Intermittent preventive treatment of malaria during pregnancy in central Mozambique.
New WHO strategies for control of malaria in pregnancy (MiP) recommend intermittent preventive treatment (IPTp), bednet use and improved case management. A pilot MiP programme in Mozambique was designed to determine requirements for scale-up. The Ministry of Health worked with a nongovernmental organization and an academic institution to establish and monitor a pilot programme in two impoverished malaria-endemic districts. Implementing the pilot programme required provision of additional sulfadoxine-pyrimethamine (SP), materials for directly observed SP administration, bednets and a modified antenatal card. National-level formulary restrictions on SP needed to be waived. The original protocol required modification because imprecision in estimation of gestational age led to missed SP doses. Multiple incompatibilities with other health initiatives (including programmes for control of syphilis, anaemia and HIV) were discovered and overcome. Key outputs and impacts were measured; 92.5% of 7911 women received at least 1 dose of SP, with the mean number of SP doses received being 2.2. At the second antenatal visit, 13.5% of women used bednets. In subgroups (1167 for laboratory analyses; 2600 births), SP use was significantly associated with higher haemoglobin levels (10.9 g/dL if 3 doses, 10.3 if none), less malaria parasitaemia (prevalence 7.5% if 3 doses, 39.3% if none), and fewer low-birth-weight infants (7.3% if 3 doses, 12.5% if none). National-level scale-up will require attention to staffing, supplies, bednet availability, drug policy, gestational-age estimation and harmonization of vertical initiatives.


An analysis of successful litigation claims in children in England.
To analyse the number of successful claims against the National Health Service (NHS) involving children, the nature and outcome of incidents leading to litigation and the costs of claims. Under the Freedom of Information Act, details were sought of claims involving children made to the National Health Service Litigation Authority (NHSLA) from 1 April 2005 to 31 March 2010 together with the claim status on 30 September 2010. Closed cases involving financial compensation were analysed in relation to the nature of the incident, outcome and total cost of litigation. 195 closed cases were examined. The commonest causes of litigation were medication or vaccination errors (10), delayed septicaemia diagnosis (8), delayed meningitis diagnosis (7), delayed unspecified sepsis diagnosis (7), extravasation (7), delayed anorectal abnormality diagnosis (6), delayed cardiological diagnosis (6), delayed appendicitis diagnosis (6), epilepsy misdiagnosis (6), psychological/psychiatric effects on parent(s) following a medical error (4), delayed fracture diagnosis (4), gastrostomy related errors (3) and delayed testicular torsion diagnosis (3). The commonest outcomes were death (74), unnecessary pain (35), unnecessary operation (16), brain damage (12), scarring (12), psychiatric/psychological morbidity in parent(s) and/or child (10) and amputation (5). Total costs of litigation ranged from £600 to £3 044,943 (mean £127,975). Delayed diagnosis of severe sepsis is the commonest adverse incident leading to successful litigation and the commonest adverse outcome is death. The cost to the NHS is considerable. A better understanding of the causes of common errors in paediatrics should inform training and help to decrease these adverse events.


Causal effect of disconnection lesions on interhemispheric functional connectivity in rhesus monkeys.
In the absence of external stimuli or task demands, correlations in spontaneous brain activity (functional connectivity) reflect patterns of anatomical connectivity. Hence, resting-state functional connectivity has been used as a proxy measure for structural connectivity and as a biomarker for brain changes in disease. To relate changes in functional connectivity to physiological changes in the brain, it is important to understand how correlations in functional connectivity depend on the physical integrity of brain tissue. The causal nature of this relationship has been called into question by patient data suggesting that decreased structural connectivity does not necessarily lead to decreased functional connectivity. Here we provide evidence for a causal but complex relationship between structural connectivity and functional connectivity: we tested interhemispheric functional connectivity before and after corpus callosum section in rhesus monkeys. We found that forebrain commissurotomy severely reduced interhemispheric functional connectivity, but surprisingly, this effect was greatly mitigated if the anterior commissure was left intact. Furthermore, intact structural connections increased their functional connectivity in line with the hypothesis that the inputs to each node are normalized. We conclude that functional connectivity is likely driven by corticocortical white matter connections but with complex network interactions such that a near-normal pattern of functional connectivity can be maintained by just a few indirect structural connections. These surprising results highlight the importance of network-level interactions in functional connectivity and may cast light on various paradoxical findings concerning changes in functional connectivity in disease states.


Virtual Patient Simulations in Health Professions Education: Systematic Review and Meta-Analysis by the Digital Health Education Collaboration.
Virtual patients are interactive digital simulations of clinical scenarios for the purpose of health professions education. There is no current collated evidence on the effectiveness of this form of education. The goal of this study was to evaluate the effectiveness of virtual patients compared with traditional education, blended with traditional education, compared with other types of digital education, and design variants of virtual patients in health professions education. The outcomes of interest were knowledge, skills, attitudes, and satisfaction. We performed a systematic review on the effectiveness of virtual patient simulations in pre- and postregistration health professions education following Cochrane methodology. We searched 7 databases from the year 1990 up to September 2018. No language restrictions were applied. We included randomized controlled trials and cluster randomized trials. We independently selected studies, extracted data, and assessed risk of bias and then compared the information in pairs. We contacted study authors for additional information if necessary. All pooled analyses were based on random-effects models. A total of 51 trials involving 4696 participants met our inclusion criteria. Furthermore, 25 studies compared virtual patients with traditional education, 11 studies investigated virtual patients as blended learning, 5 studies compared virtual patients with different forms of digital education, and 10 studies compared different design variants. The pooled analysis of studies comparing the effect of virtual patients to traditional education showed similar results for knowledge (standardized mean difference [SMD]=0.11, 95% CI -0.17 to 0.39, I2=74%, n=927) and favored virtual patients for skills (SMD=0.90, 95% CI 0.49 to 1.32, I2=88%, n=897). Studies measuring attitudes and satisfaction predominantly used surveys with item-by-item comparison. Trials comparing virtual patients with different forms of digital education and design variants were not numerous enough to give clear recommendations. Several methodological limitations in the included studies and heterogeneity contributed to a generally low quality of evidence. Low to modest and mixed evidence suggests that when compared with traditional education, virtual patients can more effectively improve skills, and at least as effectively improve knowledge. The skills that improved were clinical reasoning, procedural skills, and a mix of procedural and team skills. We found evidence of effectiveness in both high-income and low- and middle-income countries, demonstrating the global applicability of virtual patients. Further research should explore the utility of different design variants of virtual patients.


Ottowia pentelensis sp. nov., a floc-forming betaproteobacterium isolated from an activated sludge system treating coke plant effluent.
A Gram-negative-staining, short-rod-shaped, floc-forming bacterium, designated strain RB3-7(T), was isolated from a laboratory-scale activated sludge system treating coke plant effluent. Comparative analysis of the 16S rRNA gene sequence demonstrated that the novel isolate was distantly related (≤ 95.8 % similarity) to Ottowia thiooxydans K11(T) within the family Comamonadaceae. Strain RB3-7(T) was catalase- and oxidase-positive and non-motile. The predominant fatty acids were C₁₆:₀, cyclo C₁₇:₀, C₁₈:₁ω7c and C₁₆:₁ω7c, and the major respiratory quinone was Q-8. The G+C content of the genomic DNA of strain RB3-7(T) was 68.5 mol%. On the basis of phenotypic, chemotaxonomic and molecular data, strain RB3-7(T) is considered to represent a novel species of the genus Ottowia, for which the name Ottowia pentelensis sp. nov. is proposed. The type strain is RB3-7(T) ( = DSM 21699(T) = NCAIM B 02336(T)).


Global health justice and governance.
While there is a growing body of work on moral issues and global governance in the fields of global justice and international relations, little work has connected principles of global health justice with those of global health governance for a theory of global health. Such a theory would enable analysis and evaluation of the current global health system and would ethically and empirically ground proposals for reforming it to more closely align with moral values. Global health governance has been framed as an issue of national security, human security, human rights, and global public goods. The global health governance literature is essentially untethered to a theorized framework to illuminate or evaluate governance. This article ties global health justice and ethics to principles for governing the global health realm, developing a theoretical framework for global and domestic institutions and actors.


A more biologically plausible learning rule than backpropagation applied to a network model of cortical area 7a.
Area 7a of the posterior parietal cortex of the primate brain is concerned with representing head-centered space by combining information about the retinal location of a visual stimulus and the position of the eyes in the orbits. An artificial neural network was previously trained to perform this coordinate transformation task using the backpropagation learning procedure, and units in its middle layer (the hidden units) developed properties very similar to those of area 7a neurons presumed to code for spatial location (Andersen and Zipser, 1988; Zipser and Andersen, 1988). We developed two neural networks with architecture similar to Zipser and Andersen's model and trained them to perform the same task using a more biologically plausible learning procedure than backpropagation. This procedure is a modification of the Associative Reward-Penalty (AR-P) algorithm (Barto and Anandan, 1985), which adjusts connection strengths using a global reinforcement signal and local synaptic information. Our networks learn to perform the task successfully to any degree of accuracy and almost as quickly as with backpropagation, and the hidden units develop response properties very similar to those of area 7a neurons. In particular, the probability of firing of the hidden units in our networks varies with eye position in a roughly planar fashion, and their visual receptive fields are large and have complex surfaces. The synaptic strengths computed by the AR-P algorithm are equivalent to and interchangeable with those computed by backpropagation. Our networks also perform the correct transformation on pairs of eye and retinal positions never encountered before. All of these findings are unaffected by the interposition of an extra layer of units between the hidden and output layers. These results show that the response properties of the hidden units of a layered network trained to perform coordinate transformations, and their similarity with those of area 7a neurons, are not a specific result of backpropagation training. The fact that they can be obtained by a more biologically plausible learning rule corroborates the validity of this neural network's computational algorithm as a plausible model of how area 7a may perform coordinate transformations.


