg equilibrium or association of alleles of different loci in the population samples. The probability of identity values for the different populations range from 1/3.3 × 10(17) (White) to 1/1.88 × 10(18) (Coloured). The combined probability of paternal exclusion for the different population groups ranges from 0.9995858 (Coloured) to 0.9997874 (Indian).


Understanding the Community Risk Perceptions of the COVID-19 Outbreak in South Korea: Infodemiology Study.
South Korea is among the best-performing countries in tackling the coronavirus pandemic by using mass drive-through testing, face mask use, and extensive social distancing. However, understanding the patterns of risk perception could also facilitate effective risk communication to minimize the impacts of disease spread during this crisis. We attempt to explore patterns of community health risk perceptions of COVID-19 in South Korea using internet search data. Google Trends (GT) and NAVER relative search volumes (RSVs) data were collected using COVID-19-related terms in the Korean language and were retrieved according to time, gender, age groups, types of device, and location. Online queries were compared to the number of daily new COVID-19 cases and tests reported in the Kaggle open-access data set for the time period of December 5, 2019, to May 31, 2020. Time-lag correlations calculated by Spearman rank correlation coefficients were employed to assess whether correlations between new COVID-19 cases and internet searches were affected by time. We also constructed a prediction model of new COVID-19 cases using the number of COVID-19 cases, tests, and GT and NAVER RSVs in lag periods (of 1-3 days). Single and multiple regressions were employed using backward elimination and a variance inflation factor of <5. The numbers of COVID-19-related queries in South Korea increased during local events including local transmission, approval of coronavirus test kits, implementation of coronavirus drive-through tests, a face mask shortage, and a widespread campaign for social distancing as well as during international events such as the announcement of a Public Health Emergency of International Concern by the World Health Organization. Online queries were also stronger in women (r=0.763-0.823; P<.001) and age groups ≤29 years (r=0.726-0.821; P<.001), 30-44 years (r=0.701-0.826; P<.001), and ≥50 years (r=0.706-0.725; P<.001). In terms of spatial distribution, internet search data were higher in affected areas. Moreover, greater correlations were found in mobile searches (r=0.704-0.804; P<.001) compared to those of desktop searches (r=0.705-0.717; P<.001), indicating changing behaviors in searching for online health information during the outbreak. These varied internet searches related to COVID-19 represented community health risk perceptions. In addition, as a country with a high number of coronavirus tests, results showed that adults perceived coronavirus test-related information as being more important than disease-related knowledge. Meanwhile, younger, and older age groups had different perceptions. Moreover, NAVER RSVs can potentially be used for health risk perception assessments and disease predictions. Adding COVID-19-related searches provided by NAVER could increase the performance of the model compared to that of the COVID-19 case-based model and potentially be used to predict epidemic curves. The use of both GT and NAVER RSVs to explore patterns of community health risk perceptions could be beneficial for targeting risk communication from several perspectives, including time, population characteristics, and location.


Distribution and Abundance of Hopanoid Producers in Low-Oxygen Environments of the Eastern Pacific Ocean.
Hopanoids are bacterial membrane lipid biomarker molecules that feature prominently in the molecular fossil record. In the modern marine water column, recent reports implicate bacteria inhabiting low-oxygen environments as important sources of hopanoids to marine sediments. However, the preliminary biogeography reported by recent studies and the environmental conditions governing such distributions can only be confirmed when the numerical abundance of these organisms is known with more certainty. In this study, we employ two different approaches to examine the quantitative significance of phylogenetically distinct hopanoid producers in low-oxygen environments. First, we develop a novel quantitative PCR (qPCR) assay for the squalene hopene cyclase (sqhC) gene, targeting a subset of hopanoid producers previously identified to be important in the eastern North Pacific Ocean. The results represent the first quantitative gene abundance data of any kind for hopanoid producers in the marine water column and show that these putative alphaproteobacterial hopanoid producers are rare, comprising at most 0.2 % of the total bacterial community in our samples. Second, a complementary analysis of existing low-oxygen metagenomic datasets further examined the generality of the qPCR observation. We find that the dominant sqhC sequences in these metagenomic datasets are associated with phyla such as Nitrospinae rather than Proteobacteria, consistent with the qPCR finding that alphaproteobacterial hopanoid producers are not very abundant in low-oxygen environments. In fact, positive correlations between sqhC gene abundance and environmental parameters in these samples identify nitrite availability as a potentially important factor in the ecology of hopanoid producers that dominate low-oxygen environments.


A 2017 US Reference for Singleton Birth Weight Percentiles Using Obstetric Estimates of Gestation.
To provide an updated birth weight-for-gestational age (BW-for-GA) reference in the United States by using the most recent, nationally representative birth data with obstetric estimates of gestational age (GA). We abstracted 3 285 552 singleton births between 22 and 42 weeks' gestation with nonmissing race and/or ethnicity, infant sex, parity, birth weight, and obstetric estimate of GA from the 2017 US natality files. We used 2 techniques (nonlinear, resistant smoothing [4253H] and lambda-mu-sigma) to derive smoothed BW-for-GA curves and compared resulting BW-for-GA cut-points at the third, 10th, 90th, and 97th percentiles with US references from 1999 to 2009. The smoothed BW-for-GA curves from both techniques overlapped considerably with each other, with strong agreements seen between the 2 techniques (>99% agreement; κ-statistic >0.9) for BW-for-GA cut-points at the third, 10th, 90th, and 97th percentiles across all GAs. Cut-points from 2017 using the lambda-mu-sigma method captured 9.8% to 10.2% of births <10th and >90th percentiles and 2.6% to 3.3% of births below the third and above the 97th percentile across all GAs. However, cut-points from US references in 1999 and 2009 (when GA was based on last menstrual period) captured a much larger range of proportions of 2017 births at these thresholds, especially among preterm and postterm GA categories. We have provided an updated BW-for-GA reference in the United States using the most recent births with obstetric estimates of GA and information to calculate continuous measures of birth size that are sex or parity specific.


