Adsorption behavior of large plasmids on the anion-exchange methacrylate monolithic columns.
The objective of this study was to investigate the behavior of large plasmids on the monolithic columns under binding and nonbinding conditions. The pressure drop measurements under nonbinding conditions demonstrated that the flow velocities under which plasmid passing monolith became hindered by the monolithic pore structure depended on the plasmid size as well as on the average monolith pore size; however, they were all very high exceeding the values encountered when applying CIM monolithic columns at their maximal flow rate. The impact of the ligand density and the salt concentration in loading buffer on binding capacity of the monolith for different sized plasmids was examined. For all plasmids the increase of dynamic binding capacity with the increase of salt concentration in the loading solution was observed reaching maximum of 7.1 mg/mL at 0.4M NaCl for 21 kbp, 12.0 mg/mL at 0.4 M NaCl for 39.4 kbp and 8.4 mg/mL at 0.5M NaCl for 62.1 kbp. Analysis of the pressure drop data measured on the monolithic column during plasmid loading revealed different patterns of plasmid binding to the surface, showing "car-parking problem" phenomena under certain conditions. In addition, layer thickness of adsorbed plasmid was estimated and at maximal dynamic binding capacity it matched calculated plasmid radius of gyration. Finally, it was found that the adsorbed plasmid layer acts similarly as the grafted layer responding to changes in solution's ionic strength as well as mobile phase flow rate and that the density of plasmid layer depends on the plasmid size and also loading conditions.


Exposure to Adolescent Community Reinforcement Approach treatment procedures as a mediator of the relationship between adolescent substance abuse treatment retention and outcome.
Data from 399 adolescents, who participated in one of four randomly controlled trials of the Adolescent Community Reinforcement Approach (A-CRA) intervention, were used to examine the extent to which exposure to A-CRA procedures mediated the relationship between treatment retention and outcomes. Although zero-order correlations indicated that retention in treatment was a significant predictor of alcohol and other drug (AOD) use (r = -0.18, p < .001), this relationship was reduced to nonsignificance (p = .39) when exposure to A-CRA procedures was included in the model. Overall, the final model evidenced a very good fit (root mean square error of approximation = .00; comparative fit index = 1.00) and explained 29% and 43% of the variance in adolescents' posttreatment AOD use and AOD-related problems, respectively. In addition, Chi-Squared Automatic Interaction Detection analysis was used to derive a target level of A-CRA exposure, which was found to be significantly related to being in recovery at follow-up. The current findings are useful, as little research to date has identified significant mediators of the relationship between retention and treatment outcomes or identified target thresholds of treatment exposure.


A comparison of video laryngoscopy to direct laryngoscopy for the emergency intubation of trauma patients.
Direct laryngoscopy (DL) has long been the gold standard for tracheal intubation in emergency and trauma patients. Video laryngoscopy (VL) is increasingly used in many settings and the purpose of this study was to compare its effectiveness to direct laryngoscopy in trauma patients. Our hypothesis was that the success rate of VL would be higher than that of DL. Data were collected prospectively on all trauma patients, from January 2008 to June 2011, who were intubated emergently in an academic level I trauma center. After intubation, the physician that performed the intubation completed a structured data collection form that included demographics, complications, and the presence of difficult airway predictors. Our primary outcome measure was overall successful tracheal intubation, which was defined as successful intubation with the first device used. During the study period, 709 trauma patients were intubated by either VL or DL. VL was performed in 55% of cases. The overall success rate of VL was 88% compared to 83% with DL (P = 0.05). Cervical (C-Spine) immobilization was predictive of higher initial success with VL (87%) than with DL (80%) (P < 0.05). In multivariate regression analysis DL was associated with higher risk of intubation failure compared to VL (OR 1.82, CI: 1.15-2.86). In trauma patients intubated emergently, VL had a significantly higher success rate than DL. These data suggest that, in select circumstances, VL is superior to DL for the intubation of trauma patients with difficult airways.


Nrf2-mediated mucoprotective and anti-inflammatory actions of Artemisia extracts led to attenuate stress related mucosal damages.
The aim of this study was to compare biological actions between isopropanol and ethanol extracts of Artemisia including antioxidant, anti-inflammatory, and cytoprotective actions. Antioxidant activities were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) method and confocal microscopy on lipopolysaccharide-induced RGM1 cells, cytoprotection effects evaluated by detecting heme oxygenase-1 (HO-1), Nf-E2 related factor2 (Nrf2) and heat shock protein 70 (HSP70), and anti-inflammatory effects investigated by measuring inflammatory mediators. Water immersion restraint stress was imposed to provoke stress related mucosal damages (SRMD) in rats. Isopropanol extracts of Artemisia showed the higher DPPH radical scavenging activity and lesser LPS-induced reactive oxygen species productions and increased HO-1 expression through increased nuclear translocation of Nrf2 transcription factor compared to ethanol extracts. The increased expression of HSP70 and decreased expression of endothelin-1 were only increased with isopropanol extracts. A concentration-dependent inhibition of LPS-induced COX-2 and iNOS even at a rather lower concentration than ethanol extract was achieved with isopropanol extracts. Cytokine protein array revealed Artemisia extracts significantly attenuated the levels of CXCL-1, CXCL-16, and MCP-1. These orchestrated actions led to significant rescue from SRMD. Conclusively, Artemisia extracts imposed significant antioxidant and anti-inflammatory activity against SRMD and isopropanol extracts were superior to ethanol extracts in these beneficiary actions of Artemisia.


Direct label-free imaging of brain tissue using synchrotron light: a review of new spectroscopic tools for the modern neuroscientist.
The incidence of brain disease and brain disorders is increasing on a global scale. Unfortunately, development of new therapeutic strategies has not increased at the same rate, and brain diseases and brain disorders now inflict substantial health and economic impacts. A greater understanding of the fundamental neurochemistry that underlies healthy brain function, and the chemical pathways that manifest in brain damage or malfunction, are required to enable and accelerate therapeutic development. A previous limitation to the study of brain function and malfunction has been the limited number of techniques that provide both a wealth of biochemical information, and spatially resolved information (i.e., there was a previous lack of techniques that provided direct biochemical or elemental imaging at the cellular level). In recent times, a suite of direct spectroscopic imaging techniques, such as Fourier transform infrared spectroscopy (FTIR), X-ray fluorescence microscopy (XFM), and X-ray absorption spectroscopy (XAS) have been adapted, optimized and integrated into the field of neuroscience, to fill the above mentioned capability-gap. Advancements at synchrotron light sources, such as improved light intensity/flux, increased detector sensitivities and new capabilities of imaging/optics, has pushed the above suite of techniques beyond "proof-of-concept" studies, to routine application to study complex research problems in the field of neuroscience (and other scientific disciplines). This review examines several of the major advancements that have occurred over the last several years, with respect to FTIR, XFM and XAS capabilities at synchrotron facilities, and how the increases in technical capabilities have being integrated and used in the field of neuroscience.


Lens Opacities Classification System.
The Lens Opacities Classification System (LOCS) is a simple system for classifying age-related human lens opacities at the slit lamp or in retroilluminated and slit-lamp photographs. The system employs a set of standard Neitz CTR retroilluminated black-and-white photographs for classification of cortical and posterior subcapsular cataracts and a single color slit-lamp photograph for classification of nuclear color and opalescence. We present a detailed description of the system.


[Prevention of Infections of the Upper Respiratory Tract].
Because of its high prevalence acute respiratory diseases have a significant impact on the population. The focus of this review was the current state of knowledge for the prophylactic efficacy of: zinc, vitamin C, Echinacea preparations, garlic and carrying out physical measures. Furthermore, the benefits of pneumococcal and influenza vaccine were elicited. In the synopsis, the physical measures proved to be the most effective, cost-effective method to prevent infections. The intake of zinc, Echinacea preparations (for example: E. purpurea), vitamin C and garlic showed moderate success in the prevention of infection and must be elicited individually. Pneumococcal and annual influenza vaccines in family practice should be given furthermore accordingly topical STIKO-recommendation. Nevertheless, the prophylactic effect from influenza vaccines on usual cold illnesses is unsettled.


Worsening of contact dermatitis by oral hydroxyzine: a case report.
Hydroxyzine is commonly used to treat pruritic skin lesions. Although rare, hydroxyzine can sometimes be linked to worsening dermatitis in patients who have sensitivities to phenothiazines and/or ethylenediamines. Herein we describe a patient who developed papulovesicular eruptions following the use of topical neomycin. Our patient's contact dermatitis initially improved after the use of oral steroids. However, the patient's skin condition was exacerbated by the continued use of hydroxyzine to treat her pruritus. Patch testing was positive at 48 hours for neomycin sulfate, ethylenediamine dihydrochloride, and p-phenylenediamine. Given the suspected cross-reactivity between hydroxyzine and ethylenediamine, hydroxyzine was discontinued and the patient's cutaneous symptoms improved. In summary, physicians must be aware that oral hydroxyzine can worsen contact dermatitis in ethylenediamine-sensitive patients.


GaAs core--shell nanowires for photovoltaic applications.
We report the use of Te as an n-type dopant in GaAs core-shell p-n junction nanowires for use in photovoltaic devices. Te produced significant change in the morphology of GaAs nanowires grown by the vapor-liquid-solid process in a molecular beam epitaxy system. The increase in radial growth of nanowires due to the surfactant effect of Te had a significant impact on the operating characteristics of photovoltaic devices. A decrease in solar cell efficiency occurred when the Te-doped GaAs growth duration was increased.


Momordica Charantia lectin, a type II ribosome inactivating protein, exhibits antitumor activity toward human nasopharyngeal carcinoma cells in vitro and in vivo.
The incidence of nasopharyngeal carcinoma (NPC) remains high in endemic regions, including southern China, northern Africa, and North America. One of the promising therapeutic approaches on NPC is drug screening from natural products, such as components from traditional Chinese medicine. In this study, the antitumor activity of Momordica charantia lectin (MCL), a type II ribosome inactivating protein from bitter gourd, on NPC was investigated. MCL evinced potent cytotoxicity toward NPC CNE-1 (IC(50) = 6.9) and CNE-2 (IC(50) = 7.4) cells but minimally affected normal NP 69 cells. Further investigation disclosed that MCL induced apoptosis, DNA fragmentation, G(1)-phase arrest, and mitochondrial injury in both types of NPC cells. The reduction of cyclin D1 and phosphoretinoblastoma (Rb) protein expression contributed to arrest at G(1)-phase of the cell cycle. These events were associated with regulation of mitogen-activated protein kinases (MAPK; including p38 MAPK, JNK, and ERK) phosphorylation and promoted downstream nitric oxide (NO) production. Concurrent administration of the p38 MAPK inhibitor SB-203580 significantly diminished NO production and lethality of MCL toward NPC cells. Further studies revealed that MCL increased cytochrome c release into the cytosol, activated caspases-8, -9, and -3, and enhanced production of cleaved PARP, subsequently leading to DNA fragmentation and apoptosis. Finally, an intraperitoneal injection of MCL (1.0 mg/kg/d) led to an average of 45% remission of NPC xenograft tumors subcutaneously inoculated in nude mice. This is the first article that unveils the potential of a type II RIP, MCL, for prevention and therapy of NPC.