[Avian influenza: eradication from commercial poultry is still not in sight].
Avian influenza viruses are highly infectious micro-organisms that primarily affect birds. Nevertheless, they have also been isolated from a number of mammals, including humans. Avian influenza virus can cause large economic losses to the poultry industry because of its high mortality. Although there are pathogenic variants with a low virulence and which generally cause only mild, if any, clinical symptoms, the subtypes H5 and H7 can mutate from a low to a highly virulent (pathogenic) virus and should be taken into consideration in eradication strategies. The primary source of infection for commercial poultry is direct and indirect contact with wild birds, with waterfowl forming a natural reservoir of the virus. Live-poultry markets, exotic birds, and ostriches also play a significant role in the epidemiology of avian influenza. The secondary transmission (i.e., between poultry farms) of avian influenza virus is attributed primarily to fomites and people. Airborne transmission is also important, and the virus can be spread by aerosol in humans. Diagnostic tests detect viral proteins and genes. Virus-specific antibodies can be traced by serological tests, with virus isolation and identification being complementary procedures. The number of outbreaks of avian influenza seems to be increasing - over the last 5 years outbreaks have been reported in Italy, Hong Kong, Chile, the Netherlands, South Korea, Vietnam, Japan, Thailand, Cambodia, Indonesia, Laos, China, Pakistan, United States of America, Canada, South Africa, and Malaysia. Moreover, a growing number of human cases of avian influenza, in some cases fatal, have paralleled the outbreaks in commercial poultry. There is great concern about the possibility that a new virus subtype with pandemic potential could emerge from these outbreaks. From the perspective of human health, it is essential to eradicate the virus from poultry; however, the large number of small-holdings with poultry, the lack of control experience and resources, and the international scale of transmission and infection make rapid control and long-term prevention of recurrence extremely difficult. In the Western world, the renewed interest in free-range housing carries a threat for future outbreaks. The growing ethical objections to the largescale culling of birds require a different approach to the eradication of avian influenza.


Genetic variation on chromosome 6 influences F cell levels in healthy individuals of African descent and HbF levels in sickle cell patients.
Fetal haemoglobin (HbF) is a major ameliorating factor in sickle cell disease. We investigated if a quantitative trait locus on chromosome 6q23 was significantly associated with HbF and F cell levels in individuals of African descent. Single nucleotide polymorphisms (SNPs) in a 24-kb intergenic region, 33-kb upstream of the HBS1L gene and 80-kb upstream of the MYB gene, were typed in 177 healthy Afro-Caribbean subjects (AC) of approximately 7% European admixture, 631 healthy Afro-Germans (AG, a group of African and German descendents located in rural Jamaica with about 20% European admixture), 87 West African and Afro-Caribbean individuals with sickle cell anaemia (HbSS), as well as 75 Northern Europeans, which served as a contrasting population. Association with a tag SNP for the locus was detected in all four groups (AC, P = 0.005, AG, P = 0.002, HbSS patients, P = 0.019, Europeans, P = 1.5 x 10(-7)). The association signal varied across the interval in the African-descended groups, while it is more uniform in Europeans. The 6q QTL for HbF traits is present in populations of African origin and is also acting in sickle cell anaemia patients. We have started to distinguish effects originating from European and African ancestral populations in our admixed study populations.


Levels of neonatal thyroid hormone in preterm infants and neurodevelopmental outcome at 5 1/2 years: millennium cohort study.
Transient hypothyroxinemia is the commonest thyroid dysfunction of premature infants, and recent studies have found adverse associations with neurodevelopment. The validity of these associations is unclear because the studies adjusted for a differing range of factors likely to influence neurodevelopment. The aim was to describe the association of transient hypothyroxinemia with neurodevelopment at 5.5 yr corrected age. We conducted a follow-up study of a cohort of infants born in Scotland from 1999 to 2001 ≤34 wk gestation. We measured scores on the McCarthy scale adjusted for 26 influences of neurodevelopment including parental intellect, home environment, breast or formula fed, growth retardation, and use of postnatal drugs. A total of 442 infants ≤34 wk gestation who had serum T(4) measurements on postnatal d 7, 14, or 28 and 100 term infants who had serum T(4) measured in cord blood were followed up at 5.5 yr. Infants with hypothyroxinemia (T(4) level ≤ 10th percentile on d 7, 14, or 28 corrected for gestational age) scored significantly lower than euthyroid infants (T(4) level greater than the 10th percentile and less than the 90th percentile on all days) on all McCarthy scales, except the quantitative. After adjustment for confounders of neurodevelopment, hypothyroxinemic infants scored significantly lower than euthyroid infants on the general cognitive and verbal scales. Our findings do not support the view that the hypothyroxinemic state, in the context of this analysis, is harmless in preterm infants. Many factors contribute both to the etiology of hypothyroxinemia and neurodevelopment; strategies for correction of hypothyroxinemia should acknowledge its complex etiology and not rely solely on one approach.


Pet Ownership and Cardiovascular Health in the US General Population.
The 2013 American Heart Association Scientific Statement on pet ownership and cardiovascular risk suggested that dog ownership is probably associated with decreased cardiovascular risk. Several studies have been shown that pet ownership, particularly of dogs, is associated with increased physical activity levels, social support, and improved outcomes after a major cardiovascular event. We hypothesized that pet ownership is associated with a lower risk of cardiovascular disease in the US general population. Using the National Health and Nutrition Examination Survey, we identified all patients with heart failure, coronary artery disease, systemic hypertension (SH), diabetes mellitus, and stroke between 1999 and 2016. Multivariable analyses were performed to adjust for demographic factors such as age, gender, marital status, education, co-morbidities, cigarette smoking, family income, working hours, sleeping duration, depression, and lipid profiles. Of 42,038 National Health and Nutrition Examination Survey participants, 10,905 (31%) were inquired about pet ownership. Pet owners were older, less likely to be women or widowed, and more likely to be white, more educated, tobacco users, and work more than 35 hours per week than non-owners (all p values <0.05). Pet owners had higher hemoglobin, lower low-density lipoprotein cholesterol, and a lower prevalence of DM, SH, and stroke (all p values <0.05). After adjusting for potential confounders, pet ownership (either cat or dog ownership) was independently associated with a lower prevalence of SH (odds ratio 0.67; 95% confidence interval 0.49 to 0.89; p = 0.01), but not heart failure, coronary artery disease, DM, or stroke, compared with non-owners. In conclusions, using a large national database, we found that pet ownership is associated with a decreased prevalence of SH. Further longitudinal studies are needed to draw a conclusion on the protective effect of pet ownership in patients with cardiovascular disease.


A 27 kDa cysteine protease secreted by newly excysted Paragonimus westermani metacercariae induces superoxide anion production and degranulation of human eosinophils.
Eosinophil degranulation plays a crucial role in tissue inflammatory reactions associated with helminth parasitic infections and allergic diseases. Paragonimus westermani, a lung fluke causing human paragonimiasis, secretes a large amount of cysteine proteases, which are involved in nutrient uptake, tissue invasion, and modulation of hos's immune responses. There is, however, limited information about the response of eosinophils to direct stimulation by cysteine proteases (CP) secreted by P. westermani. In the present study, we tested whether degranulation and superoxide production from human eosinophils can be induced by stimulation of the 2 CP (27 kDa and 28 kDa) purified from excretory-secretory products (ESP) of P. westermani newly excysted metacercariae (PwNEM). A large quantity of eosinophil-derived neurotoxin (EDN) was detected in the culture supernatant when human eosinophils isolated from the peripheral blood were incubated with the purified 27 kDa CP. Furthermore, the 27 kDa CP induced superoxide anion production by eosinophils in time- and dose-dependent manners. In contrast, the purified 28 kDa CP did not induce superoxide production and degranulation. These findings suggest that the 27 kDa CP secreted by PwNEM induces superoxide production and degranulation of human eosinophils, which may be involved in eosinophil-mediated tissue inflammatory responses during the larval migration in human paragonimiasis.


Homelessness and Sexual Identity Among Middle School Students.
Lesbian, gay, bisexual, or questioning (LGBQ) high school students experience higher rates of homelessness than their heterosexual peers. Moreover, LGBQ high school students are more likely to stay in riskier locations (eg, with a stranger) and less likely to stay in a shelter. This study tested whether these trends also apply to middle school students. Using representative data, we examined sexual identity and homelessness among Los Angeles Unified School District middle school students. Nearly 10% of middle school students identified as LGBQ and 23.5% experienced at least 1 night of homelessness during the previous year. Contrary to high school data, LGBQ students did not experience higher rates of homelessness overall. However, when limiting the sample to students who had experienced homelessness, LGBQ students were more than 5 times as likely as heterosexual students to have stayed in a public place and 63% as likely to have stayed in a shelter. Lesbian, gay, bisexual, or questioning students are more likely to experience public homelessness. Schools must implement homelessness surveillance systems to assist in identifying early episodes of homelessness, thereby reducing the likelihood of poor physical and mental health outcomes associated with chronic homelessness.


Continuous monitoring of cerebrovascular pressure-reactivity in head injury.
Cerebrovascular vasomotor reactivity reflects changes in smooth muscle tone in the arterial wall in response to changes in transmural pressure or concentration of carbon dioxide in blood. We have investigated whether slow waves in ABP and ICP may be used to derive an index which reflects reactivity of vessels to changes in arterial blood pressure. A method for the continuous monitoring of the association between slow spontaneous waves in ICP and AP has been adopted in a group of 98 head injured patients. ABP, ICP and transcranial Doppler blood flow velocity (FV) in the middle cerebral artery was recorded daily (20 to 120 minutes time periods). A Pressure-Reactivity Index (PRx) was calculated as a moving correlation coefficient between 40 consecutive samples of values for ICP and ABP averaged over 5 seconds. A moving correlation coefficient between spontaneous fluctuations of mean FV and CPP (Mx), which was previously reported to describe cerebral blood flow autoregulation, was also calculated. In an additional 25 patients, PRx was calculated and recorded continuously along with mean ICP, ABP and parameters describing ICP waveform. A positive PRx correlated with high ICP (r = 0.366; p < 0.001), low admission GCS (r = 0.29; p < 0.01), and poor outcome at 6 months after injury (r = 0.48; p < 0.00001). During the first two days following injury, PRx was positive (p < 0.05) in patients with unfavourable outcome. The correlation between PRx and Mx (r = 0.63) was highly significant (p < 0.000001). Continuous recordings demonstrated that PRx was able to indicate individual thresholds of vascular reactivity for CPP, ICP, and ventilation parameters. Computer analysis of slow waves in ABP and ICP is able to provide a continuous index of cerebrovascular reactivity to changes in arterial pressure, which is of prognostic significance.