Isolation of a coronavirus during studies on puffinosis, a disease of the Manx shearwater (Puffinus puffinus).
A virus was isolated from 2 day-old mice inoculated with homogenates of either the lungs or blood of 2 different shearwaters affected by puffinosis. Examination of infected suckling mouse brain and infected NCTC-1469 (mouse liver) cell cultures, by electron microscopy, revealed virus particles and inclusion bodies characteristic of a coronavirus. Neutralization, complement fixation and fluorescent antibody tests showed that the virus was related to mouse hepatitis virus. The virus was not isolated from 445 control, uninfected mice. Neutralising antibodies were not detected in 39 sera from diseased shearwaters and 2 sera from apparently healthy birds. Two shearwaters inoculated with the virus did not develop clinical signs of infection. The question of whether the virus was isolated from shearwaters or from laboratory mice is discussed.


Numerical analysis of a comprehensive model of M-phase control in Xenopus oocyte extracts and intact embryos.
To contribute to a deeper understanding of M-phase control in eukaryotic cells, we have constructed a model based on the biochemistry of M-phase promoting factor (MPF) in Xenopus oocyte extracts, where there is evidence for two positive feedback loops (MPF stimulates its own production by activating Cdc25 and inhibiting Wee1) and a negative feedback loop (MPF stimulates its own destruction by indirectly activating the ubiquitin pathway that degrades its cyclin subunit). To uncover the full dynamical possibilities of the control system, we translate the regulatory network into a set of differential equations and study these equations by graphical techniques and computer simulation. The positive feedback loops in the model account for thresholds and time lags in cyclin-induced and MPF-induced activation of MPF, and the model can be fitted quantitatively to these experimental observations. The negative feedback loop is consistent with observed time lags in MPF-induced cyclin degradation. Furthermore, our model indicates that there are two possible mechanisms for autonomous oscillations. One is driven by the positive feedback loops, resulting in phosphorylation and abrupt dephosphorylation of the Cdc2 subunit at an inhibitory tyrosine residue. These oscillations are typical of oocyte extracts. The other type is driven by the negative feedback loop, involving rapid cyclin turnover and negligible phosphorylation of the tyrosine residue of Cdc2. The early mitotic cycles of intact embryos exhibit such characteristics. In addition, by assuming that unreplicated DNA interferes with M-phase initiation by activating the phosphatases that oppose MPF in the positive feedback loops, we can simulate the effect of addition of sperm nuclei to oocyte extracts, and the lengthening of cycle times at the mid-blastula transition of intact embryos.


Feasibility and Effectiveness of a Delirium Prevention Bundle in Critically Ill Patients.
Strategies for preventing delirium include early identification and avoiding or modifying patient, environmental, and iatrogenic factors. Minimal research exists on a prescriptive delirium prevention bundle that details elements or strategies for each bundle component. Even less research has been focused on nurse-driven interventions or components. To evaluate the effectiveness of a delirium prevention bundle in decreasing delirium incidence in 2 medical-surgical intensive care units in a large Texas medical center. Researchers used the Confusion Assessment Method for the Intensive Care Unit to assess delirium incidence by using a controlled interventional cohort design with 447 delirium-negative critically ill patients. Bundle components consist of sedation cessation, pain management, sensory stimulation, early mobilization, and sleep promotion. The intervention, analyzed by using a logistic regression model, reduced the odds of delirium by 78% (odds ratio, 0.22; P = .001). The delirium prevention bundle was effective in reducing the incidence of delirium in critically ill medical-surgical patients. Further validation studies are under way.


miR-21 and miR-214 are consistently modulated during renal injury in rodent models.
Transforming growth factor (TGF)-β is one of the main fibrogenic cytokines that drives the pathophysiology of progressive renal scarring. MicroRNAs (miRNAs) are endogenous non-coding RNAs that post-transcriptionally regulate gene expression. We examined the role of TGF-β-induced expression of miR-21, miRNAs in cell culture models and miRNA expression in relevant models of renal disease. In vitro, TGF-β changed expression of miR-21, miR-214, and miR-145 in rat mesangial cells (CRL-2753) and miR-214, miR-21, miR-30c, miR-200b, and miR-200c during induction of epithelial-mesenchymal transition in rat tubular epithelial cells (NRK52E). miR-214 expression was robustly modulated in both cell types, whereas in tubular epithelial cells miR-21 was increased and miR-200b and miR-200c were decreased by 58% and 48%, respectively, in response to TGF-β. TGF-β receptor-1 was found to be a target of miR-200b/c and was down-regulated after overexpression of miR-200c. To assess the differential expression of these miRNAs in vivo, we used the anti-Thy1.1 mesangial glomerulonephritis model and the unilateral ureteral obstruction model in which TGF-β plays a role and also a genetic model of hypertension, the stroke-prone spontaneously hypertensive rat with and without salt loading. The expressions of miR-214 and miR-21 were significantly increased in all in vivo models, showing a possible miRNA signature of renal damage despite differing causes.


