Plastic responses of bryozoans to ocean acidification.
Phenotypic plasticity has the potential to allow organisms to respond rapidly to global environmental change, but the range and effectiveness of these responses are poorly understood across taxa and growth strategies. Colonial organisms might be particularly resilient to environmental stressors, as organizational modularity and successive asexual generations can allow for distinctively flexible responses in the aggregate form. We performed laboratory experiments to examine the effects of increasing dissolved carbon dioxide (CO2) (i.e. ocean acidification) on the colonial bryozoan Celleporella cornuta sampled from two source populations within a coastal upwelling region of the northern California coast. Bryozoan colonies were remarkably plastic under these CO2 treatments. Colonies raised under high CO2 grew more quickly, investing less in reproduction and producing lighter skeletons when compared with genetically identical clones raised under current surface atmosphere CO2 values. Bryozoans held under high CO2 conditions also changed the Mg/Ca ratio of skeletal calcite and increased the expression of organic coverings in new growth, which may serve as protection against acidified water. We also observed strong differences between source populations in reproductive investment and organic covering reaction norms, consistent with adaptive responses to persistent spatial variation in local oceanographic conditions. Our results demonstrate that phenotypic plasticity and energetic trade-offs can mediate biological responses to global environmental change, and highlight the broad range of strategies available to colonial organisms.


ER stress-induced cell death proceeds independently of the TRAIL-R2 signaling axis in pancreatic β cells.
Prolonged ER stress and the associated unfolded protein response (UPR) can trigger programmed cell death. Studies in cancer cell lines demonstrated that the intracellular accumulation of TRAIL receptor-2 (TRAIL-R2) and the subsequent activation of caspase-8 contribute significantly to apoptosis induction upon ER stress. While this might motivate therapeutic strategies that promote cancer cell death through ER stress-induced caspase-8 activation, it could also support the unwanted demise of non-cancer cells. Here, we therefore investigated if TRAIL-R2 dependent signaling towards apoptosis can be induced in pancreatic β cells, whose loss by prolonged ER stress is associated with the onset of diabetes. Interestingly, we found that elevated ER stress in these cells does not result in TRAIL-R2 transcriptional induction or elevated protein levels, and that the barely detectable expression of TRAIL-R2 is insufficient to allow TRAIL-induced apoptosis to proceed. Overall, this indicates that apoptotic cell death upon ER stress most likely proceeds independent of TRAIL-R2 in pancreatic β cells. Our findings therefore point to differences in ER stress response and death decision-making between cancer cells and pancreatic β cells and also have implications for future targeted treatment strategies that need to differentiate between ER stress susceptibility of cancer cells and pancreatic β cells.


VEGF drives cancer-initiating stem cells through VEGFR-2/Stat3 signaling to upregulate Myc and Sox2.
Vascular endothelial growth factor-A (VEGF), a potent angiogenic factor, is also implicated in self-renewal in several normal tissue types. VEGF has been shown to drive malignant stem cells but mechanisms thereof and tumor types affected are not fully characterized. Here, we show VEGF promotes breast and lung cancer stem cell (CSC) self-renewal via VEGF receptor-2 (VEGFR-2)/STAT3-mediated upregulation of Myc and Sox2. VEGF increased tumor spheres and aldehyde dehydrogenase activity, both proxies for stem cell function in vitro, in triple-negative breast cancer (TNBC) lines and dissociated primary cancers, and in lung cancer lines. VEGF exposure before injection increased breast cancer-initiating cell abundance in vivo yielding increased orthotopic tumors, and increased metastasis from orthotopic primaries and following tail vein injection without further VEGF treatment. VEGF rapidly stimulated VEGFR-2/JAK2/STAT3 binding and activated STAT3 to bind MYC and SOX2 promoters and induce their expression. VEGFR-2 knockdown or inhibition abrogated VEGF-mediated STAT3 activation, MYC and SOX2 induction and sphere formation. Notably, knockdown of either STAT3, MYC or SOX2 impaired VEGF-upregulation of pSTAT3, MYC and SOX2 expression and sphere formation. Each transcription factor, once upregulated, appears to promote sustained activation of the others, creating a feed-forward loop to drive self-renewal. Thus, in addition to angiogenic effects, VEGF promotes tumor-initiating cell self-renewal through VEGFR-2/STAT3 signaling. Analysis of primary breast and lung cancers (>1300 each) showed high VEGF expression, was prognostic of poor outcome and strongly associated with STAT3 and MYC expression, supporting the link between VEGF and CSC self-renewal. High-VEGF tumors may be most likely to escape anti-angiogenics by upregulating VEGF, driving CSC self-renewal to re-populate post-treatment. Our work highlights the need to better define VEGF-driven cancer subsets and supports further investigation of combined therapeutic blockade of VEGF or VEGFR-2 and JAK2/STAT3.


Rheumatoid arthritis in an urban South African Negro population.
(1) An epidemiological study of an urban South African Negro community has been carried out in Johannesburg. Altogether 964 respondents were examined and in each case radiographs of the hands and feet were obtained. Rheumatoid factor tests were carried out on 404 serum samples. (2) Rheumatoid arthritis (RA) was graded 'definite' or 'probable' on the basis of a modification of the Rome criteria (Kellgren, Jeffrey, and Ball, 1963a). (3) In marked contrast to the findings in rural Africans the prevalence of RA in this community was similar to that in Caucasian populations. Five respondents (all elderly women) had 'definite' RA, giving a prevalence of 1.4% of the females and 0.9% of the total population sample over 15 years old. The prevalence of 'definite' and 'probable' RA combined was 2.6% for males, 3.7% for females, and 3.3% for all individuals over 15 years old. Prevalence increased with age, reaching a maximum in the 65- to 74-year cohort. (4) The form and severity of the clinical and radiological features were unlike the mild manifestations seen in rural African peoples and closely resembled the usual clinical picture of rheumatoid disease. (5) The latex fixation test was positive in 12.1% of the sera tested, which is similar to the high titres found in other African populations. No obvious cause for this phenomenon was found. (6) Several reasons for the marked difference in prevalence of RA between this urban African population and a rural African population are considered. Marked intraracial differences such as this point to the importance of sociological and environmental factors in the pathogenesis of rheumatoid arthritis.


An in vivo platform for translational drug development in pancreatic cancer.
Effective development of targeted anticancer agents includes the definition of the optimal biological dose and biomarkers of drug activity. Currently available preclinical models are not optimal to this end. We aimed at generating a model for translational drug development using pancreatic cancer as a prototype. Resected pancreatic cancers from 14 patients were xenografted and expanded in successive groups of nude mice to develop cohorts of tumor-bearing mice suitable for drug therapy in simulated early clinical trials. The xenografted tumors maintain their fundamental genotypic features despite serial passages and recapitulate the genetic heterogeneity of pancreatic cancer. The in vivo platform is useful for integrating drug screening with biomarker discovery. Passages of tumors in successive cohorts of mice do not change their susceptibility to anticancer agents and represent a perpetual live bank, facilitating the application of new technologies that will result in the creation of an integrated stable database of tumor-drug response data and biomarkers.


Update on the clinical utility of coronary computed tomographic angiography in stable angina pectoris.
Over the last decade, coronary computed tomographic angiography (CCTA) has emerged as a valuable non-invasive imaging modality with excellent diagnostic performance compared to invasive coronary angiography (ICA) for identifying patients with coronary artery disease (CAD). Beyond the diagnosis of CAD, CCTA also provides valuable prognostic information. While patients with normal CCTA have excellent long-term prognosis, among those with CAD, increasing CAD extent and severity is associated with increased cardiovascular event risk over both medium- and long-term follow-up in both men and women. The ability to image non-obstructive CAD is a particularly unique attribute of CCTA. Moreover, the ability to assess plaque features on CCTA has further enhanced our understanding of coronary plaque dynamics and the prediction of future cardiovascular events. The clinical impact of CCTA has been recently evaluated in two landmark prospective multicenter trials, which have provided insights into the influence of CCTA on the clinical management of symptomatic patients with suspected CAD. We review the value of CCTA in the evaluation of patients with stable chest pain including its diagnostic performance, prognostic utility and real-world clinical application.


Child work and labour among orphaned and abandoned children in five low and middle income countries.
The care and protection of the estimated 143,000,000 orphaned and abandoned children (OAC) worldwide is of great importance to global policy makers and child service providers in low and middle income countries (LMICs), yet little is known about rates of child labour among OAC, what child and caregiver characteristics predict child engagement in work and labour, or when such work infers with schooling. This study examines rates and correlates of child labour among OAC and associations of child labour with schooling in a cohort of OAC in 5 LMICs. The Positive Outcomes for Orphans (POFO) study employed a two-stage random sampling survey methodology to identify 1480 single and double orphans and children abandoned by both parents ages 6-12 living in family settings in five LMICs: Cambodia, Ethiopia, India, Kenya, and Tanzania. Regression models examined child and caregiver associations with: any work versus no work; and with working <21, 21-27, and 28+ hours during the past week, and child labour (UNICEF definition). The majority of OAC (60.7%) engaged in work during the past week, and of those who worked, 17.8% (10.5% of the total sample) worked 28 or more hours. More than one-fifth (21.9%; 13% of the total sample) met UNICEF's child labour definition. Female OAC and those in good health had increased odds of working. OAC living in rural areas, lower household wealth and caregivers not earning an income were associated with increased child labour. Child labour, but not working fewer than 28 hours per week, was associated with decreased school attendance. One in seven OAC in this study were reported to be engaged in child labour. Policy makers and social service providers need to pay close attention to the demands being placed on female OAC, particularly in rural areas and poor households with limited income sources. Programs to promote OAC school attendance may need to focus on the needs of families as well as the OAC.


Synthetically encoding 10 nm morphology in silicon nanowires.
Si nanowires (NWs) have been widely explored as a platform for photonic and electronic technologies. Here, we report a bottom-up method to break the conventional "wire" symmetry and synthetically encode a high-resolution array of arbitrary shapes, including nanorods, sinusoids, bowties, tapers, nanogaps, and gratings, along the NW growth axis. Rapid modulation of phosphorus doping combined with selective wet-chemical etching enabled morphological features as small as 10 nm to be patterned over wires more than 50 μm in length. This capability fundamentally expands the set of technologies that can be realized with Si NWs, and as proof-of-concept, we demonstrate two distinct applications. First, nanogap-encoded NWs were used as templates for Noble metals, yielding plasmonic structures with tunable resonances for surface-enhanced Raman imaging. Second, core/shell Si/SiO2 nanorods were integrated into electronic devices that exhibit resistive switching, enabling nonvolatile memory storage. Moving beyond these initial examples, we envision this method will become a generic route to encode new functionality in semiconductor NWs.


