Bimodal-hybrid heterocyclic amine targeting oxidative pathways and copper mis-regulation in Alzheimer's disease.
Oxidative stress resulting from metal-ion misregulation plays a role in the development of Alzheimer's disease (AD). This process includes the production of tissue-damaging reactive oxygen species and amyloid aggregates. Herein we describe the synthesis, characterization and protective capacity of the small molecule, lipoic cyclen, which has been designed to target molecular features of AD. This construct utilizes the biologically compatible and naturally occurring lipoic acid as a foundation for engendering low cellular toxicity in multiple cell lines, radical scavenging capacity, tuning the metal affinity of the parent cyclen, and results in an unexpected affinity for amyloid without inducing aggregation. The hybrid construct thereby shows protection against cell death induced by amyloid aggregates and copper ions. These results provide evidence for the rational design methods used to produce this fused molecule as a potential strategy for the development of lead compounds for the treatment of neurodegenerative disorders.


Beverage consumption and BMI of British schoolchildren aged 9-13 years.
Adequate fluid intake has been well documented as important for health but whether it has adverse effects on overall energy and sugar intakes remains under debate. Many dietary studies continue to refrain from reporting on beverage consumption, which the present study aimed to address. A cross-sectional survey investigated self-reported measures of dietary intake and anthropometric measurements. Primary and secondary schools in south-west London, UK. Boys and girls (n 248) aged 9–13 years. Boys consumed 10 % and girls consumed 9 % of their daily energy intake from beverages and most children had total sugar intakes greater than recommended. Beverages contributed between a quarter and a third of all sugars consumed, with boys aged 11–13 years consuming 32 % of their total sugar from beverages. There was a strong relationship between consumption of beverages and energy intake; however, there was no relationship between beverage type and either BMI or BMI Z-score. Fruit juices and smoothies were consumed most frequently by all girls and 9–10-year-old boys; boys aged 11–13 years preferred soft drinks and consumed more of their daily energy from soft drinks. Milk and plain water as beverages were less popular. Although current health promotion campaigns in schools merit the attention being given to improving hydration and reducing soft drinks consumption, it may be also important to educate children on the energy and sugar contents of all beverages. These include soft drinks, as well as fruit juices and smoothies, which are both popular and consumed regularly.


Cell-surface antigens associated with dualtropic and thymotropic murine leukemia viruses inducing thymic and nonthymic lymphomas.
Unique type-specific antigens were detected on cells infected with dualtropic and thymotropic viruses isolated and x-ray-induced T cell- and B cell-malignant lymphomas of C57BL/6 mice. These antigens were defined by membrane fluorescence with antisera made in rabbits against rabbit cells chronically infected with cloned virus. The antisera were qualitatively absorbed with a group of cells chronically infected with related dualtropic, ecotropic, and xenotropic viruses. The absorbed antisera detected type-specific, virus-related cell-surface antigens that were unique for different dualtropic virus isolates. The unabsorbed sera also reacted with antigens found specifically on ecotropic and xenotropic virus-infected cells. These findings support the contention that T cell lymphoma (TCL)-inducing and B cell lymphoma (BCL)-inducing viruses isolated from x-irradiated C57BL/6 mice are env gene recombinants in which ecotropic gene sequences have been substituted by xenotropic sequences. We found that unique antigenicities are associated with each TCL-inducing and BCL-inducing dualtropic virus, and that the thymotropic TCL-inducing virus isolates (e.g., 136.5 adn 136.7 viruses) represent a separate serologic group, different from the dualtropic TCL-inducing viruses. By using a series of absorbed antisera in microimmunofluorescence tests we could perform serologic virus mapping of dualtropic clones isolated by us or by others and relate them serologically to previously isolated clones. These virus mapping experiments indicated that many serologically different recombinant viruses can be isolated from C57BL/6 mice. It is suggested that many distinct recombinant viruses may exist in lymphomagenic C57BL/6 mice, some of which are associated with specific lymphoma induction.