End of life after stroke: A nationwide study of 42,502 deaths occurring within a year after stroke.
In the scientific literature, there is very limited empirical information on end-of-life issues after stroke in the scientific literature. The present nationwide study describes the circumstances surrounding deaths that occur within a year after a stroke. Datasets from three nationwide Swedish registers (on stroke, palliative care and cause of death) were linked. Basic information was available for 42,502 unselected cases of death that occurred within a year after a stroke and more detailed information was available for 16,408 deaths. Odds ratios for characteristics of end-of-life care were calculated by logistic regression. In the late phase after stroke (three months to one year), 46% of patients died in a nursing home, whereas 37% of patients died in a hospital after readmission and 10% of patients died at home. Eleven per cent of deaths were reported as being unexpected. A next of kin was present at 49% of deaths. The frequency of unattended deaths (neither next of kin nor staff were present at the time of death) ranged from 5% at home with specialised home care to 25% in hospitals. This is, by far, the largest study published on end-of-life issues after stroke. Major differences between countries in healthcare, community services, family structure and culture may limit direct transfer of the present results to other settings. There is considerable discordance between presumed 'good death' late after stroke (dying at home surrounded by family members) and the actual circumstances at the end of life.


Endothelium-derived factors modulate contraction of bronchial smooth muscle.
Experiments were designed to determine how changes in the ratio of specific vascular endothelium-derived factors might affect reactivity of bronchial smooth muscle. The epithelium was mechanically removed from rings of third-order canine bronchi, and the rings were suspended in organ chambers for measurement of isometric force. Rings of pulmonary artery were cut; in some the endothelium was mechanically removed. The arterial rings were everted and pairs, with and without endothelium, were incubated with control solution, NG-monomethyl-L-arginine (L-NMMA), indomethacin, or both L-NMMA and indomethacin for 90 min. They were then inserted into the lumen of the bronchial rings. Cumulative concentration-response curves to acetylcholine and 5-hydroxytryptamine were obtained. There was no significant difference in response between bronchi containing arteries with or without endothelium incubated with control solution. Contractions of bronchi containing arteries without endothelium, incubated in either L-NMMA or indomethacin, increased from control. However, in rings containing arteries with endothelium, incubation with either inhibitor decreased bronchial contractions. Incubation with both inhibitors eliminated the difference. These results suggest that the vascular endothelium produces factors that can both contract and relax bronchial smooth muscle. With inhibition of production of either nitric oxide with L-NMMA or prostanoids with indomethacin, release of other endothelium-derived factors occurs that attenuates contractions of bronchial smooth muscle. Therefore, an imbalance in the ratio of production of endothelium-derived factors may contribute to bronchospastic disorders.


Sphingosine 1-phosphate regulation of extracellular signal-regulated kinase-1/2 in embryonic stem cells.
Recent evidence suggests that sphingosine 1-phosphate (S1P) regulates self-renewal of human embryonic stem (ES) cells and differentiation of mouse embryoid bodies (derived from mouse ES cells) to cardiomyocytes. We have investigated the role of S1P in regulating ERK-1/2 signaling in mouse ES cells. In this regard, we found that both mouse ES-D3 and CGR8 cells express S1P(1), S1P(2), S1P(3), and S1P(5) but lack S1P(4). The treatment of ES cells with S1P induced the activation of ERK-1/2 via a mechanism that was not mediated by S1P(1), S1P(2), or S1P(3). This was based on: (i) the failure of S1P(1), S1P(2), or S1P(3) antagonists to inhibit S1P-stimulated ERK-1/2 activation and (ii) the failure of SEW 2871 (S1P(1) receptor agonist) to stimulate ERK-1/2 activation. The treatment of ES cells with phytosphingosine 1-phosphate (phyto-S1P), which we show here is an agonist of the S1P(5) receptor, stimulated ERK-1/2 activation. These findings therefore suggest that S1P(5) may mediate the effects of S1P in terms of regulating ERK-1/2 signaling in ES cells. The S1P-dependent activation of ERK-1/2 was sensitive to inhibition by pertussis toxin (uncouples the G-protein, G(i) from GPCR), bisindolylmaleimide I (PKC inhibitor), and PP2 (c-Src inhibitor), but was not reduced by LY29004 (PI3K inhibitor) suggesting that S1P uses G(i)-, PKC-, and c-Src-dependent mechanisms to activate the ERK-1/2 pathway in ES cells.


Treatment for adult HIV infection: 2006 recommendations of the International AIDS Society--USA panel.
Guidelines for antiretroviral therapy are important for clinicians worldwide given the complexity of the field and the varied clinical situations in which these agents are used. The International AIDS Society-USA panel has updated its recommendations as warranted by new developments in the field. To provide physicians and other human immunodeficiency virus (HIV) clinicians with current recommendations for the use of antiretroviral therapy in HIV-infected adults in circumstances for which there is relatively unrestricted access to drugs and monitoring tools. The recommendations are centered on 4 key issues: when to start antiretroviral therapy; what to start; when to change; and what to change. Antiretroviral therapy in special circumstances is also described. A 16-member noncompensated panel was appointed, based on expertise in HIV research and patient care internationally. Data published or presented at selected scientific conferences from mid 2004 through May 2006 were identified and reviewed by all members of the panel. Data that might change previous guidelines were identified and reviewed. New guidelines were drafted by a writing committee and reviewed by the entire panel. Antiretroviral therapy in adults continues to evolve rapidly, making delivery of state-of-the-art care challenging. Initiation of therapy continues to be recommended in all symptomatic persons and in asymptomatic persons after the CD4 cell count falls below 350/microL and before it declines to 200/microL. A nonnucleoside reverse transcriptase inhibitor or a protease inhibitor boosted with low-dose ritonavir each combined with 2 nucleoside (or nucleotide) reverse transcriptase inhibitors is recommended with choice being based on the individual patient profile. Therapy should be changed when toxicity or intolerance mandate it or when treatment failure is documented. The virologic target for patients with treatment failure is now a plasma HIV-1 RNA level below 50 copies/mL. Adherence to antiretroviral therapy in the short-term and the long-term is crucial for treatment success and must be continually reinforced.


Specificity of amino acid transport in renal papilla: microinfusion of Henle's loops and vasa recta.
Amino acids can be reabsorbed distal to tips of Henle's loops and may be recycled between loops and vasa recta in rat papilla. Transport specificity was examined during continuous microinfusions of ascending Henle's loops and vasa recta with radiolabeled amino acids. Percent of recovered radiolabel as intact amino acid was also determined. Previous data indicated that, relative to simultaneously microinfused inulin, 30-40% of radiolabeled L- and D-Ala, L-Glu, L-Glu(NH2), and Gly, but no taurine (Tau) or mannitol, microinfused into Henle's loops was reabsorbed. In the present study, reabsorption was shown to involve intact L- and D-Ala, D-Glu, and L-Ser. L-Phe (50 mM) in infusate had no effect on reabsorption of L-Ala (2.5 mM) or L-Glu(NH2) (42.6 microM), and D-Asp (50 mM) had no effect on reabsorption of L-Glu (1.5 mM). Thus reabsorption from Henle's loops is not stereospecific, not different for neutral and acidic amino acids, and not inhibited by competitive inhibitors of proximal tubule amino acid transport, but it was not completely nonspecific and not a simple leak. Previous vasa recta microinfusions suggested that Ala could move directly from vasa recta to tubules. These studies were extended with simultaneous collections from ipsilateral and contralateral kidneys. Relative to simultaneously microinfused inulin, 40-50% of radiolabeled L- and D-Ala, L-Glu, and L-Glu(NH2) and 30% of L-Ser microinfused into ascending vasa recta appeared intact in urine from ipsilateral kidney, whereas only 1-3% appeared in urine from contralateral kidney. Fifty percent of infused D-Glu was excreted intact by each kidney; 70% of infused Tau was excreted intact by ipsilateral kidney, and 22% was excreted by contralateral kidney. L-Phe (50 mM) in infusate inhibited appearance of L-Ala (2.5 mM) and D-Ala (10 mM) but not L-Glu(NH2) (42.6 microM) in ipsilateral urine. D-Asp (50 mM) inhibited appearance of L-Glu (1.5 mM), and beta-Ala (50 mM) inhibited appearance of Tau (78 microM) in ipsilateral urine. Thus some amino acids can move directly from vasa recta into tubules (probably descending thin limbs of Henle's loops) by a process showing significant specificity.


Intimacy versus isolation: a qualitative study of sexual practices among sexually active HIV-infected patients in HIV care in Brazil, Thailand, and Zambia.
The success of global treatment as prevention (TasP) efforts for individuals living with HIV/AIDS (PLWHA) is dependent on successful implementation, and therefore the appropriate contribution of social and behavioral science to these efforts. Understanding the psychosocial context of condomless sex among PLWHA could shed light on effective points of intervention. HPTN 063 was an observational mixed-methods study of sexually active, in-care PLWHA in Thailand, Zambia, and Brazil as a foundation for integrating secondary HIV prevention into HIV treatment. From 2010-2012, 80 qualitative interviews were conducted with PLWHA receiving HIV care and reported recent sexual risk. Thirty men who have sex with women (MSW) and 30 women who have sex with men (WSM) participated in equal numbers across the sites. Thailand and Brazil also enrolled 20 biologically-born men who have sex with men (MSM). Part of the interview focused on the impact of HIV on sexual practices and relationships. Interviews were recorded, transcribed, translated into English and examined using qualitative descriptive analysis. The mean age was 25 (SD = 3.2). There were numerous similarities in experiences and attitudes between MSM, MSW and WSM across the three settings. Participants had a high degree of HIV transmission risk awareness and practiced some protective sexual behaviors such as reduced sexual activity, increased use of condoms, and external ejaculation. Themes related to risk behavior can be categorized according to struggles for intimacy and fears of isolation, including: fear of infecting a sex partner, guilt about sex, sexual communication difficulty, HIV-stigma, and worry about sexual partnerships. Emphasizing sexual health, intimacy and protective practices as components of nonjudgmental sex-positive secondary HIV prevention interventions is recommended. For in-care PLWHA, this approach has the potential to support TasP. The overlap of themes across groups and countries indicates that similar intervention content may be effective for a range of settings.