Tetrahydrobiopterin in cardiovascular health and disease.
Tetrahydrobiopterin (BH4) functions as a cofactor for several important enzyme systems, and considerable evidence implicates BH4 as a key regulator of endothelial nitric oxide synthase (eNOS) in the setting of cardiovascular health and disease. BH4 bioavailability is determined by a balance of enzymatic de novo synthesis and recycling, versus degradation in the setting of oxidative stress. Augmenting vascular BH4 levels by pharmacological supplementation has been shown in experimental studies to enhance NO bioavailability. However, it has become more apparent that the role of BH4 in other enzymatic pathways, including other NOS isoforms and the aromatic amino acid hydroxylases, may have a bearing on important aspects of vascular homeostasis, inflammation, and cardiac function. This article reviews the role of BH4 in cardiovascular development and homeostasis, as well as in pathophysiological processes such as endothelial and vascular dysfunction, atherosclerosis, inflammation, and cardiac hypertrophy. We discuss the therapeutic potential of BH4 in cardiovascular disease states and attempt to address how this modulator of intracellular NO-redox balance may ultimately provide a powerful new treatment for many cardiovascular diseases.


Neurological and behavioral outcomes of focal cerebral ischemia in rats.
The aim of this study was to investigate the neurobehavioral consequences of focal ischemia in rats. We induced permanent occlusion of the left middle cerebral artery in 14 Sprague-Dawley rats, and used 13 sham-operated rats as controls. During surgery, brain temperature and body temperature were kept at normothermia. Neurobehavioral studies (neurological examination, passive avoidance task, Y maze test, and modified open-field test) were carried out 4 days after ischemia before killing the rats to evaluate histological damage. Ischemia induced large infarcts in the cortex (138.6 +/- 8.5 mm3) and caudate-putamen (48.8 +/- 2.6 mm3) and, compared with sham-operated rats, produced a dramatic neurological deficit (p less than 0.001) characterized by sensorimotor dysfunctions and hemiplegia. Memory retention was significantly (p less than 0.05) impaired in the passive avoidance task, but neither vigilance and exploratory behavior measured in the modified open-field test nor working memory evaluated in the Y maze test were disturbed. Infarct size was not correlated with the neurological or behavioral deficits. This lack of correlation indicates the necessity of carrying out parallel histological, neurological, and behavioral studies in any assays of new drugs using this model of focal ischemia.


Prediction of invasive activities in hepatocellular carcinomas with special reference to alpha-fetoprotein and des-gamma-carboxyprothrombin.
Hepatocellular carcinoma (HCC) is one of the world's major malignancies, especially in Asian countries. To improve the prognosis of HCC, it is essential to predict its invasive behavior in both intra- and extra-hepatic modalities. For this purpose, we examined the predictive values of two tumor markers, alpha-fetoprotein (AFP) and des-gamma-carboxyprothrombin (DCP). 194 HCC cases at the National Cancer Center Hospital were selected from the Pathology Records. Detailed information regarding the existence of extra-hepatic venous invasion (EHVI) at the portal vein and intra-hepatic multiple malignant tumors (IHMTs) were collected and combined with the preoperative AFP and DCP testing results. Furthermore, information about the viral infection status such as HBs antigen positive or HCV antibody positive or no viral hepatitis was obtained. The information was analyzed by the ROC (Receiver Operating Characteristic Curve) method. and In both the EHVI group and the IHMT group, all the combinations except the HCV-positive group of IHMT revealed a tendency for DCP to offer better diagnostic accuracies, although this was statistically significant only in the HBs-positive group of IHMT. This result indicates either (1). that in a strict sense, DCP is not necessarily better than AFP at predicting the invasive characteristics or (2). that DCP is better than AFP at reflecting the invasive characteristics of HCC although not statistically significant. In either situation, from a long-term viewpoint, it is advisable to find a new marker or to re-evaluate the existing markers in order to predict the invasive characteristics more accurately.


Prooxidative effects of aspartame on antioxidant defense status in erythrocytes of rats.
Since aspartame (L-aspartyl-L-phenylalanine methyl ester, ASP) is one of the most widely used artificial sweeteners, the aim of the present study was to investigate its effects on serum glucose and lipid levels as well as its effects on oxidative/antioxidative status in erythrocytes of rats. The experiment included two groups of animals: the control group was administered with water only, while the experimental group was orally administered with ASP (40 mg/kg b.w.) daily, for a period of six weeks. When compared with the control group, the group administrated with ASP indicated higher values of serum glucose, cholesterol and triglycerides. Significantly increased concentrations of superoxide anion (O2 .-), hydrogen peroxide (H2O2), peroxynitrite (?N??-) and lipid peroxides (LPO) were recorded in the erythrocytes of ASP treated group in comparison to the control group. In the course of chronic ASP administration, the following was observed: the concentration of reduced glutathione (GSH) and the activity of catalase (CAT) increased. Thus, these findings suggest that long-term consumption of ASP leads to hyperglycemia and hyperlipidemia, as well as to oxidative stress in erythrocytes.


Insights into the origin of carbonaceous chondrite organics from their triple oxygen isotope composition.
Dust grains of organic matter were the main reservoir of C and N in the forming Solar System and are thus considered to be an essential ingredient for the emergence of life. However, the physical environment and the chemical mechanisms at the origin of these organic grains are still highly debated. In this study, we report high-precision triple oxygen isotope composition for insoluble organic matter isolated from three emblematic carbonaceous chondrites, Orgueil, Murchison, and Cold Bokkeveld. These results suggest that the O isotope composition of carbonaceous chondrite insoluble organic matter falls on a slope 1 correlation line in the triple oxygen isotope diagram. The lack of detectable mass-dependent O isotopic fractionation, indicated by the slope 1 line, suggests that the bulk of carbonaceous chondrite organics did not form on asteroidal parent bodies during low-temperature hydrothermal events. On the other hand, these O isotope data, together with the H and N isotope characteristics of insoluble organic matter, may indicate that parent bodies of different carbonaceous chondrite types largely accreted organics formed locally in the protosolar nebula, possibly by photochemical dissociation of C-rich precursors.


Colonoscopy screening markedly reduces the occurrence of colon carcinomas and carcinoma-related death: a closed cohort study.
Colonoscopy with a possible polypectomy is an efficient and preferred screening method to reduce the incidence of colorectal cancer (CRC). However, critics argue that, to date, a reduction of incidence and mortality from CRC has not been demonstrated in a population-based setting. To compare the incidence of and mortality from CRC among individuals screened by colonoscopy and non-screened individuals. A closed cohort study. Population-based setting in a precisely defined area with a low level of population migration. This study involved 1912 screened and 20,774 control participants. CRC cases in this closed cohort study were prospectively collected during the screening period of 1 year and the follow-up period of 6 years. Follow-up data were corrected for negligible migration balance in the area. Tumor characteristics and risk or protective factors, age and sex, participation in general health screening examinations, history of CRC in a first-degree relative, smoking status, body mass index, frequency of sports activity, eating habits, and patients' professions were recorded. Overall cancer incidence was significantly lower in the screened group compared with the non-screened group (adjusted odds ratio [OR] 0.31; 95% confidence interval [CI], 0.16-0.59; P < .001). Colon cancer-associated mortality also was clearly lower (adjusted OR 0.12; 95% CI, 0.01-0.93; P = .04). Risk factors such as lifestyle, smoking, and body mass index as well as family history were similar in both groups. Blue-collar workers had a higher incidence of CRC compared with professionals. The risk factors for CRC were a positive family history and smoking. Number and ethnicity of the participants, non-randomized study. Colonoscopy with polypectomy significantly reduces CRC incidence and cancer-related mortality in the general population.


Cardiovascular and catecholamine responses to static exercise in partially curarized humans.
Neural control of the circulation was evaluated during static exercise in 19 subjects by the determination of heart rate (HR), mean arterial pressure (MAP), cardiac output (CO) and plasma catecholamines. Influence from central command was evaluated during contractions with weakened muscles following partial curarization and reflex influence from metaboreceptors was assessed by post-exercise muscle ischaemia. Static handgrip increased HR and more so MAP and CO and MAP remained elevated during post-exercise muscle ischaemia. With partial curarization plasma catecholamines were also increased (P < 0.05). Two-leg extension increased all variables and during post-exercise muscle ischaemia elevations of HR, MAP and CO were maintained (P < 0.05). With partial curarization HR, MAP and plasma noradrenaline were even greater during the contraction. With the involvement of both legs during static exercise, reflex influence from the muscles elevated blood pressure by way of HR and CO and the importance of central command was detectable for HR and MAP as plasma catecholamines became elevated. However, the results indicate a separation between a central command influence on HR and CO related to an increase in plasma catecholamines during a handgrip, while the reflex influence on blood pressure was directed towards total peripheral resistance.


Simulation of specimen-induced aberrations for objects with spherical and cylindrical symmetry.
Wavefront aberrations caused by the refractive index structure of the specimen are known to compromise signal intensity and three-dimensional resolution in confocal and multiphoton microscopy. However, adaptive optics can measure and correct specimen-induced aberrations. For the design of an adaptive optics system, information on the type and amount of the aberration is required. We have previously described an interferometric set-up capable of measuring specimen-induced aberrations and a method for the extraction of the Zernike mode content. In this paper we have modelled specimen-induced aberrations caused by spherical and cylindrical objects using a ray tracing method. The Zernike mode content of the wavefronts was then extracted from the simulated wavefronts and compared with experimental results. Aberrations for a simple model of an oocyte cell consisting of two spherical regions and for a model of a well-characterized optical fibre are calculated. This simple model gave Zernike mode data that are in good agreement with experimental results.


DWI acquisition schemes and diffusion tensor estimation: a simulation-based study.
Diffusion Tensor Imaging (DTI) is the most used paradigm among the Diffusion Weighted Magnetic Resonance Imaging techniques, due to its inner simplicity and huge application potential. Least Squares has become the standard technique to estimate the Diffusion Tensor (DT) from Diffusion Weighted Images. This approach is known to be optimal when the acquired data follows Gaussian, Rician or non-central chi distributions. In this paper we study the effect of different acquisition schemes over the quality of the DT estimation. The following cases are considered: single coil, multiple coil fully sampled, and accelerated subsampled multiple coil acquisitions with SENSE and GRAPPA reconstructions. Reconstructed subsampled data will show an acceleration in the acquisition process, but also greater variance and bias in the DT estimation.