Multioctave supercontinuum from visible to mid-infrared and bend effects on ultrafast nonlinear dynamics in gas-filled hollow-core fiber.
Broadband supercontinuum generation is numerically investigated in a Xe-filled nested hollow-core antiresonant (HC-AR) fiber pumped at 3 μm with pulses of 100 fs duration and 15 μJ energy. For a 25 cm long fiber, under 7 bar pressure, the supercontinuum spectrum spans multiple octaves from 400 nm to 5000 nm. Furthermore, the influence of bending on ultrafast nonlinear pulse propagation dynamics is investigated for two types of HC-AR fibers (nested and non-nested capillaries). Our results predict similar nonlinear dynamics for both fiber types and a significant reduction of the spectral broadening under tight bending conditions.


Comparison of prothrombin time (INR) results and main characteristics of patients on warfarin treatment in primary health care centers and anticoagulation clinics.
Oral anticoagulant therapy is used to prevent thrombosis in patients with atrial fibrillation (AF), venous thrombosis and prosthetic heart valves. The introduction of new therapies emphasizes the need to discern the best practice for the patients remaining on warfarin treatment. This study compares patient characteristics and therapeutic control in two settings managing warfarin treatment: Swedish primary health care centers (PHCC) and specialized anticoagulation clinics (ACC). Prothrombin time (PT) test results reported as International Normalized Ratio (INR) were collected for five consecutive days from patients on warfarin treatment; 564 PHCC and 927 ACC patients. Therapeutic control was calculated as PT test results in relation to intended therapeutic range (TR). Mann-Whitney Rank Sum Test and Chi2 test were used for statistical comparisons. The PHCC patients were older than the ACC patients, 76 v. 70 years (p<0.01) with a predominance of men in both groups. The reasons for treating differed between the groups. Seventy-two percent of PHCC patients and 66% of ACC patients had a PT-INR within the intended TR (p<0.05). Men generally had better results than women (72% v. 63%, p<0.001) and particularly in the PHCC group v. the ACC group (78% v. 69%, p<0.01).PT-INR above intended TR was significantly more common in the ACC setting, (p<0.05), for women overall (p<0.01), for women in the PHCC setting, and for ACC men (p<0.05). In this study both settings achieved good therapeutic control of warfarin treatment with a minor advantage for PHCC over ACC, and better results for men, especially in the PHCC setting. As patient characteristics differ between the PHCC and ACC, it is important to conduct further randomized studies to discern the best practice locally for warfarin management also after the introduction of new drugs.


Kidney-intrinsic factors determine the severity of ischemia/reperfusion injury in a mouse model of delayed graft function.
Delayed graft function due to transplant ischemia/reperfusion injury adversely affects up to 50% of deceased-donor kidney transplant recipients. However, key factors contributing to the severity of ischemia/reperfusion injury remain unclear. Here, using a clinically relevant mouse model of delayed graft function, we demonstrated that donor genetic background and kidney-intrinsic MyD88/Trif-dependent innate immunity were key determinants of delayed graft function. Functional deterioration of kidney grafts directly corresponded with the duration of cold ischemia time. The graft dysfunction became irreversible after cold ischemia time exceeded six hours. When cold ischemia time reached four hours, kidney grafts displayed histological features reflective of delayed graft function seen in clinical kidney transplantation. Notably, kidneys of B6 mice exhibited significantly more severe histological and functional impairment than kidneys of C3H or BALB/c mice, regardless of recipient strains or alloreactivities. Furthermore, allografts of B6 mice also showed an upregulation of IL-6, neutrophil gelatinase-associated lipocalin, and endoplasmic reticulum stress genes, as well as an increased influx of host neutrophils and memory CD8 T-cells. In contrast, donor MyD88/Trif deficiency inhibited neutrophil influx and decreased the expression of IL-6 and endoplasmic reticulum stress genes, along with improved graft function and prolonged allograft survival. Thus, kidney-intrinsic factors involving genetic characteristics and innate immunity serve as critical determinants of the severity of delayed graft function. This preclinical murine model allows for further investigations of the mechanisms underlying delayed graft function.


Regulation of RB transcription in vivo by RB family members.
In cancer cells, the retinoblastoma tumor suppressor RB is directly inactivated by mutation in the RB gene or functionally inhibited by abnormal activation of cyclin-dependent kinase activity. While variations in RB levels may also provide an important means of controlling RB function in both normal and cancer cells, little is known about the mechanisms regulating RB transcription. Here we show that members of the RB and E2F families bind directly to the RB promoter. To investigate how the RB/E2F pathway may regulate Rb transcription, we generated reporter mice carrying an eGFP transgene inserted into a bacterial artificial chromosome containing most of the Rb gene. Expression of eGFP largely parallels that of Rb in transgenic embryos and adult mice. Using these reporter mice and mutant alleles for Rb, p107, and p130, we found that RB family members modulate Rb transcription in specific cell populations in vivo and in culture. Interestingly, while Rb is a target of the RB/E2F pathway in mouse and human cells, Rb expression does not strictly correlate with the cell cycle status of these cells. These experiments identify novel regulatory feedback mechanisms within the RB pathway in mammalian cells.


Partial monosomy 8q and partial trisomy 9q due to the maternal translocation t(8;9(q24.3;q34.1): a case report.
Partial trisomy 9q34-qter and partial monosomy 8q24.3-qter are very rare chromosomal abnormalities. Characteristic features of partial trisomy 9q34-qter are hypotonia, developmental delay, mild intellectual disability, dolichocephaly, distinct facial phenotype, long and thin fingers, and cardiac anomalies. Unlike the partial trisomy 9q34-qter, partial monosomy 8q24.3-qter has no distinct phenotype. Here we report a four years old female patient with partial trisomy 9q34-qter and partial monosomy 8q24.3-qter due to the maternal translocation t(8;9)(q24.3;q34. I). She has developmental delay, brachycephaly, facial dysmorphism, hand and foot anomalies, bilateral hearing loss, cardiac defect and abnormal brain MRI findings. To the best of our knowledge, this is the first report of the combination of partial trisomy 9q and partial monosomy 8q.


Microbial quality of water in rural households of Ethiopia: implications for milk safety and public health.
Waterborne pathogenic agents affect the health of people either by direct consumption of contaminated water or by its indirect use in food production and/or processing. Studies on the microbiological quality of water in rural areas of Ethiopia are still limited, especially at the household level. The aim of the present study was to assess the microbial quality of water from different sources in rural households in two districts of the Ethiopian Rift Valley area. The correlation between E. coli counts in water and milk was also investigated. In total, 233 water samples (126 collected in dry and 107 in wet season) and 53 milk samples (19 from raw milk and 36 from processed milk products) were analyzed for E. coli contamination. The overall prevalence of E. coli in water samples was 54.9% (n = 233). In most of the analyzed samples, a higher prevalence of E. coli was recorded during the wet compared to the dry season. The highest load of E. coli was detected in water samples from dugouts. The quality of raw milk and traditionally-processed milk products showed variations between districts, and the traditionally-processed milk products were found to contain higher E. coli loads than raw milk. The correlation between the E. coli counts in water and milk only showed a weak but positive relationship (r = 0.1). Taking E. coli as a proxy for water quality, the microbiological quality of water consumed in the study area was found to be very poor, posing a potential food safety and health risk to the rural communities.


The complexity of role balance: support for the Model of Juggling Occupations.
This pilot study aimed to establish the appropriateness of the Model of Juggling Occupations in exploring the complex experience of role balance amongst working women with family responsibilities living in Perth, Australia. In meeting this aim, an evaluation was conducted of a case study design, where data were collected through a questionnaire, time diary, and interview. Overall role balance varied over time and across participants. Positive indicators of role balance occurred frequently in the questionnaires and time diaries, despite the interviews revealing a predominance of negative evaluations of role balance. Between-role balance was achieved through compatible role overlap, buffering, and renewal. An exploration of within-role balance factors demonstrated that occupational participation, values, interests, personal causation, and habits were related to role balance. This pilot study concluded that the Model of Juggling Occupations is an appropriate conceptual framework to explore the complex and dynamic experience of role balance amongst working women with family responsibilities. It was also confirmed that the case study design, including the questionnaire, time diary, and interview methods, is suitable for researching role balance from this perspective.


A main chain biodegradable polyurethane with anti-protein adsorption and anti-bacterial adhesion performances.
Biofilms are initially formed by substances such as proteins secreted by bacteria adhering to a surface. To achieve a durable antibacterial material, biodegradable dihydroxyl-terminated poly[(ethylene oxide)-co-(ethylene carbonate)] (PEOC(OH)2) with anti-protein adsorption properties was synthesized in this study. Further polycondensation of PEOC(OH)2 and isophorone diisocyanate (IPDI) led to biodegradable polyurethane (PEOC-PU) with PEOC as the soft segment. For comparison, polyurethanes with polyethylene glycol (PEG-PU) and polypropylene glycol (PPG-PU) as soft segments were also synthesized. The chemical structures of the polyurethanes were characterized by 1H NMR and FTIR. The biodegradation behavior of PEOC-PU promoted by lipase due to the presence of ethylene carbonate units was also studied. Their resistance to proteins was studied using quartz crystal microbalance with dissipation (QCM-D) and the results revealed that PEOC-PU exhibited excellent nonspecific resistance to proteins. The colonization of microorganisms on PU in the liquid culture medium was further examined and the results showed that PEOC-PU exhibited excellent antibacterial adhesion performance due to its protein resistance and biodegradation.


Mouse models of diabetic nephropathy.
Mice provide an experimental model of unparalleled flexibility for studying mammalian diseases. Inbred strains of mice exhibit substantial differences in their susceptibility to the renal complications of diabetes. Much remains to be established regarding the course of diabetic nephropathy (DN) in mice as well as defining those strains and/or mutants that are most susceptible to renal injury from diabetes. Through the use of the unique genetic reagents available in mice (including knockouts and transgenics), the validation of a mouse model reproducing human DN should significantly facilitate the understanding of the underlying genetic mechanisms that contribute to the development of DN. Establishment of an authentic mouse model of DN will undoubtedly facilitate testing of translational diagnostic and therapeutic interventions in mice before testing in humans.