A hypomorphic PIGA gene mutation causes severe defects in neuron development and susceptibility to complement-mediated toxicity in a human iPSC model.
Mutations in genes involved in glycosylphosphatidylinositol (GPI) anchor biosynthesis underlie a group of congenital syndromes characterized by severe neurodevelopmental defects. GPI anchored proteins have diverse roles in cell adhesion, signaling, metabolism and complement regulation. Over 30 enzymes are required for GPI anchor biosynthesis and PIGA is involved in the first step of this process. A hypomorphic mutation in the X-linked PIGA gene (c.1234C>T) causes multiple congenital anomalies hypotonia seizure syndrome 2 (MCAHS2), indicating that even partial reduction of GPI anchored proteins dramatically impairs central nervous system development, but the mechanism is unclear. Here, we established a human induced pluripotent stem cell (hiPSC) model containing the PIGAc.1234C>T mutation to study the effects of a hypomorphic allele of PIGA on neuronal development. Neuronal differentiation from neural progenitor cells generated by EB formation in PIGAc.1234C>T is significantly impaired with decreased proliferation, aberrant synapse formation and abnormal membrane depolarization. The results provide direct evidence for a critical role of GPI anchor proteins in early neurodevelopment. Furthermore, neural progenitors derived from PIGAc.1234C>T hiPSCs demonstrate increased susceptibility to complement-mediated cytotoxicity, suggesting that defective complement regulation may contribute to neurodevelopmental disorders.


A comprehensive evaluation of binning methods to recover human gut microbial species from a non-redundant reference gene catalog.
The human gut microbiota performs functions that are essential for the maintenance of the host physiology. However, characterizing the functioning of microbial communities in relation to the host remains challenging in reference-based metagenomic analyses. Indeed, as taxonomic and functional analyses are performed independently, the link between genes and species remains unclear. Although a first set of species-level bins was built by clustering co-abundant genes, no reference bin set is established on the most used gut microbiota catalog, the Integrated Gene Catalog (IGC). With the aim to identify the best suitable method to group the IGC genes, we benchmarked nine taxonomy-independent binners implementing abundance-based, hybrid and integrative approaches. To this purpose, we designed a simulated non-redundant gene catalog (SGC) and computed adapted assessment metrics. Overall, the best trade-off between the main metrics is reached by an integrative binner. For each approach, we then compared the results of the best-performing binner with our expected community structures and applied the method to the IGC. The three approaches are distinguished by specific advantages, and by inherent or scalability limitations. Hybrid and integrative binners show promising and potentially complementary results but require improvements to be used on the IGC to recover human gut microbial species.


Quantitative small-animal surrogate to evaluate drug efficacy in preventing wear debris-induced osteolysis.
Individuals who suffer from severe joint destruction caused by the various arthritidies often undergo total joint arthroplasty. A major limitation of this treatment is the development of aseptic loosening of the prosthesis in as many as 20% of patients. The current paradigm to explain aseptic loosening proposes that wear debris generated from the prosthesis initiates a macrophage-mediated inflammatory response by resident macrophages, leading to osteoclast activation and bone resorption at the implant interface. No therapeutic interventions have been proved to prevent or inhibit aseptic loosening. The development of therapeutic strategies is limited due to the absence of a quantitative surrogate in which drugs can be screened rapidly in large numbers of animals. We have previously described a model in which titanium particles implanted on mouse calvaria induce an inflammatory response with osteolysis similar to that observed in clinical aseptic loosening. Here, we present new methods by which the osteolysis in this model can be quantified. We determined that 6-8-week-old mice in normal health have a sagittal suture area of 50 (+/-6) microm2, which contains approximately five osteoclasts. As a result of the titanium-induced inflammation and osteolysis, the sagittal suture area increases to 197 (+/-27) microm2, with approximately 30 osteoclasts, after 10 days of treatment. The sagittal suture area and the number of osteoclasts in the calvaria of sham-treated mice remained unchanged during the 10 days. We also determined the effects of pentoxifylline, a drug that blocks the responses of tumor necrosis factor-alpha to wear debris, and the osteoclast inhibitor alendronate. We found that both drugs effectively block wear debris-induced osteolysis but not osteoclastogenesis. In conclusion, we found the measurements made with this model to be reproducible and to permit quantitative analysis of agents that are to be screened for their potential to prevent aseptic loosening.