A preliminary analysis of genome structure and composition in Gossypium hirsutum.
Upland cotton has the highest yield, and accounts for > 95% of world cotton production. Decoding upland cotton genomes will undoubtedly provide the ultimate reference and resource for structural, functional, and evolutionary studies of the species. Here, we employed GeneTrek and BAC tagging information approaches to predict the general composition and structure of the allotetraploid cotton genome. 142 BAC sequences from Gossypium hirsutum cv. Maxxa were downloaded http://www.ncbi.nlm.nih.gov and confirmed. These BAC sequence analysis revealed that the tetraploid cotton genome contains over 70,000 candidate genes with duplicated gene copies in homoeologous A- and D-subgenome regions. Gene distribution is uneven, with gene-rich and gene-free regions of the genome. Twenty-one percent of the 142 BACs lacked genes. BAC gene density ranged from 0 to 33.2 per 100 kb, whereas most gene islands contained only one gene with an average of 1.5 genes per island. Retro-elements were found to be a major component, first an enriched LTR/gypsy and second LTR/copia. Most LTR retrotransposons were truncated and in nested structures. In addition, 166 polymorphic loci amplified with SSRs developed from 70 BAC clones were tagged on our backbone genetic map. Seventy-five percent (125/166) of the polymorphic loci were tagged on the D-subgenome. By comprehensively analyzing the molecular size of amplified products among tetraploid G. hirsutum cv. Maxxa, acc. TM-1, and G. barbadense cv. Hai7124, and diploid G. herbaceum var. africanum and G. raimondii, 37 BACs, 12 from the A- and 25 from the D-subgenome, were further anchored to their corresponding subgenome chromosomes. After a large amount of genes sequence comparison from different subgenome BACs, the result showed that introns might have no contribution to different subgenome size in Gossypium. This study provides us with the first glimpse of cotton genome complexity and serves as a foundation for tetraploid cotton whole genomesequencing in the future.


Netrin/DCC signaling controls contralateral dendrites of octavolateralis efferent neurons.
The guidance molecule Netrin and its receptor DCC (deleted in colorectal cancer) attract commissural axons toward the midline en route to their final destination. To test whether these molecules can also guide dendrites, we studied the contralateral dendrites of zebrafish octavolateralis efferent (OLe) neurons, which are unusual in that they navigate toward and cross the midline. We found that, at the time of dendrite outgrowth, OLe neurons express dcc, and the hindbrain midline expresses netrin1. Knocking down dcc or netrin1 function by injecting antisense morpholino oligonucleotides prevented OLe contralateral dendrites from crossing the midline, showing that dcc and netrin1 are necessary for dendrite guidance or formation. Furthermore, by transplanting cells from dcc morphants into wild-type embryos and vice versa, we demonstrated that dcc acts cell autonomously in OLe dendrites. This work is the first evidence that Netrin/DCC signaling acts in dendrites in a vertebrate system.


Altered amygdala and hippocampus function in adolescents with hypercortisolemia: a functional magnetic resonance imaging study of Cushing syndrome.
Chronic elevations of endogenous cortisol levels have been shown to alter medial temporal cortical structures and to be accompanied by declarative memory impairments and depressive symptoms in human adults. These effects of elevated endogenous levels of cortisol have not been directly studied in adolescents. Because adolescents with Cushing syndrome show endogenous elevations in cortisol, they represent a unique natural model to study the effects of prolonged hypercortisolemia on brain function, and memory and affective processes during this developmental stage. Using functional magnetic resonance imaging (fMRI), we compared 12 adolescents with Cushing syndrome with 22 healthy control adolescents on amygdala and anterior hippocampus activation during an emotional faces encoding task. None of these adolescents manifested depressive symptoms. Encoding success was assessed using a memory recognition test performed after the scan. The fMRI analyses followed an event-related design and were conducted using the SPM99 platform. Compared to healthy adolescents, patients with Cushing syndrome showed greater left amygdala and right anterior hippocampus activation during successful face encoding. Memory performance for faces recognition did not differ between groups. This first study of cerebral function in adolescents with chronic endogenous hypercortisolemia due to Cushing syndrome demonstrates the presence of functional alterations in amygdala and hippocampus, which are not associated with affective or memory impairments. Such findings need to be followed by work examining the role of age and related brain maturational stage on these effects, as well as the identification of possible protective factors conferring resilience to affective and cognitive consequences in this disease and/or during this stage of cerebral development.


Effects of rhBMP-2 on Sandblasted and Acid Etched Titanium Implant Surfaces on Bone Regeneration and Osseointegration: Spilt-Mouth Designed Pilot Study.
This study was conducted to evaluate effects of rhBMP-2 applied at different concentrations to sandblasted and acid etched (SLA) implants on osseointegration and bone regeneration in a bone defect of beagle dogs as pilot study using split-mouth design. Methods. For experimental groups, SLA implants were coated with different concentrations of rhBMP-2 (0.1, 0.5, and 1 mg/mL). After assessment of surface characteristics and rhBMP-2 releasing profile, the experimental groups and untreated control groups (n = 6 in each group, two animals in each group) were placed in split-mouth designed animal models with buccal open defect. At 8 weeks after implant placement, implant stability quotients (ISQ) values were recorded and vertical bone height (VBH, mm), bone-to-implant contact ratio (BIC, %), and bone volume (BV, %) in the upper 3 mm defect areas were measured. Results. The ISQ values were highest in the 1.0 group. Mean values of VBH (mm), BIC (%), and BV (%) were greater in the 0.5 mg/mL and 1.0 mg/mL groups than those in 0.1 and control groups in buccal defect areas. Conclusion. In the open defect area surrounding the SLA implant, coating with 0.5 and 1.0 mg/mL concentrations of rhBMP-2 was more effective, compared with untreated group, in promoting bone regeneration and osseointegration.


Fine-scale differences between Accumulibacter-like bacteria in enhanced biological phosphorus removal activated sludge.
A lab-scale sequencing batch reactor (SBR) and six full-scale wastewater treatment plants (WWTPs) performing enhanced biological phosphorus removal (EBPR) were surveyed. The abundance of Accumulibacter-related organisms in the full-scale plants was investigated using fluorescent in situ hybridization. Accumulibacter-related organisms were present in all of the full-scale EBPR plants, at levels ranging from 9% to 24% of total cells. The high percentage of Accumulibacter-related organisms seemed to be associated with configurations which minimize the nitrate recycling to the anaerobic zone and low influent BOD:TP ratios. PCR-based clone libraries were constructed from the community 16S rRNA gene plus the internally transcribed spacer region amplified from the SBR and five of the full-scale WWTPs. Comparative sequence analysis was carried out using Accumulibacter-related clones, providing higher phylogenetic resolution and revealing finer-scale clustering of the sequences retrieved from the SBR and full-scale EBPR


Anatomically Precontoured Locked Plates in Pilon Fractures: A Computed Tomography Based and Cadaveric Study.
The treatment of comminuted tibia plafond fractures remains clinically challenging due to the complexity of the articular fracture pattern despite using the anatomically precontoured locked plates. This study describes the morphologic characteristics of the anterolateral fragment and to evaluate the fixability of the anterolateral fragment with the anatomically precontoured locked plate in the pilon fracture. One hundred and twenty five cases of AO 43-B and C fracture were evaluated using the computed tomography (CT) scan. The anterior-posterior distance in CT (APDc), medial-lateral distance in CT (MLDc), coronal and sagittal height, and articular surface area of the anterolateral fragment were measured in CT. Four types of anatomically precontoured locked plates were used for cadaveric measurement. Four cadaveric parameters were also evaluated; anteroposterior distance in plate (APDp), height of the screw in the medial plate, medial-lateral distance in plate (MLDp), and height of the screw in the anterolateral plate. The anterolateral fragment was described with a mean surface area of 167.13 mm2 (APDc: 10.89 ± 4.64 mm, MLDc: 15.02 ± 6.56 mm, sagittal height: 14.85 ± 6.25 mm, and coronal height: 17.27 ± 6.88 mm). The cadaveric measurement showed that the juxta-articular screw of the medial distal tibia plate was placed away from the anterolateral fragment. The anterolateral distal tibia plate did not purchase the anterolateral fragment due to the higher position of the most distal-lateral screw (Synthes 18.37 ± 1.86 mm and Zimmer 17.78 ± 2.37 mm of the height of screw in the anterolateral plate). Anatomical distal tibial locked plates did not take purchase on the anterolateral fragment in pilon fracture in the best anatomical fit. Preoperative CT measurement can be used for determining a fixation strategy for the anterolateral fragment. In addition, a newly designed anterolateral distal tibia plate can be another solution when the usual anatomically precontoured distal tibia locked plate fails to cover the anterolateral fragment.


The biomechanical role of the chondrocranium and sutures in a lizard cranium.
The role of soft tissues in skull biomechanics remains poorly understood. Not least, the chondrocranium, the portion of the braincase which persists as cartilage with varying degrees of mineralization. It also remains commonplace to overlook the biomechanical role of sutures despite evidence that they alter strain distribution. Here, we examine the role of both the sutures and the chondrocranium in the South American tegu lizard Salvator merianae We use multi-body dynamics analysis (MDA) to provide realistic loading conditions for anterior and posterior unilateral biting and a detailed finite element model to examine strain magnitude and distribution. We find that strains within the chondrocranium are greatest during anterior biting and are primarily tensile; also that strain within the cranium is not greatly reduced by the presence of the chondrocranium unless it is given the same material properties as bone. This result contradicts previous suggestions that the anterior portion (the nasal septum) acts as a supporting structure. Inclusion of sutures to the cranium model not only increases overall strain magnitudes but also leads to a more complex distribution of tension and compression rather than that of a beam under sagittal bending.


Differentiation of intervertebral notochordal cells through live automated cell imaging system in vitro.
We demonstrated the differentiation of notochordal cells by direct observation using a live automated cell imaging system. We also hypothesized that notochordal cells have characteristics of chondrocyte-like cells. To determine characteristics of notochordal cells by matrix protein expression and their differentiation using a live automated cell imager. Although notochordal cells are critical to homeostasis of intervertebral disc, their fate has not been extensively studied and there is little evidence of notochordal cells as progenitors. Notochordal cells purified from rabbit nucleus pulposus were isolated after serial filtration. Notochordal cells in 3-dimensional culture were compared to chondrocyte-like cells by S sulfate incorporation into proteoglycan and reverse transcription polymerase chain reaction for gene expression(collagen II and aggrecan). Notochordal cells in 2-D culture were used for immunocytochemical staining (collagen II, aggrecan, and SOX9) and time-lapsed cell tracking study. Notochordal cells were capable of proteoglycan production at a rate comparable to chondrocyte-like cells (108% +/- 22.6% to chondrocyte-like cells) and expressed collagen II, aggrecan, and SOX9. In time-lapsed cell tracking analysis, notochordal cells were slower in population doubling time than chondrocyte-like cells and differentiated into 3 morphologically distinct cell types: vacuolated cells (area: 2392 +/- 507.1 microm, velocity: 0.09 +/- 0.01 microm/min); giant cells (area: 12678 +/- 1637.0 microm, velocity: 0.08 +/- 0.01 microm/min) which grew rapidly without cell division; polygonal cells (area: 3053 +/- 751.2 microm, 0.14 +/- 0.01 microm/min) morphologically similar to typical differentiation type of chondrocyte-like cells (area: 2671 +/- 235.6 microm, 0.19 +/- 0.01 microm/min). Rarely, notochordal cells formed clusters analogous to that observed in vivo. These studies demonstrate a chondrocyte phenotype of notochordal cells and are the first direct evidence of notochordal cell differentiation, suggesting that they may act as progenitor cells, which has the potential to lead to their use in novel approaches to regeneration of degenerative intervertebral disc.