Accuracy of the Intra- and Extra-Oral Scanning Technique for Transferring the Intaglio Surface of a Pontic of Provisional Restorations to Definitive Restorations.
When taking the final impression for a three-unit fixed partial denture (FPD), the intaglio surface of the pontic of provisional restoration cannot be transferred accurately to that of definitive restoration. The intra- and extra-oral scanning (IEOS) technique, a method for accurately reproducing the submucosal morphology of the superstructure of an implant, has been reported using an intraoral scanner. In the present study, we evaluated the difference between the conventional impression method using impression material and the IEOS technique in reproducing the morphology of the surface of the pontic of a definitive FPD. There was a significant difference in the trueness of the intaglio surface morphology of the pontic between the conventional method and the IEOS technique; however, no significant difference in precision was observed. As a result, the intaglio surface of the pontic of the three-unit FPD could be transferred to definitive restorations more accurately with the IEOS technique than with the conventional method. These results suggest that the IEOS technique can duplicate the intaglio surface of the pontic more reproducibly to the definitive restorations compared with the conventional method.


Lens ion homeostasis relies on the assembly and/or stability of large connexin 46 gap junction plaques on the broad sides of differentiating fiber cells.
The eye lens consists of layers of tightly packed fiber cells, forming a transparent and avascular organ that is important for focusing light onto the retina. A microcirculation system, facilitated by a network of gap junction channels composed of connexins 46 and 50 (Cx46 and Cx50), is hypothesized to maintain and nourish lens fiber cells. We measured lens impedance in mice lacking tropomodulin 1 (Tmod1, an actin pointed-end capping protein), CP49 (a lens-specific intermediate filament protein), or both Tmod1 and CP49. We were surprised to find that simultaneous loss of Tmod1 and CP49, which disrupts cytoskeletal networks in lens fiber cells, results in increased gap junction coupling resistance, hydrostatic pressure, and sodium concentration. Protein levels of Cx46 and Cx50 in Tmod1(-/-);CP49(-/-) double-knockout (DKO) lenses were unchanged, and electron microscopy revealed normal gap junctions. However, immunostaining and quantitative analysis of three-dimensional confocal images showed that Cx46 gap junction plaques are smaller and more dispersed in DKO differentiating fiber cells. The localization and sizes of Cx50 gap junction plaques in DKO fibers were unaffected, suggesting that Cx46 and Cx50 form homomeric channels. We also demonstrate that gap junction plaques rest in lacunae of the membrane-associated actin-spectrin network, suggesting that disruption of the actin-spectrin network in DKO fibers may interfere with gap junction plaque accretion into micrometer-sized domains or alter the stability of large plaques. This is the first work to reveal that normal gap junction plaque localization and size are associated with normal lens coupling conductance.


Algal turf scrubber (ATS) floways on the Great Wicomico River, Chesapeake Bay: productivity, algal community structure, substrate and chemistry(1).
Two Algal Turf Scrubber (ATS) units were deployed on the Great Wicomico River (GWR) for 22 months to examine the role of substrate in increasing algal productivity and nutrient removal. The yearly mean productivity of flat ATS screens was 15.4 g · m(-2) · d(-1) . This was elevated to 39.6 g · m(-2) · d(-1) with a three-dimensional (3-D) screen, and to 47.7 g · m(-2) · d(-1) by avoiding high summer harvest temperatures. These methods enhanced nutrient removal (N, P) in algal biomass by 3.5 times. Eighty-six algal taxa (Ochrophyta [diatoms], Chlorophyta [green algae], and Cyan-obacteria [blue-green algae]) self-seeded from the GWR and demonstrated yearly cycling. Silica (SiO2 ) content of the algal biomass ranged from 30% to 50% of total biomass; phosphorus, nitrogen, and carbon content of the total algal biomass ranged from 0.15% to 0.21%, 2.13% to 2.89%, and 20.0% to 25.7%, respectively. Carbohydrate content (at 10%-25% of AFDM) was dominated by glucose. Lipids (fatty acid methyl ester; FAMEs) ranged widely from 0.5% to 9% AFDM, with Omega-3 fatty acids a consistent component. Mathematical modeling of algal produ-ctivity as a function of temperature, light, and substrate showed a proportionality of 4:3:3, resp-ectively. Under landscape ATS operation, substrate manipulation provides a considerable opportunity to increase ATS productivity, water quality amelioration, and biomass coproduction for fertilizers, fermentation energy, and omega-3 products. Based on the 3-D prod-uctivity and algal chemical composition demonstrated, ATS systems used for nonpoint source water treat-ment can produce ethanol (butanol) at 5.8× per unit area of corn, and biodiesel at 12.0× per unit area of soy beans (agricultural production US).


Proteomic profiling drug-induced apoptosis in non-small cell lung carcinoma: identification of RS/DJ-1 and RhoGDIalpha.
The growing knowledge of the tight connection between apoptosis and cancer has lead to an explosion of research revolving around apoptotic induction with chemotherapeutic agents and small molecule inhibitors. The chemotherapeutic agent paclitaxel (Taxol) activates mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase and, combined with MEK inhibition, synergistically enhances apoptosis. Here we implement a proteomic approach using two-dimensional gels coupled with mass spectrometry to identify proteins altered with this coordinated combination treatment. We found that the combined treatment of paclitaxel and MEK inhibitor uniquely altered the proteins RS/DJ-1 (RNA-binding regulatory subunit/DJ-1 PARK7) and RhoGDIalpha (Rho GDP-dissociation inhibitor alpha). Functional proteomic analysis by exogenous expression or short interfering RNA targeting confirmed a role in survival and apoptosis for these proteins. Analysis of primary lung tumors with matched adjacent normal tissue confirmed RS/DJ-1 overexpression in non-small cell lung carcinoma. This study shows the power of proteomic profiling coupled with functional analysis for the discovery of novel molecular targets and potential cancer cell-specific biomarkers.


HIV-1 Nef limits communication between linker of activated T cells and SLP-76 to reduce formation of SLP-76-signaling microclusters following TCR stimulation.
Signal initiation by engagement of the TCR triggers actin rearrangements, receptor clustering, and dynamic organization of signaling complexes to elicit and sustain downstream signaling. Nef, a pathogenicity factor of HIV, disrupts early TCR signaling in target T cells. To define the mechanism underlying this Nef-mediated signal disruption, we employed quantitative single-cell microscopy following surface-mediated TCR stimulation that allows for dynamic visualization of distinct signaling complexes as microclusters (MCs). Despite marked inhibition of actin remodeling and cell spreading, the induction of MCs containing TCR-CD3 or ZAP70 was not affected significantly by Nef. However, Nef potently inhibited the subsequent formation of MCs positive for the signaling adaptor Src homology-2 domain-containing leukocyte protein of 76 kDa (SLP-76) to reduce MC density in Nef-expressing and HIV-1-infected T cells. Further analyses suggested that Nef prevents formation of SLP-76 MCs at the level of the upstream adaptor protein, linker of activated T cells (LAT), that couples ZAP70 to SLP-76. Nef did not disrupt pre-existing MCs positive for LAT. However, the presence of the viral protein prevented de novo recruitment of active LAT into MCs due to retargeting of LAT to an intracellular compartment. These modulations in MC formation and composition depended on Nef's ability to simultaneously disrupt both actin remodeling and subcellular localization of TCR-proximal machinery. Nef thus employs a dual mechanism to disturb early TCR signaling by limiting the communication between LAT and SLP-76 and preventing the dynamic formation of SLP-76-signaling MCs.


Editorial: Focus on waste-heat harvesting via thermoelectric conversion: Materials, devices and systems for sustainable energy technologies.
The Focus collection on Waste-Heat Harvesting via Thermoelectric Conversion: Materials, Devices and Systems for Sustainable Energy Technologies collates several research articles and a Roadmap highlighting the most recent advances in the field of thermoelectricity from the viewpoint of both basic and applied research, with a special eye on the work of the Italian community.


Mutations in VIPAR cause an arthrogryposis, renal dysfunction and cholestasis syndrome phenotype with defects in epithelial polarization.
Arthrogryposis, renal dysfunction and cholestasis syndrome (ARC) is a multisystem disorder associated with abnormalities in polarized liver and kidney cells. Mutations in VPS33B account for most cases of ARC. We identified mutations in VIPAR (also called C14ORF133) in individuals with ARC without VPS33B defects. We show that VIPAR forms a functional complex with VPS33B that interacts with RAB11A. Knockdown of vipar in zebrafish resulted in biliary excretion and E-cadherin defects similar to those in individuals with ARC. Vipar- and Vps33b-deficient mouse inner medullary collecting duct (mIMDC-3) cells expressed membrane proteins abnormally and had structural and functional tight junction defects. Abnormal Ceacam5 expression was due to mis-sorting toward lysosomal degradation, but reduced E-cadherin levels were associated with transcriptional downregulation. The VPS33B-VIPAR complex thus has diverse functions in the pathways regulating apical-basolateral polarity in the liver and kidney.


Double hydrophilic polyphosphoester containing copolymers as efficient templating agents for calcium carbonate microparticles.
The use of calcium carbonate (CaCO3) microparticles is becoming more and more attractive in many fields especially in biomedical applications in which the fine tuning of the size, morphology and crystalline form of the CaCO3 particles is crucial. Although some structuring compounds, like hyaluronic acid, give satisfying results, the control of the particle structure still has to be improved. To this end, we evaluated the CaCO3 structuring capacity of novel well-defined double hydrophilic block copolymers composed of poly(ethylene oxide) and a polyphosphoester segment with an affinity for calcium like poly(phosphotriester)s bearing pendent carboxylic acids or poly(phosphodiester)s with a negatively charged oxygen atom on each repeating monomer unit. These copolymers were synthesized by a combination of organocatalyzed ring opening polymerization, thiol-yne click chemistry and protection/deprotection methods. The formulation of CaCO3 particles was then performed in the presence of these block copolymers (i) by the classical chemical pathway involving CaCl2 and Na2CO3 and (ii) by a process based on supercritical carbon dioxide (scCO2) technology in which CO3 2- ions are generated in aqueous media and react with Ca2+ ions. Porous CaCO3 microspheres composed of vaterite nanocrystals were obtained. Moreover, a clear dependence of the particle size on the structure of the templating agent was emphasized. In this work, we show that the use of the supercritical process and the substitution of hyaluronic acid for a carboxylic acid containing copolymer decreases the size of the CaCO3 particles by a factor of 6 (∼1.5 μm) while preventing their aggregation.


