A novel filter for accurate estimation of fluid pressure and fluid velocity using poroelastography.
Fluid pressure and fluid velocity carry important information for cancer diagnosis, prognosis and treatment. Recent work has demonstrated that estimation of these parameters is theoretically possible using ultrasound poroelastography. However, accurate estimation of these parameters requires high quality axial and lateral strain estimates from noisy ultrasound radio frequency (RF) data. In this paper, we propose a filtering technique combining two efficient filters for removal of noise from strain images, i.e., Kalman and nonlinear complex diffusion filters (NCDF). Our proposed filter is based on a novel noise model, which takes into consideration both additive and amplitude modulation noise in the estimated strains. Using finite element and ultrasound simulations, we demonstrate that the proposed filtering technique can significantly improve image quality of lateral strain elastograms along with fluid pressure and velocity elastograms. Technical feasibility of the proposed method on an in vivo set of data is also demonstrated. Our results show that the CNRe of the lateral strain, fluid pressure and fluid velocity as estimated using the proposed technique is higher by at least 10.9%, 51.3% and 334.4% when compared to the results obtained using a Kalman filter only, by at least 8.9%, 27.6% and 219.5% when compared to the results obtained using a NCDF only and by at least 152.3%, 1278% and 742% when compared to the results obtained using a median filter only for all SNRs considered in this study.


Baseline Cognitive Impairment in Patients With Asymptomatic Carotid Stenosis in the CREST-2 Trial.
Studies of carotid artery disease have suggested that high-grade stenosis can affect cognition, even without stroke. The presence and degree of cognitive impairment in such patients have not been reported and compared with a demographically matched population-based cohort. We studied cognition in 1000 consecutive CREST-2 (Carotid Revascularization and Medical Management for Asymptomatic Carotid Stenosis Trial) patients, a treatment trial for asymptomatic carotid disease. Cognitive assessment was after randomization but before assigned treatment. The cognitive battery was developed in the general population REGARDS Study (Reasons for Geographic and Racial Differences in Stroke), involving Word List Learning Sum, Word List Recall, and Word List fluency for animal names and the letter F. The carotid stenosis patients were >45 years old with ≥70% asymptomatic carotid stenosis and no history of prevalent stroke. The distribution of cognitive performance for the patients was standardized, accounting for age, race, and education using performance from REGARDS, and after further adjustment for hypertension, diabetes, dyslipidemia, and smoking. Using the Wald Test, we tabulated the proportion of Z scores less than the anticipated deviate for the population-based cohort for representative percentiles. There were 786 baseline assessments. Mean age was 70 years, 58% men, and 52% right-sided stenosis. The overall Z score for patients was significantly below expected for higher percentiles (P<0.0001 for 50th, 75th, and 95th percentiles) and marginally below expected for the 25th percentile (P=0.015). Lower performance was attributed largely to Word List Recall (P<0.0001 for all percentiles) and for Word List Learning (50th, 75th, and 95th percentiles below expected, P≤0.01). The scores for left versus right carotid disease were similar. Baseline cognition of patients with severe carotid stenosis showed below normal cognition compared to the population-based cohort, controlling for demographic and cardiovascular risk factors. This cohort represents the largest group to date to demonstrate that poorer cognition, especially memory, in this disease. Registration: URL: https://www.clinicaltrials.gov; Unique identifier: NCT02089217.


Optimization of fibronectin-assisted retroviral gene transfer into human CD34+ hematopoietic cells.
Efficient retroviral gene transfer into hematopoietic stem and progenitor cells can be achieved by co-localizing retrovirus and target cells on specific adhesion domains of recombinant fibronectin (FN) fragments. In this paper, we further optimize this technology for human CD34+ cells. Investigating the role of cytokine prestimulation in retrovirus-mediated gene transfer on plates coated with the recombinant FN CH-296 revealed that prestimulation of granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood (PB) CD34+ cells was essential to achieve efficient gene transfer into clonogenic cells. The highest gene transfer occurred by prestimulating PB CD34+ cells for 40 hr with a combination of stem cell factor (SCF), G-CSF, and megakaryocyte growth and development factor (MGDF) prior to retroviral infection on CH-296. Surprisingly, a prolonged simultaneous exposure of primary CD34+ PB cells to retrovirus and cytokines in the presence of CH-296 lowered the gene transfer efficiency. Gene transfer into cytokine prestimulated CD34+ bone marrow (BM) cells was not influenced by increasing the coating concentrations of a recombinant FN fragment, CH-296, nor was it adversely influenced by increasing the number of CD34+ target cells, suggesting that the amount of retroviral particles present in the supernatant was not a limiting factor for transduction of CD34+ BM cells on CH-296-coated plates. The polycation Polybrene was not required for efficient transduction of hematopoietic cells in the presence of CH-296. Furthermore, we demonstrated that repeated exposure of CH-296 to retrovirus containing supernatant, called preloading, can be employed to concentrate the amount of retroviral particles bound to CH-296. These findings establish a simple and short clinically applicable transduction protocol that targets up to 68% of BM or G-CSF-mobilized PB CD34+ cells and is capable of genetically modifying up to 17% of CD34+CD38-/dim PB cells.


Chaperonin overproduction and metabolic erosion caused by mutation accumulation in Escherichia coli.
Bacterial cells adapting to a constant environment tend to accumulate mutations in portions of their genome that are not maintained by selection. This process has been observed in bacteria evolving under strong genetic drift, and especially in bacterial endosymbionts of insects. Here, we study this process in hypermutable Escherichia coli populations evolved through 250 single-cell bottlenecks on solid rich medium in a mutation accumulation experiment that emulates the evolution of bacterial endosymbionts. Using phenotype microarrays monitoring metabolic activity in 95 environments distinguished by their carbon sources, we observe how mutation accumulation has decreased the ability of cells to metabolize most carbon sources. We study if the chaperonin GroEL, which is naturally overproduced in bacterial endosymbionts, can ameliorate the process of metabolic erosion, because of its known ability to buffer destabilizing mutations in metabolic enzymes. Our results indicate that GroEL can slow down the negative phenotypic consequences of genome decay in some environments.


Risk factors for dental caries in the five-year-old South Australian population.
This study tested the hypothesis that risk behaviours in disadvantaged groups would explain socio-economic inequality in dental caries prevalence among preschool children. Using a case-control study, children with caries experience (one or more decayed, missing or filled primary tooth surfaces) and with no caries experience were sampled with known probabilities from among five year olds attending the South Australian Dental Service (SADS). Dental caries experience of primary teeth was recorded by SADS clinicians. Social and behavioural information was collected using a questionnaire mailed to parents. Prevalence rates, prevalence ratios (PR) and 95 per cent confidence intervals (95% CI) were computed, taking into account sampling probabilities. Questionnaires were obtained for 64.6 per cent of sampled children (n = 1398) and 40.2 per cent (95% CI = 37.8-42.6) of them had caries experience. Five statistically significant risk factors were identified relating to previous feeding, current oral hygiene and parent's own oral health perceptions. The prevalence of four risk factors was greater in low-income households compared with high-income households (P < or = 0.01). In multivariate analysis, after adjusting for age of tooth cleaning onset, age at which toothpaste was introduced was not significantly associated with caries prevalence. Behavioural risk factors did not explain income-related gradients in caries prevalence but modified the level of risk associated with delayed onset of tooth cleaning. Children who delayed tooth cleaning until the age of 24 months or more and who were from low-income households had a 2.7-fold increase in caries prevalence (95% CI = 2.1-3.4). Caries prevention efforts need to target behaviours in infancy and non-behavioural risk factors among preschoolers in low-income households.


Effects of arginine and lysine addition to casein and soya-bean protein on serum lipids, apolipoproteins, insulin and glucagon in rats.
1. The effect of variation in arginine: lysine on the relative cholesterolaemic effects of dietary soya-bean protein and casein was studied. Male rats received semi-purified diets containing soya-bean protein isolate or casein supplemented respectively with varying amounts of lysine or arginine for 40 d and blood samples were taken after a 5 h fast. 2. Neither the addition of arginine to casein nor lysine to soya-bean protein modified the intrinsic effect of these proteins on serum cholesterol. 3. Serum triglyceride levels tended to rise with increasing amounts of lysine supplementation. The opposite trend was obtained with arginine supplementation. 4. Casein caused an increase in the concentration of serum insulin, but not glucagon. The glucagon level was increased proportionately with increasing amounts of arginine, while the addition of lysine showed no effect. The effects of added amino acids on serum insulin were inconclusive. 5. There was a parallel increase in serum apo E and glucagon in response to arginine supplementation, while lysine supplementation increased serum apo E. 6. Thus, arginine: lysine was more effective in regulating serum triglyceride than serum cholesterol. Insulin was associated with different effects of these proteins on serum lipids.


Are above- and below-ground phenology in sync?
Globally, root production accounts for 33-67% of terrestrial net primary productivity and influences decomposition via root production and turnover, carbon (C) allocation to mycorrhizal fungi and root exudation. As recognized above ground, the timing of phenological events affects terrestrial C balance, yet there is no parallel understanding for below-ground phenology. In this paper we examine the phenology of root production and its relationship to temperature, soil moisture, and above-ground phenology. Synthesizing 87 observations of whole-plant phenology from 40 studies, we found that, on average, root growth occurs 25 ± 8 d after shoot growth but that the offset between the peak in root and shoot growth varies > 200 d across biomes (boreal, temperate, Mediterranean, and subtropical). Root and shoot growth are positively correlated with median monthly temperature and mean monthly precipitation in boreal, temperate, and subtropical biomes. However, a temperature hysteresis in these biomes leads to the hypothesis that internal controls over C allocation to roots are an equally, if not more, important driver of phenology. The specific mechanisms are as yet unclear but they are likely mediated by some combination of photoassimilate supply, hormonal signaling, and growth form.


[Endoscopic evaluation of the status of the stomach in patients after ulcer surgery].
On the basis of the investigation of nearest and long-term clinical results of the operative treatment of acute complications of gastroduodenal ulcers and results of endoscopic examinations of 309 patients a conclusion is made of the expediency of the endoscopic examination in the nearest postoperative period. The group of patients with good results should include patients having no complaints, with normal functions of the operated stomach, without duodeno-jejunogastric reflux and pronounced gastric changes.


[Epidermoid carcinoma of the stomach].
A case of pure squamous cell carcinoma of the stomach in a 63-year-old woman has been reported. The tumor was located in pyloric antrum and histologic examination disclosed a well differentiated squamous cell carcinoma, with many horn pearls. Histogenesis of similar cases of this unusual tumor have been briefly discussed.