Melanoma cells homing to the brain: an in vitro model.
We developed an in vitro contact through-feet blood brain barrier (BBB) model built using type IV collagen, rat astrocytes, and human umbilical vein endothelial cells (HUVECs) cocultured through Transwell porous polycarbonate membrane. The contact between astrocytes and HUVECs was demonstrated by electron microscopy: astrocytes endfeet pass through the 8.0 μm pores inducing HUVECs to assume a cerebral phenotype. Using this model we evaluated transmigration of melanoma cells from two different patients (M1 and M2) selected among seven melanoma primary cultures. M2 cells showed a statistically significant higher capability to pass across the in vitro BBB model, compared to M1. Expression of adhesion molecules was evaluated by flow cytometry: a statistically significant increased expression of MCAM, αvβ3, and CD49b was detected in M1. PCR array data showed that M2 had a higher expression of several matrix metalloproteinase proteins (MMPs) compared to M1. Specifically, data suggest that MMP2 and MMP9 could be directly involved in BBB permeability and that brain invasion by melanoma cells could be related to the overexpression of many MMPs. Future studies will be necessary to deepen the mechanisms of central nervous system invasion.


Mechanism of Radiation Damage Reduction in Equiatomic Multicomponent Single Phase Alloys.
Recently a new class of metal alloys, of single-phase multicomponent composition at roughly equal atomic concentrations ("equiatomic"), have been shown to exhibit promising mechanical, magnetic, and corrosion resistance properties, in particular, at high temperatures. These features make them potential candidates for components of next-generation nuclear reactors and other high-radiation environments that will involve high temperatures combined with corrosive environments and extreme radiation exposure. In spite of a wide range of recent studies of many important properties of these alloys, their radiation tolerance at high doses remains unexplored. In this work, a combination of experimental and modeling efforts reveals a substantial reduction of damage accumulation under prolonged irradiation in single-phase NiFe and NiCoCr alloys compared to elemental Ni. This effect is explained by reduced dislocation mobility, which leads to slower growth of large dislocation structures. Moreover, there is no observable phase separation, ordering, or amorphization, pointing to a high phase stability of this class of alloys.


[Preliminary Study on the Relationship between the Developmental Stage of Multiple Myeloma Disease and the Results of Bone Marrow Whole Exome Sequencing].
To investigate the genetic results of whole exome sequencing of bone marrow from new onset multiple myeloma (MM) patients to analyze the process of genetic clonal evolution in MM patients. Genomic DNA was extracted from bone marrow samples of 15 MM patients and the whole exomes sequencing was performed using next generation sequencing technology. Using own buccal cells as germline controls, combinated with clinical information, the mutation profile of genes from high-risk asymptomatic myeloma to symptomatic myeloma were analyzed, and genes that may be associated with the efficacy and side effects of bortezomib were screened. Except for two patients in whom no peripheral neuropathy was observed after a short treatment period, other patients peripheral neuropathy developed of various degrees during treatment with bortezomib containing chemotherapy, and the vast majority of patients achieved remission after receiving this bortezomib-related chemotherapy regimen. All patients had comparable levels of the inherited mutations number, but the somatic mutations was correlated with disease evolution. different gene "mutational spectra" exist in myeloma patients at different stages and are associated with progression through all stages of the disease.


[Effects of Metarhizium anisopliae (Metsch.) Sorok. and Beauveria bassiana (Bals.) Vuill. on the predatory stinkbug Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae)].
Entomopathogenic fungi and the predatory stinkbug Podisus nigrispinus (Dallas) can target simultaneously the same or different pests in agroecosystems. Topical contact during fungal dispersion or spraying, walking on plant surfaces and ingestion of contaminated prey are some of possible ways of interactions between fungi and the predatory stinkbug. The impact of three isolates of Metarhizium anisopliae (Metsch.) Sorok. (866, 1022 and 1189) and Beauveria bassiana (Bals.) Vuill. (604, 634 and 561) against nymphs and adults of P. nigrispinus was investigated under laboratory conditions. M. anisopliae caused higher mortality compared to B. bassiana either by topical application or by dry residue on cotton leaves. Topic contact with both fungi caused higher mortality to the predator. No mortality confirmation was reported for the adults. Nymphs fed with cotton leafworm larvae Alabama argillacea (Hübner) (Lepidoptera: Noctuidae) contaminated by the fungi had their reproduction affected, but this was not observed when adult predators fed on contaminated larvae. These findings suggest that isolates of M. anisopliae can cause mortality of P. nigrispinus nymphs by topical contact, while isolates of B. bassiana were less harmful by all ways of infection as compared to M. anisopliae.