SU-E-J-96: Investigation into the Accuracy of Deformable Registration Algorithms for Use in Dose Summation: A Case Study in Combined Modalities.
To better understand the accuracy and limitation of deformable registration algorithms for dose summation by studying the results produced through two different deformable registration techniques for a clinical case of combining a Cyberknife radiosurgery plan and an electron boost to the scar tissues. A patient was treated with a Cyberknife plan with supine CT and an electron boost plan with prone CT concurrently. The CT, RT structures and dose grids were exported from the Cyberknife MultiPlan system and the Pinnacle system and imported into two commercial DICOM viewing systems equipped with deformable registration algorithms. One of the systems uses intensity-based free-form deformable registration while the other uses B-spline free-form deformable registration. The electron boost plan was first registered to the CK plan using rigid registration, then secondly using deformable registration in each system. The region of interest used for registration was chosen to encompass the whole CT volumes due to the supine vs. prone positions. The summation doses for the Cyberknife CTV and critical structures do not differ between rigid registration and deformable registration for both systems. The electron boost volume does show higher mean dose received for the deformable registration compared with rigid registration for both systems (12.90 Gy vs. 11.71 Gy and 12.39 Gy vs. 11.53 Gy). There are slight variations between the doses produced by the two systems for all the structures, with an averaged difference of approximately 0.02% to 2.63%. These results show that for cases like this one, where the two treatment volumes do not overlap, there will not be significant differences between rigid and deformable registration, and that the only significant difference in summation dose between the different deformable registration algorithms is where the volume is deformed the most, in this case, the electron boost volume.


Genome-wide expression analyses establish dendritic cells as a new osteoclast precursor able to generate bone-resorbing cells more efficiently than monocytes.
Dendritic cells (DCs), mononuclear cells that initiate immune responses, and osteoclasts (OCs), multinucleated bone-resorbing cells, are hematopoietic cells derived from monocytic precursor cells. Using in vitro generated dendritic cells, we previously showed that human and murine DCs could transdifferentiate into resorbing osteoclasts in the presence of macrophage colony-stimulating factor (M-CSF) and receptor activator of NF-kappaB ligand (RANKL). In this study we globally compared by transcriptomic profiling this new osteoclast differentiation pathway from DCs with the canonical differentiation pathway from monocytes. DNA chip data revealed that starting from two very distinct cell types, treatment with M-CSF and RANKL generated two highly similar types of osteoclast. In particular, DC-derived osteoclasts expressed all the characteristic marker genes of monocyte-derived osteoclasts. Two major molecular events could be observed during osteoclastogenesis: downregulation of a large set of monocyte or DC specific markers, together with upregulation of characteristic osteoclast marker genes. Most interestingly, our transcriptomic data showed a closer molecular profile between DCs and OCs than between monocytes and OCs. Our data establish DCs as a new osteoclast precursor able to generate OCs more efficiently than monocytes.


Susceptibility of raccoons (Procyon lotor) to infection with Mycobacterium bovis.
Tuberculosis due to Mycobacterium bovis infection is endemic in white-tailed deer (Odocoileus virginianus) in the northeastern portion of the lower Michigan peninsula (USA). Various wild carnivores and omnivores, including raccoons (Procyon lotor), are infected with M. bovis within the endemic area. To investigate the pathogenesis of tuberculosis in raccoons and the likelihood of M. bovis transmission from infected raccoons to other susceptible hosts, we experimentally inoculated raccoons with single oral doses of M. bovis (ranging from 30 to 1.7 x 10(5) colony forming units [CFU]), five daily oral doses of M. bovis (ranging from 10 to 1 x 10(5) CFU), or a single intravenous (i.v.) dose of 1 x 10(5) CFU of M. bovis, from November 1998 through December 2000. Granulomatous lesions consistent with tuberculosis, or tissue colonization with M. bovis, were seen in one of five raccoons in the single low oral dose group, one of five raccoons in the multiple low oral dose group, two of five raccoons in the multiple medium oral dose group, five of five raccoons in the multiple high oral dose group, and five of five raccoons in the i.v. inoculated group. In oral inoculated raccoons, lesions were most common in the tracheobronchial and mesenteric lymph nodes and lung. Excretion of M. bovis in saliva or nasal secretions was noted in all i.v. inoculated raccoons and two of five multiple low oral dose raccoons. Mycobacterium bovis was not isolated from urine or feces from any experimentally inoculated raccoons. The need for multiple large oral doses to establish infection, and the low number of orally inoculated raccoons that excreted M. bovis in nasal secretions or saliva, suggest that wide-spread tuberculosis among raccoons is unlikely.


Newborn screening for high-risk human leukocyte antigen markers associated with insulin-dependent diabetes mellitus: the ABIS study.
Type 1 (insulin-dependent) diabetes mellitus is associated with specific high-risk HLA DQ and DR haplotypes and islet cell antibodies. IDDM susceptibility in Caucasians is more strongly associated with DQ2/DQ8 (DQA1*0501-DQB1*0201/DQA1*0301-DQB1*0302) and DQ6 (B1*0604) than with DRB1*03/DRB1*04, while a single copy of DQ6 (B1*0602) gives sufficient protection against type 1 diabetes. As a part of the ABIS (All Babies in Southeast Sweden) study we have done typing of DQA1, DQB1, and DRB1 by polymerase chain reaction (PCR) amplification of the second exon of the genes, manually dot-blotting onto nylon membranes synthetic sequence-specific oligonucleotide (SSO) probes, 3' end-labeling with (32)P-dCTP, and hybridization followed by stringency washes and autoradiography using the SSO probe. Among 3756 newborns born in southeast Sweden we have found the high-risk genotype DQ2/DR3-DO8/DR4 to be present in 1%, haplotype DQ8/DR4 in 7.8%, and haplotype DQ2/DR3 in 9.6%. DQ2/DR3 or DQ8/DR4 was carried by 16.4% of newborns; the low-risk DQ6 molecule was carried by newborns as follows: DQ2/DR3-DQ6/DR15, 1.3%; DQ8/DR4-DQ6/DR15, 1.3%; and DQ6/DR15, 9.4%. We conclude from our results that the high incidence of IDDM in Sweden is at least in part due to increased prevalence of high-risk HLA haplotypes compared to protective haplotypes (20% vs. 13%) in the general population.


Association of neonatal serum immunoglobulin G1 concentration with health and performance in beef calves.
To evaluate associations between neonatal serum IgG1 concentration and pre- and postweaning morbidity and mortality rates and average daily gains (ADGs) in beef calves and define a cutoff point for serum IgG1 concentration necessary for optimal health and performance of beef calves. Nonconcurrent cohort study. 1,568 crossbred beef calves. Single radial immunodiffusion was used to quantitate IgG1 concentration in sera collected from calves between 24 and 72 hours after birth. Logistic regression, ANCOVA, and likelihood ratios were used to analyze data. In the preweaning period, lower perinatal IgG1 concentrations were significantly associated with higher morbidity rates, higher mortality rates, and lower ADGs. Calves with serum IgG1 concentration < 2,400 mg/dL were 1.6 times as likely to become ill before weaning and 2.7 times as likely to die before weaning as calves with higher serum IgG1 concentrations. Calves with serum IgG1 concentration of at least 2,700 mg/dL weighed an estimated 3.35 kg (7.38 lb) more at 205 days of age than calves with lower serum IgG1 concentration. No significant association of serum IgG1 concentration with feedlot morbidity, death, or ADG was identified. By use of likelihood ratios, the threshold of serum IgG1 concentration for optimal health and performance of calves was higher than values reported previously. Implementation and maintenance of management and intervention strategies designed for early detection and treatment of calves at risk for failure of passive transfer will likely result in increases in preweaning health and performance parameters.


Analytical method for total chromium and nickel in urine using an inductively coupled plasma-universal cell technology-mass spectrometer (ICP-UCT-MS) in kinetic energy discrimination (KED) mode.
Biomonitoring and emergency response measurements are an important aspect of the Division of Laboratory Sciences of the National Center for Environmental Health, Centers for Disease Control and Prevention (CDC). The continuing advancement in instrumentation allows for enhancements to existing analytical methods. Prior to this work, chromium and nickel were analyzed on a sector field inductively coupled plasma-mass spectrometer (SF-ICP-MS). This type of instrumentation provides the necessary sensitivity, selectivity, accuracy, and precision but due to the higher complexity of instrumentation and operation, it is not preferred for routine high throughput biomonitoring needs. Instead a quadrupole based method has been developed on a PerkinElmer NexION™ 300D ICP-MS. The instrument is operated using 6.0 mL min-1 helium as the collision cell gas and in kinetic energy discrimination mode, interferences are successfully removed for the analysis of 52Cr (40Ar12C and 35Cl16O1H) and 60Ni (44Ca16O). The limits of detection are 0.162 μg L-1 Cr and 0.248 μg L-1 Ni. Method accuracy using NIST SRM 2668 level 1 (1.08 μg L-1 Cr and 2.31μg L-1 Ni) and level 2 (27.7 μg L-1 Cr and 115 μg L-1 Ni) was within the 95% confidence intervals reported in the NIST certificate. Among-run precision is less than 10% RSDs (N = 20) for in house quality control and NIST SRM urine samples. While the limits of detection (LOD) for the new quadrupole ICP-UCT-MS with KED method are similar to the SF-ICP-MS method, better measurement precision is observed for the quadrupole method. The new method presented provides fast, accurate, and more precise results on a less complex and more robust ICP-MS platform.