Environmental risk management for radiological accidents: integrating risk assessment and decision analysis for remediation at different spatial scales.
The consequences of the Tohuku earthquake and subsequent tsunami in March 2011 caused a loss of power at the Fukushima Daiichi nuclear power plant, in Japan, and led to the release of radioactive materials into the environment. Although the full extent of the contamination is not currently known, the highly complex nature of the environmental contamination (radionuclides in water, soil, and agricultural produce) typical of nuclear accidents requires a detailed geospatial analysis of information with the ability to extrapolate across different scales with applications to risk assessment models and decision making support. This article briefly summarizes the approach used to inform risk-based land management and remediation decision making after the Chernobyl, Soviet Ukraine, accident in 1986.


[Diaphragmatic rupture in abdominal trauma].
The aim of the study is to evaluate the patients with diaphragmatic rupture due to penetrating or blunt abdominal trauma. Thirty-eight patients with diaphragmatic rupture due to penetrating or blunt abdominal trauma were investigated retrospectively. The average age was 41,72 and there were 31 male and seven female patients. The injury forms were penetrating trauma in 22 (58%) and blunt trauma in 16 (42%) cases. Associated abdominal organ injuries were found in 27 (71%) cases. Among 47 diaphragmatic ruptures, 27 (57%) were on the left and 20 (43%) were on the right side. The average diameter of the rupture was 5,45 (1-20) cm. Management of the diaphragmatic rupture and other associated organ injuries were accomplished through laparotomy. Morbidity was developed in 18 cases and mortality in four cases with associated abdominal organ injuries. Diaphragmatic rupture results in high morbidity and mortality due to associated organ injuries.


The Relationship Between Eyebrow and Eyelid Position in Patients With Ptosis, Dermatochalasis and Controls.
The purpose of this study is to explore mechanical and co-innervational factors involved in both voluntary and involuntary brow elevation among people affected by ptosis and dermatochalasis. In this prospective cohort study of normal controls and eyelids with ptosis or dermatochalasis, marginal reflex distance (MRD1) and brow height were measured under the following conditions: neutral position, involuntary mechanical brow elevation, voluntary brow elevation, and maximal eyelid opening. The primary outcome measure was change in MRD1. Secondary outcome measures included brow height and coupling (mm brow height change per mm MRD1 change). Analysis of variance and t tests were performed for intra- and intercondition comparisons, respectively. Mechanical (involuntary) brow elevation significantly raised MRD1 in control eyelids and eyelids with dermatochalasis, but not in eyelids with ptosis. Voluntary brow elevation produced significantly greater brow height than maximal eyelid opening in controls and eyelids with dermatochalasis, but not in eyelids with ptosis. Maximal eyelid opening increased MRD1 greater than voluntary brow elevation significantly in control eyelids, but not in eyelids with dermatochalasis or ptosis. Coupling of the brow and eyelid margin during maximal eyelid opening was significantly greater in eyelids with ptosis relative to controls. In eyelids with ptosis, mechanical brow elevation does not change eyelid position; however, voluntary brow elevation raises eyelid position to a similar position as maximal eyelid opening. These results argue against the contention that the brow is elevated to mechanically lift the eyelid in ptosis and instead suggest that the brow elevation is driven by efforts to raise the eyelid, possibly via co-innervation.


Dynamic contrast-enhanced MR imaging and dual-phase helical CT in the preoperative assessment of suspected pancreatic cancer: a comparative study with receiver operating characteristic analysis.
The purpose of this study was to compare dynamic contrast-enhanced MR imaging and dual-phase helical CT in the preoperative assessment of patients with suspected pancreatic carcinoma. Thirty-three consecutive surgical candidates (20 men, 13 women; 39-81 years old) were included. MR imaging comprised fast spin-echo (TR/TE 4000/91), fat-suppressed T1-weighted spin-echo (500/15), and T1-weighted breath-hold gradient-echo fast low-angle shot (100/4; flip angle, 80 degrees) images before and after the administration of gadopentetate dimeglumine. Helical CT used 5-mm collimation with a pitch of 1:1.5-1.7; images were obtained 20 and 65-70 sec after injection of 150 ml of contrast material. Two pairs of interpreters who were unaware of the results of the other imaging method independently scored each examination for the presence of a lesion and for surgical resectability using a five-point scale. Results were correlated with surgery (n = 25) or consensus review (n = 8). Receiver operating characteristic methodology was used to analyze the results for resectability, and positive predictive values were calculated. Both MR imaging and helical CT revealed 29 of 31 lesions. In determining lesion resectability, the mean areas under the receiver operating characteristic curves were 0.96 and 0.81 (p = .01) and the positive predictive values were 86.5% and 76% (p = .02) for MR imaging and helical CT, respectively. MR imaging and helical CT performed equally well in lesion detection. MR imaging was significantly better in the assessment of resectability of pancreatic tumors.


HSF1 as a mitotic regulator: phosphorylation of HSF1 by Plk1 is essential for mitotic progression.
Previously, heat shock factor 1 (HSF1) had been reported to induce genomic instability and aneuploidy by interaction with Cdc20. Here, we have further examined the functions of HSF1 in the regulation of mitosis. A null mutant or knockdown of HSF1 caused defective mitotic progression. By monitoring chromosomes in living cells, we determined that HSF1 was localized to the centrosome in mitosis and especially to the spindle poles in metaphase. HSF1 was phosphorylated by Plk1 at Ser(216) of the DSGXXS motif during the timing of mitosis and a phospho-defective mutant form of HSF1 inhibited mitotic progression. Phosphorylated HSF1 during spindle pole localization underwent ubiquitin degradation through the SCF(beta-TrCP) pathway. However, binding of HSF1 with Cdc20 stabilized the phosphorylation of HSF1. Moreover, SCF(beta-TrCP)-mediated degradation only occurred when phosphorylated HSF1 was released from Cdc20. HSF1 phosphorylation at Ser(216) occurred in the early mitotic period with simultaneous binding of Cdc20. The interaction of HSF1 with SCF(beta-TrCP) was followed and then the interaction of APC/Cdc20 was subsequently observed. From these findings, it was shown that Plk1 phosphorylates HSF1 in early mitosis and that the binding of phosphorylated HSF1 with Cdc20 and ubiquitin degradation by SCF(beta-TrCP) regulates mitotic progression.


Genotypes and their interaction effects on reproduction and mating-induced immune activation in Drosophila melanogaster.
Mating causes considerable alterations in female physiology and behaviour, and immune gene expression, partly due to proteins transferred from males to females during copulation. The magnitude of these phenotypic changes could be driven by the genotypes of males and females, as well as their interaction. To test this, we carried out a series of genotype-by-genotype (G × G) experiments using Drosophila melanogaster populations from two distant geographical locations. We expected lines to have diverged in male reproductive traits and females to differ in their responses to these traits. We examined female physiological and behavioural post-mating responses to male mating traits, that is behaviour and ejaculate composition, in the short to mid-term (48 hr) following mating. We then explored whether a sexually transferred molecule, sex peptide (SP), is the mechanism behind our observed female post-mating responses. Our results show that the genotypes of both sexes as well as the interaction between male and female genotypes affect mating and post-mating reproductive traits. Immune gene expression of three candidate genes increased in response to mating and was genotype-dependent but did not show a G × G signature. Males showed genotype-dependent SP expression in the 7 days following eclosion, but female genotypes showed no differential sensitivity to the receipt of SP. The two genotypes demonstrated clear divergence in physiological traits in short- to mid-term responses to mating, but the longer-term consequences of these initial dynamics remain to be uncovered.


Construction and Characterization of a Novel Sustained-Release Delivery System for Hydrophobic Pesticides Using Biodegradable Polydopamine-Based Microcapsules.
Microcapsule formulations have been highly desirable and widely developed for effective utilization of pesticides and environmental pollution reduction. However, commercial and traditional microcapsule formulations of λ-cyhalothrin (LC) were prepared by complicated synthesis procedures and thereby specific organic solvents were needed. In this work, LC was encapsulated into versatile, robust, and biodegradable polydopamine (PDA) microcapsules by self-polymerization of dopamine. LC-loaded PDA microcapsules were characterized by transmission electron microscopy (TEM), small-angle X-ray scattering (SAXS), and thermogravimetric analysis measurements (TGA). LC-loaded PDA microcapsules have uniform morphology with nanoscale, decent LC loading content (>50.0% w/w), and good physicochemical stability and sustained release properties. The bioassay against housefly ( Musca domestica) showed that the bioactivity and long-term efficiency of LC-loaded PDA microcapsules was superior to that of the commercial formulation. All of these results demonstrated that LC-loaded PDA microcapsules could be applied as a commercial LC microcapsule formulation with fewer environmental side effects and higher effective delivery.


Conjugation of the mycotoxins alternariol and alternariol monomethyl ether in tobacco suspension cells.
The mycotoxins alternariol (AOH) and alternariol-9-O-methyl ether (AME) carry three and two phenolic hydroxyl groups, respectively, which makes them candidates for the formation of conjugated metabolites in plants. Such conjugates may escape routine methods of analysis and have therefore been termed masked or, more recently, modified mycotoxins. We report now that AOH and AME are extensively conjugated in suspension cultures of tobacco BY-2 cells. Five conjugates of AOH were identified by MS and NMR spectroscopy as β-D-glucopyranosides (attached in AOH 3- or 9-position) as well as their 6'-malonyl derivatives, and as a gentiobiose conjugate. For AME, conjugation resulted in the d-glucopyranoside (mostly attached in the AME 3-position) and its 6'- and 4'-malonyl derivatives. Pronounced differences were noted for the quantitative pattern of AOH and AME conjugates as well as for their phytotoxicity. Our in vitro study demonstrates for the first time that masked mycotoxins of AOH and AME can be formed in plant cells.