Simultaneous expansion and harvest of hematopoietic stem cells and mesenchymal stem cells derived from umbilical cord blood.
The simultaneous expansion and harvest of hematopoietic stem cells and mesenchymal stem cells derived from umbilical cord blood were carried out using bioreactors. The co-culture of umbilical cord blood (UCB)-derived hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs) was performed within spinner flasks and a rotating wall vessel (RWV) bioreactor using glass-coated styrene copolymer (GCSC) microcarriers. The medium used was composed of serum-free IMDM containing a cocktail of SCF 15 ng·mL(-1), FL 5 ng·mL(-1), TPO 6 ng·mL(-1), IL-3 15 ng·mL(-1), G-CSF 1 ng·mL(-1) and GM-CSF 5 ng·mL(-1). Accessory stromal cells derived from normal allogeneic adipose tissue were encapsulated in alginate-chitosan (AC) beads and used as feeding cells. The quality of the harvested UCB-HSCs and MSCs was assessed by immunophenotype analysis, methylcellulose colony and multi-lineage differentiation assays. After 12 days of culture, the fold-expansion of total cell numbers, colony-forming units (CFU-C), CD34(+)/CD45(+)/CD105(-) (HSCs) cells and CD34(-)/CD45(-)/CD105(+) (MSCs) cells using the RWV bioreactor were (3.7 ± 0.3)- , (5.1 ± 1.2)- , (5.2 ± 0.4)- , and (13.9 ± 1.2)-fold respectively, significantly better than those obtained using spinner flasks. Moreover, UCB-HSCs and UCB-MSCs could be easily separated by gravity sedimentation after the co-culture period as only UCB-MSCs adhered on to the microcarriers. Simultaneously, we found that the fibroblast-like cells growing on the surface of the GCSC microcarriers could be induced and differentiated towards the osteoblastic, chondrocytic and adipocytic lineages. Phenotypically, these cells were very similarly to the MSCs derived from bone marrow positively expressing the MSCs-related markers CD13, CD44, CD73 and CD105, while negatively expressing the HSCs-related markers CD34, CD45 and HLA-DR. It was thus demonstrated that the simultaneous expansion and harvest of UCB-HSCs and UCB-MSCs is possible to be accomplished using a feasible bioreactor culture system such as the RWV bioreactor with the support of GCSC microcarriers.


Hawkmoths produce anti-bat ultrasound.
Bats and moths have been engaged in aerial warfare for nearly 65 Myr. This arms race has produced a suite of counter-adaptations in moths, including bat-detecting ears. One set of defensive strategies involves the active production of sound; tiger moths' ultrasonic replies to bat attack have been shown to startle bats, warn the predators of bad taste and jam their biosonar. Here, we report that hawkmoths in the Choerocampina produce entirely ultrasonic sounds in response to tactile stimulation and the playback of biosonar attack sequences. Males do so by grating modified scraper scales on the outer surface of the genital valves against the inner margin of the last abdominal tergum. Preliminary data indicate that females also produce ultrasound to touch and playback of echolocation attack, but they do so with an entirely different mechanism. The anti-bat function of these sounds is unknown but might include startling, cross-family acoustic mimicry, warning of unprofitability or physical defence and/or jamming of echolocation. Hawkmoths present a novel and tractable system to study both the function and evolution of anti-bat defences.


Use of screening phase data to evaluate observer variation of sputum cytodiagnosis as an outcome measure in a chemoprevention trial.
Sputum samples obtained during the screening phase of a chemoprevention trial in heavy smokers were evaluated independently by trained cytotechnologists and classified for degree of cellular atypia according to the method of Saccommano et al. (G. Saccommano et al., Cancer (Phila.), 33: 256-270, 1974). The level of agreement within and between Observers A and B was calculated as the percentage of agreement and, in addition, a statistic was used (kappa) to correct for chance-expected agreement. Between observer agreement on 300 specimens from 130 subjects was 68% (204 of 300) (kappa = 0.58). Of the 96 disagreements, only 17 were of more than one category in the six-category classification. Within observer agreement for both Observers A and B was evaluated on a subset of 60 specimens from 49 subjects examined on two separate occasions by each observer. The percentage of within observer agreement was 80% for Observer A (kappa = 0.73) and 62% for Observer B (kappa = 0.49) (P less than 0.04). Altogether, 71% (25 of 35) of within observer discordant readings were confined to only one category. These data, obtained from the screening phase of the study, will allow us to refine the outcome measure for the trial without introducing bias that could result from knowledge of the actual study results. The failure of conventional cancer treatments to impact significantly on overall cancer mortality has led to an emphasis on prevention. If premalignant changes can be reliably detected at a time when the malignant process is reversible, then it may be feasible to intervene to prevent the occurrence of some cancers. The appropriate selection of subjects for such intervention trials and the subsequent demonstration of the efficacy of chemoprevention strategies are therefore crucial.


Zif268/Egr1 gain of function facilitates hippocampal synaptic plasticity and long-term spatial recognition memory.
It is well established that Zif268/Egr1, a member of the Egr family of transcription factors, is critical for the consolidation of several forms of memory; however, it is as yet uncertain whether increasing expression of Zif268 in neurons can facilitate memory formation. Here, we used an inducible transgenic mouse model to specifically induce Zif268 overexpression in forebrain neurons and examined the effect on recognition memory and hippocampal synaptic transmission and plasticity. We found that Zif268 overexpression during the establishment of memory for objects did not change the ability to form a long-term memory of objects, but enhanced the capacity to form a long-term memory of the spatial location of objects. This enhancement was paralleled by increased long-term potentiation in the dentate gyrus of the hippocampus and by increased activity-dependent expression of Zif268 and selected Zif268 target genes. These results provide novel evidence that transcriptional mechanisms engaging Zif268 contribute to determining the strength of newly encoded memories.


Training the Workforce: Description of a Longitudinal Interdisciplinary Education and Mentoring Program in Palliative Care.
The rapid increase in demand for palliative care (PC) services has led to concerns regarding workforce shortages and threats to the resiliency of PC teams. To describe the development, implementation, and evaluation of a regional interdisciplinary training program in PC. Thirty nurse and physician fellows representing 22 health systems across the Chicago region participated in a two-year PC training program. The curriculum was delivered through multiple conferences, self-directed e-learning, and individualized mentoring by expert local faculty (mentors). Fellows shadowed mentors' clinical practices and received guidance on designing, implementing, and evaluating a practice improvement project to address gaps in PC at their institutions. Enduring, interdisciplinary relationships were built at all levels across health care organizations. Fellows made significant increases in knowledge and self-reported confidence in adult and pediatric PC and program development skills and frequency performing these skills. Fellows and mentors reported high satisfaction with the educational program. This interdisciplinary PC training model addressed local workforce issues by increasing the number of clinicians capable of providing PC. Unique features include individualized longitudinal mentoring, interdisciplinary education, on-site project implementation, and local network building. Future research will address the impact of the addition of social work and chaplain trainees to the program.


A novel function for the N-terminal nucleophile hydrolase fold demonstrated by the structure of an archaeal inosine monophosphate cyclohydrolase.
Inosine 5'-monophosphate (IMP) cyclohydrolase catalyzes the cyclization of 5-formaminoimidazole-4-carboxamide ribonucleotide (FAICAR) to IMP in the final step of de novo purine biosynthesis. Two major types of this enzyme have been discovered to date: PurH in Bacteria and Eukarya and PurO in Archaea. The structure of the MTH1020 gene product from Methanothermobacter thermoautotrophicus was previously solved without functional annotation but shows high amino acid sequence similarity to other PurOs. We determined the crystal structure of the MTH1020 gene product in complex with either IMP or 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) at 2.0 and 2.6 A resolution, respectively. On the basis of the sequence analysis, ligand-bound structures, and biochemical data, MTH1020 is confirmed as an archaeal IMP cyclohydrolase, thus designated as MthPurO. MthPurO has a four-layered alphabeta betaalpha core structure, showing an N-terminal nucleophile (NTN) hydrolase fold. The active site is located at the deep pocket between two central beta-sheets and contains residues strictly conserved within PurOs. Comparisons of the two types of IMP cyclohydrolase, PurO and PurH, revealed that there are no similarities in sequence, structure, or the active site architecture, suggesting that they are evolutionarily not related to each other. The MjR31K mutant of PurO from Methanocaldococcus jannaschii showed 76% decreased activity and the MjE102Q mutation completely abolished enzymatic activity, suggesting that these highly conserved residues play critical roles in catalysis. Interestingly, green fluorescent protein (GFP), which has no structural homology to either PurO or PurH but catalyzes a similar intramolecular cyclohydrolase reaction required for chromophore maturation, utilizes Arg96 and Glu222 in a mechanism analogous to that of PurO.


[Development of new peptide-based chelating agents for site-specific radiolabeling with 64Cu].
Radiolabeled monoclonal antibodies (mAbs) are very useful molecular probes for nuclear imaging technique due to their high-target specificity and high-stability in the bloodstream. MAbs are generally labeled by indirect method using the combination of relatively long-lived metallic radionuclides (including (64)Cu, (89)Zr, and (111)In) and bifunctional chelating agents which are capable of binding both antibodies and radio metals. The indirect radiolabeling method has some advantages such as high labeling efficiency and long-term retention ability within their target cells. However, this conventional labeling method can potentially lead to low-target affinity of the mAb probes, because of the non-site-specific introduction of the bifunctional chelators into the active site of the mAbs. To overcome the shortcoming, we proposed a new direct labeling method utilizing fusion proteins comprising mAbs linked to metal binding peptides at the N- or C-terminus. In this study, we synthesized new peptide derivatives possessing an N-terminal tripeptide sequence (Xaa-Yaa-His) called the amino terminal Cu(2+)- and Ni(2+)-binding (ATCUN) motif as (64)Cu binding peptides for the proposed labeling method. Moreover, we studied the stability constants of Cu(2+)-ATCUN peptide complexes by pH titration. From these studies, we found that a low basicity of the N-terminal amine in the peptide resulted in a high stability constant of the complex. This finding may provide valuable guidelines in designing the ATCUN peptide with high-binding affinity toward (64)Cu.