Infliximab therapy downregulation of basic fibroblast growth factor/syndecan 1 link: a possible molecular pathway of mucosal healing in ulcerative colitis.
It is known that syndecan 1 in inflammatory bowel diseases is able to migrate from epithelial basolateral site to the stromal area and apical surface of epithelium with a consequent activation and modulation of basic fibroblast growth factor (bFGF), and this process sustains mucosal healing of ulcers. On the other hand, tumour necrosis factor (TNF) α mucosal levels are directly related to the entity of the damage in these disorders. Aim of the study A 'post-hoc' retrospective study was performed to estimate mucosal TNF α in rectal biopsies of subjects with ulcerative colitis (UC) before and after effective infliximab therapy and its relationship with syndecan 1, bFGF and endoscopic mucosal healing. Paraffin-embedded rectal samples from 12 patients with UC responders to infliximab were analysed for TNF α, syndecan 1 and bFGF before and 6 months after therapy using a real-time reverse transcriptase polymersase chain reaction. Additionally, syndecan 1 location was evaluated by immunohistochemistry. Samples from 12 subjects with irritable bowel symptoms without endoscopic/histological abnormalities represented the control group. Mucosal healing induced by the treatment was defined by an endoscopic Mayo subscore changing from 2-3 to 0. ANOVA plus Student-Newman-Keuls was used for statistical analysis. The authors found that in the active disease, an increase in TNF α (p<0.001) is accompanied by raised levels of both syndecan 1 (p<0.005) and bFGF (p<0.005) compared with the control group. Infliximab-induced TNF α decrease to levels similar to controls is associated with both endoscopic mucosal healing and adhesion molecule/growth factor significant reduction. Additionally, syndecan 1 location, which is predominant in the stromal cells and apical epithelium in the active disorder, is quite exclusively located at the basolateral epithelial area in both healed mucosa and controls. Balanced interaction among TNF α inhibition by infliximab, syndecan 1 migration, bFGF repair modulation and final adhesion molecule reversal to its normal location might represent a suitable molecular pathway of endoscopic mucosal healing in UC.


Biological Functions and Current Advances in Isolation and Detection Strategies for Exosome Nanovesicles.
Exosomes are nanoscale (≈30-150 nm) extracellular vesicles of endocytic origin that are shed by most types of cells and circulate in bodily fluids. Exosomes carry a specific composition of proteins, lipids, RNA, and DNA and can work as cargo to transfer this information to recipient cells. Recent studies on exosomes have shown that they play an important role in various biological processes, such as intercellular signaling, coagulation, inflammation, and cellular homeostasis. These functional roles are attributed to their ability to transfer RNA, proteins, enzymes, and lipids, thereby affecting the physiological and pathological conditions in various diseases, including cancer and neurodegenerative, infectious, and autoimmune diseases (e.g., cancer initiation, progression, and metastasis). Due to these unique characteristics, exosomes are considered promising biomarkers for the diagnosis and prognosis of various diseases via noninvasive or minimally invasive procedures. Over the last decade, a plethora of methodologies have been developed for analyzing disease-specific exosomes using optical and nonoptical tools. Here, the major biological functions, significance, and potential role of exosomes as biomarkers and therapeutics are discussed. Furthermore, an overview of the most commonly used techniques for exosome analysis, highlighting the major technical challenges and limitations of existing techniques, is presented.


[Breast self-examination by nursing professionals].
Most breast tumors are detected by women, not health professionals, therefore breast self-examination (BSE) continues to be an efficacious strategy. The study objective was to analyze BSE performed by nursing professionals and factors that hinder their perseverance in this practice. This descriptive study was performed with 159 professionals: 40 nurses, 48 nurses aides, and 71 health agents from 19 Basic Health Units located in Fortaleza (Ceará, Brazil). Data collection was performed using a self-administered questionnaire, and the analysis was based on the Self Care Theory. Of the 159 professionals, 86 (54%) performed BSE on a monthly basis. Of the 73 professionals who did not perform BSE, 60 (82%) reported the reason for that was forgetting, 38 (52%) for not trusting the technique/did not know the correct technique, and 35 (48%) due to lack of health care. It was found that although most women reported performing BSE, the professionals felt insecure and would like to learn better about this technique.


Switching HIV treatment in adults based on CD4 count versus viral load monitoring: a randomized, non-inferiority trial in Thailand.
Viral load (VL) is recommended for monitoring the response to highly active antiretroviral therapy (HAART) but is not routinely available in most low- and middle-income countries. The purpose of the study was to determine whether a CD4-based monitoring and switching strategy would provide a similar clinical outcome compared to the standard VL-based strategy in Thailand. The Programs for HIV Prevention and Treatment (PHPT-3) non-inferiority randomized clinical trial compared a treatment switching strategy based on CD4-only (CD4) monitoring versus viral-load (VL). Consenting participants were antiretroviral-naïve HIV-infected adults (CD4 count 50-250/mm(3)) initiating non-nucleotide reverse transcriptase inhibitor (NNRTI)-based therapy. Randomization, stratified by site (21 public hospitals), was performed centrally after enrollment. Clinicians were unaware of the VL values of patients randomized to the CD4 arm. Participants switched to second-line combination with confirmed CD4 decline >30% from peak (within 200 cells from baseline) in the CD4 arm, or confirmed VL >400 copies/ml in the VL arm. Primary endpoint was clinical failure at 3 years, defined as death, new AIDS-defining event, or CD4 <50 cells/mm(3). The 3-year Kaplan-Meier cumulative risks of clinical failure were compared for non-inferiority with a margin of 7.4%. In the intent to treat analysis, data were censored at the date of death or at last visit. The secondary endpoints were difference in future-drug-option (FDO) score, a measure of resistance profiles, virologic and immunologic responses, and the safety and tolerance of HAART. 716 participants were randomized, 356 to VL monitoring and 360 to CD4 monitoring. At 3 years, 319 participants (90%) in VL and 326 (91%) in CD4 were alive and on follow-up. The cumulative risk of clinical failure was 8.0% (95% CI 5.6-11.4) in VL versus 7.4% (5.1-10.7) in CD4, and the upper-limit of the one-sided 95% CI of the difference was 3.4%, meeting the pre-determined non-inferiority criterion. Probability of switch for study criteria was 5.2% (3.2-8.4) in VL versus 7.5% (5.0-11.1) in CD4 (p=0.097). Median time from treatment initiation to switch was 11.7 months (7.7-19.4) in VL and 24.7 months (15.9-35.0) in CD4 (p=0.001). The median duration of viremia >400 copies/ml at switch was 7.2 months (5.8-8.0) in VL versus 15.8 months (8.5-20.4) in CD4 (p=0.002). FDO scores were not significantly different at time of switch. No adverse events related to the monitoring strategy were reported. The 3-year rates of clinical failure and loss of treatment options did not differ between strategies although the longer-term consequences of CD4 monitoring would need to be investigated. These results provide reassurance to treatment programs currently based on CD4 monitoring as VL measurement becomes more affordable and feasible in resource-limited settings. ClinicalTrials.govNCT00162682 Please see later in the article for the Editors' Summary.


Secondary structure models for the internal transcribed spacer (ITS) region 1 from symbiotic dinoflagellates.
Ribosomal genes and their spacers have been extensively utilized to examine the biodiversity and phylogenetics of protists. Among these, the internal transcribed spacer regions 1 and 2 (ITS1 and ITS2) are known to form secondary structures that are critically important for proper processing of the pre-rRNA into mature ribosomes. Although the secondary structure of ITS2 has been widely investigated, considerably less is known about ITS1 and its secondary structure. Here, secondary structures of the ITS1 were modeled for 46 ITS "types" from Symbiodinium, a diverse dinoflagellate genus that forms symbioses with many protists and metazoans, using comparative phylogenetic and minimum free energy approaches. The predicted ITS1 secondary structures for each Symbiodinium "type" were highly stable (DeltaG=-46.40 to -85.30 kcal mol(-1) at 37 degrees C) and consisted of an open loop with five helices separated by single-stranded regions. Several structural characteristics were conserved within monophyletic sub-groups, providing additional support for the predicted structures and the relationships within this genus. Finally, the structures were applied to identify potential pseudogenes from five Symbiodinium ITS1 datasets. Consequently, ITS1 secondary structures are useful in understanding the biology and phylogenetics, as well as recognizing and excluding questionable sequences from datasets, of protists such as Symbiodinium.


The effect of Tc-99m-labeled MDM2 antisense oligonucleotide on gene expression in human breast cancer MCF-7 cells.
To investigate the effect of radiolabed mouse double minute 2 (MDM2) antisense oligonucleotide on gene expression in human breast cancer MCF-7 cells, an antisense oligonucleotide (ASON) targeting MDM2 mRNA was synthesized and radiolabeled with 99Tcm. The labeling efficiency, radiochemical purity, and the ability of labeled ASON to hybridize to the sense oligonucleotides (SON) were investigated. To study whether the antisense probe hybridizes to respective sequence on MDM2 mRNA strand after radiolabeling, cells were incubated with radiolabeling oligonucleotides antisense oligonucleotide (0, 100, 500 nm/L) or mismatch oligonucleotide (ASONM) (500 nm/L) for 24 h, in the presence of Lipofectin 2000. RT-PCR and Western blotting was carried out to measure the MDM2 mRNA and protein levels. The antisense oligonucleotide was radiolabeled with the bifunctional chelator HYNIC at the labeling efficiency of 57.2 +/- 2.98% (n = 5) and the mismatch oligonucleotide was 56.3 +/- 3.01% (n = 5). The radiochemical purity was above 95% and labeled antisense oligonucleotide has the ability to hybridize to the sense oligonucleotide. The levels of mRNA and protein have significant differences in different concentration groups. The oligonucleotide can be successfully radiolabeled, and specially hybridized to the MDM2 mRNA and inhibit gene expression intensively as compared to mismatch oligonucleotide. This method will be very useful in the in vivo investigation of tumor targeting.


Surface Effect on 2D Hybrid Perovskite Crystals: Perovskites Using an Ethanolamine Organic Layer as an Example.
Despite the remarkable progress of optoelectronic devices based on hybrid perovskites, there are significant drawbacks, which have largely hindered their development as an alternative of silicon. For instance, hybrid perovskites are well-known to suffer from moisture instability which leads to surface degradation. Nonetheless, the dependence of the surface effect on the moisture stability and optoelectronic properties of hybrid perovskites has not been fully investigated. In this work, the influence of the surface effect of 2D layered perovskites before and after mechanical exfoliation, representing rough and smooth surfaces of perovskite crystals, are studied. It is found that the smooth 2D perovskite is less sensitive to ambient moisture and exhibits a considerably low dark current, which outperforms the rough perovskites by 23.6 times in terms of photodetectivity. The superior moisture stability of the smooth perovskites over the rough perovskites is demonstrated. Additionally, ethanolamine is employed as an organic linker of the 2D layered perovskite, which further improves the moisture stability. This work reveals the strong dependence of the surface conditions of 2D hybrid perovskite crystals on their moisture stability and optoelectronic properties, which are of utmost importance to the design of practical optoelectronic devices based on hybrid perovskite crystals.