New glycoprotein-associated amino acid transporters.
The L-type amino acid transporter LAT1 has recently been identified as being a disulfide-linked "light chain" of the ubiquitously expressed glycoprotein 4F2hc/CD98. Several LAT1-related transporters have been identified, which share the same putative 12-transmembrane segment topology and also associate with the single transmembrane domain 4F2hc protein. They display differing amino acid substrate specificities, transport kinetics and localizations such as, for instance, y(+)LAT1 which is localized at the basolateral membrane of transporting epithelia, and the defect of which causes lysinuric protein intolerance. The b(0,+)AT transporter which associates with the 4F2hc-related rBAT protein to form the luminal high-affinity diamino acid transporter defective in cystinuria, belongs to the same family of glycoprotein-associated amino acid transporters (gpaATs). These glycoprotein-associated transporters function as amino acid exchangers. They extend the specificity range of vectorial amino acid transport when located in the same membrane as carriers that unidirectionally transport one of the exchanged substrates. gpaATs belong to a phylogenetic cluster within the amino acid/polyamine/choline (APC) superfamily of transporters. This cluster, which we designate the LAT family (named after its first vertebrate member), includes some members from nematodes, yeast and bacteria. The latter of these proteins presumably lack association with a second subunit. In this review, we focus on the animal members of the LAT cluster that form, together with some of the nematode members, the family of glycoprotein-associated amino acid transporters (gpaAT family).


Galleria mellonella as an in vivo model for assessing the protective activity of probiotics against gastrointestinal bacterial pathogens.
The antagonistic activity against gastrointestinal bacterial pathogens is an important property of probiotic bacteria and a desirable feature for pre-selection of novel strains with probiotic potential. Pre-screening of candidate probiotics for antibacterial activity should be based on in vitro and in vivo tests. This study investigated whether the protective activity of probiotic bacteria against gastrointestinal bacterial pathogens can be evaluated using Galleria mellonella larvae as an in vivo model. Larvae were pre-inoculated with either of two widely used probiotic bacteria, Lactobacillus rhamnosus GG or Clostridium butyricum Miyairi 588, and then challenged with Salmonella enterica Typhimurium, enteropathogenic Escherichia coli or Listeria monocytogenes. Survival rates increased in the probiotic pretreated larvae compared with control larvae inoculated with pathogens only. The hemocyte density increased as well in the probiotic pretreated larvae, indicating that both probiotics induce an immune response in the larvae. The antibacterial activity of probiotics against the pathogens was also assayed by an in vitro agar spot test: results were partially consistent with those obtained by the G. mellonella protection assay. The results obtained, as a whole, suggest that G. mellonella larvae are a potentially useful in vivo model that can complement in vitro assays for pre-screening of candidate probiotics.


Erythrocyte receptors for (Plasmodium knowlesi) malaria: Duffy blood group determinants.
Duffy blood group negative human erythrocytes (FyFy) are resistant to infection by Plasmodium knowlesi, a simian malaria that infects Duffy positive human erythrocytes. The P. knowlesi resistance factor, Duffy negative erythrocytes, occurs in high frequency in West Africa, where the people are resistant to vivax malaria. This suggests that Duffy blood group determinants (Fya or Fyb) may be erythrocyte receptors for P. vivax.


Serologic Monitoring of Public Health Interventions against Strongyloides stercoralis.
Northwestern Argentina is endemic for soil-transmitted helminths, and annual deworming programs are carried out in prioritized areas. High prevalence of Strongyloides stercoralis was reported in this area; therefore, control programs including ivermectin are being evaluated. The NIE-enzyme linked immunosorbent assay (ELISA) was used for this purpose. In this community trial, two groups of patients, classified according to housing and living conditions were evaluated. Simultaneous with baseline survey, Group 1 was moved to new households with access to improved water and sanitation facilities (W and S), where deworming (MDA, massive drug administration) took place within 1 month; whereas Group 2 received MDA but remained living with unimproved W and S. The mean time interval between baseline and the follow-up was 331 days for Group 1 and 508 for Group 2. Anti-NIE levels were measured for each individual before and after interventions and follow-up optical density (OD) ratios were calculated to quantify the variation. A significant decrease of the anti-NIE levels between baseline and follow-up was observed in both groups. Nonetheless, the number of patients that achieved the cure criteria (OD ratio < 0.6) was higher in Group 1 than Group 2 with values of 72.7% (24/33) and 45.0% (18/40), respectively (P = 0.0197). Our results support the conclusion that a combined intervention including deworming and improvements in life conditions is more effective, in terms of the proportion of subjects cured than deworming alone. Furthermore, we found that NIE-ELISA is a useful test for assessing the response to treatment and to evaluate the outcome of control intervention programs.


Tweener Fractures in Children and Adolescents.
Tweener fractures are defined as long bone or metadiaphyseal fractures in a pediatric population for which multiple treatment options may be used. It is important to focus specifically on patients nearing skeletal maturity who present with length-unstable femoral shaft fractures, both-bone forearm fractures, distal radius metadiaphyseal fractures, and adolescent tibial shaft fractures. Although there is no gold standard of treatment for any of these fracture patterns, it is important to discuss the risks and benefits, proper application, and important technical aspects of each treatment method to allow surgeons to make an informed decision and optimize surgical outcomes in this patient population.


DNA Nanogels To Snare Carcinogens: A Bioinspired Generic Approach with High Efficiency.
Polycyclic aromatic hydrocarbons (PAHs) are combustion-related pollutants and are ubiquitous in the environment, including in sources of drinking water. Upon contact with DNA, stable PAH-DNA adducts form rapidly as the first step towards their toxic effects. In this work, we prepared hydrophilic DNA nanogels to exploit this generic complexation process as a biomimetic scavenging method. This approach relies on interaction between PAHs and the complete network that constitutes the water-swollen nanogels, and is not restricted to interfacial adsorption. Up to 720 μg of PAH per gram of DNA nanogel are taken up, meaning that 1 mg of DNA nanogel is sufficient to purify a liter of water containing the critical PAH concentration for cancer risk (600 ng L(-1) ). As a result of short diffusion pathways, PAH uptake is rapid, reaching 50 % loading after 15 minutes. Beyond PAHs, DNA nanogels may be useful for the generic detoxification of water containing genotoxins, since most known molecules that strongly associate with DNA are mutagenic.


Late life cognitive control deficits are accentuated by white matter disease burden.
Recent evidence suggests that age-related impairments in cognition may be mediated by a specific deficit in the ability to maintain goal-relevant information, a critical component of cognitive control dependent on the dorsolateral prefrontal cortex, although the underlying neural mechanism of these deficits remains unclear. To examine white matter hyperintensities as a neurobiological mechanism of these impairments, older individuals with severe white matter hyperintensity burden, older individuals with low white matter hyperintensity burden, and young adults were assessed in an event-related functional imaging scan while performing the 'AX'-continuous performance task. Individuals with severe white matter hyperintensity burden showed a significant reduction in dorsolateral prefrontal cortex activity during the high cognitive control cue condition relative to the low white matter hyperintensity group and young individuals. Conversely, those with severe white matter hyperintensity burden showed greater activity in rostral anterior cingulate cortex compared to young individuals. These results are consistent with impaired cognitive control and a possible failure to deactivate default-mode regions in these subjects. Additionally, those with severe white matter hyperintensity burden showed reduced functional connectivity between dorsolateral prefrontal cortex and task-relevant brain regions including middle frontal gyrus, and supramarginal gyrus relative to young subjects and those with minimal white matter hyperintensity burden. These results suggest that age-related goal maintenance impairments and associated dorsolateral prefrontal cortex dysfunction may partly reflect incipient white matter disease of interconnected cognitive networks.


Hyaluronidase 2 negatively regulates RON receptor tyrosine kinase and mediates transformation of epithelial cells by jaagsiekte sheep retrovirus.
The candidate tumor-suppressor gene hyaluronidase 2 (HYAL2) encodes a glycosylphosphatidylinositol-anchored cell-surface protein that serves as an entry receptor for jaagsiekte sheep retrovirus, a virus that causes contagious lung cancer in sheep that is morphologically similar to human bronchioloalveolar carcinoma. The viral envelope (Env) protein alone can transform cultured cells, and we hypothesized that Env could bind and sequester the HYAL2 receptor and thus liberate a potential oncogenic factor bound and negatively controlled by HYAL2. Here we show that the HYAL2 receptor protein is associated with the RON receptor tyrosine kinase (also called MST1R or Stk in the mouse), rendering it functionally silent. In human cells expressing a jaagsiekte sheep retrovirus Env transgene, the Env protein physically associates with HYAL2. RON liberated from the association with HYAL2 becomes functionally active and consequently activates the Akt and mitogen-activated protein kinase pathways leading to oncogenic transformation of immortalized human bronchial epithelial cells. We find activated RON in a subset of human bronchioloalveolar carcinoma tumors, suggesting RON involvement in this type of human lung cancer.


Interaction between ouabain and the phosphorylated intermediate of Na,K-ATPase.
After phosphorylation of electric eel Na,K-ATPase by Na+, Mg2+, and ATP was terminated by removing the unbound Mg2+, the phosphoenzyme was able to bind ouabain upon the addition of 2 mM ouabain under certain conditions. This binding was demonstrated by a 50% inhibition of ATPase after the removal of unbound ouabain by a Sephadex G-50 column (ouabain trapping method). At 4 degrees, this ouabain binding was observed on the K+- sensitive phosphoprotein (E2P) formed in the presence of 10 mM Na+ but was not observed on the ADP-sensitive phosphoprotein (E1P) formed in the presence of 1000 mM Na+. The increase in the dephosphorylation rate of E2P with various concentrations of K+ paralleled the decrease in inhibition by the addition of 2 mM ouabain after the termination of phosphorylation. In 50-200 mM Na+, the eel enzyme used here formed the E1P-rich phosphoprotein, but this phosphoprotein could bind with ouabain, even though the presence of ADP or oligomycin, which prevents the conversion of E1P to E2P, partially interfered with this ouabain binding. At 25 degrees, ouabain binding with E1P-rich phosphoprotein was observed in higher yield (up to 71%), but in each of these cases ADP or oligomycin strongly inhibited ouabain binding. Moreover, ouabain binding with E2P-rich phosphoprotein did not significantly change with temperature, but ouabain binding with E1P-rich phosphoprotein increased more than 6 times at temperatures from 4 degrees to 25 degrees. From these results, it can be concluded that E2P can bind with ouabain in the absence of free Mg2+ whereas E1P cannot, and that the interconversion between E1P and E2P can be stimulated with ouabain binding and accelerated with elevation of temperature. ADP- and K+ -insensitive phosphoprotein probably is only a minor intermediate for ouabain binding.