Early tracheal stenosis causing extubation failure and prolonged ventilator dependency.
Postintubation stenosis is the most frequent cause of benign tracheal stenosis and may cause reintubation and delay in weaning of intensive care unit patients. This case study describes typical patients with tracheal stenosis and the management of these patients. Five patients requiring reintubation and mechanical ventilation due to early intubation-related stenosis are discussed. Stridor developed in three cases after extubation. In these cases, bronchoscopy revealed tracheal stenosis. Dilatation and silicone stent placement were performed using rigid bronchoscopy. The other two patients were on ventilators when they were admitted to the intensive care unit and their stenoses were also treated by rigid bronchoscopy. Hypercapnia and hypoxia resolved after intervention in three cases. Of the remaining two patients, one had the tracheostomy closed and in the other patient ventilation was stopped but the tracheostomy was maintained. Tracheal stenosis developing in the subglottic region after extubation, especially after exposure to cuff pressure, may lead to reintubation. A tracheostomy may hinder the diagnosis of progressive stenosis and may lead to unnecessary maintenance of ventilator treatment. Early intubation-related tracheal stenosis should therefore be considered in cases of weaning or extubation failure and prompt appropriate investigation and treatment.


Circulating pEGFR is a candidate response biomarker of cetuximab therapy in colorectal cancer.
The lack of secreted biomarkers measurable by noninvasive tests hampers the development of effective targeted therapies against cancer. Our hypothesis is that cetuximab (an anti-EGFR mAb) induces a specific secretome in colorectal cancer cells that could be exploited for biomarker discovery. Considering the strong correlation between mutated KRAS and a lack of response to cetuximab therapy, we addressed whether performing secretome-based proteomics on isogenic colorectal cancer cells sharing the KRAS mutations found on patients would yield candidate-secreted biomarkers useful in the clinical setting. Because 2D culture did not optimally model the sensitivity/resistance to cetuximab observed in colorectal cancer patients, we moved to 3D spheroids, developing a methodology for both cell-based assays and quantitative proteomics. A large comparative quantitative proteomic analysis of the 3D secretomes of colorectal cancer isogenic cells treated with cetuximab uncovered an EGFR pathway-centric secretome found only when cells grow in 3D. The validation of the secretome findings in plasma of colorectal cancer patients, suggests that phosphorylated-EGFR (pEGFR) is a candidate-secreted biomarker of response to cetuximab. We have proved that 3D spheroids from colorectal cancer cells generate secretomes with a drug-sensitivity profile that correlates well with patients with colorectal cancer, illustrating molecular connections between intracellular and extracellular signaling. Furthermore, we show how the secretion of pEGFR is associated with the sensitivity of colorectal cancer cells to cetuximab and the response of patients with colorectal cancer to the drug. Our work could allow the noninvasive monitoring of anti-EGFR treatment in patients with colorectal cancer.


Phosphorylation of collapsin response mediator protein-2 disrupts neuronal maturation in a model of adult neurogenesis: Implications for neurodegenerative disorders.
Recent studies suggest that the pathogenic process in neurodegenerative disorders may disrupt mature neuronal circuitries and neurogenesis in the adult brain. Abnormal activation of CDK5 is associated with neurodegenerative disorders, and recently a critical role for CDK5 in adult neurogenesis has been identified. We have developed an in vitro model of abnormal CDK5 activation during adult hippocampal neurogenesis, and here we used this model to investigate aberrantly phosphorylated downstream targets of CDK5. Abnormal CDK5 activation in an in vitro model of adult neurogenesis results in hyperphosphorylation of collapsin-response mediator protein-2 (CRMP2) and impaired neurite outgrowth. Inhibition of CDK5, or expression of a non-phosphorylatable (S522A) CRMP2 construct reduced CRMP2 hyperphosphorylation, and reversed neurite outgrowth deficits. CRMP2 plays a role in microtubule dynamics; therefore we examined the integrity of microtubules in this model using biochemical and electron microscopy techniques. We found that microtubule organization was disrupted under conditions of CDK5 activation. Finally, to study the relevance of these findings to neurogenesis in neurodegenerative conditions associated with HIV infection, we performed immunochemical analyses of the brains of patients with HIV and transgenic mice expressing HIV-gp120 protein. CDK5-mediated CRMP2 phosphorylation was significantly increased in the hippocampus of patients with HIV encephalitis and in gp120 transgenic mice, and this effect was rescued by genetic down-modulation of CDK5 in the mouse model. These results reveal a functional mechanism involving microtubule destabilization through which abnormal CDK5 activation and CRMP2 hyperphosphorylation might contribute to defective neurogenesis in neurodegenerative disorders such as HIV encephalitis.


Functional analysis of N-linking oligosaccharyl transferase enzymes encoded by deep-sea vent proteobacteria.
Bacterial N-linking oligosaccharyl transferases (OTase enzymes) transfer lipid-linked glycans to selected proteins in the periplasm and were first described in the intestinal pathogen Campylobacter jejuni, a member of the ε-proteobacteria-subdivision of bacteria. More recently, orthologues from other ε-proteobacterial Campylobacter and Helicobacter species and a δ-proteobacterium, Desulfovibrio desulfuricans, have been described, suggesting that these two subdivisions of bacteria may be a source of further N-linked protein glycosylation systems. Whole-genome sequencing of both ε- and δ-proteobacteria from deep-sea vent habitats, a rich source of species from these subdivisions, revealed putative ORFs encoding OTase enzymes and associated adjacent glycosyltransferases similar to the C. jejuni N-linked glycosylation locus. We expressed putative OTase ORFs from the deep-sea vent species Nitratiruptor tergarcus, Sulfurovum lithotrophicum and Deferribacter desulfuricans in Escherichia coli and showed that they were able to functionally complement the C. jejuni OTase, CjPglB. The enzymes were shown to possess relaxed glycan specificity, transferring diverse glycan structures and demonstrated different glycosylation sequon specificities. Additionally, a permissive D. desulfuricans acceptor protein was identified, and we provide evidence that the N-linked glycan synthesized by N. tergarcus and S. lithotrophicum contains an acetylated sugar at the reducing end. This work demonstrates that deep-sea vent bacteria encode functional N-glycosylation machineries and are a potential source of biotechnologically important OTase enzymes.