Examining the role of positive and negative affect in recovery from spine surgery.
Consistent evidence supports a significant association between lower positive affect and higher negative affect and increased pain and disability in adults with chronic pain. However, examining this relation in surgical populations has received little empirical consideration. The primary purpose of this study was to determine whether preoperative and postoperative positive and negative affect predict pain, disability, and functional status after spine surgery. A secondary objective was to assess the relation of depression to postoperative outcomes compared with positive and negative affect. Participants were 141 patients treated by spine surgery for lumbar or cervical degeneration. Data collection occurred at baseline and 6 weeks and 3 months postoperatively. Affect was measured with the Positive and Negative Affect Schedule. Multivariable mixed-model linear regression analyses found that preoperative variables were not predictive of postoperative pain, disability and functional status. However, multivariable postoperative analysis found that 6-week positive affect predicted functional status, and 6-week negative affect predicted pain interference and pain-related disability at 3 months following surgery. Postoperative depression demonstrated statistically significant and stronger associations with pain intensity, pain interference, and pain-related disability at 3-month follow-up, as compared with negative affect. Results suggest that positive affect and depression are important variables to target when seeking to improve postoperative outcomes in a spine surgery population. Recommendations include postoperative screening for positive affect and depression, and treating depression as well as focusing on rehabilitation strategies to bolster positive affect so as to improve functional outcomes after spine surgery.


Hypersensitivity reaction to human papillomavirus vaccine due to polysorbate 80.
A 17-year-old girl reported generalised urticaria, eyelid angioedema, rhino-conjunctivitis, dyspnoea and wheezing 1 h after third intramuscular administration of quadrivalent human papilloma virus vaccine (Gardasil). She was treated with antihistamine, and corticosteroids with prompt relief of rhinitis and dyspnoea, while urticaria and angioedema lasted 24 h. Intradermal test with Gardasil, which contains polysorbate 80 (PS80), resulted positive, while skin tests with the bivalent vaccine were negative. Prick test performed with PS80 resulted positive in the patient and negative in ten healthy controls. The CD203 basophil activation test result was negative for PS80 at all the tested dilutions and specific IgE was not found. As flu vaccine was recommended, the authors skin tested two flu vaccine, one containing PS80 (Fluarix, GSK), which resulted positive and another flu vaccine with no adjuvant or preservative (Vaxigrip, Sanofi Pasteur MSD), which gave negative results. The patient then received Vaxigrip without adverse reactions.


Vitamin D metabolites regulate osteocalcin synthesis and proliferation of human bone cells in vitro.
The effects of six natural vitamin D metabolites of potential biological and therapeutic interest, 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3), 25-hydroxyvitamin D3 (25-OH-D3), 24R,25-dihydroxyvitamin D3 (24R,25-(OH)2D3), 1,24R,25-trihydroxyvitamin D3 (1,24R,25-(OH)3D3), 25S,26-dihydroxyvitamin D3 (25S,26-(OH)2D3) and 1,25S,26-trihydroxyvitamin D3 (1,25S,26-(OH)3D3) on cell replication and expression of the osteoblastic phenotype in terms of osteocalcin production were examined in cultured human bone cells. At a dose of 5 X 10(-12) mol/1, 1,25-(OH)2D3 stimulated cell proliferation, whereas at higher doses (5 X 10(-9)-5 X 10(-6) mol/1) cell growth was inhibited in a dose-dependent manner. The same pattern of effects was seen for the other metabolites in a rank order of potency: 1,25-(OH)2D3 greater than 1,25S,26-(OH)3D3 = 1,24R,25-(OH)3D3 greater than 25S,26-(OH)2D3 = 24R,25-(OH)2D3 = 25-OH-D3. Synthesis of osteocalcin was induced by 1,25-(OH)2D3 in doses similar to those required to inhibit cell proliferation. Biphasic responses were observed for some of the metabolites in terms of osteocalcin synthesis, inhibitory effects becoming apparent at 5 X 10(-6) mol/1. The cells did not secrete osteocalcin spontaneously. These results indicate that vitamin D metabolites may regulate growth and expression of differentiated functions of normal human osteoblasts.


Computer-navigated versus conventional total knee arthroplasty a prospective randomized trial.
The literature lacks studies that confirm whether the improved radiographic alignment that can be achieved with computer-navigated total knee arthroplasty improves patients' activities of daily living or the durability of total knee prostheses. The purpose of this study was to determine whether computer-navigated total knee arthroplasty improves the clinical function, alignment, and survivorship of the components. We prospectively compared the results of 520 patients with osteoarthritis who underwent computer-navigated total knee arthroplasty for one knee and conventional total knee arthroplasty for the other. The assignment of the knee to navigation or not was done randomly. There were 452 women (904 knees) and sixty-eight men (136 knees) with a mean age of sixty-eight years (range, forty-nine to eighty-eight years) at the time of the index arthroplasty. The mean follow-up period was 10.8 years (range, ten to twelve years). The patients were assessed clinically and radiographically with the rating system of the Knee Society and with the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score at three months, one year, and annually thereafter. Total knee scores, knee function scores, pain scores, WOMAC scores, knee motion, and activity scores did not show statistically significant differences between the two groups preoperatively or at the time of the final follow-up. Alignment and the survivorship of the components were not significantly different between the two groups. The Kaplan-Meier survivorship with revision as the end point at 10.8 years was 98.8% (95% confidence interval [CI], 0.96 to 1.00) in the computernavigated total knee arthroplasty group and 99.2% (95% CI, 0.96 to 1.00) in the conventional total knee arthroplasty group. Our data demonstrated no difference in clinical function or alignment and survivorship of the components between the knees that underwent computer-navigated total knee arthroplasty and those that underwent conventional total knee arthroplasty.


Assessment of the effect of pulsed electromagnetic field application on the healing of bone defects in rats with heparin-induced osteoporosis.
Osteoporosis is a systemic skeletal disease characterized by an increase in bone fragility and fracture risk due to low bone mass and deterioration of bone tissue. Application of pulsed electromagnetic fields (PEMF), a non-invasive method with a low complication risk, is known to stimulate bone formation. The present study examines the histomorphometric and biochemical effects of PEMF application on the healing of bone defects in rats with heparin-induced secondary osteoporosis. Briefly, 12-month-old male Sprague-Dawley rats were examined in a prospective, randomized, single-blind study. Osteoporosis was induced by administering a daily dose of 2 IU/g heparin for 33 days. Bone defects were created on the right femur on Day 35. PEMF of an average intensity of 0.8 ± 0.2 mT and a frequency of 7.3 Hz, was applied for 1 h/day, for 28 days following surgery. Bone healing was evaluated by histomorphometric and biochemical analyses. The heparin + PEMF group displayed the largest amount of new bone area (P = .002) and the lowest mean CTx on Day 63 (P = .05). This study demonstrates that heparin administration leads to bone loss and osteoporosis, whereas the application of PEMF decreases this effect.


Meta-omic analyses of Baltic Sea cyanobacteria: diversity, community structure and salt acclimation.
Cyanobacteria are important phytoplankton in the Baltic Sea, an estuarine-like environment with pronounced north to south gradients in salinity and nutrient concentrations. Here, we present a metagenomic and -transcriptomic survey, with subsequent analyses targeting the genetic identity, phylogenetic diversity, and spatial distribution of Baltic Sea cyanobacteria. The cyanobacterial community constituted close to 12% of the microbial population sampled during a pre-bloom period (June-July 2009). The community was dominated by unicellular picocyanobacteria, specifically a few highly abundant taxa (Synechococcus and Cyanobium) with a long tail of low abundance representatives, and local peaks of bloom-forming heterocystous taxa. Cyanobacteria in the Baltic Sea differed genetically from those in adjacent limnic and marine waters as well as from cultivated and sequenced picocyanobacterial strains. Diversity peaked at brackish salinities 3.5-16 psu, with low N:P ratios. A shift in community composition from brackish to marine strains was accompanied by a change in the repertoire and expression of genes involved in salt acclimation. Overall, the pre-bloom cyanobacterial population was more genetically diverse, widespread and abundant than previously documented, with unicellular picocyanobacteria being the most abundant clade along the entire Baltic Sea salinity gradient.


Phase behaviour and structure of a model biomolecular condensate.
Phase separation of immiscible fluids is a common phenomenon in polymer chemistry, and is recognized as an important mechanism by which cells compartmentalize their biochemical reactions. Biomolecular condensates are condensed fluid droplets in cells that form by liquid-liquid phase separation of intrinsically-disordered proteins. They have a wide range of functions and are associated with chronic neurodegenerative diseases in which they become pathologically rigid. However, it remains unclear how their material properties depend on the molecular structure of the proteins. Here we explore the phase behaviour and structure of a model biomolecular condensate composed of semi-flexible polymers with attractive end-caps using coarse-grained simulations. The model contains the minimal molecular features that are sufficient to observe liquid-liquid phase separation of soluble polymers into a porous, three-dimensional network in which their end-caps reversibly bind at junctions. The distance between connected junctions scales with the polymer length as a self-avoiding random walk over a wide range of concentration with a weak affinity-dependent prefactor. By contrast, the average number of polymers that meet at the junctions depends on the end-cap affinity but only weakly on the polymer length. The structured porosity of the condensed phase suggests a mechanism for cells to regulate biomolecular condensates. Protein interaction sites may be turned on or off to modulate the condensate's porosity and therefore the diffusion and interaction of additional proteins.


Expression of rat DNA (cytosine-5) methyltransferase (DNA MTase) in rodent trophoblast giant cells: molecular cloning and characterization of rat DNA MTase.
Methylation of genomic DNA is involved in the basic methanism of gene inactivation, chromatin organization, X chromosome inactivation and genomic imprinting. A pattern of DNA methylation is maintained in mitotic cells by DNA (cytosine-5) methytransferase (DNA MTase). The DNA MTase has been shown to be also expressed in postmitotic cells such as neurons. In the present report, as an approch to analyzing mechanisms underlying regulation of DNA MTase expression, we first isolated rat DNA MTase cDNA. The isolated cDNA encoded a protein of 1,622 amino acid residues showing 88.3% and 64.2% of homology with mouse and human DNA MTase, respectively. Northern blot analysis showed that DNA MTase mRNA was highly expressed in placenta during mid- to late- pregnancy. We then analyzed the expression of DNA MTase in Rcho-1 cells, a rat choriocarcinoma-derived cell line, which cease cell division but keep replicating genomic DNA when differentiated in vitro. We found that the expression of DNA MTase protein was decreased in terminally differentiated Rcho-1 cells whereas DNA MTase mRNA was consistently expressed. This result suggested posttranscriptional regulation of DNA MTase activity in Rcho-1 cells. The Rcho-1 cells would be a valuable model for studying the regulation of gene expression and function of DNA MTase in postmitotic, differentiated cells.