A pharmacokinetic study of injectable testosterone undecanoate in hypogonadal men.
Testosterone undecanoate (TU) provides testosterone (T) replacement for hypogonadal men when administered orally but requires multiple doses per day and produces widely variable serum T levels. We investigated the pharmacokinetics of a newly available TU preparation administered by intramuscular injection to hypogonadal men. Eight patients with Klinefelter's syndrome received either 500 mg or 1,000 mg of TU by intramuscular injection; 3 months later, the other dose was given to each man (except to one, who did not receive the 1,000-mg dose). Serum levels of reproductive hormones were measured at regular intervals before and after the injections. Mean serum T levels increased significantly at the end of the first week, from less than 10 nmol/L to 47.8+/-10.1 and 54.2+/-4.8 nmol/ L for the lower and higher doses, respectively. Thereafter, serum T levels decreased progressively and reached the lower-normal limit for adult men by day 50 to 60. Pharmacokinetic analysis showed a terminal elimination half-life of 18.3+/-2.3 and 23.7+/-2.7 days and showed a mean residence time of 21.7+/-1.1 and 23.0+/-0.8 days for the lower and higher doses, respectively. The area under the serum T concentration-time curve and the T-distribution value related to serum T concentration were significantly higher following the 1,000-mg dose than following the 500-mg dose. The 500-mg dose, when given as the second injection, yielded optimal pharmacokinetics (defined as mean peak T values not exceeding the normal range and persistence of normal levels for at least 7 weeks), suggesting that repeated injections of 500 mg at 6-8-week intervals may provide optimal T replacement. The mean serum levels of estradiol were normalized following the injections, and prolactin levels were normal throughout the study. Significant decrease of serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels was observed, with the decrease in LH levels being more pronounced. There were no significant differences in serum LH and FSH levels between the two doses. Sex hormone-binding globulin (SHBG) levels before any T therapy were near the upper limit of normal for adult men and were reduced by approximately 50% just prior to the second dose of TU. The decreased SHBG levels produced by the first TU injection could have led to lower peak total T levels and to a more rapid clearance of T following the second TU injection. We conclude that single-dose injections of TU to hypogonadal men can maintain serum T concentration within the normal range for at least 7 weeks without immediately apparent side effects. It is likely that this form of T would require injections only at 6-8-week or longer intervals, not at the 2-week intervals necessary with currently used T esters (enanthate and cypionate). This injectable TU preparation may provide improved substitution therapy for male hypogonadism and, in addition, may be developed as an androgen component of male contraceptives.


Induction of specific transcription by RNA polymerase III in transformed cells.
RNA polymerase III (pol III) transcripts of the highly repeated mouse B2 gene family are increased in many oncogenically transformed murine cell lines. In cells transformed by simian virus 40, the small, cytoplasmic B2 RNAs are present at 20-fold-higher levels than in normal cells (M. R. D. Scott, K. Westphal, and P. W. J. Rigby, Cell 34:557-567, 1983; K. Singh, M. Carey, S. Saragosti, and M. Botchan, Nature [London] 314:553-556). We found that transcripts of the highly repeated B1 gene family are also increased 20-fold upon simian virus 40 transformation and showed that these RNAs result from pol III transcription. In contrast, transcripts from less highly repeated pol III templates such as the 5S, 7SL, 7SK, 4.5SI, tRNAMet, and tRNAPro genes are unaffected. The expression of the B2 RNAs in isolated nuclei shows that the augmentation is due mainly to an increased rate of transcription by pol III. There is thus specific transformation-inducible pol III transcription. We developed an in vitro transcription assay which utilizes genomic DNA as a template to study the transcription of all members of a repetitive gene family in their native context. This assay reproduces the low cytoplasmic levels of B1 compared with B2 RNAs suggesting that this ratio is dictated by intrinsic signals in the DNA.


Pregnancy late in life: a hospital-based study of birth outcomes.
Pregnancy in older women is of great relevance, particularly in developed countries where many women experience pregnancy late in the childbearing age. A hospital-based data analysis of 9506 delivery records from 1998 to 2003 at the Liverpool Women's Hospital was undertaken to assess pregnancy outcomes in older women of reproductive age. Overall, 2.4 % of mothers were >40 years of age (advanced), 5.6% were <20 years (adolescents), and 92% were between 20 and 40 years. The prevalence of low birthweight (LBW), preterm birth, and small for gestational age by maternal age category followed a U-shaped curve with nadirs in the middle age classes. The gestational age of older mothers was 1 week shorter than that for women aged 26-30 years (p = 0.005). Primiparaes >40 years were at higher risk for delivering a LBW (9.4% vs. 5.3%, p = 0.005) or a very preterm baby (8.9% vs. 4.4%, p = 0.001) than were multiparous mothers of the same age. There was an association between maternal advanced age and LBW (adjusted OR [AOR], 1.7, 95% CI 1.4-2.5, p = 0.001), preterm birth (AOR 1.4, 95% CI 1.1-2.4, p = 0.04), or very preterm birth (AOR 1.6, 95% CI 1.2-3.5, p = 0.002) after controlling for prenatal alcohol and smoking exposure, household deprivation, maternal anemia, obesity, parity, and single parenthood. Pregnancy in older women is associated with adverse birth outcomes, particularly in primigravidas. Increased health promotion is required to highlight the risk of adverse birth outcomes in women who become pregnant for the first time in the late childbearing years.


Hirschsprung disease: Insights on genes, penetrance, and prenatal diagnosis.
The objective of this mini-review is to provide insights on the advances in the understanding of the genetic variants associated with different manifestations of Hirschsprung disease, which may present with a range of denervation from a short segment of colon to total colonic and small bowel or extensive aganglionosis. A recent article in this journal documented potential gene variants involved in long-segment Hirschsprung disease in 23 patients. Gene variants were identified using a 31-gene panel of genes related to Hirschsprung disease or enteric neural crest cell development, as previously reported in the literature. The study identified potentially harmful variants in eight genes across 13 patients, with a detection rate of 56.5% (13/23 patients). Five patients had pathologic variants in RET, NRG1, and L1CAM, and the remainder were considered variants of unknown significance. The authors attempted prenatal diagnosis of Hirschsprung disease utilizing an amniocentesis sample obtained for advanced maternal age in a family with a known deleterious RET mutation, manifested in the father (long-segment Hirschsprung disease) and older daughter (total colonic aganglionosis). The fetus had the same RET variant but, after several years of follow-up, has not developed any symptoms of Hirschsprung disease, supporting the conclusion that this RET mutation is an autosomal dominant gene with incomplete penetrance. This experience suggests that genetic counseling is appropriate to carefully assess the justification of prenatal testing, especially, when the phenotype of long-segment Hirschsprung disease is so variable and the disease is potentially curable with surgery.


Molecular screening of the human melanocortin-4 receptor gene: identification of a missense variant showing no association with obesity, plasma glucose, or insulin.
Disruption of the melanocortin-4 (MC-4) receptor gene in mice results in maturity-onset obesity, hyperinsulinaemia and hyperglycaemia. These phenotypes are characteristic of human obesity that frequently accompanies non-insulin-dependent diabetes. It is therefore possible that human MC-4 receptor gene mutations contribute to human obesity. To test this possibility, we examined by DNA sequencing the entire coding region of the human MC-4 receptor gene in 40 morbidly obese (BMI > 35 kg/m2) white British males and examined the 5'- and 3'-flanking regions in 20 out of these obese subjects. We also sequenced all these regions in 10 lean (BMI < 18 kg/m2) white British males for a reference. We identified a single nucleotide substitution that replaces valine with isoleucine at codon 103, in two obese subjects in the heterozygous state. No other nucleotide alterations were found. The prevalence of this missense variant was studied in 322 white British males (190 with BMI > 28 kg/m2 and 132 with BMI < 22 kg/m2) selected from a population-based epidemiological survey. In these subjects, no homozygotes for the isoleucine allele were found. The frequency of heterozygotes was similar (4.2 vs 4.5%) in the two groups and there was no significant difference in BMI, total skinfold thickness, plasma insulin and glucose levels between heterozygotes and codon-103 valine homozygotes in either group. These results suggest that coding sequence mutations in the MC-4 receptor gene are unlikely to be a major cause of human obesity, at least in white British males.


Cancer risk in hyperprolactinemia patients: a population-based cohort study.
Experimental evidence indicates that prolactin might play a role in tumorigenesis of several human cancers, but data on cancer risk in hyperprolactinemia patients are sparse. The aim of this study was to investigate cancer risk in hyperprolactinemia patients. Design A population-based matched cohort study in Sweden. The hyperprolactinemia cohort consisted of patients hospitalized for hyperprolactinemia from 1987 to 1995 identified in the National Patient Register (n=585) and a hospital cohort of prolactinoma patients at Karolinska University Hospital (n=384). For each patient, ten matched individuals were identified via the Register of Population. Cancer occurrence was ascertained via the Swedish Cancer Registry. Hazard ratios (HRs) were estimated by Cox proportional hazards regression. Seventy-three malignant tumors were identified in the hyperprolactinemia patients and 660 tumors in the comparison group (HR 1.31; 95% confidence interval (CI): 1.02-1.68), mainly attributed to an increased risk of upper gastrointestinal cancer in both males and females (HR 3.69; 95% CI: 1.70-8.03) and hematopoietic cancer in females (HR 3.51; 95% CI: 1.06-11.6). Twelve breast cancers occurred in the female patients, corresponding to an HR of 1.09 (95% CI: 0.60-1.99). Prostate cancer risk in hyperprolactinemia men was reduced (HR 0.40; 95% CI: 0.16-0.99). An increased overall cancer risk was found in hyperprolactinemia patients, but no increased risk of breast cancer in women and a reduced risk of prostate cancer in men. These findings warrant further investigations and to be confirmed in larger studies but may indicate the importance of an active treatment strategy and follow-up of hyperprolactinemia patients.


Redefining the maximum sustainable yield for the Schaefer population model including multiplicative environmental noise.
The focus of this article is to investigate the biological reference points, such as the maximum sustainable yield (MSY), in a common Schaefer (logistic) surplus production model in the presence of a multiplicative environmental noise. This type of model is used in fisheries stock assessment as a first-hand tool for biomass modelling. Under the assumption that catches are proportional to the biomass, we derive new conditions on the environmental noise distribution such that stationarity exists and extinction is avoided. We then get new explicit results about the stationary behavior of the biomass distribution for a particular specification of the noise, namely the biomass distribution itself and a redefinition of the MSY and related quantities that now depend on the value of the variance of the noise. Consequently, we obtain a more precise vision of how less optimistic the stochastic version of the MSY can be than the traditionally used (deterministic) MSY. In addition, we give empirical conditions on the error variance to approximate our specific noise by a lognormal noise, the latter being more natural and leading to easier inference in this context. These conditions are mild enough to make the explicit results of this paper valid in a number of practical applications. The outcomes of two case-studies about northwest Atlantic haddock [Spencer, P.D., Collie, J.S., 1997. Effect of nonlinear predation rates on rebuilding the Georges Bank haddock (Melanogrammus aeglefinus) stock. Can. J. Fish. Aquat. Sci. 54, 2920-2929] and South Atlantic albacore tuna [Millar, R.B., Meyer, R., 2000. Non-linear state space modelling of fisheries biomass dynamics by using Metropolis-Hastings within-Gibbs sampling. Appl. Stat. 49, 327-342] are used to illustrate the impact of our results in bioeconomic terms.