Risk factors for inpatient violence and self-harm in forensic psychiatry: the role of head injury, schizophrenia and substance misuse.
To investigate factors relevant to violence and self-harm in forensic psychiatric inpatients the cross-sectional association between four potential contributory factors; head injury, schizophrenia, drug and alcohol misuse, and self-harm or violence-related outcomes was examined. Data were extracted from an existing dataset of routinely collected data on all patients under the care of Scotland's Forensic Mental Health Managed Care Network, of whom (n = 432) met inclusion criteria. A Factorial MANOVA and Pearson's chi-square tests were conducted to assess the relationship between potential contributory factors and self-harm and violence. Forty-seven individuals had a documented head injury (10.9%). The presence of head injury was significantly associated with inpatient violence and assessed violence risk. Number of historic violent offences was significantly associated with a history of drug misuse and co-morbid alcohol misuse and schizophrenia. Self-harm was significantly associated with drug misuse and a diagnosis of schizophrenia. These findings highlight a significant association between head injury and actual/assessed risk in forensic psychiatry, over and above that of substance misuse and a diagnosis of schizophrenia, emphasising the need for routine assessment of head injury in clinical practice. Further examination of the impact of head injury in forensic psychiatric populations is needed.


Monoclonal antibodies against EGFRvIII are tumor specific and react with breast and lung carcinomas and malignant gliomas.
Despite molecular biological advances in understanding human cancers, translation into therapy has been less forthcoming; targeting neoplastic cells still requires that tumor-specific markers, preferably those on the cell surface, be identified. The epidermal growth factor receptor (EGFR) exists in a deletion-mutant form, EGFRvIII, which has been identified by genetic and immunological means in a subset of gliomas and non-small cell lung carcinomas. Specific polyvalent antisera to the extracellular portion of the variant were readily induced, but immunization using a synthetic linear peptide representing the unique EGFRvIII primary sequence has been unsuccessful in mice or macaques. We report here five specific monoclonal antibodies (mAbs) developed through long-term immunization protocols using the EGFRvIII-specific synthetic peptide and the intact variant in different formats that maintained secondary and tertiary conformation. These mAbs identify the EGFRvIII on the cell surface with relatively high affinity (KA range, 0.13 to 2.5 x 10(9) M-1) by live cell Scatchard analysis. These mAbs are specific for EGFRvIII as determined by RIA, ELISA, Western blot, analytical flow cytometry, autophosphorylation, and immunohistochemistry. Isolating specific mAbs enabled us to analyze normal and neoplastic human tissue and establish that EGFRvIII is truly tumor specific for subsets of breast carcinomas and for previously reported non-small cell lung carcinomas and gliomas. Also, this receptor is not expressed by any normal human tissues thus far examined, including elements of the peripheral, central nervous, and lymphoid systems. With mAbs, we identified a higher incidence of EGFRvIII positivity in gliomas than previously described and identified an EGFRvIII-positive subset of breast tumors; also, we observed that the EGFRvIII epitope is not expressed in normal tissues, and we demonstrated the localizing and therapeutic potential of the mAbs for tumors expressing this epitope. Our observations strongly warrant development of this mAb-antigen system as therapy for breast, lung, and central nervous system tumors.


Children with pertussis inform the investigation of other pertussis cases among contacts.
The number of reported pertussis has increased in the last two decades. However, many cases of pertussis may be underreported or not diagnosed. The World Health Organization estimates that pertussis causes 200,000-400,000 deaths each year, most deaths are in infants and in developing countries. Infants with pertussis can indicate an undetected source cases in the community. At a University Hospital in Brazil individuals that had frequent contacts with a child with confirmed pertussis (the index case) and had recent history of cough were enrolled into the study. Nasopharyngeal swabs were collected from every contact that had cough within the last 21 days. Cases confirmation followed the guidelines of the Center for Disease Control and Prevention-Atlanta, USA. Pertussis diagnosis was confirmed in 51 children, (considered the index cases). Among the index cases, 72.5% (37/51) were under 6 months of age; culture for Bordetella pertussis was positive in 78.4% (40/51). Pertussis was confirmed in 39% (107/276) of the contacts of 51 index cases. Among these contacts identified as a pertussis case, 40.2% (43/107) were between 6 months and 111/2 years of age and 59.8% (64/107) were older than 111/2 years of age. Pertussis was confirmed by culture in 11.2% (12/107) of them and by epidemiologic linkage in 88.8% (95/107). Each index case allowed identifying two new cases of pertussis. Public health authorities should consider implementing early recognition of pertussis index cases and searching for pertussis cases among the contacts. Treatment of the cases and prophylaxis of the contacts is fundamental to control outbreaks in the community.