Serum concentration comparisons of amino acids, fatty acids, lipoproteins, vitamins A and E, and minerals between zoo and free-ranging giraffes (Giraffa camelopardalis).
Serum concentrations of amino acids, fatty acids, lipoproteins, vitamins A and E, and minerals in zoo giraffes (Giraffa camelopardalis) were compared to values obtained from free-ranging giraffes in an effort to identify potential nutritional differences in the zoo population. Zoo giraffes have a specific set of maladies that may be nutritionally related, including peracute mortality, energy malnutrition, pancreatic disease, urolithiasis, hoof disease, and severe intestinal parasitism. Dietary requirements for giraffes are not known; invasive studies used with domestic animals cannot be performed on zoo animals. Though domestic animal standards are often used to evaluate nutritional health of exotic animals, they may not be the most appropriate standards to use. Serum samples from 20 zoo giraffes at 10 zoological institutions in the United States were compared to previously collected samples from 24 free-ranging giraffes in South Africa. Thirteen of the zoo animal samples were collected from animals trained for blood collection, and seven were banked samples obtained from a previous serum collection. Dietary information was also collected on each zoo giraffe; most zoo giraffe diets consisted of alfalfa-based pellets (acid detergent fiber-16), alfalfa hay, and browse in varying quantities. Differences between zoo and free-ranging giraffes, males and females, and adults and subadults were analyzed with the use of a 2 x 2 x 2 factorial and Fisher's Least Significant Difference (LSD) for mean separation. Of the 84 parameters measured, 54 (60%) were significantly different (P < or = 0.05) between zoo and free-ranging giraffes. Nine (11%) items were significantly different (P < or = 0.05) between adult and subadult animals. Only one parameter, sodium concentration, was found to be significantly different (P < or = 0.05) between genders. Further investigation in zoo giraffe diets is needed to address the differences seen in this study and the potentially related health problems.


The NIDA Methamphetamine Clinical Trials Group: a strategy to increase clinical trials research capacity.
In order to increase the number of investigative teams and sites conducting research on pharmacological treatments for methamphetamine use disorders, the National Institute on Drug Abuse (NIDA) established an infrastructure of clinical sites in areas where methamphetamine addiction is prevalent. This multi-site infrastructure would serve to run multiple Phases II and III protocols effectively and expeditiously. NIDA collaborated with investigators from the University of California at Los Angeles (UCLA) to set up the Methamphetamine Clinical Trials Group (MCTG). This paper describes the development process, as well as data from a test trial to assess the capability of research-naive sites to recruit research participants and conduct study procedures according to research protocol. Subsequent trials are also described. A total of 151 candidates signed consent; 65 individuals were enrolled and 35 (53.8%) completed the 12 weeks' behavioral trial. Self-reported substance use report (SUR) showed comparable use of methamphetamine across sites with the individual site means ranging from 59% (site 5) to 80% (site 3). Drug use as measured by urinalysis was greatly reduced at week 13 compared to the baseline measure; the average rate of methamphetamine-free urine samples across all participants in sites at week 13 was 53%. The highest percentage of methamphetamine-free samples was 85% at site 5; the lowest was at site 1 (40%). Addiction severity index (ASI) composite scores at baseline and protocol completion for all participants demonstrated improvement in all categories over time, except for the medical composite score. The largest composite score reduction in baseline-protocol completion was in the drug domain (0.23 versus 0.15). The changes in the ASI scores from baseline to week 13 were consistent across all five sites. Outcomes of the behavioral trial indicated that the MCTG recruited well; collected study data accurately and reliably; and created a vehicle that can assess promising pharmacotherapies for methamphetamine addiction treatment medications. The MCTG strategy appears to be a feasible approach to increase NIDA's capacity to conduct clinical trials to evaluate potential pharmacotherapies for methamphetamine addiction.


Long-term impact of a preclinical endovascular skills course on medical student career choices.
Surging interest in the 0 + 5 integrated vascular surgery (VS) residency and successful recruitment of the top students in medical school requires early exposure to the field. We sought to determine the impact of a high-fidelity simulation-based preclinical endovascular skills course on medical student performance and ultimate career specialty choices. Fifty-two preclinical medical students enrolled in an 8-week VS elective course from 2007 to 2009. Students completed a baseline and postcourse survey and performed a renal angioplasty/stent procedure on an endovascular simulator (pretest). A curriculum consisting of didactic teaching covering peripheral vascular disease and weekly mentored simulator sessions concluded with a final graded procedure (posttest). Long-term follow-up surveys 1 to 3 years after course completion were administered to determine ultimate career paths of participants as well as motivating factors for career choice. Objective and subjective performance measured on the simulator and through structured global assessment scales improved in all students from pre- to posttest, particularly with regard to technical skill and overall procedural competency (P < .001). Prior to enrolling in the course, 9% of the students expressed high interest in VS, and after completing the course, this response nearly tripled in terms of seriously considering VS as a career option (P = .03). Overall interest postcourse in VS and procedural-based surgical specialties was nearly 90%. In long-term follow-up, 25% were still strongly considering integrated VS residencies, with other top career choices including surgical subspecialties (64%), radiology (10%), and cardiology (6%). Most respondents indicated major reasons for continued interest in VS were the ability to practice endovascular procedures on the simulator (92%) and mentorship from VS faculty (70%). Basic endovascular skills can be efficiently introduced through a simulation-based curriculum and lead to improved novice performance. Early exposure of preclinical medical students provides an effective teaching and recruitment tool for procedural-based fields, particularly surgical subspecialties. Mentored exposure to endovascular procedures on the simulator positively impacts long-term medical student attitudes toward vascular surgery and ultimate career choices.


Modeling Atomistic Dynamic Fracture Mechanisms Using a Progressive Transformer Diffusion Model.
Dynamic fracture is an important area of materials analysis, assessing the atomic-level mechanisms by which materials fail over time. Here, we focus on brittle materials failure and show that an atomistically derived progressive transformer diffusion machine learning model can effectively describe the dynamics of fracture, capturing important aspects such as crack dynamics, instabilities, and initiation mechanisms. Trained on a small dataset of atomistic simulations, the model generalizes well and offers a rapid assessment of dynamic fracture mechanisms for complex geometries, expanding well beyond the original set of atomistic simulation results. Various validation cases, progressively more distinct from the data used for training, are presented and analyzed. The validation cases feature distinct geometric details, including microstructures generated by a generative neural network used here to identify novel bio-inspired material designs for mechanical performance. For all cases, the model performs well and captures key aspects of material failure.


Continuous polyelectrolyte adsorption under an applied electric potential.
Interactions between charged macromolecules (e.g., proteins, nucleic acids, polyelectrolytes) and charged surfaces govern many natural and industrial processes. We investigate here the influence of an applied electric potential on the adsorption of charged polymers, and report the following significant result: the adsorption of certain amine side chain-containing polycations may become continuous, i.e., asymptotically linear (or nearly linear) in time over hours, upon the application of a modest anodic potential. Employing optical waveguide lightmode spectroscopy (OWLS) and an indium tin oxide (ITO) substrate, we show that asymptotic kinetics, and the adsorbed mass at the onset of the asymptotic regime, depend sensitively on polymer chemistry (in particular, side chain volume and charge location), increase with applied potential and ionic strength (conditions favoring a thicker initial layer), and are independent of bulk polymer concentration (suggesting postadsorption events to be rate limiting). X-ray photoelectron spectra reveal a suppressed polymer charge within layers formed via continuous adsorption, but no evidence of electrochemical reactions. We propose a mechanism based on polymer-polymer binding within the adsorbed layer, enabled by suppressed electrostatic repulsion and/or enhanced ionic correlations near the conducting surface, and stabilized by short-range attractive interactions. Continuous adsorption under an applied electric potential offers the possibility of nanoscale films of tailored polymer content realized in a single step.


Construction of an infectious rhesus rhadinovirus bacterial artificial chromosome for the analysis of Kaposi's sarcoma-associated herpesvirus-related disease development.
Rhesus rhadinovirus (RRV) is closely related to Kaposi's sarcoma-associated herpesvirus (KSHV)/human herpesvirus 8 (HHV-8) and causes KSHV-like diseases in immunocompromised rhesus macaques (RM) that resemble KSHV-associated diseases including multicentric Castleman's disease and non-Hodgkin's lymphoma. RRV retains a majority of open reading frames (ORFs) postulated to be involved in the pathogenesis of KSHV and is the closest available animal model to KSHV infection in humans. Here we describe the generation of a recombinant clone of RRV strain 17577 (RRV(17577)) utilizing bacterial artificial chromosome (BAC) technology. Characterization of the RRV BAC demonstrated that it is a pathogenic molecular clone of RRV(17577), producing virus that behaves like wild-type RRV both in vitro and in vivo. Specifically, BAC-derived RRV displays wild-type growth properties in vitro and readily infects simian immunodeficiency virus-infected RM, inducing B cell hyperplasia, persistent lymphadenopathy, and persistent infection in these animals. This RRV BAC will allow for rapid genetic manipulation of the RRV genome, facilitating the creation of recombinant versions of RRV that harbor specific alterations and/or deletions of viral ORFs. This system will provide insights into the roles of specific RRV genes in various aspects of the viral life cycle and the RRV-associated pathogenesis in vivo in an RM model of infection. Furthermore, the generation of chimeric versions of RRV containing KSHV genes will allow analysis of the function and contributions of KSHV genes to viral pathogenesis by using a relevant primate model system.


Unveiling the Peptide Motifs of HLA-C and HLA-G from Naturally Presented Peptides and Generation of Binding Prediction Matrices.
The classical HLA-C and the nonclassical HLA-E and HLA-G molecules play important roles both in the innate and adaptive immune system. Starting already during embryogenesis and continuing throughout our lives, these three Ags exert major functions in immune tolerance, defense against infections, and anticancer immune responses. Despite these important roles, identification and characterization of the peptides presented by these molecules has been lacking behind the more abundant HLA-A and HLA-B gene products. In this study, we elucidated the peptide specificities of these HLA molecules using a comprehensive analysis of naturally presented peptides. To that end, the 15 most frequently expressed HLA-C alleles as well as HLA-E*01:01 and HLA-G*01:01 were transfected into lymphoblastoid C1R cells expressing low endogenous HLA. Identification of naturally presented peptides was performed by immunoprecipitation of HLA and subsequent analysis of HLA-bound peptides by liquid chromatographic tandem mass spectrometry. Peptide motifs of HLA-C unveil anchors in position 2 or 3 with high variances between allotypes, and a less variable anchor at the C-terminal end. The previously reported small ligand repertoire of HLA-E was confirmed within our analysis, and we could show that HLA-G combines a large ligand repertoire with distinct features anchoring peptides at positions 3 and 9, supported by an auxiliary anchor in position 1 and preferred residues in positions 2 and 7. The wealth of HLA ligands resulted in prediction matrices for octa-, nona-, and decamers. Matrices were validated in terms of their binding prediction and compared with the latest NetMHC prediction algorithm NetMHCpan-3.0, which demonstrated their predictive power.