Interleukin-1, inflammasomes and the skin.
Interleukin (IL)-1 is a highly active and pleiotropic pro-inflammatory cytokine. Recent data impressively demonstrate that activating mutations in a human gene involved in proIL-1beta maturation or loss-of-function mutations in the gene encoding IL-1 receptor antagonist (IL-1Ra) cause excessive activity of this cytokine. This can result in life-threatening systemic and local inflammation, particularly in the skin. Interestingly, experiments in mice revealed that epidermal keratinocytes can secrete large amounts of IL-1alpha, which induces an inflammatory response in the skin. Secretion of IL-1 requires caspase-1 activity, and activation of the protease takes place in innate immune complexes, called inflammasomes. As keratinocytes express and activate caspase-1 in an inflammasome-dependent manner, these epithelial cells might be critically involved in the innate immunity of the skin. In this review we summarize the current knowledge on IL-1 and inflammasomes in the skin, particularly their involvement in skin homeostasis and disease. In addition, we discuss the hypothesis that keratinocytes are not only static bricks of the epidermal wall, but immunologically active cells critically involved in different (auto)-inflammatory (skin) diseases.


High levels of household food insecurity on the Navajo Nation.
To assess levels of and identify factors associated with food insecurity on the Navajo Nation. A cross-sectional study was conducted utilizing the ten-item Radimer/Cornell food insecurity instrument. Sociodemographic, psychosocial and anthropometric data were collected. Navajo Nation, USA. Two hundred and seventy-six members of the Navajo Nation were randomly selected at food stores and other community locations. Of the sample, 76·7% had some level of food insecurity. Less education (mean years of schooling: P = 0·0001; non-completion of higher education: P = 0·0003), lower full-time employment rates (P = 0·01), and lower material style of life (P = 0·0001), food knowledge (P = 0·001) and healthy eating self-efficacy (P < 0·0001) scores were all positively associated with food insecurity. Perceived expensiveness (P < 0·0001) and perceived inconvenience (P = 0·0001) of healthy choices were also positively associated with food insecurity. Food insecurity rates on the Navajo Nation are the highest reported to date in the USA and are likely attributable to the extremely high rates of poverty and unemployment. Reducing food insecurity on the Navajo Nation will require increasing the availability of affordable healthy foods, addressing poverty and unemployment, and providing nutrition programmes to increase demand.


Patients with multiple synchronous colonic cancer hepatic metastases benefit from enrolment in a "liver first" approach protocol.
To assess a protocol for treating patients with multiple synchronous colonic cancer liver metastases, which are unresectable in one stage. Patients enrolled in the "liver first" protocol presented with colon-only (not rectal) cancer and multiple synchronous hepatic metastases (type II or III). All patients showed good performance status (ECOG PS 0-1) and were treated with curative intent. Complete oncologic staging including positron emission tomography-computed tomography was performed in order to rule out extrahepatic disease. If bowel obstruction was imminent, an intraluminal colonic stent was placed endoscopically. Subsequently, all patients received standardised neo-adjuvant chemotherapy, that is, FOLFOX or XELOX regimens combined with an antiangiogenic agent (bevacizumab or cetuximab). Provided that a response to chemotherapy was observed, patients underwent either one or two hepatectomies with or without portal vein embolization followed by the indicated colectomy. Further chemotherapy was administered after each procedure. Re-staging was performed after each chemotherapeutic treatment. Disease progression at any stage resulted in discontinuation of the protocol and conversion to palliative disease management. Prospectively recorded data from 11 consecutive patients (8 men) were analysed for this study. Their mean age at the time of their first assessment was 65.7 (SD ± 15.3) years. Six (54.6%) patients presented with type III metastatic disease. The minimum and maximum follow-up periods were 7.3 and 39.6 mo, respectively. The mean overall survival of all patients was 16.5 (95%CI: 10.0-23.2) mo. A colonic stent had to be placed in 5 (45.5%) patients due to the onset of an intraluminal obstruction. Four (36.4%) patients succeeded in completing all planned surgical operations. Their mean overall survival was 27.2 (95%CI: 15.1-39.3) mo and the mean disease-free survival was 7.7 (95%CI: 3.0-12.5) mo. Patients, who were obliged to shift to palliative treatment due to disease progression, had a mean overall survival of 10.5 (95%CI: 8.6-12.4) mo. None of these patients underwent palliative colectomy. No postoperative mortality was recorded. The implementation of a structured "liver first" approach protocol for the treatment of patients with extensive, liver-limited colon cancer metastatic disease may be beneficial.


Apparent genetic complexity generated by developmental thresholds: the apterous locus in Drosophila melanogaster.
Mutations at the apterous (ap) locus in Drosophila melanogaster give rise to three distinct phenotypes: aberrant wings, female sterility and precocious adult death. The wing phenotype includes five types of abnormality: blistering, deficiencies, duplications, high-order repetitions and transformation of structures. The mildest phenotype is seen with homozygous apblt animals which have either normal or slightly blistered wings. Most alleles produce, in the homozygote, a deficient wing in which part or all of the wing margin and wing blade is missing, but wing hinge and notum regions are normal. Animals hemizygous for each of 20 ap alleles, as well as apID/apXa heterozygotes, show duplication of parts of the notum associated with complete wing deficiency. Animals heterozygous for apc and the other tested ap alleles show repetitions of parts of the anterior wing margin, an engrailed-like transformation of posterior wing margin into anterior margin or both. Both apblt and apc show similar phenotypes in homozygotes and hemizygotes, yet both produce a less extreme phenotype than that of the other hemizygotes, suggesting that neither mutation causes loss of the entire ap+ function. The 15 alleles that cause precocious death and female sterility occur in six complementation groups based on complementation for these phenotypes. This supports the previous conclusion that the effects of apterous mutations on the wing do not correlate with their effects on viability and fertility. We propose an explanation for the effects of apterous mutations on the wing in which quantitative reductions in the activity of gene product give rise to qualitatively different phenotypes because of different threshold requirements of the ap+ function for critical events in wing disc development.


Acanthamoeba infection in a patient with chronic graft-versus-host disease occurring during treatment with voriconazole.
We report a case of disseminated infection with Acanthamoeba in a patient with graft-versus-host disease after hematopoietic stem cell transplant (HSCT) for acute lymphocytic leukemia. The infection involved the brain, skin, and lungs and occurred despite treatment with voriconazole for mold prophylaxis, and did not respond to treatment with multiple other agents reported to have activity against Acanthamoeba. To our knowledge, infection with Acanthamoeba has been reported in 4 other patients after HSCT or bone marrow transplant, and our case is the first to be diagnosed ante-mortem.


Scaling up the global reef restoration activity: Avoiding ecological imperialism and ongoing colonialism.
The health and condition of the world's reefs are in steep decline. This has triggered the development of fledgling micro-scale coral reef restoration projects along many reef coastlines. However, it is increasingly recognised that the scale and productivity of micro-scale coral gardening projects will be insufficient to meet the growing global threats to reefs. More recently, efforts to develop and implement restoration techniques for application at regional scales have been pursued by research organisations. Coral reefs are mostly located in the unindustrialised world. Yet, most of the funding, and scientific and engineering method development for larger-scale methods will likely be sourced and created in the industrialised world. Therefore, the development of the emerging at-scale global reef restoration sector will inevitably involve the transfer of methods, approaches, finances, labour and skills from the industrialised world to the unindustrialised world. This opens the door to the industrialised world negatively impacting the unindustrialised world and, in some cases, First Nations peoples. In Western scientific parlance, ecological imperialism occurs when people from industrialised nations seek to recreate environments and ecosystems in unindustrialised nations that are familiar and comfortable to them. How a coral reef 'should' look depends on one's background and perspective. While predominately Western scientific approaches provide guidance on the ecological principles for reef restoration, these methods might not be applicable in every scenario in unindustrialised nations. Imposing such views on Indigenous coastal communities without the local technical and leadership resources to scale-up restoration of their reefs can lead to unwanted consequences. The objective of this paper is to introduce this real and emerging risk into the broader reef restoration discussion.


Monoclonal antibodies to human transglutaminase 4.
Transglutaminase 4 (TG4) is a member of the enzyme family that catalyzes the calcium-dependent post-translational modification of proteins via cross-linking, polyamination, or deamidation. TG4 exhibits prostate-specific expression pattern and plays a crucial role in the formation of the copulatory plug in rodents. However, the physiological function(s) of human TG4 remains speculative. Human TG4 has been postulated to participate in the maturation process of sperm by modifying its cell surface, which results in suppression of sperm antigenicity in the female genital tract. To better understand the pathophysiological role of TG4 in prostate tissue, we generated monoclonal antibodies (MAb) against human TG4 in mice by repeated injections with the recombinant human TG4. Western blot analysis demonstrated that the selected MAbs react specifically with TG4, but not with other isoenzymes of the TG family. Immunocytochemical and immunohistochemical analyses showed that specific staining is observed with the cells overexpressing TG4 and with the paraffin-embedded prostate tissue specimens obtained from the benign prostate hyperplasia and prostate cancer patients, respectively. Our results indicate that these MAbs are suitable for detecting TG4 in the cultured cells or prostate tissues for investigating the biological functions of human TG4.