A large eddy simulation study to assess low-speed wind and baffle orientation effects in a water treatment sedimentation basin.
Hydraulic performance of clarifiers in water and wastewater treatment plants significantly affects the settling efficiency of suspended particles. Structural and ambient parameters can deteriorate this performance. Through a verified three dimensional numerical study, we evaluated hydraulic performance and settling efficiency in a rectangular clarifier with a nominal hydraulic retention time (HRT) of 1 h and options for structural baffles with angles of 20°, 30°, 45° and 70°. Large eddy simulation and Lagrangian particle tracing were used to trace particles 80 to 850 μm in diameter. A passive scalar tracer study was conducted to reveal discrepancies in nominal and real HRT. By posing a 5 m/s wind, ten different scenarios were simulated. The wind caused 17% and 6% reduction in HRT and settling efficiency, respectively. Baffles improved these indicators with the 45° baffle showing the best performance with an approximate settling efficiency of 93%. The study highlighted the importance of using baffles, in particular for small size particles for which influencing factors such as wind deteriorate their settling efficiency.


Contribution of nitric oxide to beta2-adrenoceptor mediated vasodilatation in human forearm arterial vasculature.
beta2-adrenoceptor agonists are generally considered to produce endothelium independent vasodilatation through adenylate cyclase. We determined whether nitric oxide contributes to beta2-adrenoceptor vasodilatation in human arterial vasculature. Forearm blood flow responses to brachial intra-arterial infusions of ritodrine (2.5-50 microg min(-1)), a selective beta2-adrenoceptor agonist, were determined in 24 healthy, normotensive subjects (mean age 22 years, 5F) on two occasions with initial and concomitant administration of L-NMMA (800 microg min(-1)), an NO synthase inhibitor, or noradrenaline (5-30 ng min(-1)), a control constrictor not affecting basal NO activity. Responses to the endothelium dependent vasodilator scrotonin (n = 6) and an endothelium independent vasodilator GTN (n = 9) were also determined. Maximal dilatation to ritodrine during L-NMMA infusion (310+/-32%; mean+/-s.e.mean) was reduced compared to that during noradrenaline infusion (417+/-41%, P<0.05), as were summary responses (1023+/-101 vs 1415+/-130; P<0.05). Responses to GTN were unaffected by L-NMMA compared to noradrenaline; max 177+/-26 vs 169+/-20%, 95% CI for difference -33,48; P=0.68; summary response 361+/-51 vs 396+/-37, 95% CI -142,71; P=0.46. Dilator responses to serotonin were reduced by L-NMMA; max 64+/-20 vs 163+/-26%, P<0.01; summary response 129+/-36 vs 293+/-60; P<0.05) and to a greater extent than ritodrine (58+/-7 vs 25+/-14%, P<0.05). beta2-adrenoceptor mediated vasodilatation in the human forearm has an NO mediated component. The underlying mechanism for this effect is unclear, but flow mediated vasodilatation is unlikely to be responsible.


Bioavailability of sediment-associated Cu and Zn to Daphnia magna.
Exposures to mining-impacted, field-collected sediment (Clear Creek, CO, USA) contaminated with Cu (2.4 mg/g) and Zn (5.2 mg/g) were acutely toxic to juvenile Daphnia magna. Dissolved Cu and Zn in the overlying water (sediment+reference water) were at levels that could cause acute toxicity. To reduce dissolved metals below toxic levels, the sediment was repeatedly rinsed to remove any easily mobilized metals. Washing the sediment reduced dissolved Cu by 60% and Zn by 80%. D. magna exposed to washed sediment experienced higher survival (95%) compared to those exposed to the original sediment (<50%). Cu and Zn that remained associated with suspended sediment after washing were not bioavailable, since survival and tissue metal concentrations in D. magna exposed to both filtered (>0.45 microm) and unfiltered overlying water were statistically similar. Multiple regression analysis indicated that only dissolved Cu significantly contributed to mortality of D. magna whereas particulate Cu, particulate Zn, and dissolved Zn did not. Regression analysis on a combined dataset from all Clear Creek exposures (washed and unwashed), revealed a significant (p < 0.0001, r(2) = 0.76) relationship between the concentration of dissolved copper in the overlying water and the mortality of exposed Daphnia, yielding an estimated LC50 of 26 microg/L dissolved copper (hardness approximately 140 mg/L). The results of this study indicate that if the sediment of Clear Creek was subjected to a resuspension event that there would be a significant efflux of metals from the sediment into the water column, resulting in potentially toxic levels in the water column.


Does cataract surgery alleviate poverty? Evidence from a multi-centre intervention study conducted in Kenya, the Philippines and Bangladesh.
Poverty and blindness are believed to be intimately linked, but empirical data supporting this purported relationship are sparse. The objective of this study is to assess whether there is a reduction in poverty after cataract surgery among visually impaired cases. A multi-centre intervention study was conducted in three countries (Kenya, Philippines, Bangladesh). Poverty data (household per capita expenditure--PCE, asset ownership and self-rated wealth) were collected from cases aged ≥50 years who were visually impaired due to cataract (visual acuity<6/24 in the better eye) and age-sex matched controls with normal vision. Cases were offered free/subsidised cataract surgery. Approximately one year later participants were re-interviewed about poverty. 466 cases and 436 controls were examined at both baseline and follow-up (Follow up rate: 78% for cases, 81% for controls), of which 263 cases had undergone cataract surgery ("operated cases"). At baseline, operated cases were poorer compared to controls in terms of PCE (Kenya: $22 versus £35 p = 0.02, Bangladesh: $16 vs $24 p = 0.004, Philippines: $24 vs 32 p = 0.0007), assets and self-rated wealth. By follow-up PCE had increased significantly among operated cases in each of the three settings to the level of controls (Kenya: $30 versus £36 p = 0.49, Bangladesh: $23 vs $23 p = 0.20, Philippines: $45 vs $36 p = 0.68). There were smaller increases in self-rated wealth and no changes in assets. Changes in PCE were apparent in different socio-demographic and ocular groups. The largest PCE increases were apparent among the cases that were poorest at baseline. This study showed that cataract surgery can contribute to poverty alleviation, particularly among the most vulnerable members of society. This study highlights the need for increased provision of cataract surgery to poor people and shows that a focus on blindness may help to alleviate poverty and achieve the Millennium Development Goals.


Psychosocial Predictors and Outcomes of Delayed Breast Reconstruction in Mastectomized Women in Mainland China: An Observational Study.
The aim of the present study was to evaluate potential psychosocial factors that impact Chinese female breast cancer patients to select breast reconstruction (BR), and potential connection of psychosocial outcomes with their satisfaction with BR. A total of 264 female breast cancer patients with mastectomy were recruited from 2012 to 2014. All patients were informed with BR options at their first visit. Personal and medical profiles were collected. Body image, self-esteem, depression and anxiety were assessed using validated scales. Patients who were selected to undergo BR after the first visit were followed up for six months. The same assessment was performed at 6 months post BR, and their satisfaction with BR was evaluated using the Alderman scale. Multivariate linear and logistic regressions were performed. Forty-seven percent of the patients (126/264) opted to undergo BR within six months after the initial visit. Multivariate logistic regression analysis revealed that self-esteem (P < 0.05), body image (P < 0.01), education level (P < 0.05), and their husband's recommendation (P < 0.05) were highly related to the patients' decision to undergo BR. In addition, multivariate linear regression analysis showed that patient satisfaction with BR was significantly associated with preoperative body image (P < 0.01), postoperative improvement in self-esteem (P < 0.01), improvement in body image (P < 0.01), reduction in depression (P < 0.05), pain (P < 0.05), and scarring (P < 0.01). The psychosocial factors including self-esteem and body image are highly related to selecting the BR option and post-BR satisfaction in Chinese female breast cancer patients.


Myricetin Ameliorates Ethanol-Induced Lipid Accumulation in Liver Cells by Reducing Fatty Acid Biosynthesis.
Alcoholic liver disease is a serious threat to human health. The development of drug candidates from complementary and alternative medicines is an attractive approach. Myricetin can be found in fruit, vegetables, and herbs. This study investigates the protective effect of myricetin on ethanol-induced injury in mouse liver cells. Oil-red O staining, assays of oxidative stress and measurements of inflammatory markers in mouse AML12 liver cells collectively demonstrate that myricetin elicits a curative effect on ethanol-induced injury. Next, the role of myricetin in the metabolic regulation of ethanol pathology in liver cells is assessed by gas chromatography coupled with mass spectrometry. Myricetin inhibits ethanol-stimulated fatty acid biosynthesis. Additionally, dodecanoic acid may be proposed as a potential biomarker related to ethanol pathology or myricetin therapy. It is also observed that myricetin enhances ethanol-induced inhibition of the mitochondrial electron transport chain. Moreover, fumaric acid is found to be a candidate biomarker related to ethanol toxicity or myricetin therapy. Quantitative reverse-transcription-PCR shows that ethanol-induced fatty acid synthase and sterol regulatory element-binding protein-1c mRNA levels are alleviated by myricetin. Finally, myricetin increases ethanol-induced inhibition of phosphorylation of AMP-activated protein kinase. These results elucidate the pharmacological mechanism of myricetin on ethanol-induced lipid accumulation.


Extracellular proteins in pea root tip and border cell exudates.
Newly generated plant tissue is inherently sensitive to infection. Yet, when pea (Pisum sativum) roots are inoculated with the pea pathogen, Nectria haematococca, most newly generated root tips remain uninfected even though most roots develop lesions just behind the tip in the region of elongation. The resistance mechanism is unknown but is correlated spatially with the presence of border cells on the cap periphery. Previously, an array of >100 extracellular proteins was found to be released while border cell separation proceeds. Here we report that protein secretion from pea root caps is induced in correlation with border cell separation. When this root cap secretome was proteolytically degraded during inoculation of pea roots with N. haematococca, the percentage of infected root tips increased from 4% +/- 3% to 100%. In control experiments, protease treatment of conidia or roots had no effect on growth and development of the fungus or the plant. A complex of >100 extracellular proteins was confirmed, by multidimensional protein identification technology, to comprise the root cap secretome. In addition to defense-related and signaling enzymes known to be present in the plant apoplast were ribosomal proteins, 14-3-3 proteins, and others typically associated with intracellular localization but recently shown to be extracellular components of microbial biofilms. We conclude that the root cap, long known to release a high molecular weight polysaccharide mucilage and thousands of living cells into the incipient rhizosphere, also secretes a complex mixture of proteins that appear to function in protection of the root tip from infection.