Cumulative radiation exposure during follow-up after curative surgery for gastric cancer.
To quantify the cumulative effective dose (cED) of radiation due to repeated CT and PET/CT examinations after curative resection of gastric cancer and to assess the lifetime attributable risk (LAR) estimates based on Biological Effects of Ionizing Radiation VII models. Patients who underwent a curative resection for gastric cancer between January 2006 and December 2006 and were followed-up until May 2010 were included in this study. The cED was calculated by using the dose-length product values and conversion factors for quantitative risk assessment of radiation exposure. cED and LAR were compared between early and advanced gastric cancer patients and among American Joint Committee on Cancer TNM stage groups (stage I, II, and III). The nonparametric Mann-Whitney U and Kruskal-Wallis tests, followed by a post-hoc analysis with Bonferroni adjustment, were employed as part of the statistical analysis. The overall median cED was 57.8 mSv (interquartile range [IQR], 43.9-74.7). The cED was significantly higher in the advanced (median, 67.0; IQR, 49.1-102.3) than in the early gastric cancer group (median, 52.3; IQR, 41.5-67.9) (p < 0.001), and increased as the TNM stage increased. For radiation exposure, 62% of all patients received an estimated cED of over 50 mSv, while 11% of patients received over 100 mSv. The median LAR of cancer incidence was 0.28% (IQR, 0.20-0.40) and there were significant differences between the early gastric cancer and advanced gastric cancer group (p < 0.001) as well as among the three TNM stage groups (p = 0.015). The LAR of cancer incidence exceeded 1% in 2.4% of the patients. The cED increases proportionally along with tumor stage and, even in early gastric cancer or stage I patients, cED is much higher than that found among the general population. Considering the very good prognosis of early gastric cancer after curative surgery, the cED should be considered when designing a postoperative follow-up CT protocol.


PFKFB3 inhibition reprograms malignant pleural mesothelioma to nutrient stress-induced macropinocytosis and ER stress as independent binary adaptive responses.
The metabolic signatures of cancer cells are often associated with elevated glycolysis. Pharmacological (PFK158 treatment) and genetic inhibition of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3), a critical control point in the glycolytic pathway, decreases glucose uptake, ATP production, and lactate dehydrogenase activity and arrests malignant pleural mesothelioma (MPM) cells in the G0/G1 phase to induce cell death. To overcome this nutrient stress, inhibition of PFKFB3 activity led to an escalation in endoplasmic reticulum (ER) activity and aggravated ER stress mostly by upregulating BiP and GADD153 expression and activation of the endocytic Rac1-Rab5-Rab7 pathway resulting in a unique form of cell death called "methuosis" in both the sarcomatoid (H28) and epithelioid (EMMeso) cells. Transmission electron microscopy (TEM) analysis showed the formation of nascent macropinocytotic vesicles, which rapidly coalesced to form large vacuoles with compromised lysosomal function. Both immunofluorescence microscopy and co-immunoprecipitation analyses revealed that upon PFKFB3 inhibition, two crucial biomolecules of each pathway, Rac1 and Calnexin interact with each other. Finally, PFK158 alone and in combination with carboplatin-inhibited tumorigenesis of EMMeso xenografts in vivo. Since most cancer cells exhibit an increased glycolytic rate, these results provide evidence for PFK158, in combination with standard chemotherapy, may have a potential in the treatment of MPM.


Defining Seropositivity Thresholds for Use in Trachoma Elimination Studies.
Efforts are underway to eliminate trachoma as a public health problem by 2020. Programmatic guidelines are based on clinical signs that correlate poorly with Chlamydia trachomatis (Ct) infection in post-treatment and low-endemicity settings. Age-specific seroprevalence of anti Ct Pgp3 antibodies has been proposed as an alternative indicator of the need for intervention. To standardise the use of these tools, it is necessary to develop an analytical approach that performs reproducibly both within and between studies. Dried blood spots were collected in 2014 from children aged 1-9 years in Laos (n = 952) and Uganda (n = 2700) and from people aged 1-90 years in The Gambia (n = 1868). Anti-Pgp3 antibodies were detected by ELISA. A number of visual and statistical analytical approaches for defining serological status were compared. Seroprevalence was estimated at 11.3% (Laos), 13.4% (Uganda) and 29.3% (The Gambia) by visual inspection of the inflection point. The expectation-maximisation algorithm estimated seroprevalence at 10.4% (Laos), 24.3% (Uganda) and 29.3% (The Gambia). Finite mixture model estimates were 15.6% (Laos), 17.1% (Uganda) and 26.2% (The Gambia). Receiver operating characteristic (ROC) curve analysis using a threshold calibrated against external reference specimens estimated the seroprevalence at 6.7% (Laos), 6.8% (Uganda) and 20.9% (The Gambia) when the threshold was set to optimise Youden's J index. The ROC curve analysis was found to estimate seroprevalence at lower levels than estimates based on thresholds established using internal reference data. Thresholds defined using internal reference threshold methods did not vary substantially between population samples. Internally calibrated approaches to threshold specification are reproducible and consistent and thus have advantages over methods that require external calibrators. We propose that future serological analyses in trachoma use a finite mixture model or expectation-maximisation algorithm as a means of setting the threshold for ELISA data. This will facilitate standardisation and harmonisation between studies and eliminate the need to establish and maintain a global calibration standard.


Value of air trapping in detection of small airways disease in smokers.
To test the hypothesis that diffuse and/or focal air trapping are sensitive indicators of airflow obstruction in smoker's small airways disease, when age, gender and presence of emphysematous lesions were allowed for. Fifty-eight smokers and 34 never smokers, recruited from a randomized population study of men born in 1933, were investigated by HRCT and by extended pulmonary function tests, including a sensitive test for small airways disease (N2 slope). Diffuse air trapping was evaluated by calculating a quotient of mean lung density at expiration and inspiration. Focal air trapping was scored visually by consensus. Diffuse air trapping did not differ between non-emphysematous smokers and never smokers. Furthermore, diffuse air trapping correlated well to the quotient between the residual volume and total lung capacity (RV/TLC, p = 0.01) and was consequently higher in emphysematous smokers than in never smokers. Focal air trapping was found as frequently in smokers without emphysema as in never smokers. Smokers with emphysema showed significantly less focal air trapping. Neither the N2 slope nor any of the other lung function variables differed between those with and without focal air trapping among non-emphysematous smokers. Neither diffuse nor focal air trapping are sensitive indicators of smoker's small airways disease.


Hornerin Is Involved in Breast Cancer Progression.
The S100 gene family, which comprises over 20 members, including S100A1, S100A2, S100A8, S100A9, profilaggrin, and hornerin encodes low molecular weight calcium-binding proteins with physiological and pathological roles in keratinization. Recent studies have suggested a link between S100 proteins and human cancer progression. The purpose of the present study was to determine the expression levels of hornerin, S100A8, and S100A9 and evaluate their roles in the progression of invasive ductal carcinoma (IDC). Seventy cases of ductal carcinoma in situ (DCIS), IDC, and metastatic carcinoma in lymph nodes (MCN) were included. Tissue microarrays were constructed from lesions of DCIS, IDC, and MCN from the same patients. Expression of hornerin, S100A8, and S100A9 was analyzed using immunohistochemistry. The expression of hornerin was associated with the estrogen receptor-negative (p=0.003) and the human epidermal growth factor receptor 2-positive (p=0.002) groups. The expression of S100A8 was associated with a higher pT stage (p=0.017). A significant (p<0.001) correlation between the expression of S100A9 and S100A8 was also found. The mean percentages of hornerin-positive tumor cells in DCIS, IDC, and MCN were 1.0%±3.3% (mean±standard deviation), 12.0%±24.0%, and 75.3%± 27.6%, respectively. The expression of hornerin significantly (p<0.001) increased with the progression of carcinoma. The mean levels of S100A8 and S100A9 in DCIS, IDC, and MCN were not significantly (p>0.050) different. The expression of hornerin increased in a stepwise manner (DCIS<IDC<MCN). Our data suggest that hornerin is involved in breast cancer progression and malignant transformation from preinvasive lesions.


ST segment monitoring for myocardial ischemia.
Many patients in the critical care unit (CCU) are at risk for myocardial ischemia and acute coronary artery reocclusion. The use of continuous ST segment monitoring detects transient and sustained ischemia, despite the absence of symptoms, more completely than rate and rhythm monitoring alone. The accuracy and sensitivity of the ST segment for detection of ischemia is dependent on the number and location of the electrocardiographic leads used in conjunction with the site of obstruction. Thus, when using ST segment monitoring for detection of ischemia, one of the most important decisions for the nurse to make is lead selection. In addition, both ischemic and nonischemic ST segment changes must be considered.


A Combinatorial Library of Lipid Nanoparticles for RNA Delivery to Leukocytes.
Lipid nanoparticles (LNPs) are the most advanced nonviral platforms for small interfering RNA (siRNA) delivery that are clinically approved. These LNPs, based on ionizable lipids, are found in the liver and are now gaining much attention in the field of RNA therapeutics. The previous generation of ionizable lipids varies in linker moieties, which greatly influences in vivo gene silencing efficiency. Here novel ionizable amino lipids based on the linker moieties such as hydrazine, hydroxylamine, and ethanolamine are designed and synthesized. These lipids are formulated into LNPs and screened for their efficiency to deliver siRNAs into leukocytes, which are among the hardest to transfect cell types. Two potent lipids based on their in vitro gene silencing efficiencies are also identified. These lipids are further evaluated for their biodistribution profile, efficient gene silencing, liver toxicity, and potential immune activation in mice. A robust gene silencing is also found in primary lymphocytes when one of these lipids is formulated into LNPs with a pan leukocyte selective targeting agent (β7 integrin). Taken together, these lipids have the potential to open new avenues in delivering RNAs into leukocytes.