Flat Epithelial Atypia in Breast Core Needle Biopsies With Radiologic-Pathologic Concordance: Is Excision Necessary?
Flat epithelial atypia (FEA) is an alteration of terminal duct lobular units by a proliferation of ductal epithelium with low-grade atypia. No consensus exists on whether the diagnosis of FEA in core needle biopsy (CNB) requires excision (EXC). We retrospectively identified all in-house CNBs obtained between January 2012 and July 2018 with FEA. We reviewed all CNB slides and assessed radiologic-pathologic concordance. An upgrade was defined as invasive carcinoma (IC) and/or ductal carcinoma in situ in the EXC. The EXC slides of all upgraded cases were rereviewed. Out of ∼15,700 consecutive CNBs in the study period, 106 CNBs from 106 patients yielded FEA alone or with classic lobular neoplasia (LN). We excluded 52 CNBs (40 patients with prior/concurrent carcinoma and 12 without EXC). After rereview, we reclassified 14 cases (2 marked nuclear atypia, 10 focal atypical ductal hyperplasia, 2 benign). The final FEA study cohort consisted of 40 CNBs from 40 women. The CNB targeted mammographic calcifications in 36 (90%) cases, magnetic resonance imaging nonmass enhancement in 3 (8%), and 1 (2%) sonographic mass. All CNBs were deemed radiologic-pathologic concordant. FEA was present alone in 34 CNBs and with LN in 6. EXC yielded 2 low-grade IC, each spanning <2 mm, identified in tissue sections without biopsy site changes. The remaining 38 cases had no upgrade. Classic LN did not affect the upgrade. The upgrade rate of FEA was 5%; both minute, low-grade "incidental" IC. We conclude that nonsurgical management may be considered in patients without prior/concurrent carcinoma and radiologic-pathologic concordant CNB diagnosis of FEA.


Programmed Death-Ligand 1 on Antigen-presenting Cells Facilitates the Induction of Antigen-specific Cytotoxic T Lymphocytes: Application to Adoptive T-Cell Immunotherapy.
Programmed death-ligand 1 (PD-L1) binds to programmed death-1 (PD-1) on activated T cells and contributes to T-cell exhaustion. PD-L1 expressed on antigen-presenting cells (APCs) could be thought to inhibit the induction of Ag-specific cytotoxic T lymphocytes (CTLs) by transducing negative signal into T cells; however, the roles of PD-L1 on APCs have not yet been well examined. Therefore, we evaluated the roles of PD-L1 on APCs in the induction of Ag-specific CTLs. CD3 T cells isolated from cytomegalovirus (CMV)-seropositive healthy donors were stimulated with mature dendritic cells pulsed with CMV pp65-derived HLA-restricted peptides in the presence of anti-PD-L1 blocking antibody. Unexpectedly, PD-L1 blockade resulted in a less efficient induction of CMV-specific CTLs, suggesting that PD-L1 play a positive role in the induction of Ag-specific CTLs. For further evaluations and application to adoptive immunotherapy, we generated K562-based artificial APCs, which were retrovirally transduced with HLA class I molecules and various combinations of CD80/86 and PD-L1. K562/HLA+CD80/86+PD-L1 cells produced significantly higher induction of CMV-specific CTLs than K562/HLA or K562/HLA+CD80/86 cells without causing excessive differentiation or functional exhaustion of the induced CTLs, whereas PD-L1 itself did not have a stimulatory effect. Furthermore, only K562/HLA+CD80/86+PD-L1 cells pulsed with HLA-A*24:02-restricted Wilms tumor 1 (WT1) peptide clearly expanded WT1-specific CTLs from healthy donors. Our findings presumed that PD-L1 expressed on APCs along with CD80/86 enhanced the induction of Ag-specific CTLs probably depending on fine-tuning excessive stimulation of CD80/86, and that K562/HLA+CD80/86+PD-L1 cells has therapeutic potential as a novel type of artificial APCs for adoptive immunotherapy.


Unknown Extracellular and Bioactive Metabolites of the Genus Alexandrium: A Review of Overlooked Toxins.
Various species of Alexandrium can produce a number of bioactive compounds, e.g., paralytic shellfish toxins (PSTs), spirolides, gymnodimines, goniodomins, and also uncharacterised bioactive extracellular compounds (BECs). The latter metabolites are released into the environment and affect a large range of organisms (from protists to fishes and mammalian cell lines). These compounds mediate allelochemical interactions, have anti-grazing and anti-parasitic activities, and have a potentially strong structuring role for the dynamic of Alexandrium blooms. In many studies evaluating the effects of Alexandrium on marine organisms, only the classical toxins were reported and the involvement of BECs was not considered. A lack of information on the presence/absence of BECs in experimental strains is likely the cause of contrasting results in the literature that render impossible a distinction between PSTs and BECs effects. We review the knowledge on Alexandrium BEC, (i.e., producing species, target cells, physiological effects, detection methods and molecular candidates). Overall, we highlight the need to identify the nature of Alexandrium BECs and urge further research on the chemical interactions according to their ecological importance in the planktonic chemical warfare and due to their potential collateral damage to a wide range of organisms.


Stretchable Sensors with Tunability and Single Stimuli-Responsiveness through Resistivity Switching Under Compressive Stress.
The fascinating human somatosensory system with its complex structure is composed of numerous sensory receptors possessing distinct responsiveness to stimuli. It is a continuous source of inspiration for tactile sensors that mimic its functions. However, to achieve single stimulus-responsiveness with mechanical decoupling is particularly challenging in the light of structural design and has not been fully addressed to date. Here we propose a novel structural design inspired by combining the characteristics of electronic skin (e-skin) and electronic textile (e-textile) into a hybrid interface to achieve a stretchable single stimuli-responsive tactile sensor. The stencil printable biocarbon composite/silver-plated nylon hybrid interface possesses an extraordinary resistance switching (ΔR/R0 up to ∼104) under compressive stress which is controllable by the composite film-thickness. It achieves a very high normal pressure sensitivity (up to 60.8 kPa-1) in a wide dynamic range (up to ∼50 kPa) in the piezoresistive operation mode and can effectively decouple stresses induced by stretching or bending. In addition, the device is capable of high accuracy strain sensing in its capacitive operation mode through dimensional change dominant response. Because of these intriguing features, it has potential for the next-generation Internet of Things devices and user-interactive systems capable of providing visual feedback and more advanced robotics or even prosthetics.


Predictors of mortality in patients with flail chest: a systematic review.
To summarise the risk factors for mortality in patients with flail chest based on available evidence in the literature. A systematic review was completed using articles from PubMed, EMBASE, the Centre for Review and Dissemination database and the Cochrane Library. Additional studies were identified by hand-searching bibliographies, and grey literature was sought by searching abstracts from all relevant Emergency Medicine Conferences. All published and unpublished observational studies were included if they investigated estimates of association between a risk factor and mortality for patients with flail chest. This review found seven studies that matched the inclusion criteria, with a total of 944 patients. Patient age of ≥65 years was reported as a predictor of mortality when controlling for injury severity score (ISS) (p<0.02, OR 2.1, 95% CI 1.0 to 4.6). An ISS of ≥31 was reported to be a predictor of mortality in two studies; however, neither controlled for patient age. Pulmonary contusion, bilateral flail chest and hospital length of stay were not consistently found to be associated with mortality. The main independent predictors of mortality in patients with flail chest were reported to be increased age and ISS. More data are needed regarding the association of hospital length of stay, presence of pulmonary contusion and bilateral flail chest.


Anterior cruciate ligament tunnel placement.
The purpose of this cadaveric study was to analyze variation in anterior cruciate ligament (ACL) tunnel placement between surgeons and the influence of preferred surgical technique and surgeon experience level using three-dimensional (3D) computed tomography (CT). In this study, 12 surgeons drilled ACL tunnels on six cadaveric knees each. Surgeons were divided by experience level and preferred surgical technique (two-incision [TI], medial portal [MP], and transtibial [TT]). ACL tunnel aperture locations were analyzed using 3D CT scans and compared with radiographic ACL footprint criteria. The femoral tunnel location from front to back within the notch demonstrated a range of means of 16% with the TI tunnels the furthest back. A range of means of only 5% was found for femoral tunnel low to high positions by technique. The anterior to posterior tibial tunnel measure demonstrated wider variation than the medial to lateral position. The mean tibial tunnel location drilled by TT surgeons was more posterior than surgeons using the other techniques. Overall, 82% of femoral tunnels and 78% of tibial tunnels met all radiographic measurement criteria. Slight (1-7%) differences in mean tunnel placement on the femur and tibia were found between experienced and new surgeons. The location of the femoral tunnel aperture in the front to back plane relative to the notch roof and the anterior to posterior position on the tibia were the most variable measures. Surgeon experience level did not appear to significantly affect tunnel location. This study provides background information that may be beneficial when evaluating multisurgeon and multicenter collaborative ACL studies.


Inverse Kinematics of Concentric Tube Robots in the Presence of Environmental Constraints.
Inverse kinematics (IK) of concentric tube continuum robots (CTRs) is associated with two main problems. First, the robot model (e.g., the relationship between the configuration space parameters and the robot end-effector) is not linear. Second, multiple solutions for the IK are available. This paper presents a general approach to solve the IK of CTRs in the presence of constrained environments. It is assumed that the distal tube of the CTR is inserted into a cavity while its proximal end is placed inside a tube resembling the vessel enabling the entry to the organ cavity. The robot-tissue interaction at the beginning of the organ-cavity imposed displacement and force constraints to the IK problem to secure a safe interaction between the robot and tissue. The IK in CTRs has been carried out by treating the problem as an optimization problem. To find the optimized IK of the CTR, the cost function is defined to be the minimization of input force into the body cavity and the occupied area by the robot shaft body. The optimization results show that CTRs can keep the safe force range in interaction with tissue for the specified trajectories of the distal tube. Various simulation scenarios are conducted to validate the approach. Using the IK obtained from the presented approach, the tracking accuracy is achieved as 0.01 mm which is acceptable for the application.