Identification and characterization of IS2404 and IS2606: two distinct repeated sequences for detection of Mycobacterium ulcerans by PCR.
Molecular analysis of Mycobacterium ulcerans has revealed two new insertion sequences (ISs), IS2404 and IS2606. IS2404 was identified by complete sequencing of a previously described repetitive DNA segment from M. ulcerans. This element is 1,274 bp long, contains 12-bp inverted repeats and a single open reading frame (ORF) potentially encoding a protein of 327 amino acids (aa), and apparently generates 7-bp direct repeats upon transposition. Amino acid similarity was found between the putative transposase and those encoded by ISs in other bacterial sequences from Aeromonas salmonicida (AsIs1), Escherichia coli (H repeat element), Vibrio cholerae (VcIS1), and Porphyromonas gingivalis (PGIS2). The second IS, IS2606, was discovered by sequence analysis of a HaeIII fragment of M. ulcerans genomic DNA containing a repetitive sequence. This element is 1,404 bp long, with 12-bp inverted repeats and a single ORF potentially encoding a protein of 445 aa. Database searches revealed a high degree of amino acid identity (70%) with the putative transposase of IS1554 from M. tuberculosis. Significant amino acid identity (40%) was also observed with transposases from several other microorganisms, including Rhizobium meliloti (ISRm3), Burkholderia cepacia (IS1356), Corynebacterium diphtheriae, and Yersinia pestis. PCR screening of DNA from 45 other species of mycobacteria with primers for IS2404 confirm that this element is found only in M. ulcerans. However, by PCR, IS2606 was also found in Mycobacterium lentiflavum, another slow-growing member of the genus Mycobacterium that is apparently genetically distinct from M. ulcerans. Testing the sensitivity of PCR based on IS2404 and IS2606 primers demonstrated the ability to detect 0.1 and 1 M. ulcerans genome equivalents, respectively. The ability to detect small numbers of cells by using two gene targets will be particularly useful for analyzing environmental samples, where there may be low concentrations of M. ulcerans among large numbers of other environmental mycobacteria.


High fidelity: Assessing zebrafish (Danio rerio) responses to social stimuli across several levels of realism.
Behavioral assays of zebrafish shoaling have recently been employed to investigate social behavior in zebrafish models of psychiatric disease. Many studies have developed simulated models of conspecifics to serve as alternatives to live shoals in order to examine specific cues that contribute to shoaling behavior. However, no studies have investigated the extent to which zebrafish prefer one stimulus over another when given the choice between two conspecific alternatives (live or simulated). In the present study, we employed a new, four-quadrant choice preference task that allowed zebrafish to swim freely between a live shoal and a motorized mobile shoal, a live shoal and playback of a video-recorded shoal, or a motorized mobile shoal and playback of a video-recorded shoal. Behavior tracking software was used to track subjects' movements in upper and lower quadrants on either side of the test arena. Subjects spent more time near the live shoal, especially in the lower quadrant, and exhibited different swim patterns in response to each simulated conspecific alternative, suggesting that zebrafish prefer a live shoal over models of lower fidelity.