Hepatitis B virus reactivation in HBsAg-negative, anti-HBc-positive patients receiving immunosuppressive therapy: a systematic review.
The optimal management of HBsAg-negative, anti-HBc-positive patients who receive immunosuppression remains unclarified. We systematically reviewed the available data on potential predictors of the risk of hepatitis B virus (HBV) reactivation in such patients. A literature search identified 55 studies with 3640 HBsAg-negative, anti-HBc-positive patients who received immunosuppressive regimens. HBV reactivation was reported in 236 (6.5%) patients. The pooled HBV reactivation rates did not differ between patients with detectable or undetectable HBV DNA in studies with hematological diseases or regimens containing rituximab, but it was higher in patients with detectable than in those with undetectable HBV DNA who were taking rituximab-free regimens (14% vs. 2.6%; risk ratio [RR] 12.67, 95% CI: 95%CI 2.39-67.04, P=0.003) or had non-hematological diseases, although the latter was not confirmed by sensitivity analysis (RR 8.80, 95%CI 0.71-109.00, P=0.09). The pooled HBV reactivation rates were lower in patients with positive than in those with negative anti-HBs in studies with hematological (7.1% vs. 21.8%; RR 0.29, 95%CI 0.19-0.46, P<0.001) or non-hematological (2.5% vs. 10.7%; RR 0.28, 95%CI 0.11-0.76, P=0.012) diseases, and rituximab-containing (6.6% vs. 19.8%; RR 0.32, 95%CI 0.15-0.69, P=0.003) or rituximab-free (3.3% vs. 9.2%; RR 0.36, 95%CI 0.14-0.96, P=0.042) regimens. The risk of HBV reactivation is high; therefore, anti-HBV prophylaxis should be recommended in HBsAg-negative, anti-HBc-positive patients with hematological diseases and/or rituximab-containing regimens, regardless of HBV DNA and anti-HBs status. In contrast, patients with non-hematological diseases or rituximab-free regimens have a low risk of HBV reactivation and may not require anti-HBV prophylaxis if they have undetectable HBV DNA and positive anti-HBs.


AOSSM Early Sport Specialization Consensus Statement.
Early sport specialization is not a requirement for success at the highest levels of competition and is believed to be unhealthy physically and mentally for young athletes. It also discourages unstructured free play, which has many benefits. To review the available evidence on early sports specialization and identify areas where scientific data are lacking. Think tank, roundtable discussion. The primary outcome of this think tank was that there is no evidence that young children will benefit from early sport specialization in the majority of sports. They are subject to overuse injury and burnout from concentrated activity. Early multisport participation will not deter young athletes from long-term competitive athletic success. Youth advocates, parents, clinicians, and coaches need to work together with the sport governing bodies to ensure healthy environments for play and competition that do not create long-term health issues yet support athletic competition at the highest level desired.


A single-institution retrospective cases series of childhood undifferentiated embryonal liver sarcoma (UELS): success of combined therapy and the use of orthotopic liver transplant.
Undifferentiated embryonal liver sarcoma (UELS) makes up 9% to 15% of all malignant liver tumors in children. UELS is characteristically diagnosed between the ages of 6 and 10 years and presents with abdominal pain, vomiting, and an abdominal mass. There is currently no standardized treatment for UELS except attempt at complete surgical resection. There have been only about 150 cases of UELS reported in the literature all with historically poor overall survival of <37.5% at 5 years. This report is one of the largest single-institution reports of UELS consisting of 5 patients over 2 decades. The purpose of this study is to characterize presentation and to report treatment success in UELS in children, adolescents, and young adults and the use of liver transplantation and, lastly, to suggest a use of positron emission tomography/computed tomography (PET/CT) in monitoring of this disease process. We conducted an Institutional Review Board-approved retrospective chart review. Data were collected from UELS patients younger than 21 years seen at the University of California Los Angeles over the past 20 years (January 2001 to September 2011). Descriptive analysis was conducted including multiple parameters of patient demographics, tumor characteristics, treatment modalities, and morbidity and mortality. Five patients with UELS were identified. Patients initially presented with fever, abdominal pain, or nausea. Ages ranged from 10 to 19 years old (median age 13 y old), and there was a 4:1 male-to-female predominance. Tumor size ranged from 6 to 22 cm in largest diameter. One patient presented with metastatic disease to the lungs and heart and 1 patient recurred 2 years from diagnosis with bilateral paraspinal masses. Treatment included local control surgery with neoadjuvant and adjuvant chemotherapy with an anthracycline/alkylating agent combination. One patient with recurrent and refractory disease achieved local control with an orthotopic liver transplantation (OLT). Metastatic disease was controlled with surgery and radiation therapy. 18-Fluorodeoxyglucose PET/CT was a useful imaging tool for judging response to therapy with complete loss of metabolic activity in tumor after neoadjuvant chemotherapy in 2 representative cases. Although follow-up is short for some patients, overall survival in these 5 patients was 100% with follow-up ranging from 21 to 68 months. Disease-free survival ranged from 8 to 46 months with no patients with residual disease. UELS is an aggressive high-grade primary liver sarcoma with high metastatic potential. This report represents one of the largest single-institution studies of UELS. Using multimodality therapy, patients have achieved 100% overall survival even in the setting of extensive disease, metastases, and recurrence. In cases of unresectable primary tumor or recurrent and refractory disease isolated to the liver, OLT is a potential therapeutic option. We report success with adjuvant chemotherapy and complete surgical resection with OLT as an alternative in unresectable or refractory cases. We also suggest a possible utility of PET/CT in monitoring treatment response in this disease.


Pattern and factors leading to the diffusion of magnetic resonance imaging in Korean hospitals.
The purpose of this study was to examine the diffusion patterns of new medical technologies in Korean hospitals. We also sought to identify critical factors leading to the decision to acquire capital-intensive medical technology. The rationale and timing of magnetic resonance imaging (MRI) acquisitions were retrospectively evaluated according to a "whether, when, and why" paradigm. We analyzed data pertaining to 232 hospitals with active medical residency programs. Of these, 185 hospitals owned or leased an MRI unit, and 47 had not acquired units as of June 2004. Data were collected from the Ministry of Health and Welfare, Korean National Statistical Office, and Korean Hospital Association databases, and variables were identified and classified as predisposing, enabling, or reinforcing factors. The MRI diffusion rate curve was linear for two types of hospital but was S-shaped for tertiary hospitals, which were early adopters of MRI. Significant predictors for MRI adoption included the per capita number of regional physicians (+), total number of existing regional MRI units (-), percentage of the regional population over 65 years of age (+), private ownership of the hospital, presence of a radiology residency program, number of beds (+), and regional per capita taxable income (+). Diffusion of MRI technology is occurring rapidly across Korean hospitals. The factors affecting MRI adoption in Korea are similar to the factors documented in other countries, namely regional population over age 65, regional income per capita, large hospitals, and teaching hospitals. This study provides baseline information for predicting diffusion patterns of other new and/or expensive medical technologies.


Episodic simulation of future events: concepts, data, and applications.
This article focuses on the neural and cognitive processes that support imagining or simulating future events, a topic that has recently emerged in the forefront of cognitive neuroscience. We begin by considering concepts of simulation from a number of areas of psychology and cognitive neuroscience in order to place our use of the term in a broader context. We then review neuroimaging, neuropsychological, and cognitive studies that have examined future-event simulation and its relation to episodic memory. This research supports the idea that simulating possible future events depends on much of the same neural machinery, referred to here as a core network, as does remembering past events. After discussing several theoretical accounts of the data, we consider applications of work on episodic simulation for research concerning clinical populations suffering from anxiety or depression. Finally, we consider other aspects of future-oriented thinking that we think are related to episodic simulation, including planning, prediction, and remembering intentions. These processes together comprise what we have termed "the prospective brain," whose primary function is to use past experiences to anticipate future events.


Modular gene expression in Poplar: a multilayer network approach.
By applying a multilayer network approach to an extensive set of Poplar microarray data, a genome-wide coexpression network has been detected and explored. Multilayer networks were generated from minimum spanning trees (MSTs) using Kruskal's algorithm from random jack-knife resamplings of half of the full data set. The final network is obtained from the union of all the generated MSTs. The gene expression correlations display a highly clustered topology, which is more pronounced when introducing links appearing in relatively few of the generated MSTs. The network also reveals a modular architecture, reflecting functional groups with relatively frequent gene-to-gene communication. Furthermore, the observed modular structure overlaps with different gene activities in different tissues, and closely related tissues show similar over- and/or under-expression patterns at the modular scale. It is shown that including links that appear in a few of the generated MSTs increases the information quality of the network. In other words, a link may be 'weak' because it reflects rare signaling events rather than merely a signal weakened by noise. The method allows, from comparisons of random 'null networks', tuning to maximize the information obtainable.


Lens-free endoscopy probe for optical coherence tomography.
We present an ultrathin fiber-optic endoscopy probe for optical coherence tomography (OCT), which is made of a series of fused optical fibers instead of the conventional scheme based on an objective lens. The large-core fiber with a core diameter of 20 μm was utilized for the probe, while a single-mode fiber of core diameter 8.2 μm mainly delivered the OCT light. Those fibers were spliced with a bridge fiber of an intermediate core size. The guided light was stepwise converted to a beam of a large mode-field diameter to be radiated with a larger depth of focus. We obtained a 125 μm thick all-fiber endoscopy probe with a side-viewing capability implemented by an angled fiber end. Successful OCT imaging was demonstrated with a swept-source OCT system and showed the practical applicability of our lens-free endoscopy probe.


Diet and the frequency of the alanine:glyoxylate aminotransferase Pro11Leu polymorphism in different human populations.
The intermediary metabolic enzyme alanine:glyoxylate aminotransferase (AGT) contains a Pro11Leu polymorphism that decreases its catalytic activity by a factor of three and causes a small proportion to be mistargeted from its normal intracellular location in the peroxisomes to the mitochondria. These changes are predicted to have significant effects on the synthesis and excretion of the metabolic end-product oxalate and the deposition of insoluble calcium oxalate in the kidney and urinary tract. Based on the evolution of AGT targeting in mammals, we have previously hypothesised that this polymorphism would be advantageous for individuals who have a meat-rich diet, but disadvantageous for those who do not. If true, the frequency distribution of Pro11Leu in different extant human populations should have been shaped by their dietary history so that it should be more common in populations with predominantly meat-eating ancestral diets than it is in populations in which the ancestral diets were predominantly vegetarian. In the present study, we have determined frequency of Pro11Leu in 11 different human populations with divergent ancestral dietary lifestyles. We show that the Pro11Leu allelic frequency varies widely from 27.9% in the Saami, a population with a very meat-rich ancestral diet, to 2.3% in Chinese, who are likely to have had a more mixed ancestral diet. FST analysis shows that the differences in Pro11Leu frequency between some populations (particularly Saami vs Chinese) was very high when compared with neutral loci, suggesting that its frequency might have been shaped by dietary selection pressure.