Factors associated with skills of health visitors in maternal-infant mental health in Japan.
This study is a formative evaluation of a training seminar for health visitors, who visit mothers to provide them with support in terms of postpartum mental health, and was performed to examine factors that relate to the skills of these health visitors. Subjects were all health visitors (n = 232) from around Japan who participated in a 2-day training seminar. One-hundred and thirty-three valid responses (57.3%) were received and written consent to participate in the research was obtained. Results of statistical analyses indicated that a health visitor's skill at supporting a mother in terms of postpartum mental health had two domains, such as interpersonal health care skills and skill at formulating measures. In addition to the length of experience (p < 0.001), the level of expertise (p < 0.001) and the total score on the Generalized Self-Efficacy Scale (p < 0.1 for interpersonal health care skills) was related to a higher level of the health visitor's skill at supporting mothers in terms of their postpartum mental health. In contrast, having a university degree (p < 0.1) was related to a lower level of the health visitor's interpersonal health care skills. Therefore, a training seminar aimed at promoting the skills of health visitors must provide them with the latest expertise and encourage their self-efficacy by helping them successfully envision supporting mothers in terms of their postpartum mental health. In addition, careful instruction of health visitors with less experience and a university degree is crucial.


Tachykinin neurokinin 3 receptor antagonists: a new treatment for cardiovascular disease?
Great progress has been made in reducing cardiovascular mortality over the past 50 years. Nevertheless, prevalence is rising in some settings and remains higher in men than in women, even with the same level of established risk factors. To gain new insights, researchers are now considering cardiovascular disease in relation to the well known evolutionary biology model of growth and reproduction trading off against longevity, with trials of calorie restriction underway. However, calorie restriction has not been as successful as expected in primates and it is increasingly realised that effects on the reproductive axis might also be important. In this paper, the modulation of the reproductive axis using existing agents that have such properties-tachykinin neurokinin 3 receptor antagonists-is proposed as a way of reducing cardiovascular disease and combating a leading cause of global morbidity and mortality.


[Abdominal injuries in polytraumatized adults : Systematic review].
Abdominal injuries are potentially life-threatening and occur in 20-25% of all polytraumatized patients. Blunt trauma is the main mechanism. The liver and spleen are most commonly injured and much less often the intestines. The clinical evaluation proves equivocal in many cases; therefore, the gold standard is computed tomography (CT), which has been increasingly used even in hemodynamically weakly stable or sometimes even unstable patients because it promptly provides precise diagnostic findings, which present the basis for successful therapy. Hemodynamically unstable patients always need an exploratory laparotomy (EL). An EL should also be carried out with a positive focused assessment with sonography for trauma (FAST) or CT for severe parenchymal lesions, hollow organ lesions, intraperitoneal bladder lesions, peritonitis and organ evisceration, impalement injuries and lesions of the abdominal fascia. Hemodynamically stable patients without signs of peritonitis and a lack of such findings can often be treated conservatively irrespective of the extent of an injury. Angiography (and if needed embolization) can additionally be diagnostically and therapeutically utilized.


Comparative effectiveness of treatment strategies for severe splenic trauma in the pediatric population.
Splenic angioembolization (SAE) is increasingly used in the management of splenic injuries in adults, although its value in pediatric trauma is unclear. We sought to assess outcomes related to splenectomy vs SAE. The National Trauma Data Bank was queried for patients 0 to 15 years of age from 2007 to 2011. Subgroup analysis of splenectomy vs SAE was performed for high-grade injuries using propensity analysis and inverse probability weighting. Of 11,694 children presenting with splenic trauma, over 90% were treated nonoperatively. Adjusted analysis of high-grade injuries included 265 children who underwent splenectomy and 199 who underwent SAE. The Injury Severity Score, number of transfusions, and complications rates were not significantly different between the 2 groups. Overall adjusted mortality for children with high-grade injuries was 13.4% following splenectomy and 10.0% following SAE (P = .31) CONCLUSION: Patients undergoing SAE for high-grade splenic trauma have comparable morbidity and mortality with splenectomy.


Family resources moderate the relationship between seizure control and health-related quality of life in children with drug-resistant epilepsy.
Pediatric drug-resistant epilepsy (DRE) is associated with poor health-related quality of life (HRQOL). Achieving seizure control, however, does not improve HRQOL in all children. This study sought to evaluate whether (1) baseline caregiver and family factors are associated with child HRQOL at 1-year follow-up over and above epilepsy characteristics, treatment, and seizure outcome; and (2) baseline family factors moderate the association between seizure outcome and child HRQOL at 1-year follow-up. This multicenter longitudinal cohort study recruited 152 children with DRE who were being evaluated for surgical candidacy. Child HRQOL was rated by caregivers using the Quality of Life in Childhood Epilepsy Questionnaire at baseline and 1-year follow-up. Additional data collected at baseline included child epilepsy characteristics, caregiver demographics, caregiver mood, and family environment. Seizure freedom was achieved in 68% and 28% of patients 1 year after surgery and medical treatment, respectively. Caregiver and family factors were not associated with higher child HRQOL at follow-up after accounting for epilepsy characteristics, treatment, seizure outcome, and baseline child HRQOL, F11, 131 = 1.34, P = .21, ΔR2 = .04. Family resources moderated the association between seizure outcome and child HRQOL at follow-up (b = .41, 95% confidence interval [CI] = 0.09-0.74, P = .03); seizure freedom was strongly associated with higher HRQOL when family resources were high (b = 13.50, 95% CI = 8.35-18.54, P < .001), relative to when family resources were low (b = 5.17, 95% CI = -0.18 to 10.52, P = .06). Family relationships and demands did not moderate the relationship between seizure outcome and HRQOL. Achieving seizure freedom was associated with better HRQOL in children with DRE, but this association was reduced for those children who presented with limited family resources. These results highlight the importance of assessing the family environment during presurgical evaluation and implementing early family-based intervention and supports to promote better outcomes for children in the long term.


The benzamide MS-275 is a potent, long-lasting brain region-selective inhibitor of histone deacetylases.
The association of the histone deacetylase (HDAC) inhibitor valproate (VPA) with atypical antipsychotics has become a frequent treatment strategy for schizophrenia and bipolar disorder. Because the VPA doses administered are elevated, one cannot assume that the benefits of the VPA plus antipsychotic treatment are exclusively related to the covalent modifications of nucleosomal histone tails. We compared the actions of N-(2-aminophenyl)-4-[N-(pyridin-3-yl-methoxycarbonyl)aminomethyl]benzamide derivative (MS-275), which is a potent HDAC inhibitor in vitro, with the actions of VPA for their ability to (i) increase the acetylated status of brain nucleosomal histone tail domains and (ii) to regulate brain histone-RELN and histone-GAD(67) promoter interactions. MS-275 increases the content of acetylhistone 3 (Ac-H3) in the frontal cortex. Whereas this response peaks after a s.c. injection of 15 micromol/kg, the increase in Ac-H3 content in the hippocampus becomes significant only after an injection of 60 micromol/kg, suggesting that MS-275 is 30- to 100-fold more potent than VPA in increasing Ac-H3 in these brain regions. In contrast to VPA, MS-275, in doses up to 120 micromol/kg, fails to increase Ac-H3 content in the striatum. Chromatin immunoprecipitation shows that MS-275 increases Ac-H3-RELN and Ac-H3-GAD(67) promoter interaction in the frontal cortex. These results suggest that MS-275 is a potent brain region-selective HDAC inhibitor. It is likely that, in addition to MS-275, other benzamide derivatives, such as sulpiride, are brain-region selective inhibitors of HDACs. Hence, some benzamide derivatives may express a greater efficacy than VPA as an adjunctive to antipsychotics in the treatment of epigenetically induced psychiatric disorders.


Unique transcription pattern of Epstein-Barr virus (EBV) in EBV-carrying gastric adenocarcinomas: expression of the transforming BARF1 gene.
Approximately 10% of gastric adenocarcinomas worldwide are associated with human EBV. These carcinomas generally do not express the latent membrane protein 1 (LMP1), the major known EBV oncogene. Recently, another EBV gene [ie., BARF1 (BamHI A rightward open reading frame)] was shown to have transforming and immortalizing capacities. Therefore, in this study, we investigated the expression of BARF1 in EBV-carrying gastric adenocarcinomas in relation to the expression of other latent EBV transcripts. In the present study, 10 of 132 gastric adenocarcinomas tested positive for EBV using EBER1/2-RNA in situ hybridization. We demonstrate BARF1 gene transcription in nine EBV-carrying gastric adenocarcinomas (with sufficient RNA quality) using the BARF1-specific nucleic acid sequence-based amplification assay. In addition, we also detected other latent EBV transcripts (ie., BARF0-, LMP2A-, and Q/K-driven EBNA1 transcripts in these carcinomas using reverse transcription-PCR analysis. No expression of LMP1, EBNA2, and ZEBRA (either at transcription or protein level) was found. In addition, two cases were positive for BHRF1 transcripts, the viral bcl-2 homologue. Thus, together with BARF1 transcription, a unique and distinct EBV latency type has been found in EBV-associated gastric adenocarcinomas. Because BARF1 exerts immortalizing effects on human epithelial cells in vitro and EBV-carrying gastric adenocarcinomas lack the expression of LMP1, the BARF1 gene might act as the viral oncogene in EBV-carrying gastric carcinomas. The BARF1 gene offers an alternative way for EBV-mediated oncogenesis other than LMP1.


Training negative connectivity patterns between the dorsolateral prefrontal cortex and amygdala through fMRI-based neurofeedback to target adolescent socially-avoidant behaviour.
Social anxiety is prevalent in adolescence. Given its role in maintaining fears, reducing social avoidance through cognitive reappraisal may help attenuate social anxiety. We used fMRI-based neurofeedback (NF) to increase 'adaptive' patterns of negative connectivity between the dorsolateral prefrontal cortex (DLPFC) and the amygdala to change reappraisal ability, and alter social avoidance and approach behaviours in adolescents. Twenty-seven female participants aged 13-17 years with varying social anxiety levels completed a fMRI-based NF training task where they practiced cognitive reappraisal strategies, whilst receiving real-time feedback of DLPFC-amygdala connectivity. All participants completed measures of cognitive reappraisal and social approach-avoidance behaviour before and after NF training. Avoidance of happy faces was associated with greater social anxiety pre-training. Participants who were unable to acquire a more negative pattern of connectivity through NF training displayed significantly greater avoidance of happy faces at post-training compared to pre-training. These 'maladaptive' participants also reported significant decreases in re-appraisal ability from pre to post-training. In contrast, those who were able to acquire a more 'adaptive' connectivity pattern did not show these changes in social avoidance and re-appraisal. Future research could consider using strategies to improve the capacity of NF training to boost youth social-approach behaviour.