Perioperative result of carotid endarterectomies with venous patch angioplasty.
Carotid endarterectomy (CEA) is an effective treatment of carotid stenosis to reduce the risk of stroke. The purpose of the current study is to investigate the result of surgical treatment of carotid stenosis in Taipei Veterans General Hospital. The medical records of 103 patients receiving 113 CEA with or without patch angioplasty in our service between January 1993 and July 1999 were reviewed retrospectively. The clinical and operative variables were collected for analysis. According to the method of carotid arteriotomy closure, all operations were categorized into 3 groups: 76 CEA with venous patch angioplasty, 27 CEA with synthetic angioplasty and 10 primary closure without patch angioplasty. The outcomes of CEA among 3 groups were compared. There was no significant difference of patient characteristics between 3 groups. Four patients died in the same admission. The incidence of hospital mortality was 0% for venous patch group (p = 0.01), 11.1% for synthetic patch group and 10% for primary closure group. There were 5 perioperative complications including 3 with ipsilateral stroke and 2 with wound hematoma. The incidence of perioperative complication rate was 2.6% for venous patch group, 7.4% for synthetic patch group and 10% for primary closure group (p = 0.87). The incidence of perioperative CVA was 1.3% for venous patch group, 3.7% for synthetic patch group, 10% for primary closure group (p = 0.43). Our study showed that CEA could be performed with low risk and that venous angioplasty carried lower hospital mortality than other methods of carotid arteriotomy closure.


Purification and photobiochemical profile of photosystem 1 from a high-salt tolerant, oleaginous Chlorella (Trebouxiophycaea, Chlorophyta).
The eukaryotic green alga Chlamydomonas reinhardtii has been studied extensively within the biofuel industry as a model organism, as researchers look towards algae to provide chemical feedstocks (i.e., lipids) for the production of liquid transportation fuels. C. reinhardtii, however, is unsuitable for high-level production of such precursors due to its relatively poor lipid accumulation and fresh-water demand. In this study we offer insight into the primary light harvesting and electron transfer reactions that occur during phototropic growth in a high-salt tolerant strain of Chlorella (a novel strain introduced here as NE1401), a single-celled eukaryotic algae also in the phylum Chlorophyta. Under nutrient starvation many eukaryotic algae increase dramatically the amount of lipids stored in lipid bodies within their cell interiors. Microscopy and lipid analyses indicate that Chlorella sp. NE1401 may become a superior candidate for algal biofuels production. We have purified highly active Photosystem 1 (PS1) complexes to study in vitro, so that we may understand further the photobiochemisty of this promising biofuel producer and how its characteristics compare and contrast with that of the better understood C. reinhardtii. Our findings suggest that the PS1 complex from Chlorella sp. NE1401 demonstrates similar characteristics to that of C. reinhardtii with respect to light-harvesting and electron transfer reactions. We also illustrate that the relative extent of the light state transition performed by Chlorella sp. NE1401 is smaller compared to C. reinhardtii, although they are triggered by the same dynamic light stresses.


ISIS-2: 10 year survival among patients with suspected acute myocardial infarction in randomised comparison of intravenous streptokinase, oral aspirin, both, or neither. The ISIS-2 (Second International Study of Infarct Survival) Collaborative Group.
To assess effects of intravenous streptokinase, one month of oral aspirin, or both, on long term survival after suspected acute myocardial infarction. Randomised, "2 x 2 factorial," placebo controlled trial. 417 hospitals in 16 countries. 17 187 patients with suspected acute myocardial infarction randomised between March 1985 and December 1987. Follow up of vital status complete to at least 1 January 1990 for 95% of all patients and to mid-1997 for the 6213 patients in United Kingdom. Intravenous streptokinase (1.5 MU in 1 hour) and oral aspirin (162 mg daily for 1 month) versus matching placebos. Mortality from all causes during up to 10 years' follow up, with subgroup analyses based on 4 year follow up. After randomisation, 1841 deaths were recorded in days 0-35, 991 from day 36 to end of year 1, 1478 in years 2-4, and 1230 in years 5-10. Allocation to streptokinase was associated with 29 (95% confidence interval 20 to 38) fewer deaths per 1000 patients during days 0-35. This early benefit persisted (death rate ratio 0.98 (0.92 to 1.04) for additional deaths between day 36 and end of year 10), so that there were 28 (14 to 42) and 23 (2 to 44) fewer deaths per 1000 patients treated with streptokinase after 4 years and 10 years respectively. There was no evidence that absolute survival benefit increased with prolonged follow up among any category of patient, including those presenting early after symptoms started or with anterior ST elevation. Nor did the early benefits seem to be lost in any category (including those aged over 70). Allocation to one month of aspirin was associated with 26 (16 to 35) fewer deaths per 1000 during first 35 days, with little further benefit or loss during subsequent years (death rate ratio 0.99 (0.93 to 1.06) between day 36 and end of year 10). The early benefit obtained with combination of streptokinase and one month of aspirin also seemed to persist long term. The early survival advantages produced by fibrinolytic therapy and one month of aspirin started in acute myocardial infarction seem to be maintained for at least 10 years.