Mortality after Renal Allograft Failure and Return to Dialysis.
The outcomes of patients who fail their kidney transplant and return to dialysis (RTD) has not been investigated in a nationally representative sample. We hypothesized that variations in management of transplant chronic kidney disease stage 5 leading to kidney allograft failure (KAF) and RTD, such as access, nutrition, timing of dialysis, and anemia management predict long-term survival. We used an incident cohort of patients from the United States Renal Data System who initiated hemodialysis between January 1, 2003 and December 31, 2008, after KAF. We used Cox regression analysis for statistical associations, with mortality as the primary outcome. We identified 5,077 RTD patients and followed them for a mean of 30.9 ± 22.6 months. Adjusting for all possible confounders at the time of RTD, the adjusted hazards ratio (AHR) for death was increased with lack of arteriovenous fistula at initiation of dialysis (AHR 1.22, 95% CI 1.02-1.46, p = 0.03), albumin <3.5 g/dL (AHR 1.33, 95% CI 1.18-1.49, p = 0.0001), and being underweight (AHR 1.30, 95% CI 1.07-1.58, p = 0.006). Hemoglobin <10 g/dL (AHR 0.96, 95% CI 0.86-1.06, p = 0.46), type of insurance, and zip code-based median household income were not associated with higher mortality. Glomerular filtration rate <10 mL/min/1.73 m2 at time of dialysis initiation (AHR 0.83, 95% CI 0.75-0.93, p = 0.001) was associated with reduction in mortality. Excess mortality risk observed in patients starting dialysis after KAF is multifactorial, including nutritional issues and vascular access. Adequate preparation of patients with failing kidney transplants prior to resuming dialysis may improve outcomes.


Development and evaluation of a CEACAM6-targeting theranostic nanomedicine for photoacoustic-based diagnosis and chemotherapy of metastatic cancer.
Metastasis is the leading cause of cancer-related deaths. A number of chemotherapeutic and early diagnosis strategies, including nanomedicine, have been developed to target metastatic tumor cells. However, simultaneous inhibition and imaging of metastasis is yet to be fully achieved. Methods: To overcome this limitation, we have developed human serum albumin-based nanoparticles (tHSA-NPs) with photoacoustic imaging capability, which target carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6). CEACAM6 is highly expressed in metastatic anoikis-resistant tumor cells. Results:In vitro, the CEACAM6-targeting tHSA-NPs efficiently targeted CEACAM6-overexpressing metastatic anoikis-resistant tumor cells. In vivo, CEACAM6-targeting tHSA-NPs administered intravenously to BALB/c nude mice efficiently inhibited lung metastasis in circulating anoikis-resistant tumor cells compared to the controls. In addition, anoikis-resistant tumor cells can be successfully detected by photoacoustic imaging, both in vitro and in vivo, using the intrinsic indocyanine green-binding affinity of albumin. Conclusion: In summary, the CEACAM6-targeting albumin-based nanoparticles allowed the delivery of drugs and photoacoustic imaging to metastatic anoikis-resistant tumor cells in vitro and in vivo. Based on the expression of CEACAM6 in a variety of tumors, CEACAM6-targeting nanomedicine might be used to target various types of metastatic tumor cells.


Developmental changes in purine phosphoribosyltransferases in human and rat tissues.
1. The hypoxanthine/guanine and adenine phosphoribosyltransferase activities in a wide variety of human tissues were studied during their growth and development from foetal life onward. A wide range of activities develop after birth, with especially high values in the central nervous system and testes. 2. Postnatal development of hypoxanthine/guanine phosphoribosyltransferase was also defined in the rat. Although there were increases in the central nervous system and testes, there was also a rise in activity in the liver, which was less marked in man. 3. A sensitive radiochemical assay method, using dTTP to inhibit 5'-nucleotidase activity, suitable for tissue extracts, was developed. 4. No definite evidence of the existence of tissue-specific isoenzymes of hypoxanthine/guanine or adenine phosphoribosyltransferase was found. Hypoxanthine/guanine phosphoribosyltransferase in testes, however, had a significantly different thermal-denaturation rate constant. 5. The findings are discussed in an attempt to relate activity of hypoxanthine/guanine phosphoribosyltransferase to biological function. Growth as well as some developmental changes appear to be related to increase in the activity of this enzyme.


Bilirubin reduces visceral obesity and insulin resistance by suppression of inflammatory cytokines.
Although previous studies have reported a negative relationship between serum bilirubin concentration and the development of diabetes mellitus (DM), the relationship between bilirubin and insulin resistance has not been thoroughly assessed. This study was designed to determine the relationships between bilirubin, body fat distribution, and adipose tissue inflammation in patients with type 2 DM and the effect of bilirubin in an obese animal model. Body fat distribution was measured using an abdominal dual bioelectrical impedance analyzer in patients with type 2 DM. We also measured glycemic control, lipid profile, serum bilirubin concentration and other clinical characteristics, and determined their relationships with body fat distribution. In the animal study, biliverdin (20 mg/kg daily) was orally administered to high-fat diet (HFD)-induced obese (DIO) mice for 2 weeks, after which intraperitoneal insulin tolerance testing was performed. Then, adipocyte area, adipocytokine expression, and macrophage polarization were evaluated in epididymal adipose tissues. In the clinical study, univariate analysis showed that a lower bilirubin concentration was significantly correlated with higher body mass index, waist circumference, triglyceride, uric acid, creatinine, visceral fat area and lower HDL-C. In multivariate analyses, bilirubin concentration significantly correlated with diastolic blood pressure, creatinine, and visceral fat area. However, there was no association between bilirubin concentration and subcutaneous fat area. In the animal study, DIO mice treated with biliverdin had smaller adipocytes than untreated DIO mice and biliverdin improved HFD-induced insulin resistance. Biliverdin treatment reversed the higher gene expression of Cd11c, encoding an M1 macrophage marker, and Tnfa, encoding the proinflammatory cytokine tumor necrosis factor-α, in the adipose tissues of DIO mice. These data suggest biliverdin administration alleviates insulin resistance by ameliorating inflammation and the dysregulation of adipocytokine expression in adipose tissues of DIO mice. Bilirubin may protect against insulin resistance by ameliorating visceral obesity and adipose tissue inflammation.


Functional and esthetic rehabilitation of a patient with amelogenesis imperfecta.
Amelogenesis imperfecta (AI) is a hereditary disorder that causes developmental alterations in the structure of enamel. In addition, tooth sensitivity, missing or impacted teeth, taurodontism, altered dental esthetics and anterior open bite can also be associated with AI. This clinical report presents the diagnosis, treatment planning and prosthetic rehabilitation of a 19-year-old female patient with AI associated with a group of dental anomalies. Following clinical and radiographic examination, histologic evaluation of the teeth confirmed the diagnosis of rough pattern hypoplastic AI. The patient was rehabilitated with full-mouth zirconium oxide ceramic fixed bridges. Adaptation of the temporomandibular joints and masticatory muscles to the bridges was carefully observed over 3 years. At the end of this follow-up period, the patient was satisfied with the esthetics, function and phonation of her prostheses.


Use of a hierarchical oligonucleotide primer extension approach for multiplexed relative abundance analysis of methanogens in anaerobic digestion systems.
In this study, we established a rapid multiplex method to detect the relative abundances of amplified 16S rRNA genes from known cultivatable methanogens at hierarchical specificities in anaerobic digestion systems treating industrial wastewater and sewage sludge. The method was based on the hierarchical oligonucleotide primer extension (HOPE) technique and combined with a set of 27 primers designed to target the total archaeal populations and methanogens from 22 genera within 4 taxonomic orders. After optimization for their specificities and detection sensitivity under the conditions of multiple single-nucleotide primer extension reactions, the HOPE approach was applied to analyze the methanogens in 19 consortium samples from 7 anaerobic treatment systems (i.e., 513 reactions). Among the samples, the methanogen populations detected with order-level primers accounted for >77.2% of the PCR-amplified 16S rRNA genes detected using an Archaea-specific primer. The archaeal communities typically consisted of 2 to 7 known methanogen genera within the Methanobacteriales, Methanomicrobiales, and Methanosarcinales and displayed population dynamic and spatial distributions in anaerobic reactor operations. Principal component analysis of the HOPE data further showed that the methanogen communities could be clustered into 3 distinctive groups, in accordance with the distribution of the Methanosaeta, Methanolinea, and Methanomethylovorans, respectively. This finding suggested that in addition to acetotrophic and hydrogenotrophic methanogens, the methylotrophic methanogens might play a key role in the anaerobic treatment of industrial wastewater. Overall, the results demonstrated that the HOPE approach is a specific, rapid, and multiplexing platform to determine the relative abundances of targeted methanogens in PCR-amplified 16S rRNA gene products.


Chronic graft-versus-host syndrome in man. A long-term clinicopathologic study of 20 Seattle patients.
This study of chronic graft-versus-host disease (GVHD) describes the clinical, pathologic and laboratory features, and the causes of morbidity and mortality in 20 patients who received allogeneic marrow transplants from HLA identical sibling donors. Chronic GVHD is a pleiotrophic syndrome with variability in the time of onset, organ systems involved and rate of progression. The clinical-pathologic features resemble an overlap of several collagen vascular diseases with frequent involvement of the skin, liver, eyes, mouth, upper respiratory tract, esophagus and less frequent involvement of the serosal surfaces, lower gastrointestinal tract and skeletal muscles. Major causes of morbidity are scleroderma with contractures and ulceration, dry eyes and mouth, pulmonary insufficiency and wasting. Chronic GVHD has features of immune dysregulation with elevated levels of eosinophils, circulating autoantibodies, hypergammaglobulinemia and plasmacytosis of viscera and lymph nodes. In this study, three patients had limited chronic GVHD with relatively favorable prognosis characterized by localized skin involvement and/or hepatic disease without chronic aggressive histology. Most patients, however, had extensive disease with a progressive course. Survival was largely determined by the presence or absence of serious recurrent bacterial infections. The over-all severity of disease was best assessed by using the Karnofsky performance rating.


Feasibility of therapeutic endoscopic ultrasound in the bridge-to-surgery scenario: The example of pancreatic adenocarcinoma.
Upfront resection is becoming a rarer indication for pancreatic ductal adenocarcinoma, as biologic behavior and natural history of the disease has boosted indications for neoadjuvant treatments. Jaundice, gastric outlet obstruction and acute cholecystitis can frequently complicate this window of opportunity, resulting in potentially deleterious chemotherapy discontinuation, whose resumption relies on effective, prompt and long-lasting management of these complications. Although therapeutic endoscopic ultrasound (t-EUS) can potentially offer some advantages over comparators, its use in potentially resectable patients is primal and has unfairly been restricted for fear of potential technical difficulties during subsequent surgery. This is a narrative review of available evidence regarding EUS-guided choledochoduodenostomy, gastrojejunostomy and gallbladder drainage in the bridge-to-surgery scenario. Proof-of-concept evidence suggests no influence of t-EUS procedures on outcomes of eventual subsequent surgery. Moreover, the very high efficacy-invasiveness ratio over comparators in managing pancreatic cancer-related symptoms or complications can provide a powerful weapon against chemotherapy discontinuation, potentially resulting in higher subsequent resectability. Available evidence is discussed in this short paper, together with technical notes that might be useful for endoscopists and surgeons operating in this scenario. No published evidence supports restricting t-EUS in potential surgical candidates, especially in the setting of pancreatic cancer patients undergoing neoadjuvant chemotherapy. Bridge-to-surgery t-EUS deserves further prospective evaluation.