Frictional characteristics of atomically thin sheets.
Using friction force microscopy, we compared the nanoscale frictional characteristics of atomically thin sheets of graphene, molybdenum disulfide (MoS2), niobium diselenide, and hexagonal boron nitride exfoliated onto a weakly adherent substrate (silicon oxide) to those of their bulk counterparts. Measurements down to single atomic sheets revealed that friction monotonically increased as the number of layers decreased for all four materials. Suspended graphene membranes showed the same trend, but binding the graphene strongly to a mica surface suppressed the trend. Tip-sample adhesion forces were indistinguishable for all thicknesses and substrate arrangements. Both graphene and MoS2 exhibited atomic lattice stick-slip friction, with the thinnest sheets possessing a sliding-length-dependent increase in static friction. These observations, coupled with finite element modeling, suggest that the trend arises from the thinner sheets' increased susceptibility to out-of-plane elastic deformation. The generality of the results indicates that this may be a universal characteristic of nanoscale friction for atomically thin materials weakly bound to substrates.


First Reported Case of Primary Intrahepatic Cholangiocarcinoma with Pure Squamous Cell Histology: A Case Report.
In the United States, approximately 2500 cases of cholangiocarcinoma occur each year. The average incidence is 1 case/100 000 persons each year. Surgical resection is the mainstay for the treatment of cholangiocarcinoma. The result of surgery depends on location of the tumor, extent of tumor penetration of the bile duct, tumor-free resection margins, and lymph node and distant metastases. There has been an increase in the incidence of intrahepatic cholangiocarcinoma (IHCC) globally over a period of 30 years from 0.32/100 000 to 0.85/100 000 persons each year. Epidemiologically, the incidence of IHCC has been increasing in the U.S. from year 1973 to 2010. We are reporting a first case of primary intrahepatic cholangiocarcinoma of pure squamous cell histology. A 64-year-old man presented with right upper-quadrant pain, jaundice, and weight loss. Imaging studies revealed a large hepatobiliary mass, intrahepatic bile duct dilation, normal common duct, and absence of choledocholithiasis. Delayed-contrast magnetic resonance imaging of the abdomen showed peripheral enhancement of the central lesion, which is typical of cholangiocarcinoma in contrast to hepatocellular carcinoma or metastasis. Cancer antigen 19-9 was markedly elevated. Liver function tests were deranged. Endoscopic retrograde cholangiopancreatography showed high degree of left hepatic duct stricture. Brush cytopathology was positive for atypia. The patient underwent exploratory laparotomy for en-bloc resection of the hepatobiliary mass with colon resection, liver resection, and cholecystectomy. Histology revealed keratinizing squamous cell carcinoma. Based on these findings, a definitive diagnosis of well-differentiated squamous cell carcinoma of the intrahepatic bile duct was made. Squamous cell carcinoma of the biliary tree is very rare and the majority of tumors are adenocarcinomas. Cholangiocarcinomas containing a squamous cell component have a poor prognosis due to its aggressive behavior. However, prognosis of cholangiocarcinoma with pure SCC histology is unknown because this is the first case in the literature.


Laparoscopic gastric bypass with remnant gastrectomy in a super-super obese patient with gastric metaplasia: a surgical hazard?
The endoscopic inaccessibility of the gastric remnant after Roux-en-Y gastric bypass (RYGBP) for morbid obesity represents an important issue for patients with familiar history of gastric cancer (GC) or affected by premalignant lesions, such as intestinal metaplasia. If a different bariatric procedure is contraindicated, RYGBP with remnant gastrectomy represents a reasonable alternative, significantly reducing the risk of GC but potentially increasing postoperative morbidity. For this reason, only few cases have been reported in the recent Literature and none regarding a super-super obese patient. We present the case of a 55-year-old super-super obese man with a family history of GC and antral gastritis with extensive intestinal metaplasia at preoperative upper endoscopy, who underwent laparoscopic RYGBP with remnant gastrectomy.


Genome-Wide Binding of Posterior HOXA/D Transcription Factors Reveals Subgrouping and Association with CTCF.
Homeotic genes code for key transcription factors (HOX-TFs) that pattern the animal body plan. During embryonic development, Hox genes are expressed in overlapping patterns and function in a partially redundant manner. In vitro biochemical screens probing the HOX-TF sequence specificity revealed largely overlapping sequence preferences, indicating that co-factors might modulate the biological function of HOX-TFs. However, due to their overlapping expression pattern, high protein homology, and insufficiently specific antibodies, little is known about their genome-wide binding preferences. In order to overcome this problem, we virally expressed tagged versions of limb-expressed posterior HOX genes (HOXA9-13, and HOXD9-13) in primary chicken mesenchymal limb progenitor cells (micromass). We determined the effect of each HOX-TF on cellular differentiation (chondrogenesis) and gene expression and found that groups of HOX-TFs induce distinct regulatory programs. We used ChIP-seq to determine their individual genome-wide binding profiles and identified between 12,721 and 28,572 binding sites for each of the nine HOX-TFs. Principal Component Analysis (PCA) of binding profiles revealed that the HOX-TFs are clustered in two subgroups (Group 1: HOXA/D9, HOXA/D10, HOXD12, and HOXA13 and Group 2: HOXA/D11 and HOXD13), which are characterized by differences in their sequence specificity and by the presence of cofactor motifs. Specifically, we identified CTCF binding sites in Group 1, indicating that this subgroup of HOX-proteins cooperates with CTCF. We confirmed this interaction by an independent biological assay (Proximity Ligation Assay) and demonstrated that CTCF is a novel HOX cofactor that specifically associates with Group 1 HOX-TFs, pointing towards a possible interplay between HOX-TFs and chromatin architecture.


Psychological outcomes following maternal serum screening: a cohort study.
Maternal serum screening is used to assist in the prenatal detection of congenital anomalies. Its use is controversial, and one concern that has been expressed is its psychological effects on women. The authors examined whether this test leads to an increase in anxiety and depression among women who have a false-positive result as compared with those who have a true-negative result or do not undergo testing. A prospective cohort study with baseline assessment at 15 to 18 weeks' gestation and follow-up at 24 weeks' gestation was conducted. Pregnant women at 8 geographically diverse sites across Ontario were recruited. The main outcome measures were the state portion of the State--Trait Anxiety Inventory and the Center for Epidemiologic Studies Depression Scale. Of the 2418 potential subjects 2020 (83.5%) were enrolled and eligible; 1741 (86.2%) completed the follow-up. A total of 1177 women (67.6%) underwent maternal serum screening. No overall adverse psychological effects as a result of testing were found at 24 weeks' gestation. Women with a false-positive result had a mean increase in anxiety score of 1.6 (95% confidence interval [CI] -1.7 to 4.9), whereas women with a true-negative result had a mean decrease of 1.1 (95% CI -1.8 to -0.3) and those not tested had a mean decrease of 0.4 (95% CI -1.3 to 0.5). The mean depression score increased by 0.5 (95% CI -0.9 to 2.0) in the false-positive group, was unchanged (95% CI -0.3 to 0.4) in the true-negative group and increased by 0.2 (95% CI -1.7 to 1.2) in the not tested group. Of the women who underwent testing, 87 (7.6%) were unsure of their result at the time of follow-up. The results suggest that maternal screening in Ontario is not causing serious psychological harm to women. Communication regarding test results could be improved, since a substantial proportion of women were unsure of their test result.


Discovery of permuted and recently split transfer RNAs in Archaea.
As in eukaryotes, precursor transfer RNAs in Archaea often contain introns that are removed in tRNA maturation. Two unrelated archaeal species display unique pre-tRNA processing complexity in the form of split tRNA genes, in which two to three segments of tRNAs are transcribed from different loci, then trans-spliced to form a mature tRNA. Another rare type of pre-tRNA, found only in eukaryotic algae, is permuted, where the 3' half is encoded upstream of the 5' half, and must be processed to be functional. Using an improved version of the gene-finding program tRNAscan-SE, comparative analyses and experimental verifications, we have now identified four novel trans-spliced tRNA genes, each in a different species of the Desulfurococcales branch of the Archaea: tRNA(Asp(GUC)) in Aeropyrum pernix and Thermosphaera aggregans, and tRNA(Lys(CUU)) in Staphylothermus hellenicus and Staphylothermus marinus. Each of these includes features surprisingly similar to previously studied split tRNAs, yet comparative genomic context analysis and phylogenetic distribution suggest several independent, relatively recent splitting events. Additionally, we identified the first examples of permuted tRNA genes in Archaea: tRNA(iMet(CAU)) and tRNA(Tyr(GUA)) in Thermofilum pendens, which appear to be permuted in the same arrangement seen previously in red alga. Our findings illustrate that split tRNAs are sporadically spread across a major branch of the Archaea, and that permuted tRNAs are a new shared characteristic between archaeal and eukaryotic species. The split tRNA discoveries also provide new clues to their evolutionary history, supporting hypotheses for recent acquisition via viral or other mobile elements.


A novel rat model of foreign body osteomyelitis for evaluation of antimicrobial efficacy.
The most common organism-type causing orthopedic foreign body infection is the staphylococci, of which Staphylococcus aureus and Staphylococcus epidermidis are especially common. These organisms form biofilms on orthopedic foreign body surfaces, rendering such infections challenging and time consuming to treat. Our group evaluates novel therapeutics for orthopedic foreign body infection in animal models. A current limitation of most animal models is that that they only allow for the removal of one sample per animal, at the time of sacrifice. Herein, we describe a novel rat model of foreign body osteomyelitis that allows removal of foreign bodies at different time points, from the same infected animal. We demonstrate that this model can be used for both S. aureus and S. epidermidis orthopedic foreign body infection, with 3.56, 3.60 and 5.51 log10 cfu/cm2 S. aureus recovered at four, five and six weeks, respectively, after infection, and 2.08, 2.17 and 2.62 log10 cfu/cm2 S. epidermidis recovered at four, five and six weeks, respectively, after infection We evaluated the model with S. aureus infection treated with rifampin 25 mg/kg twice daily for 21 days. Using quantitative cultures, we were no longer able to detect bacteria as of the 14th day of treatment with bacteria becoming detectable again 7 days following die discontinuation of rifampin a period. This novel model allows monitoring of evolution of infection at the infection site in the same animal.