Non-invasive mitochondrial DNA quantification on Day 3 predicts blastocyst development: a prospective, blinded, multi-centric study.
In ART, embryo quality evaluation is routinely based on morphological criteria. We previously demonstrated that the mitochondrial DNA (mtDNA)/genomic DNA (gDNA) ratio in culture medium was significantly associated with embryo quality and viability potential. The purpose of this prospective, blinded, multi-centric study was to validate the use of mtDNA/gDNA ratio in Day 3 spent medium as a predictor of human embryo developmental competence. The mtDNA/gDNA ratio was assessed in Day 3 culture media (n=484) of embryos from 143 patients by quantitative PCR. A mixed effect logistic regression model was applied. We found that mtDNA/gDNA ratio in Day 3 culture medium combined with embryo morphology improves the prediction upon blastulation compared to morphology alone (P < 0.0001), independent of patient and cycle characteristics. With regard to routine use in clinics, we evaluated the ability of the novel, combined grading score to improve selection of developmentally competent embryos of a single cohort. Including embryos from 44 patients, the sensibility and specificity of the scoring system based on Day 3 morphological stage were 92% and 13%, respectively. Integration with the culture medium mtDNA/gDNA ratio increased the performance of the method (sensibility: 95%; specificity: 65%). The results of this study suggest the possibility of carrying out a non-invasive evaluation of embryonic mtDNA content through the culture medium. When combined with embryo morphology, it has the potential to help embryologists rank embryos and choose which embryo(s) has the greater development potential, and thus should be transferred on Day 3, among sibling embryos with the same morphological grade.


An ethnic comparison of eating attitudes and associated body image concerns in adolescent South African schoolgirls.
The purpose of this study was to investigate whether differences exist in eating attitudes and body shape concerns amongst adolescent schoolgirls representing South Africa's ethnically and culturally diverse population currently undergoing epidemiological transition. A questionnaire survey, including the Eating Attitudes Test, Body Shape Questionnaire and a Body Silhouette Chart, was administered to 228 South African schoolgirls (60 black, 83 mixed race and 85 white) aged 15-18 years from five secondary schools in the greater Cape Town area. Black girls had significantly higher mean BMI values (24.1 (3.3)) than either white (21.9 (3.0)) or mixed race girls (22.1 (3.7)) (P < 0.05). Controlling for differences in BMI, white subjects scored significantly higher on the Body Shape Questionnaire than did mixed race or black subjects, whereas no ethnic differences were found for Eating Attitude Test scores. A comparable percentage (mean = 18.8%) of black, mixed race and white girls had scores indicative of eating disorder pathology on the Eating Attitudes Test, while a higher percentage of white, compared to mixed race and black, girls had abnormal scores on the Body Shape Questionnaire (33%, 26% and 20%, respectively; P < 0.05). The ideal body size desired by white girls was significantly smaller than that of the mixed race or black samples. Dissatisfaction with present body size was significantly higher in white, compared to black or mixed race girls (P < 0.001). These findings suggest that the prevalence of abnormal eating attitudes is equally common in South African schoolgirls from different ethnic backgrounds. White girls exhibit greater body image concerns and body image dissatisfaction than mixed race or black individuals. These findings reinforce the notion that eating disorders are culture-reactive rather than culture-bound phenomena and provide insight into the extent of eating-related problems and body image issues in developing societies.


Biologically effective dose and prediction of obliteration of unruptured arteriovenous malformations treated by upfront Gamma Knife radiosurgery: a series of 149 consecutive cases.
Arteriovenous malformations (AVMs) present no pathologic tissue, and radiation dose is confined in a clear targeted volume. The authors retrospectively evaluated the role of the biologically effective dose (BED) after Gamma Knife radiosurgery (GKRS) for brain AVMs. A total of 149 consecutive cases of unruptured AVMs treated by upfront GKRS in Lille University Hospital, France, were included. The mean length of follow-up was 52.9 months (median 48, range 12-154 months). The primary outcome was obliteration, and the secondary outcome was complication appearance. The marginal dose was 24 Gy in a vast majority of cases (n = 115, 77.2%; range 18-25 Gy). The mean BED was 220.1 Gy2.47 (median 229.9, range 106.7-246.8 Gy2.47). The mean beam-on time was 32.3 minutes (median 30.8, range 9-138.7 minutes). In the present series, the mean radiation dose rate was 2.259 Gy/min (median 2.176, range 1.313-3.665 Gy/min). The Virginia score was 0 in 29 (19.5%), 1 in 61 (40.9%), 2 in 41 (27.5%), 3 in 18 (12.1%), and 4 in 0 (0%) patients, respectively. The mean Pollock-Flickinger score was 1.11 (median 1.52, range 0.4-2.9). Univariate (for obliteration and complication appearance) and multivariate (for obliteration only) analyses were performed. A total of 104 AVMs (69.8%) were obliterated at the last follow-up. The strongest predictor for obliteration was BED (p = 0.03). A radiosurgical obliteration score is proposed, derived from a fitted multivariable model: (0.018 × BED) + (1.58 × V12) + (-0.013689 × beam-on time) + (0.021 × age) - 4.38. The area under the receiver operating characteristic curve was 0.7438; after internal validation using bootstrap methods, it was 0.7088. No statistically significant relationship between radiation dose rate and obliteration was found (p = 0.29). Twenty-eight (18.8%) patients developed complications after GKRS; 20 (13.4%) of these patients had transient adverse radiological effects (perilesional edema developed). Predictors for complication appearance were higher prescription isodose volume (p = 0.005) and 12-Gy isodose line volume (V12; p = 0.001), higher Pollock-Flickinger (p = 0.02) and Virginia scores (p = 0.003), and lower beam-on time (p = 0.03). The BED was the strongest predictor of obliteration of unruptured AVMs after upfront GKRS. A radiosurgical score comprising the BED is proposed. The V12 appears as a predictor for both efficacy and toxicity. Beam-on time was illustrated as statistically significant for both obliteration and complication appearance. The radiation dose rate did not influence obliteration in the current analysis. The exact BED threshold remains to be established by further studies.