Involvement of AlgQ in transcriptional regulation of pyoverdine genes in Pseudomonas aeruginosa PAO1.
In response to iron limitation, Pseudomonas aeruginosa produces the fluorescent siderophore pyoverdine. Transcription of pyoverdine biosynthetic (pvd) genes is driven by the iron starvation sigma factor PvdS, which is negatively regulated by the Fur-Fe(II) holorepressor. We studied the effect of AlgQ, the Escherichia coli Rsd orthologue, on pyoverdine production by P. aeruginosa PAO1. AlgQ is a global regulatory protein which activates alginate, ppGpp, and inorganic polyphosphate synthesis through a cascade involving nucleoside diphosphate kinase (Ndk). AlgQ is also capable of interacting with region 4 of RpoD. In a reconstituted E. coli system, PvdS-dependent transcription from the pvdA promoter was doubled by the multicopy algQ gene. The P. aeruginosa DeltaalgQ mutant exhibited a moderate but reproducible reduction in pyoverdine production compared with wild-type PAO1, as a result of a decline in transcription of pvd genes. PvdS expression was not affected by the algQ mutation. Single-copy algQ fully restored pyoverdine production and expression of pvd genes in the DeltaalgQ mutant, while ndk did not. An increased intracellular concentration of RpoD mimicked the DeltaalgQ phenotype, whereas PvdS overexpression suppressed the algQ mutation. E. coli rsd could partially substitute for algQ in transcriptional modulation of pvd genes. We propose that AlgQ acts as an anti-sigma factor for RpoD, eliciting core RNA polymerase recruitment by PvdS and transcription initiation at pvd promoters. AlgQ provides a link between the pyoverdine and alginate regulatory networks. These systems have similarities in responsiveness and physiological function: both depend on alternative sigma factors, respond to nutrient starvation, and act as virulence determinants for P. aeruginosa.


Lipid metabolism in cancer progression and therapeutic strategies.
Dysregulated lipid metabolism represents an important metabolic alteration in cancer. Fatty acids, cholesterol, and phospholipid are the three most prevalent lipids that act as energy producers, signaling molecules, and source material for the biogenesis of cell membranes. The enhanced synthesis, storage, and uptake of lipids contribute to cancer progression. The rewiring of lipid metabolism in cancer has been linked to the activation of oncogenic signaling pathways and cross talk with the tumor microenvironment. The resulting activity favors the survival and proliferation of tumor cells in the harsh conditions within the tumor. Lipid metabolism also plays a vital role in tumor immunogenicity via effects on the function of the noncancer cells within the tumor microenvironment, especially immune-associated cells. Targeting altered lipid metabolism pathways has shown potential as a promising anticancer therapy. Here, we review recent evidence implicating the contribution of lipid metabolic reprogramming in cancer to cancer progression, and discuss the molecular mechanisms underlying lipid metabolism rewiring in cancer, and potential therapeutic strategies directed toward lipid metabolism in cancer. This review sheds new light to fully understanding of the role of lipid metabolic reprogramming in the context of cancer and provides valuable clues on therapeutic strategies targeting lipid metabolism in cancer.


Legal rights of children with musculoskeletal disabilities.
As market forces increasingly control medicine, we must not forget the disabled, especially children. Although the costs of appropriate comprehensive services are substantial, it is well worth the cost to facilitate an adulthood in which these patients are on their own, possibly earning their livelihood, and experiencing the best quality of life our society can provide.


Shock Wave Mediated Plume Chemistry for Molecular Formation in Laser Ablation Plasmas.
Although it is relatively straightforward to measure the ionic, atomic, molecular, and particle emission features from laser ablation plumes, the associated kinetic and thermodynamic development leading to molecular and nanocluster formation remain one of the most important topics of analytical chemistry and material science. Very little is known, for instance, about the evolutionary paths of molecular and nanocluster formation and its relation to laser plume hydrodynamics. This is, to a large extent; due to the complexity of numerous physical processes that coexist in a transient laser-plasma system. Here, we report the formation mechanisms of molecules during complex interactions of a laser-produced plasma plume expanding from a high purity aluminum metal target into ambient air. It is found that the plume hydrodynamics plays a great role in redefining the plasma thermodynamics and molecular formation. Early in the plasma expansion, the generated shock wave at the plume edge acts as a barrier for the combustion process and molecular formation is prevalent after the shock wave collapse. The temporally and spatially resolved contour mapping of atoms and molecules in laser ablation plumes highlight the formation routes and persistence of species in the plasma and their relation to plume hydrodynamics.


Physiological response properties of cells labeled intracellularly with horseradish peroxidase in cat ventral cochlear nucleus.
To determine the correspondence between anatomical and physiological cell types in the ventral cochlear nucleus of the cat, intracellular injections of horseradish peroxidase were made into cells whose extracellular and intracellular responses to sound had been studied. Three identified cells responded to a short tone burst at their characteristic frequencies with an onset pattern. This pattern is characterized by a strong response to the onset of the stimulus. One was an octopus cell. The second cell, located in the octopus-cell area, was a giant cell with a few somatic spines and thin tapering dendrites; the intracellular record revealed that even in the absence of sound it received continuous synaptic input, while tones at characteristic frequency produced a sustained depolarization. A third cell, which had an onset response at low intensities and a chopper response at high intensities, was a stellate cell located in the intermediate acoustic stria with dendrites oriented parallel to the fiber tract. This cell had an unusually broad dynamic range in response to changes in intensity. Two cells with transient chopper response patterns were stellate cells in the posteroventral cochlear nucleus with many branched and beaded dendrites. Three cells with more sustained chopper response patterns were stellate cells in the anteroventral cochlear nucleus with fewer, less-branched, smooth dendrites. Two cells with primarylike responses to tones were bushy cells with numerous short, thin, highly branched dendrites in the posterior division of the anteroventral cochlear nucleus. Intracellular responses to tones at the characteristic frequency consisted of large brief depolarizations, which were not sustained. Another cell, which responded to tones in a phase-locked fashion, was also located in the anteroventral cochlear nucleus. It was a small, stellate cell with relatively few, smooth dendrites. The labeled cells largely support previous attempts at physiological-morphological correlations: (1) bushy cells exhibit primarylike pattern; (2) stellate cells exhibit chopper patterns; and (3) octopus cells exhibit an onset pattern. It was also demonstrated that more than one cell type can exhibit a particular response pattern.


Probing the Active Site of Deubiquitinase USP30 with Noncanonical Tryptophan Analogues.
Methanosarcina mazei pyrrolysyl-tRNA synthetase (PylRS) and its cognate tRNA have been evolved to generate genetically encoded noncanonical amino acids (ncAAs). Use of tryptophan (Trp) analogues with pyrrole ring modification for their spatial and polarity tuning in enzyme activity and substrate specificity is still limited. Herein, we report the application of an evolved PylRS, FOWRS2, for efficient incorporation of five Trp analogues into the deubiquitinase USP30 to decipher the role of W475 for diubiquitin selectivity. Structures of the five FOWRS-C/Trp analogue complexes at 1.7-2.5 Å resolution showed multiple ncAA binding modes. The W475 near the USP30 active site was replaced with Trp analogues, and the effect on the activity as well as the selectivity toward diubiquitin linkage types was examined. It was found that the Trp analogue with a formyl group attached to the nitrogen atom of the indole ring led to an improved activity of USP30 likely due to enhanced polar interactions and that another Trp analogue, 3-benzothienyl-l-alanine, induced a unique K6-specificity. Collectively, genetically encoded noncanonical Trp analogues by evolved PylRS·tRNACUAPyl pair unravel the spatial role of USP30-W475 in its diubiquitin selectivity.


Design and implementation of an imprinted material for the extraction of the endocrine disruptor bisphenol A from milk.
This paper describes the determination of bisphenol A (BPA) in milk samples, using a novel molecularly imprinted polymer. The imprinted polymer was developed using a rational design approach, and pre-polymerization interactions were investigated using molecular dynamics simulations and X-ray crystallography. A hydroquinone-imprinted polymer was used for solid phase extraction (SPE) clean-up of samples. BPA was quantified by high performance liquid chromatography (HPLC) and fluorescence (FLD) detection. Following validation, the method described was capable of determining bisphenol A in milk down to a limit of detection of 1.32μgkg(-1). The method was applied to a survey (n=27) of commercial milk products; BPA was detected in one of the samples, at a level of 176μgkg(-1). Test results were confirmed by a parallel UHPLC-MS/MS analytical method. This demonstrates the utility of the hydroquinone-imprinted polymer for application to selective sample clean-up and analysis of bisphenol A in milk, avoiding possible detrimental affects associated with template bleeding and without the need for expensive or difficult-to-obtain template.


Forward masking in a bottlenose dolphin Tursiops truncatus: dependence on azimuthal positions of the masker and test sources.
Forward masking was investigated by the auditory evoked potentials (AEP) method in a bottlenose dolphin Tursiops truncatus using stimulation by two successive acoustic pulses (the masker and test) projected from spatially separated sources. The positions of the two sound sources either coincided with or were symmetrical relative to the head axis at azimuths from 0 to ± 90°. AEPs were recorded either from the vertex or from the lateral head surface next to the auditory meatus. In the last case, the test source was ipsilateral to the recording side, whereas the masker source was either ipsi- or contralateral. For lateral recording, AEP release from masking (recovery) was slower for the ipsi- than for the contralateral masker source position. For vertex recording, AEP recovery was equal both for the coinciding positions of the masker and test sources and for their symmetrical positions relative to the head axis. The data indicate that at higher levels of the auditory system of the dolphin, binaural convergence makes the forward masking nearly equal for ipsi- and contralateral positions of the masker and test.


Linearized kinetic theory of spin-1/2 particles in magnetized plasmas.
We have considered linear kinetic theory, including the electron-spin properties in a magnetized plasma. The starting point is a mean-field Vlasov-like equation, derived from a fully quantum-mechanical treatment, where effects from the electron-spin precession and the magnetic dipole force are taken into account. The general conductivity tensor is derived, including both the free current contribution and the magnetization current associated with the spin contribution. We conclude the paper with an extensive discussion of the quantum-mechanical boundary where we list parameter conditions that must be satisfied for various quantum effects to be influential.