Innovative therapy, monoclonal antibodies, and beyond: Highlights from the eighth annual meeting.
The eighth annual conference of "Innovative therapy, monoclonal antibodies, and beyond" was held in Milan on Jan. 26, 2018, and hosted by Fondazione IRCCS-Istituto Nazionale dei Tumori (Fondazione IRCCS INT). The conference was divided into two main scientific sessions, of i) pre-clinical assays and novel biotargets, and ii) clinical translation, as well as a third session of presentations from young investigators, which focused on recent achievements within Fondazione IRCCS INT on immunotherapy and targeted therapies. Presentations in the first session addressed the issue of cancer immunotherapy activity with respect to tumor heterogeneity, with key topics addressing: 1) tumor heterogeneity and targeted therapy, with the definition of the evolutionary Index as an indicator of tumor heterogeneity in both space and time; 2) the analysis of cancer evolution, with the introduction of the TRACERx Consortium-a multi-million pound UK research project focused on non-small cell lung cancer (NSCLC); 3) the use of anti-estrogen agents to boost immune recognition of breast cancer cells; and 4) the high degree of functional plasticity within the NK cell repertoire, including the expansion of adaptive NK cells following viral challenges. The second session addressed: 1) the effectiveness of radiotherapy to enhance the proportion of patients responsive to immune-checkpoint blockers (ICBs); 2) the use of MDSC scores in selecting melanoma patients with high probability to be responsive to ICBs; and 3) the relevance of the gut microbiome as a predictive factor, and the potential of its perturbation in increasing the immune response rate to ICBs. Overall, a picture emerged of tumor heterogeneity as the main limitation that impairs the effectiveness of anti-cancer therapies. Thus, the choice of a specific therapy based on reproducible and selective predictive biomarkers is an urgent unmet clinical need that should be addressed in order to increase the proportion of long-term responding patients and to improve the sustainability of novel drugs.


What are the Predictors of Death in Patients With Cranio-Cervical Artery Dissection?
Few studies have reported the predictive factors related to mortality in patients with cranio-cervical artery dissections (CCAD). Our aim was to investigate the predictors related to in-hospital mortality in patients with CCAD and its subgroups. Sixty-seven patients diagnosed with carotid artery dissection (CAD) or vertebral artery dissection (VAD), admitted to our clinic between 2000 and 2013, were retrospectively reviewed. Age, gender, modified Rankin Scale scores (pre-stroke and at admission), clinical presentation type, location of the dissection, risk factors, and treatments were analyzed as mortality-related prognostic factors. Of the 67 patients, 12 (17.9%) died, five (7.46%) with CAD and seven (10.44%) with VAD. We compared the prognostic characteristics of the surviving versus deceased patients with CCAD and in the subgroups with CAD and VAD. Age above 45 years, severe disability at admission, presentation with stroke, and intracranial VAD occurred more frequently in deceased patients and were independent variables related to mortality in patients with CCAD and its subgroup with VAD. Severe disability at admission alone was related to mortality in patients with CAD. Hypertension and hypercholesterolemia were independent variables related to mortality in patients with CCAD. Severe disability at admission was a mortality predictor in both CAD and VAD. Although the initial severity of stroke is reportedly related to poor outcomes in patients with CCAD, it has not previously been directly identified as a predictor of mortality in patients with CAD or VAD.


[Transparent nutrient medium for culturing Francisella tularensis].
A transparent nutrient medium T (tularemic) for cultivation of F. tularensis was developed on the basis of heart-brain infusion, yeast extract, bactotryptone, technical casamino acids, bactoagar, salts, L-cysteine and glucose. Introduction of an antibiotic (polymyxin B, 100 units/ml) to the medium composition provided selective conditions for growth of F. tularensis. Comparative study of the medium growth properties revealed that medium T by this parameter was not inferior to the Emel'ianova and Antsiferov media and by the yield of the bacterial mass was superior to them. A liquid variant of the medium (broth T) may be successfully used in various experiments requiring broth cultures of F. tularensis (interspecies conjugation, transformation, mutagenesis, etc.). The medium provides the use of wider temperature ranges for cultivation of F. tularensis. Because of transparency the medium may be used in identification of strains and their description by certain biochemical features. Medium T may be also used in studies with field materials (dead rodents, water).


Development of new microsatellite loci and multiplex reactions for muskellunge (Esox masquinongy).
The muskellunge (Esox masquinongy) is a valued fisheries species throughout its native range. Numerous studies have documented performance and phenotypic differences among muskellunge populations, but genetic markers for assessment have been lacking. We characterized 14 microsatellite loci and developed five multiplex polymerase chain reactions. Successful amplification of northern pike (Esox lucius) was observed for seven loci. These microsatellites will be useful for analysing population structure, performance characteristics of propagated strains, and helping to develop and monitor hatchery management guidelines for muskellunge.


Calculating the Tumor Nuclei Content for Comprehensive Cancer Panel Testing.
Comprehensive genetic panel testing generally requires that the analyzed tissues have a percent tumor nuclei (%TN) content of 20% or more to achieve assay performance comparable to the validated specifications. Pathologists play a crucial role in ensuring that the optimal results are achieved by accurately assigning %TN content of the available specimens and selecting the best material to submit for sequencing. This study addresses the issues in evaluating %TN, such as intraobserver variability, and examines whether focused training and feedback can improve pathologist performance. Nine referring institution pathologists (all board-certified and working at the core institute and the alignment hospitals under the National Cancer Genome scheme) evaluated 18 tumors that had been subjected to comprehensive genetic panel testing with the FoundationOne CDx assay. The %TN estimates provided by referring institution pathologists were compared with two standards: %TN assigned by the tumor sequencing institution's pathologist (a board-certified pathologist at Foundation Medicine, Inc.) and the computational %TN estimated from the mutant allele frequencies after sequencing was completed. The pathologists generally overestimated %TN in the first pretraining round of the evaluation, and the differences in the averaged %TN from the tumor sequencing institution and computational standards were statistically significant. However, the posttraining second-round results became significantly concordant with the standards. This study suggests that %TN content is empirically overestimated but the evaluation skill can be improved by providing a training and feedback program.


Pulmonary and systemic vascular effects of SRS-A blockade in conscious lambs.
There is preliminary evidence suggesting that hypoxic pulmonary vasoconstriction may be mediated by slow-reacting substance of anaphylaxis (SRS-A), which is comprised of leukotrienes C4, D4, and E4. We studied the effects of the SRS-A antagonist FPL 57231 (FPL) on the hypoxic pulmonary vasoconstrictor response and on systemic vascular resistance in awake, chronically instrumented young lambs. Two other studies were performed to ascertain whether FPL's vasodilation was specific for hypoxic pulmonary vasoconstriction: the effect of FPL infusion in pulmonary and systemic vascular resistance was measured in six normoxic lambs, and the effect of FPL on 5-hydroxytryptamine (5-HT)-mediated vasoconstriction was determined. In seven lambs, mean pulmonary arterial pressure was 21 mmHg in room air and 28 mmHg in hypoxia (Po2 = 43 Torr). During hypoxia, FPL infusion (2 mg X kg-1 X min-1) reversibly decreased pulmonary arterial pressure to 15 mmHg; pulmonary arteriolar resistance also fell below normoxia levels with FPL. FPL also caused a fall in aortic pressure and systemic vascular resistance in these hypoxic lambs, but the decrease in systemic resistance was less than the fall in pulmonary resistance. beta-Adrenergic blockade using propranolol (1 mg/kg) did not affect the pulmonary vasodilation caused by FPL. In six normoxic lambs, FPL infusion also significantly decreased pulmonary and systemic vascular resistance (29% in each case). These data are consistent with the idea that leukotrienes may be involved in adjusting both pulmonary and systemic vascular tone, but further work is necessary to establish whether FPL's vasodilation is mediated via its leukotriene antagonism or is a nonspecific effect of FPL.


Clinical value of prostacyclin and its analogs in the management of pulmonary arterial hypertension.
Pulmonary arterial hypertension (PAH) is associated with changes in vascular tone as well as vascular structure, with the relative contribution of each dependent upon the etiology of the increased vascular resistance. Prostacyclin treatment has markedly improved both the therapeutic options and the prognosis of PAH. The beneficial effect of prostacyclin in PAH is linked to the powerful vasodilating capacity and, even more importantly, to the inhibition of platelet aggregation, smooth muscle proliferation and antiinflammatory actions. Since prostacyclin has a very short plasma half-life, continuous intravenous administration via a portable infusion pump must be carried out. Because of the complexity of the delivery and the associated complications, intravenous prostacyclin (epoprostenol) therapy is restricted to patients with late NYHA Class III and Class IV and thus alternative modes of delivery in less advanced PAH are desirable. Recent clinical studies with more stable prostacyclin analogs such as subcutaneously delivered treprostinil, orally active beraprost and aerosolized iloprost have demonstrated beneficial effects of each of these prostanoids, especially in NYHA Class II and III patients and, therefore, these agents should be considered first for prostanoid therapy in the early stages of PAH. Prostaglandins may be more effective in conjunction with endothelin receptor antagonists or phosphodiesterase inhibitors and an increasing number of studies are now addressing the combined efficiency and safety of these combinations. This update will focus on the current development status of PAH therapy with prostacyclin and its analogs. Special attention will be accorded to the selection of patients for prostanoid therapy, therapy monitoring and improvement of therapeutic efficacy by addition of other new therapeutic agents to prostaglandins. Survival benefits and special aspects of bridging-to-transplant therapy are also important aspects of the review.


The evolution of migration in a seasonal environment.
Despite the fact that migration occurs in a wide variety of taxa worldwide, little is known about the conditions under which migration is expected to evolve from an ancestral resident population. We develop a model that focuses on ecological factors affecting the evolution of migration in a seasonal environment within a genetically explicit framework. We model the evolution of migration for two common types of migration: 'shared breeding' where migrants share a breeding ground with residents and migrate to a separate non-breeding area, versus 'shared non-breeding', where migrants share a non-breeding ground with residents and migrate to a separate breeding area. Ecologically, migration is more easily established in the shared-breeding case versus the shared-non-breeding case. Genetically, the additive effect of a migratory allele affects its establishment more in the shared-non-breeding case versus the shared-breeding case, whereas the dominance effect of the allele affects its establishment more in the shared-breeding case versus the shared-non-breeding case. Generally, migratory alleles can invade even when residents are competitively superior to migrants during the shared season. Partial migration occurs when the population is polymorphic for migratory and non-migratory alleles, and is dependent upon which season is shared and the additive and dominance behaviour of the migratory allele.


[Adult onset Langerhans cell histiocytosis with progressive cerebellar ataxia and spastic paraparesis].
A 44-year-old woman with progressive cerebellar ataxia and spastic paraparesis was referred to our hospital. Brain MRI showed bilateral high signals in superior, middle, and inferior cerebellar peduncles on the T2 weighted images. After 3 years, her symptoms progressively worsened in spite of various therapies including whole brain irradiation and high dose oral prednisone. No evidence of diabetes insipidus was noted. In MRI, brainstem lesions expanded to both hemispheres of the cerebellum without enhancement by contrast medium. We confirmed diagnosis of LCH by skin biopsy of intractable truncal rash which emerged after neurological symptoms.