A cytological study of the centrifuged whole, half, and quarter eggs of the sea urchin, Arbacia punctulata.
While the ooplasmic components of centrifuged eggs of Arbacia punctulata do not stratify in homogeneous layers, we have obtained the following strata beginning with the centripetal end: lipid droplets, pronucleus, clear zone, mitochondria, yolk, and pigment. Whereas mitochondria may be found mingled with yolk bodies, we have never observed lipid droplets nor pigment bodies among any of the other inclusions. The so-called clear zone contains a heterogeneous population of inclusions: annulate lamellae, heavy bodies, Golgi complexes, and rod-containing vacuoles. The peripheral cortical granules of immature (germinal vesicle stage) and of mature eggs are not dislodged from the cortical ooplasm with the centrifugal force utilized. When the eggs are treated with urethane, prior to centrifugation, the cortical granules of mature eggs abandon their peripheral position. Further centrifugation of the initially stratified eggs produces nucleated and nonnucleated halves and the centrifugation of the halves results in quarters. The cytology of the halves and quarters is discussed. The halves and quarters have been activated with either sperm or hypertonic sea water. With the exception of the nucleated halves, we were unable to obtain plutei larvae from the other fractions (red halves and quarters). We believe that the lack of development of the various fragments is a function of the balance of particular inclusions necessary for differentiation.


Neonatal sepsis of vertical transmission: an epidemiological study from the "Grupo de Hospitales Castrillo".
A prospective multicenter study was designed to assess the epidemiology of neonatal sepsis of vertical transmission in Spain. The study was carried out by the "Grupo de Hospitales Castrillo" that included the neonatal services of 19 tertiary care (reference) hospitals and 9 secondary care hospitals. Prospective data from infants with culture-proved neonatal sepsis, clinical sepsis and bacteremia were recorded for 1995 to 1997. In a total of 203,288 neonates, proven sepsis was diagnosed in 515 (rate of 2.5 per 1000 live births), clinical sepsis in 724 (rate of 3.6 per 1000 live births), and bacteremia of vertical transmission in 155 (rate of 0.76 per 1000 live births). Very low birth weight (VLBW) infants (< or = 1500 g) showed a significantly higher incidence of confirmed sepsis (26.5 per 1000 live births) and clinical sepsis (32.4 per 1000 live births) than infants weighing > 1500 g. Streptococcus agalactiae was the most frequent causative pathogen in cases of proven sepsis (51%) and bacteremia (33%), but Escherichia coli was the most frequently recovered organism in the VLBW group. The mortality rate of proven sepsis was significantly higher than that of clinical sepsis (8.7% versus 4.3%) (P < 0.01). In the VLBW cohort, there were no significant differences in the mortality rate between proven sepsis and clinical sepsis. In conclusion, clinical sepsis was the most frequent diagnosis, probably related to intrapartum chemoprophylaxis. Streptococcus agalactiae was the most frequent causative pathogen of culture-positive sepsis and bacteremia, whereas E. coli was the most significant in VLBW infants.


Von Hippel-Lindau Disease: Genetics and Role of Genetic Counseling in a Multiple Neoplasia Syndrome.
Von Hippel-Lindau disease (VHL) is one of the most common inherited neoplasia syndromes and is characterized by highly vascular tumors of the eyes, brain, and spine, as well as benign and malignant tumors and/or cysts of the kidneys, adrenal medullae and sympathetic paraganglia, endolymphatic sac, epididymis, and broad ligament. Since the discovery of the VHL gene in 1993, more than 900 families with VHL have been identified and examined. Genetic testing for VHL is widely available and will detect a disease-causing mutation in rate 95% to 100% of individuals who have a clinical diagnosis of VHL, making it the standard of care for diagnosis of VHL. Furthermore, genetic testing for VHL is indicated in some individuals with seemingly sporadic VHL-related tumor types, as ≤ 10% of pheochromocytoma or early-onset renal cell carcinoma and ≤ 40% of CNS hemangioblastoma harbor germline VHL mutations without a family history or additional features of VHL disease. The majority of VHL mutations are private, but there are also well-characterized founder mutations. VHL is a complex, multiorgan disease that spans the breadth of oncology subspecialties, and, as such, providers in these subspecialties should be aware of when to consider a diagnosis of VHL, when to refer a patient to a genetics specialist for consideration of gene testing, and, perhaps most importantly, how to communicate this sensitive information in an age-appropriate manner to at-risk families. This review will provide state-of-the-art information regarding the genetics of VHL and will serve as a key reference for nongenetics professionals who encounter patients with VHL.