Mass spectrometry screening reveals peptides modulated differentially in the medial prefrontal cortex of rats with disparate initial sensitivity to cocaine.
To better understand why certain individuals are more vulnerable to cocaine abuse and addiction, we identify peptide markers associated with individual variation in sensitivity to the behavioral effects of cocaine. Previous studies in rats show that low, compared to high, cocaine responders are more sensitive to cocaine-induced behavioral plasticity (sensitization), exhibit enhanced conditioning to cocaine's rewarding effects, and are more motivated to self administer cocaine. In the current study, we combine matrix-assisted laser desorption/ionization mass spectrometry with multivariate statistical methods to analyze tissue extracts from rat dorsal striatum, nucleus accumbens, and medial prefrontal cortex (mPFC) to examine trends in peptide changes that coincide with behavioral phenotype. Peptide profiles of these three regions from individual animals were characterized via mass spectrometry. Resulting mass peaks that were statistically different between these groups were identified using principal component analysis. The mass peaks were then identified in pooled samples via multistage liquid chromatography mass spectrometry. A total of 74 peptides from 28 proteins were sequenced from defined brain regions. Statistically significant changes in peak intensities for seven peptides were found in the mPFC of rats given a single injection of 10 mg/kg cocaine, with low cocaine responders showing approximately 2-fold increase in peak intensities for the acetylated N terminus peptides of stathmin and Hint 1, as well as truncated ATP synthase. These results suggest that distinct peptide profiles in the mPFC are associated with individuals that exhibit reduced sensitivity to the behavioral effects of cocaine.


Design and evaluation of an RF front-end for 9.4 T human MRI.
At the field strength of 9.4 T, the highest field currently available for human MRI, the wavelength of the MR signals is significantly shorter than the size of the examined structures. Even more than at 7 T, constructive and destructive interferences cause strong inhomogeneities of the B1 field produced by a volume coil, causing shading over large parts of the image. Specialized radio frequency hardware and B1 management methods are required to obtain high-quality images that take full advantage of the high field strength. Here, the design and characteristics of a radio frequency front-end especially developed for proton imaging at 9.4 T are presented. In addition to a 16-channel transceiver array coil, capable of volume transmit mode and independent signal reception, it consists of custom built low noise preamplifiers and TR switches. Destructive interference patterns were eliminated, in virtually the entire brain, using a simple in situ radio frequency phase shimming technique. After mapping the B1+ profile of each transmit channel, a numerical algorithm was used to calculate the appropriate transmit phase offsets needed to obtain a homogeneous excitation field over a user defined region. Between two and three phase settings are necessary to obtain homogeneous images over the entire brain.


Perinatal Nutritional Reprogramming of the Epigenome Promotes Subsequent Development of Nonalcoholic Steatohepatitis.
With the epidemic of obesity, nonalcoholic fatty liver disease (NAFLD) has become the most common pediatric liver disease. The influence of a perinatal obesity-inducing diet (OID) on the development and progression of NAFLD in offspring is important but incompletely studied. Hence, we fed breeding pairs of C57BL/6J mice during gestation and lactation (perinatally) either chow or an OID rich in fat, fructose, and cholesterol (FFC). The offspring were weaned to either chow or an FFC diet, generating four groups: perinatal (p)Chow-Chow, pChow-FFC, pFFC-Chow, and pFFC-FFC. Mice were sacrificed at 10 weeks of age. We examined the whole-liver transcriptome by RNA sequencing (RNA-seq) and whole-liver genome methylation by reduced representation bisulfite sequencing (RRBS). Our results indicated that the pFFC-FFC mice had a significant increase in hepatic steatosis, injury, inflammation, and fibrosis, as assessed histologically and biochemically. We identified 189 genes that were differentially expressed and methylated in the pFFC-FFC mice versus the pChow-FFC mice. Gene set enrichment analysis identified hepatic fibrosis/hepatic stellate cell activation as the top canonical pathway, suggesting that the differential DNA methylation events in the mice exposed to the FFC diet perinatally were associated with a profibrogenic transcriptome. To verify that this finding was consistent with perinatal nutritional reprogramming of the methylome, we exposed pFFC-Chow mice to an FFC diet in adulthood. These mice developed significant hepatic steatosis, injury, inflammation, and more importantly fibrosis when compared to the appropriate controls. Conclusion: Perinatal exposure to an OID primes the immature liver for an accentuated fibrosing nonalcoholic steatohepatitis (NASH) phenotype, likely through nutritional reprogramming of the offspring methylome. These data have potential clinical implications for monitoring children of obese mothers and risk stratification of children with NAFLD.


Proteomic analysis of the regulatory networks of ClpX in a model cyanobacterium Synechocystis sp. PCC 6803.
Protein homeostasis is tightly regulated by protein quality control systems such as chaperones and proteases. In cyanobacteria, the ClpXP proteolytic complex is regarded as a representative proteolytic system and consists of a hexameric ATPase ClpX and a tetradecameric peptidase ClpP. However, the functions and molecular mechanisms of ClpX in cyanobacteria remain unclear. This study aimed to decipher the unique contributions and regulatory networks of ClpX in the model cyanobacterium Synechocystis sp. PCC 6803 (hereafter Synechocystis). We showed that the interruption of clpX led to slower growth, decreased high light tolerance, and impaired photosynthetic cyclic electron transfer. A quantitative proteomic strategy was employed to globally identify ClpX-regulated proteins in Synechocystis cells. In total, we identified 172 differentially expressed proteins (DEPs) upon the interruption of clpX. Functional analysis revealed that these DEPs are involved in diverse biological processes, including glycolysis, nitrogen assimilation, photosynthetic electron transport, ATP-binding cassette (ABC) transporters, and two-component signal transduction. The expression of 24 DEPs was confirmed by parallel reaction monitoring (PRM) analysis. In particular, many hypothetical or unknown proteins were found to be regulated by ClpX, providing new candidates for future functional studies on ClpX. Together, our study provides a comprehensive ClpX-regulated protein network, and the results serve as an important resource for understanding protein quality control systems in cyanobacteria.


Low force decelerates L-selectin dissociation from P-selectin glycoprotein ligand-1 and endoglycan.
Selectin-ligand interactions mediate the tethering and rolling of circulating leukocytes on vascular surfaces during inflammation and immune surveillance. To support rolling, these interactions are thought to have rapid off-rates that increase slowly as wall shear stress increases. However, the increase of off-rate with force, an intuitive characteristic named slip bonds, is at odds with a shear threshold requirement for selectin-mediated cell rolling. As shear drops below the threshold, fewer cells roll and those that do roll less stably and with higher velocity. We recently demonstrated a low force regime where the off-rate of P-selectin interacting with P-selectin glycoprotein ligand-1 (PSGL-1) decreased with increasing force. This counter-intuitive characteristic, named catch bonds, might partially explain the shear threshold phenomenon. Because L-selectin-mediated cell rolling exhibits a much more pronounced shear threshold, we used atomic force microscopy and flow chamber experiments to determine off-rates of L-selectin interacting with their physiological ligands and with an antibody. Catch bonds were observed at low forces for L-selectin-PSGL-1 interactions coinciding with the shear threshold range, whereas slip bonds were observed at higher forces. These catch-slip transitional bonds were also observed for L-selectin interacting with endoglycan, a newly identified PSGL-1-like ligand. By contrast, only slip bonds were observed for L-selectin-antibody interactions. These findings suggest that catch bonds contribute to the shear threshold for rolling and are a common characteristic of selectin-ligand interactions.


Pfmdr1 Asn1042Asp and Asp1246Tyr polymorphisms, thought to be associated with chloroquine resistance, are present in chloroquine-resistant and -sensitive Brazilian field isolates of Plasmodium falciparum.
Parasite resistance to antimalarial drugs, particularly chloroquine, is the most disturbing problem of malaria chemotherapy. There is evidence that the codon 86Tyr polymorphism of the Pfmdr1 gene is associated with chloroquine resistance in West African Plasmodium falciparum. The association of this and four other coding alterations of the Pfmdr1 gene with chloroquine resistance has not been extensively investigated in South American isolates. In this study, we examined 51 Brazilian P. falciparum isolates for the presence or absence of Asn86Tyr, Asn1042Asp, and Asp1246Tyr polymorphisms. While these isolates were all sensitive in vitro to mefloquine, amodiaquine, and quinine, only 2 (4%) were chloroquine-sensitive. The findings reported here provide the first observations of this kind on a large number of field parasite samples from South America. We show that in vitro chloroquine-resistant and -sensitive strains carry the Asn1042Asp and Asp1246Tyr polymorphisms and provide support for earlier suggestions that Asn86Tyr may be rare or absent in South American P. falciparum.


[Ultrasonography of the inferior vena cava (IVC) in the diagnosis and monitoring of therapy in patients with chronic congestive heart failure].
This prospective clinical study investigated whether the relatively simple and cost-effective ultrasonography of the inferior vena cava (IVC) represents a valid clinical tool to assess the effectiveness of diuretic therapy in patients with chronic congestive heart failure (CHF). Measurement of the resting and inspiratory diameter of the VCI repeatedly during diuretic therapy in 23 consecutive patients (11 women, 70 +/- 10 years) with CHF and comparing the results to the daily measured body weight and serum creatinine in these patients. Results were compared with the IVC diameter obtained in 33 healthy controls (16 women, 42 +/- 15 years). In addition, the IVC collapse index was calculated to assess inspiratory movements of the IVC in patients on day 1 and 10 of therapy. The IVC diameter at rest was 2.4 +/- 0.6 cm and decreased to 2.0 +/- 0.7 cm at inspiration, which was significantly greater than in healthy controls (1.4 +/- 0.4 cm at rest and 1.05 +/- 0.5 cm at inspiration; p = 0.008 and p = 0.01, respectively). The IVC diameter decreased continuously and significantly (p < 0.003) from day 1 to day 10 during diuretic therapy without a concomitant rise in serum creatinine. At beginning of therapy, the collapse-index of the IVC was significantly greater in patients than in controls. However, after 10 days of therapy this index reached similar values to those observed in controls. Ultrasonographic measurements of IVC diameter and inspiratory movements are a quantifiable and reliable approach to assess the hypervolemia associated with CHF. Normalization of inspiratory IVC collapse movement correlates with successful diuretic therapy and can be reliably used for bedside assessment and monitoring treatment in CHF patients.