Occludin protein expression in human cervical cancer and its association with patient's clinical characteristics.
The objective of the study was to investigate the expression of the tight junction protein occludin (encoded by OCLN gene) in human cervical cancer and its association with clinical features of patients. Sixty-one patients with cervical cancer were included in this study from June 30, 2015 to April 30, 2017. Immuno-histochemical assay was applied to examine the expression of occludin protein in 61 cervical cancer tissues and matched adjacent cancer normal tissues. The association of occludin protein expression with clinical pathology characteristics was analyzed. Occludin protein was mainly expressed in cell membranes and cytoplasm of both the cervical cancer cell and the normal cells. The protein was manifested with brownish-yellow granules. In cervical cancer tissues, the positive rate of occludin protein was 77.05% (47/61), whereas, in adjacent normal tissues of the cancer, the positive rate was 96.72% (59/61). Therefore, the positive rate of occludin in cervical cancer tissues was significantly lower than that of the adjacent cancer tissues (P < 0.05). Occludin protein expression level was not significantly correlated with the age (P > 0.05), tumor size (P > 0.05), International Federation of Gynecology and Obstetrics staging (P > 0.05), pathological grades (P > 0.05), and lymph node metastasis (P > 0.05) of the patients. Occludin protein may contribute to the development of cervical cancer. However, it was not correlated with the clinical features.


Incidence of and survival from Wilms' tumour in adults in Europe: data from the EUROCARE study.
Wilms' tumour, or nephroblastoma, is an embryonal cancer of the kidney that occurs mainly in young children. This is a very rare tumour among adults, with an incidence rate of less than 0.2 per million per year. The aims of this study were to report the survival of adults diagnosed with nephroblastoma in Europe and to analyse time trends and geographic variations in survival. All the adults (age range 15-99 years) diagnosed with a Wilms' tumour during 1983-1994 and registered by one of the 22 cancer registries in 16 countries contributing to the EUROCARE (European cancer registries study on cancer patients' survival and care) database were analysed. Relative survival at 1 and 5 years after diagnosis was estimated by age, sex, geographic area, period of diagnosis and tumour stage. A total of 143 patients, with a median age of 34 years, were included in the analysis. Crude annual incidence rates varied geographically between 0.17 and 0.27 per million. Overall relative survival was 69.9% (95% confidence interval (CI) 61.8-78.0%) at 1 year and 47.3% (38.2-56.4%) at 5 years. Survival was 2.1-fold higher for women than for men (95% CI 1.3-3.5). There was a non-significant trend for better survival for younger patients and localised tumours, but no improvement in survival by period of diagnosis. Survival was not different between geographic areas. Our results suggest a poorer outcome of nephroblastoma in adults compared with published results in children. This may, at least partly, be explained by the rarity of this diagnosis. Prognosis may be improved by the use of specific treatment guidelines for nephroblastoma in adults.


A mutation in the human heme A:farnesyltransferase gene (COX10 ) causes cytochrome c oxidase deficiency.
Cytochrome c oxidase (COX) defects are found in a clinically and genetically heterogeneous group of mitochondrial disorders. To date, mutations in only two nuclear genes causing COX deficiency have been described. We report here a genetic linkage study of a consanguineous family with an isolated COX defect and subsequent identification of a mutation in a third nuclear gene causing a deficiency of the enzyme. A genome-wide search for homozygosity allowed us to map the disease gene to chromosome 17p13.1-q11.1 (Z (max)= 2.46; theta = 0.00 at the locus D17S799). This region encompasses two genes, SCO1 and COX10, encoding proteins involved in COX assembly. Mutation analysis followed by a complementation study in yeast permitted us to ascribe the COX deficiency to a homozygous missense mutation in the COX10 gene. This gene encodes heme A:farnesyltransferase, which catalyzes the first step in the conversion of protoheme to the heme A prosthetic groups of the enzyme. All three nuclear genes now linked to isolated COX deficiency are involved in the maturation and assembly of COX, emphasizing the major role of such genes in COX pathology.


Chronic kidney disease in India: challenges and solutions.
Chronic diseases have become a major cause of global morbidity and mortality even in developing countries. The burden of chronic kidney disease (CKD) in India cannot be assessed accurately. The approximate prevalence of CKD is 800 per million population (pmp), and the incidence of end-stage renal disease (ESRD) is 150-200 pmp. The most common cause of CKD in population-based studies is diabetic nephropathy. India currently has 820+ nephrologists, 710+ hemodialysis units with 2,500+ dialysis stations and 4,800+ patients on CAPD. There are 172+ transplant centers, two-thirds of which are in South India and mostly privately run. Nearly 3,500 transplants are done annually, the total number of cadaver donors being approximately 700 till now. Thus, taken together, nearly 18,000-20,000 patients (10% of new ESRD cases) in India get renal replacement therapy. The cost of single hemodialysis varies between USD 15 and 40 with an additional cost of erythropoietin being USD 150-200/month. The cost of CAPD using a 'Y' set with 3 exchanges/week is USD 400/month. The cost of the transplant procedure in a state-run hospital is USD 800-1,000, and the cost of immunosuppression using tacrolimus, steroid and mycophenolate is USD 350-400/month. Until recently, the government did not recognize CKD/ESRD as a significant problem in India. However, some illustrious activities in relation to CKD brought attention of the media and policymakers to this very common but till now deprived group of diseases. On the one side the government has initiated a process by which it is planning to establish stand-alone hemodialysis units in the country to increase the facilities at an affordable cost, and on the transplant side it had launched a National Organ Transplant Program to facilitate transplantation on a national scale. Hemodialysis program is halfway to being implemented. Thus, in India there is still a long way to go with respect to CKD. Until then, in a country like India, screening of high-risk individuals for CKD and the risk factors is the best bet.


The significance of small intestinal epithelium in gastric antral biopsies in children.
Intestinal metaplasia of the gastric antrum is common in adults with chronic gastritis and occurs in Helicobacter -associated gastritis in children. This study examined the frequency and clinical correlates of intestinal epithelium in 1690 consecutive antral biopsies obtained from children over a 2-year period in a tertiary pediatric care facility. Intestinal epithelium in gastric glands not associated with overlying villi was present in 22 (1.3%) biopsies. These came from 20 patients, 2-17 years of age, none of whom had clinical or histologic evidence of Helicobacter infection or significant chronic gastritis. Eight (40%) had an antral pancreatic rest, 8 had some other localized antral abnormality, and 4 were endoscopically normal. Four additional patients with a pancreatic rest had no intestinal epithelium. Six surgically resected rests and 2 rests found at autopsy were also reviewed. Heterotopic intestinal epithelium was present in 1 of the 2 postmortem specimens but was absent from all 6 surgically resected lesions. No intestinal epithelium was present in 67 antral biopsies with Helicobacter gastritis observed during this same period. Although the intestinal epithelium in these patients could be metaplastic, it more likely represents inadvertent sampling of the gastroduodenal junction induced by a lesion in the distal antrum or a focus of heterotopic epithelium and might best be addressed in the surgical pathology report by a comment to this effect. The distinction from metaplasia is more than semantic, because a diagnosis of intestinal metaplasia can have adverse clinical implications and should be made with caution in a child.


Slowing of sodium channel opening kinetics in squid axon by extracellular zinc.
The interaction of Zn ion on Na channels was studied in squid giant axons. At a concentration of 30 mM Zn2+ slows opening kinetics of Na channels with almost no alteration of closing kinetics. The effects of Zn2+ can be expressed as a "shift" of the gating parameters along the voltage axis, i.e., the amount of additional depolarization required to overcome the Zn2+ effect. In these terms the mean shifts caused by 30 mM Zn2+ were +29.5 mV for Na channel opening (on) kinetics (t1/2 on), +2 mV for closing (off) kinetics (tau off), and +8.4 mV for the gNa-V curve. Zn2+ does not change the shape of the instantaneous I-V curve for inward current, but reduces it in amplitude by a factor of or approximately 0.67. Outward current is unaffected. Effects of Zn2+ on gating current (measured in the absence of TTX) closely parallel its actions on gNa. On gating current kinetics are shifted by +27.5 mV, off kinetics by +6 mV, and the Q-V distribution by +6.5 mV. Kinetic modeling shows that Zn2+ slows the forward rate constants in activation without affecting backward rate constants. More than one of the several steps in activation must be affected. The results are not compatible with the usual simple theory of uniform fixed surface charge. They suggest instead that Zn2+ is attracted by a negatively charged element of the gating apparatus that is present at the outer membrane surface at rest, and migrates inward on activation.


[Differential analgesic treatment in arthrosis and arthritis].
The leading symptom of arthrosis and arthritis is pain. As in the case of pharmacotherapy fortumor pain, a stepped approach is also recommended for rheumatic complaints. Mild-to-moderate pain in noninflammatory arthrosis can be ameliorated by paracetamol or low-dose ibuprofen. If inflammation is present, nonsteroidal anti-inflammatory drugs (NSAIDs) must be employed. If this treatment does not suffice to manage systemic arthritis, oral short-acting corticosteroids are applied. Intra-articular corticosteroid injections can be used to individual inflamed active joints. For chronic pain, opioids may be necessary in addition to NSAID treatment. The use of NSAIDs is limited by gastrointestinal side effects. In the case of risk patients, therefore, preventive measures must be taken, and PPI or, instead of NSAIDs, coxibs employed in addition.


[Perception of physicians and nurses on presence of family during invasive procedures].
The purpose of this descriptive study was to determine the perception of physicians and nurses on family presence during invasive procedures. The study population consisted of 100 physicians and 100 nurses from five hospitals which have more than 500 beds in B city. Data collection was done from August 20 to September 19, 2007. The Family Presence During Resuscitation (FPDR) Inventory developed by Fulbrook, Albarran and Latour (2005) was used as the study instrument. The collected data was analyzed by t-test, ANOVA and Duncan's multiple range test using SPSS/WIN 14.0 version. Nurses had more positive attitudes to allowing family members to be present during invasive procedures than physicians did. However, compared to physicians, nurses had more concern about problems of confidentiality, arguing with medical team, and increased rate of legal action if family members were present during invasive procedure. The results of this study indicate that educational programs and policy for family presence be required within the hospital to enhance the perception of physicians and nurses to the family presence.