A pollen-specific calmodulin-binding protein, NPG1, interacts with putative pectate lyases.
Previous genetic studies have revealed that a pollen-specific calmodulin-binding protein, No Pollen Germination 1 (NPG1), is required for pollen germination. However, its mode of action is unknown. Here we report direct interaction of NPG1 with pectate lyase-like proteins (PLLs). A truncated form of AtNPG1 lacking the N-terminal tetratricopeptide repeat 1 (TPR1) failed to interact with PLLs, suggesting that it is essential for NPG1 interaction with PLLs. Localization studies with AtNPG1 fused to a fluorescent reporter driven by its native promoter revealed its presence in the cytosol and cell wall of the pollen grain and the growing pollen tube of plasmolyzed pollen. Together, our data suggest that the function of NPG1 in regulating pollen germination is mediated through its interaction with PLLs, which may modify the pollen cell wall and regulate pollen tube emergence and growth.


Industrial Thermal Insulation Properties above Sintering Temperatures.
Processing highly flammable products, the oil and gas (O&G) industry can experience major explosions and fires, which may expose pressurized equipment to high thermal loads. In 2020, oil fires occurred at two Norwegian O&G processing plants. To reduce the escalation risk, passive fire protection may serve as a consequence-reducing barrier. For heat or cold conservation, equipment and piping often require thermal insulation, which may offer some fire protection. In the present study, a representative thermal insulation (certified up to 700 °C) was examined with respect to dimensional changes and thermal transport properties after heat treatment to temperatures in the range of 700 °C to 1200 °C. Post heat treatment, the thermal conductivity of each test specimen was recorded at ambient temperature and up to 700 °C, which was the upper limit for the applied measurement method. Based on thermal transport theory for porous and/or amorphous materials, the thermal conductivity at the heat treatment temperature above 700 °C was estimated by extrapolation. The dimensional changes due to, e.g., sintering, were also analyzed. Empirical equations describing the thermal conductivity, the dimensional changes and possible crack formation were developed. It should be noted that the thermal insulation degradation, especially at temperatures approaching 1200 °C, is massive. Thus, future numerical modeling may be difficult above 1150 °C, due to abrupt changes in properties as well as crack development and crack tortuosity. However, if the thermal insulation is protected by a thin layer of more robust material, e.g., passive fire protection to keep the thermal insulation at temperatures below 1100 °C, future modeling seems promising.


The medicinal mushroom Ganoderma lucidum prevents lung tumorigenesis induced by tobacco smoke carcinogens.
Ganoderma lucidum (GL), commonly known as "Lingzhi", is a well-known medicinal mushroom with antioxidant and anti-cancer activity. This study examined the effects of a commercial GL product (GLSF) containing the spore and fruiting body in a 30:8 ratio on tobacco smoke carcinogen-induced lung toxicity and carcinogenesis. The potential chemopreventive effect of GLSF was evaluated in vitro and in vivo. The non-tumorous human bronchial epithelial cells (BEAS-2B cells) were treated with GLSF extract (0.025 and 0.05 mg/mL), which significantly blocked malignant transformation induced by benzo[a]pyrene diol epoxide (BPDE) in a dose-dependent manner. To confirm its anti-carcinogenic activity in vivo, the mice were pre-treated with GLSF (2.0 g/kg of body weight) or curcumin (100 mg/kg of body weight) by oral gavage daily for 7 days and then exposed to a single dose of benzo[a]pyrene (B[a]P) (125 mg/kg of body weight). The GLSF-treated mice showed a significant reduction in B[a]P-induced lung toxicity, as indicated by decreased lactate dehydrogenase activity, malondialdehyde levels, inflammatory cell infiltration, and improved lung histopathology. We next determined the chemopreventive activity of GLSF in mice which were exposed to two weekly doses of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK, 100 mg/kg, on the 1st and 8th days) and fed with control or a modified diet containing GLSF (2.0 g/kg) or metformin (250 mg/kg) for 33 weeks. The GLSF and metformin treatments blocked NNK-induced lung tumor development by decreasing the lung weight, tumor area, and tumor burden compared to the mice exposed to NNK only. GLSF treatment also attenuated the expression of inflammatory, angiogenic, and apoptotic markers in lung tumors. Therefore, GLSF may be used for ameliorating tobacco smoke carcinogens-induced lung toxicity and carcinogenesis.