Examining non-response bias in substance use research--are late respondents proxies for non-respondents?
Non-response is a major concern among substance use epidemiologists. When differences exist between respondents and non-respondents, survey estimates may be biased. Therefore, researchers have developed time-consuming strategies to convert non-respondents to respondents. The present study examines whether late respondents (converted former non-participants) differ from early respondents, non-consenters or silent refusers (consent givers but non-participants) in a cohort study, and whether non-response bias can be reduced by converting former non-respondents. 6099 French- and 5720 German-speaking Swiss 20-year-old males (more than 94% of the source population) completed a short questionnaire on substance use outcomes and socio-demographics, independent of any further participation in a cohort study. Early respondents were those participating in the cohort study after standard recruitment procedures. Late respondents were non-respondents that were converted through individual encouraging telephone contact. Early respondents, non-consenters and silent refusers were compared to late respondents using logistic regressions. Relative non-response biases for early respondents only, for respondents only (early and late) and for consenters (respondents and silent refusers) were also computed. Late respondents showed generally higher patterns of substance use than did early respondents, but lower patterns than did non-consenters and silent refusers. Converting initial non-respondents to respondents reduced the non-response bias, which might be further reduced if silent refusers were converted to respondents. Efforts to convert refusers are effective in reducing non-response bias. However, converted late respondents cannot be seen as proxies of non-respondents, and are at best only indicative of existing response bias due to persistent non-respondents.


The roles of NADPH-oxidase and nNOS for the increased oxidative stress and the oxygen consumption in the diabetic kidney.
Sustained hyperglycaemia induces increased renal oxygen consumption resulting in reduced oxygen availability in the diabetic kidney. We investigated the roles of the nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase and the neuronal nitric oxide synthase (nNOS) for the increased oxygen consumption in streptozotocin-diabetic rats. Oxygen consumption was measured in isolated proximal tubular cells (PTC) from streptozotocin-induced diabetic rats (n = 7-9 per group) with and without chronic treatment with apocynin, a NADPH-oxidase inhibitor, or S-methyl-L-thiocitrulline (SMTC), a selective nNOS inhibitor, or a combination of the two and the results were compared to normoglycaemic controls (n = 10). Oxidative stress was estimated from thiobarbituric acid reactive substances and protein expression measured by Western blot. Proximal tubular cells from untreated diabetic rats had increased oxygen consumption compared to controls (40.6 +/- 7.9 versus 10.9 +/- 2.0 nmol/mg protein/min). All treatments reduced the diabetes-induced increase in oxygen consumption (apocynin 10.5 +/- 1.7, SMTC 19.7 +/- 3.0 and apocynin + SMTC 21.6 +/- 3.6 nmol/mg protein/min). Neither apocynin nor SMTC had any effect on the oxygen consumption in cells pre-incubated with ouabain, an inhibitor of active electrolyte transport. Oxidative stress was elevated in the diabetic kidney and inhibited by all treatments. The increased oxygen consumption by diabetic proximal tubular cells correlated with increased protein expressions of p47(phox) and nNOS and the treatments prevented these increases. Diabetes induces oxidative stress, which increases oxygen consumption in proximal tubular cells. Inhibition of either NADPH-oxidase or nNOS prevented the increased oxygen consumption. The effect of blocking both these enzymes was less than additive suggesting overlapping pathways which warrant further studies.


Carbon black and soot: two different substances.
Carbon blacks are manufactured under controlled conditions for commercial use primarily in the rubber, painting, and printing industries. In contrast, soots are unwanted byproducts from the combustion of carbon-based materials for the generation of energy or heat, or for the disposal of waste. Unfortunately, the terms carbon black and soot often have been used interchangeably; however, carbon black is physically and chemically distinct from soot. Greater than 97% of carbon black consists of elemental carbon arranged as aciniform particulate. Depending on the type of soot, the relative amount of carbon (< 60% of the total particle mass), the type of particulate carbon, and particle characteristics (size, shape, and heterogeneity) can vary considerably. For both carbon black and soot, other elements and chemical compounds are associated with the particulate carbon. Total inorganics (ash) represent < 1% of the carbon black particle mass. Organic compounds can be extracted from particle surfaces (solvent extractable fraction [SOF]), and for carbon black, also are < 1% of the particle mass. Soots have much greater percentages of ash, SOF, or both, than carbon black. There has been concern about the adsorbed organic compounds because of potential biologic activity. For carbon black, the SOF is strongly adherent to carbon surfaces and is not released by biological fluids. The types of organic compounds consist primarily of unsubstituted polycyclic aromatic hydrocarbons and are not as biologically potent as those compounds present in soot. Thus, carbon black is distinctly different from soot, and when discussing potential health effects, care must be taken to differentiate between the two types of carbon-based particles.


In vivo structure-activity relationship study of dorsomorphin analogues identifies selective VEGF and BMP inhibitors.
The therapeutic potential of small molecule signaling inhibitors is often limited by off-target effects. Recently, in a screen for compounds that perturb the zebrafish embryonic dorsoventral axis, we identified dorsomorphin, the first selective inhibitor of bone morphogenetic protein (BMP) signaling. Here we show that dorsomorphin has significant "off-target" effects against the VEGF (vascular endothelial growth factor) type-2 receptor (Flk1/KDR) and disrupts zebrafish angiogenesis. Since both BMP and VEGF signals are known to be involved in vascular development, we sought to determine whether dorsomorphin's antiangiogenic effects are due to its impact on the BMP or VEGF signals through the development of analogues that target BMP but not VEGF signaling and vice versa. In a structure-activity relationship (SAR) study of dorsomorphin analogues based primarily on their effects on live zebrafish embryos, we identified highly selective and potent BMP inhibitors as well as selective VEGF inhibitors. One of the BMP inhibitors, DMH1, which exclusively targets the BMP but not the VEGF pathway, dorsalized the embryonic axis without disrupting the angiogenic process, demonstrating that BMP signaling was not involved in the angiogenic process. This is one of the first full-scale SAR studies performed in vertebrates and demonstrates the potential of zebrafish as an attractive complementary platform for drug development that incorporates an assessment of in vivo bioactivity and selectivity in the context of a living organism.