Mortality Outcomes of Patients with Chronic Kidney Disease and Chronic Obstructive Pulmonary Disease.
Chronic obstructive pulmonary disease (COPD) is associated with higher mortality in the general population. We studied the associations between COPD and death among chronic kidney disease (CKD) patients along with reporting cause-specific death data. We included 56,960 patients with stages 3 and 4 CKD who were followed in a large health care system. Associations between COPD and all-cause mortality and various causes of death (respiratory deaths, cardiovascular deaths, malignancy-related deaths and deaths due to other reasons) were studied using the Cox proportional hazards and competing risk models. Out of 56,960 CKD patients, 4.7% (n = 2,667) had underlying COPD. Old age, presence of diabetes, hypertension, coronary artery disease, congestive heart failure, and smoking were associated with higher risk for COPD. During a median follow-up of 3.7 years, 15,969 patients died. After covariate adjustment, COPD was associated with a 41% increased risk (95% CI 1.31-1.52) for all-cause mortality, and fourfold increased risk (sub-hazard ratio 4.36, 95% CI 3.54-5.37) for respiratory-related deaths. In a sensitivity analysis that was performed by defining COPD as the use of relevant International Classification of Diseases-9 codes and medications used to treat COPD, similar results were noted. COPD is associated with higher risk for death among those with CKD, and an underlying lung disease accounts for significant proportion of deaths. These data highlight the need for further prospective studies to understand the underlying mechanisms and potential interventions to improve outcomes in this population.


Risk stratification of patients with locally aggressive differentiated thyroid cancer. Results of the MSDS trial.
The Multicentre Study Differentiated Thyroid Cancer (MSDS) collective represents a well defined group of patients with locally aggressive thyroid carcinomas (pT4; AJCC/UICC 1997). The aim of the present study was to compare the survival of patients with minimum and extensive extrathyroidal growth according to the new AJCC/UICC TNM staging system 2009. The follow-up data of 347 patients were analysed. Patients were reclassified according to the current AJCC/UICC 2009 classification. The event-free and overall survival was evaluated using Kaplan-Meier analysis. In addition, postoperative complications and status of disease were documented. 327 patients were assigned to stage pT3 and 20 patients to stage pT4a, respectively. Median follow-up was 6.1 years (range 0.04-9.8 years). 92.5% of patients reached complete remission. There were 7.8 % recurrences in the thyroid bed, in locoregional lymph nodes and/or in distant sites. The overall survival was >98% both in pT3 and pT4a patients (p = n. s.). In contrast, the event-free survival was significantly less favourable in pT4a patients (p < 0.001). Using multivariate analysis the following parameters were significant predictors of event-free survival: histological tumour type, degree of extrathyroidal extension and nodal metastasis (p < 0.05). The MSDS patients with locally aggressive differentiated thyroid cancer showed an excellent overall survival during a median follow-up of 6.1 years. According to the current AJCC/UICC 2009 classification, pT3 patients with minimal extrathyroidal extension revealed a significantly better event-free survival than pT4a patients with extensive extrathyroidal growth.


Dual vs single protease inhibitor therapy following antiretroviral treatment failure: a randomized trial.
Management of antiretroviral treatment failure in patients receiving protease inhibitor (PI)-containing regimens is a therapeutic challenge. To assess whether adding a second PI improves antiviral efficacy of a 4-drug combination in patients with virologic failure while taking a PI-containing regimen. Multicenter, randomized, 4-arm trial, double-blind and placebo-controlled for second PI, conducted between October 1998 and April 2000, for which there was a 24-week primary analysis with extension to 48 weeks. Thirty-one participating AIDS (acquired immunodeficiency syndrome) Clinical Trials Units in the United States. A total of 481 human immunodeficiency virus (HIV)-infected persons with prior exposure to a maximum of 3 PIs and viral load above 1000 copies/mL. Selectively randomized assignment (per prior PI exposure) to saquinavir (n = 116); indinavir (n = 69); nelfinavir (n = 139); or placebo twice per day (n = 157); in combination with amprenavir, abacavir, efavirenz, and adefovir dipivoxil. Primary efficacy analysis involved the proportion with viral load below 200 copies/mL at 24 weeks. Other measures were changes in viral load and CD4 cell count from baseline, adverse events, and HIV drug susceptibility. Of 481 patients, 148 (31%) had a viral load below 200 copies/mL at week 24. The proportions of patients with a viral load below 200 copies/mL in the saquinavir, indinavir, nelfinavir, and placebo arms were 34% (40/116), 36% (25/69), 34% (47/139), and 23% (36/157), respectively. The proportion in the combined dual-PI arms was higher than in the amprenavir-plus-placebo arm (35% [112/324] vs 23% [36/157], respectively; P =.002). Overall, a higher proportion of nonnucleoside reverse transcriptase inhibitor (NNRTI)-naive patients had a viral load below 200 copies/mL compared with NNRTI-experienced patients (43% [115/270] vs 16% [33/211], respectively; P<.001). Baseline HIV-1 hypersusceptibility to efavirenz (< or = 0.4-fold difference in susceptibility compared with reference virus) was associated with suppression of viral load at 24 weeks to below 200 copies/mL (odds ratio [OR], 3.49; 95% confidence interval [CI], 1.62-7.33; P =.001), and more than 10-fold reduction in efavirenz susceptibility, with less likelihood of suppression at 24 weeks (OR, 0.28; 95% CI, 0.09-0.87; P =.03). In this study of antiretroviral-experienced patients with advanced immunodeficiency, viral load suppression to below 200 copies/mL was achieved in 31% of patients with regimens containing 4 or 5 new drugs. Use of 2 PIs, being naive to NNRTIs, and baseline hypersusceptibility to efavirenz were associated with a favorable outcome.


Dual Effect of Glucuronidation of a Pyrogallol-Type Phytophenol Antioxidant: A Comparison between Scutellarein and Scutellarin.
To explore whether and how glucuronidation affects pyrogallol-type phytophenols, scutellarein and scutellarin (scutellarein-7-O-glucuronide) were comparatively investigated using a set of antioxidant analyses, including spectrophotometric analysis, UV-vis spectra analysis, and ultra-performance liquid chromatography coupled with electrospray ionization-quadrupole time-of-flight tandem mass spectrometry (UPLC-ESI-Q-TOF-MS/MS) analysis. In spectrophotometric analyses of the scavenging of 1,1-diphenyl-2-picrylhydrazyl (DPPH•), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+•), and 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide radicals (PTIO•) and the reduction of Cu2+ ions, scutellarein showed lower IC50 values than scutellarin. However, in •O₂--scavenging spectrophotometric analysis, scutellarein showed higher IC50 value than scutellarin. The analysis of UV-Vis spectra obtained after the Fe2+-chelating reaction of scutellarin showed a typical UV-Vis peak (λmax = 611 nm), while scutellarein showed no typical peak. In UPLC-ESI-Q-TOF-MS/MS analysis, mixing of scutellarein with DPPH• yielded MS peaks (m/z 678, 632, 615, 450, 420, 381, 329, 300, 288, 227, 196, 182, 161, and 117) corresponding to the scutellarein-DPPH adduct and an MS peak (m/z 570) corresponding to the scutellarein-scutellarein dimer. Scutellarin, however, generated no MS peak. On the basis of these findings, it can be concluded that glucuronidation of pyrogallol-type phytophenol antioxidants has a dual effect. On the one hand, glucuronidation can decrease the antioxidant potentials (except for •O₂- scavenging) and further lower the possibility of radical adduct formation (RAF), while on the other hand, it can enhance the •O₂--scavenging and Fe2+-chelating potentials.


Drug-induced hypothermia in stroke models: does it always protect?
Ischemic stroke is a common neurological disorder lacking a cure. Recent studies show that therapeutic hypothermia is a promising neuroprotective strategy against ischemic brain injury. Several methods to induce therapeutic hypothermia have been established; however, most of them are not clinically feasible for stroke patients. Therefore, pharmacological cooling is drawing increasing attention as a neuroprotective alternative worthy of further clinical development. We begin this review with a brief introduction to the commonly used methods for inducing hypothermia; we then focus on the hypothermic effects of eight classes of hypothermia-inducing drugs: the cannabinoids, opioid receptor activators, transient receptor potential vanilloid, neurotensins, thyroxine derivatives, dopamine receptor activators, hypothermia-inducing gases, adenosine, and adenine nucleotides. Their neuroprotective effects as well as the complications associated with their use are both considered. This article provides guidance for future clinical trials and animal studies on pharmacological cooling in the setting of acute stroke.


Abundantly expressed class of noncoding RNAs conserved through the multicellular evolution of dictyostelid social amoebas.
Aggregative multicellularity has evolved multiple times in diverse groups of eukaryotes, exemplified by the well-studied development of dictyostelid social amoebas, for example, Dictyostelium discoideum However, it is still poorly understood why multicellularity emerged in these amoebas while the majority of other members of Amoebozoa are unicellular. Previously, a novel type of noncoding RNA, Class I RNAs, was identified in D. discoideum and shown to be important for normal multicellular development. Here, we investigated Class I RNA evolution and its connection to multicellular development. We identified a large number of new Class I RNA genes by constructing a covariance model combined with a scoring system based on conserved upstream sequences. Multiple genes were predicted in representatives of each major group of Dictyostelia and expression analysis confirmed that our search approach identifies expressed Class I RNA genes with high accuracy and sensitivity and that the RNAs are developmentally regulated. Further studies showed that Class I RNAs are ubiquitous in Dictyostelia and share highly conserved structure and sequence motifs. In addition, Class I RNA genes appear to be unique to dictyostelid social amoebas because they could not be identified in outgroup genomes, including their closest known relatives. Our results show that Class I RNA is an ancient class of ncRNAs, likely to have been present in the last common ancestor of Dictyostelia dating back at least 600 million years. Based on previous functional analyses and the presented evolutionary investigation, we hypothesize that Class I RNAs were involved in evolution of multicellularity in Dictyostelia.


Preliminary report of psychiatric disorders in survivors of a severe earthquake.
The authors' goal was to study psychiatric morbidity after a natural disaster in rural India. As members of a volunteer medical team assigned to 23 households in three villages in India affected by an earthquake, the authors examined the results of semi-structured interviews used to interview all of the adults (older than 14 years) in these households (N=56). DSM-III-R diagnoses were assigned on the basis of these interviews, and non-parametric tests were applied to comparisons of subjects who were or were not given a psychiatric diagnosis. Thirty-three (59%) of the subjects received a psychiatric diagnosis; the most common diagnoses were posttraumatic stress disorder (13 subjects [23%]) and major depression (12 subjects [21%]). Psychiatric morbidity was associated with female sex, destruction of house, and destruction of possessions. Attention to the mental health needs of disaster survivors in third world countries is indicated.


Decreased ketonaemia in the monosodium glutamate-induced obese rats.
Plasma concentrations of total ketone bodies, acetoacetate (AcAc) and 3-hydroxybutyrate (3-OHBA) in monosodium glutamate (MSG)-induced obese rats were measured. MSG-treated rats showed higher Lee's indices, shorter naso-anal and tail length, and a more marked intraperitoneal fat deposition than control rats. Plasma concentrations of glucose, free fatty acid, triglyceride and phospholipids were significantly increased in the MSG-treated rats as compared to the control rats (24 weeks-old). Plasma levels of total ketone bodies, AcAc and 3-OHBA were all decreased in the MSG-treated rats as compared to control rats. The ratio, 3-OHBA/AcAc in the MSG-treated rats were not different from those in the control rats.


2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)-elicited effects on bile acid homeostasis: Alterations in biosynthesis, enterohepatic circulation, and microbial metabolism.
2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is a persistent environmental contaminant which elicits hepatotoxicity through activation of the aryl hydrocarbon receptor (AhR). Male C57BL/6 mice orally gavaged with TCDD (0.01-30 µg/kg) every 4 days for 28 days exhibited bile duct proliferation and pericholangitis. Mass spectrometry analysis detected a 4.6-fold increase in total hepatic bile acid levels, despite the coordinated repression of genes involved in cholesterol and primary bile acid biosynthesis including Cyp7a1. Specifically, TCDD elicited a >200-fold increase in taurolithocholic acid (TLCA), a potent G protein-coupled bile acid receptor 1 (GPBAR1) agonist associated with bile duct proliferation. Increased levels of microbial bile acid metabolism loci (bsh, baiCD) are consistent with accumulation of TLCA and other secondary bile acids. Fecal bile acids decreased 2.8-fold, suggesting enhanced intestinal reabsorption due to induction of ileal transporters (Slc10a2, Slc51a) and increases in whole gut transit time and intestinal permeability. Moreover, serum bile acids were increased 45.4-fold, consistent with blood-to-hepatocyte transporter repression (Slco1a1, Slc10a1, Slco2b1, Slco1b2, Slco1a4) and hepatocyte-to-blood transporter induction (Abcc4, Abcc3). These results suggest that systemic alterations in enterohepatic circulation, as well as host and microbiota bile acid metabolism, favor bile acid accumulation that contributes to AhR-mediated hepatotoxicity.


Bone Saw for Calcified Peyronie's Disease Plaques.
To determine the incidence of calcified Peyronie's disease plaque that cannot be cut with a blade in a 100-case series and to describe the use of a soft tissue-protecting bone saw for plaque incision. Chart reviews were done of all surgically treated Peyronie's disease patients at our center between October 1996 and December 2012. 100 cases were included. We evaluated our novel technique of tissue-protecting bone saw surgical use. 100 consecutive patients underwent surgery for Peyronie's disease, and 6 required transverse bone saw plaque incision due to the severity of calcification that could not be cut with a blade. Four of those 6 underwent grafting procedures with porcine submucosal intestinal substance (SIS) and 2 underwent placement of inflatable penile prosthesis (IPP) after plaque incision. There were no surgical complications. Both IPP patients had functioning prosthesis 4 and 7.3 years after surgery. One SIS-graft patient required re-operation for more proximal curvature 11 months later and ultimately required multiple plaque incisions and an IPP. Densely calcified plaques occurred in 6% of a surgical series of Peyronie's disease patients. The vibrating bone saw is a novel technique to incise calcified plaques before grafting or IPP placement.


Biphasic insulin aspart 30 for the treatment of type 1 diabetes mellitus.
Type 1 diabetes mellitus is associated with acute and long-term complications, to which pre- and postprandial hyperglycemia are independent contributors. The objective of this review was to evaluate evidence-based information using biphasic insulin aspart 30 in the treatment of type 1 diabetes mellitus. The study reviewed the Cochrane Database and scientific literature (PubMed) published until January 2008 using the words biphasic insulin aspart 30 insulin or premixed aspart insulin. Biphasic insulin aspart 30 is similar in efficacy to biphasic human insulin in improving hemoglobin A(1c) levels, with the advantage of a better postprandial glucose profile. There is evidence supporting the efficacy and safety of biphasic insulin aspart 30 insulin. However, the need for well-designed clinical trials aimed at understanding the potential differences in safety and efficacy between patients with type 1 and type 2 diabetes is crucial.


Conformational and Interaction Landscape of Histone H4 Tails in Nucleosomes Probed by Paramagnetic NMR Spectroscopy.
The fundamental repeat unit of chromatin, the nucleosome, consists of approximately 147 base pairs of double-stranded DNA and a histone protein octamer containing two copies each of histones H2A, H2B, H3, and H4. Each histone possesses a dynamically disordered N-terminal tail domain, and it is well-established that the tails of histones H3 and H4 play key roles in chromatin compaction and regulation. Here we investigate the conformational ensemble and interactions of the H4 tail in nucleosomes by means of solution NMR measurements of paramagnetic relaxation enhancements (PREs) in recombinant samples reconstituted with 15N-enriched H4 and nitroxide spin-label tagged H3. The experimental PREs, which report on the proximities of individual H4 tail residues to the different H3 spin-label sites, are interpreted by using microsecond time-scale molecular dynamics simulations of the nucleosome core particle. Collectively, these data enable improved localization of histone H4 tails in nucleosomes and support the notion that H4 tails engage in a fuzzy complex interaction with nucleosomal DNA.


Adjuvant immunotherapy with nonviable Mycobacterium smegmatis in resected primary lung carcinoma. A randomized clinical trial of 219 patients.
Two hundred and nineteen patients with resected lung carcinoma were randomized 3 weeks after surgery between two treatment arms: a control group (110 cases) and an immunotherapy group (109 cases). The immunostimulant was a nonviable saprophytic mycobacterium, M. smegmatis, given monthly by subcutaneous injection in four sites. The two groups were equivalent in terms of prognostic factors, including a nonsignificant difference favoring the control group based on the N (node) classification. This interim analysis was carried out on June 1, 1981. Treatment comparison by the log-rank test did not show any significant differences between these two groups in regards to disease-free interval and overall survival. There was no significant difference between the two groups after stratification of the comparison according to the N classification or adjustment with a subset of eight prognostic parameters through the Cox model. The initially expected difference (20% 1-year survival) will probably not be achieved, given these interim results, but patients will continue to be treated and followed-up according to the protocol as to allow further evaluation of this nonspecific immunotherapy.


Collaborative regulation of development but independent control of metabolism by two epidermis-specific transcription factors in Caenorhabditis elegans.
Cell fate specification is typically initiated by a master regulator, which is relayed by tissue-specific regulatory proteins (usually transcription factors) for further enforcement of cell identities, but how the factors are coordinated among each other to "finish up" the specification remains poorly understood. Caenorhabditis elegans epidermis specification is initiated by a master regulator, ELT-1, that activates its targets, NHR-25 and ELT-3, two epidermis-specific transcription factors that are important for development but not for initial specification of epidermis, thus providing a unique paradigm for illustrating how the tissue-specific regulatory proteins work together to enforce cell fate specification. Here we addressed the question through contrasting genome-wide in vivo binding targets between NHR-25 and ELT-3. We demonstrate that the two factors bind discrete but conserved DNA motifs, most of which remain in proximity, suggesting formation of a complex between the two. In agreement with this, gene ontology analysis of putative target genes suggested differential regulation of metabolism but coordinated control of epidermal development between the two factors, which is supported by quantitative analysis of expression of their specific or common targets in the presence or absence of either protein. Functional validation of a subset of the target genes showed both activating and inhibitory roles of NHR-25 and ELT-3 in regulating their targets. We further demonstrated differential control of specification of AB and C lineage-derived epidermis. The results allow us to assemble a comprehensive gene network underlying C. elegans epidermis development that is likely to be widely used across species and provides insights into how tissue-specific transcription factors coordinate with one another to enforce cell fate specification initiated by its master regulator.


Role of choline in the nutrition of the rumen protozoon Entodinium caudatum.
A requirement of choline for growth of Entodinium caudatum in a simplified culture medium has been demonstrated. Ethanolamine, N-methylethanolamine, or N-dimethylethanolamine were ineffective as substitutes. In the rumen, the normal environment of this organism, levels of free choline were virtually zero even after ingestion of pasture containing phosphatidylcholine which was rapidly catabolized. Free [Me-14C]choline is very rapidly cleared from rumen fluid, a little being incorporated into the phosphatidylcholine of protozoa, but the clearance also occurs in animals with defaunated rumens. It is suggested that E. caudatum obtains choline for growth mainly from plant membrane material which it has ingested, rather than from the free base in the rumen liquor.


Heat shock does not attenuate low-frequency fatigue.
Whole-body hyperthermia or heat shock confers protection to myocardial contractility against reperfusion-induced injury. The purpose of this study was to determine whether heat shock could provide similar protection to skeletal muscle contractility against low-frequency fatigue. Male Sprague-Dawley rats (6 rats/group) were heat shocked at 41.5 degrees C for 15 min either 24 h or 4 days prior to fatiguing stimulation to compare the contractile responses of the plantaris muscle with those of a nonheated group. Both 24 h and 4 days after heat shock, the 72-kDa heat shock protein (HSP72) was elevated above control levels. There were no differences between the heat-shocked and non-heat-shocked animals in measures of contractility prior to fatiguing contractions or in resistance to fatigue. Heat-shock preconditioning did not lead to improved postfatigue force recovery above control responses and, in fact, delayed the recovery of force. This study does not support the use of heat-shock therapy to improve skeletal muscle contractile performance under fatiguing conditions.


Alterations in gastric mucosal lineages before or after acute oxyntic atrophy in gastrin receptor and H2 histamine receptor-deficient mice.
Spasmolytic polypeptide (SP/TFF2)-expressing metaplasia (SPEM) is induced by oxyntic atrophy and is known as a precancerous or paracancerous lesion. We seek to determine whether the gastrin receptor or H(2) histamine receptor influence the development of SPEM. DMP-777 was administered to gastrin receptor and/or H(2) receptor-deficient mice and wild-type mice. Gastric mucosal lineage changes were analyzed. The mucosa from double knockout mice and H(2) receptor knockout mice contained elevated numbers of dual TFF2 and intrinsic factor immunoreactive cells even before DMP-777 treatment. All genotypes of mice showed SPEM after 7-day treatment. In all types of knockout mice, the number of TFF2 immunoreactive cells remained elevated after cessation of treatment. The H(2) receptor and gastrin receptor do not affect emergence of SPEM. However, it is suggested that the absence of H(2) receptor signaling causes a delay in the maturation of chief cells from mucous neck cells.