Contrasted responses of Ruditapes decussatus (filter and deposit feeding) and Loripes lacteus (symbiotic) exposed to polymetallic contamination (Port-Camargue, France).
The use of symbiotic bivalve species to assess the effect of anthropogenic metal pollution was rarely investigated whereas data on filter feeding bivalves are common. The aim of this study was the exposure of two bivalve species, Ruditapes decussatus and Loripes lacteus to polymetallic pollution gradient, originating from harbor activities (Port-Camargue, south of France). Both bivalves differ by their trophic status, filter and deposit feeder for Ruditapes and symbiotic for Loripes that underlies potential differences in metal sensibility. The bivalves were immerged in July (for Ruditapes during 2 and 8 days) and in August 2012 (for Loripes during 2, 6 and 8 days) in the water column of the harbor, at 3 stations according to pollution gradient. Metal concentrations (Cu, Mn, Zn) in the water column were quantified as dissolved metals (measured by ICP-MS) and as labile metals (measured by ICP-MS using DGT technique). For each exposure time, accumulation of metals in the soft tissue of bivalves ("bioaccumulation") was measured for both species. In addition, specific parameters, according to the trophic status of each bivalve, were investigated: filtering activity (specific clearance rate, SCR) for Ruditapes, and relative cell size (SSC) and genomic content (FL1) of bacterial symbionts hosted in the gills of Loripes. The SCR of Ruditapes drops from 100% (control) to 34.7% after 2 days of exposure in the less contaminated site (station 8). On the other hand, the relative cell size (SSC) and genomic content (FL1), measured by flow cytometry were not impacted by the pollution gradient. Bioaccumulation was compared for both species, showing a greater capability of Cu accumulation for Loripes without lethal effect. Mn, Fe and Zn were generally not accumulated by any of the species according to the pollution gradient. The trophic status of each species may greatly influence their respective responses to polymetallic pollution.


Scene Segmentation-Based Luminance Adjustment for Multi-Exposure Image Fusion.
We propose a novel method for adjusting luminance for multi-exposure image fusion. For the adjustment, two novel scene segmentation approaches based on luminance distribution are also proposed. Multi-exposure image fusion is a method for producing images that are expected to be more informative and perceptually appealing than any of the input ones, by directly fusing photos taken with different exposures. However, existing fusion methods often produce unclear fused images when input images do not have a sufficient number of different exposure levels. In this paper, we point out that adjusting the luminance of input images makes it possible to improve the quality of the final fused images. This insight is the basis of the proposed method. The proposed method enables us to produce high-quality images, even when undesirable inputs are given. Visual comparison results show that the proposed method can produce images that clearly represent a whole scene. In addition, multi-exposure image fusion with the proposed method outperforms state-of-the-art fusion methods in terms of MEF-SSIM, discrete entropy, tone mapped image quality index, and statistical naturalness.


Severe intergenerational reproductive conflict and the evolution of menopause.
Human menopause is ubiquitous among women and is uninfluenced by modernity. In addition, it remains an evolutionary puzzle: studies have largely failed to account for diminishing selection on reproduction beyond 50 years. Using a 200-year dataset on pre-industrial Finns, we show that an important component is between-generation reproductive conflict among unrelated women. Simultaneous reproduction by successive generations of in-laws was associated with declines in offspring survivorship of up to 66%. An inclusive fitness model revealed that incorporation of the fitness consequences of simultaneous intergenerational reproduction between in-laws, with those of grandmothering and risks of dying in childbirth, were sufficient to generate selection against continued reproduction beyond 51 years. Decomposition of model estimates suggested that the former two were most influential in generating selection against continued reproduction. We propose that menopause evolved, in part, because of age-specific increases in opportunities for intergenerational cooperation and reproductive competition under ecological scarcity.


Heart applications of 4D flow.
Four-dimensional (4D) flow sequences are an innovative type of MR sequences based upon phase contrast (PC) sequences which are a type of application of Angio-MRI together with the Time of Flight (TOF) sequences and Contrast-Enhanced Magnetic Resonance Acquisition (CE-MRA). They share the basic principles of PC, but unlike PC sequences, 4D flow has velocity encoding along all three flow directions and three-dimensional (3D) anatomic coverage. They guarantee the analysis of flow with multiplanarity on a post-processing level, which is a unique feature among MR sequences. Furthermore, this technique provides a completely new level to the in vivo flow analysis as it allows measurements in never studied districts such as intracranial applications or some parts of the heart never studied with echo-color-doppler, which is its sonographic equivalent. Furthermore, this technique provides a completely new level to the in vivo flow analysis as it allows accurate measurement of the flows in different districts (e.g., intracranial, cardiac) that are usually studied with echo-color-doppler, which is its sonographic equivalent. Of note, the technique has proved to be affected by less inter and intra-observer variability in several application. 4D-flow basic principles, advantages, limitations, common pitfalls and artefacts are described. This review will outline the basis of the formation of PC image, the construction of a 4D-flow and the huge impact the technique is having on the cardiovascular non-invasive examination. It will be then studied how this technique has had a huge impact on cardiovascular examinations especially on a central heart level.