Chromatic fading following complete adaptation to unique hues.
Profound vision loss occurs after prolonged exposure to an unchanging featureless visual environment. The effect is sometimes called visual fade. Here we investigate this phenomenon in the color domain using two different experiments. In the first experiment we determine the time needed for a colored background to appear achromatic. Four backgrounds were tested. Each represented the observers' four unique hues. This adaptation time was compared with time to recover after adaptation Hue shifts at the end of the adaptation period were also measured. There were wide individual differences in adaptation times and recovery times. Overall recovery was faster than adaptation (p < 0.02). There were minimal shifts in hue. In the second experiment the changes in saturation (Munsell chroma) and lightness (Munsell value) of the background were monitored at six time intervals during the adapting process. Again asymmetric matching with Munsell samples was used. There were two distinct components to both the adaptation and recovery phases; one fast with time constant <1s, the other slow with time constant between 40 and 160s. The experiments show that the special case of visual fade involving color represents the sensory basis for many color-related effects involving adaptation.


Methodological Considerations for Advancing Research on the Health and Wellbeing of Sexual and Gender Minority Youth.
Continued research with sexual and gender minority (SGM) youth is essential both to understand health disparities and to develop interventions targeting those disparities, but conducting rigorous, ethical research with these populations remains a substantial challenge. In addition to considerations for research with adolescents in general, such as utilizing developmentally appropriate measures and obtaining parental permission, factors unique to SGM youth must be addressed at every step of the research process. Defining the study population is complex, as is recruiting a sample once it is defined. Measurement is another challenge, given the paucity of measures developed for or validated with SGM samples. Key constructs, such as sexual orientation, gender identity, and family acceptance, are not amenable to randomization and involving minor participants' parents poses ethical concerns given the precarious home and safety situations that can arise from employing typical study procedures with youth who have a stigmatized identity. In this article, we examine some of these unique methodological challenges. Informed by theoretical and empirical literature, practical experience, and an ongoing dialogue with SGM youth themselves, we present a guide to best practices for ethical, productive research with SGM youth. By discussing existing approaches to studying SGM youth and suggesting innovative ways to approach the questions that remain, we hope to assist the research community in addressing methodological gaps to advance research on SGM youth in relation to families and schools.


Choice of analgesia/anaesthesia during labour and delivery.
Considerations for analgesic or anaesthetic intervention during labour and delivery are reviewed. The amelioration of maternal discomfort must be safe for both mother and fetus. The origin and consequences of labour pain are discussed. Maternal stress resulting from pain can adversely affect fetal oxygen supply. The pros and cons of systemic and regional analgesia are discussed.


An economic model of adverse events and costs for oral anticoagulants used for atrial fibrillation.
To construct a semi-Markov model to compare health outcomes and medical costs associated with warfarin and a second anticoagulant over 1- and 5-year periods. We posited a hypothetical cohort of 10,000 identical 70-year-old patients with atrial fibrillation. We posited 20 scenarios for events that included four possibilities for ischemic strokes (mild, moderate, severe, death) and 16 possibilities for hemorrhages. The model allowed for four levels of International Normalized Ratio. Event rates were based on outcomes in clinical trials and observational studies. Costs were estimated from the perspective of the third-party payer. The greatest cost-generating events were virtually the same for the two drugs and included severe stroke ($1,758,548 for 1 year for both drugs), moderate stroke ($380,355 for 1 year for both drugs), and severe lower gastrointestinal (GI) hemorrhage ($193,804 for 1 year for warfarin and $193,474 for second drug). The least costly events for both drugs were mild intracranial or intracerebral hemorrhage ($7584 for warfarin and $4314 for second drug) and fatal upper GI hemorrhage ($16,781 and $16,752). Total costs for adverse events over 5 years were similar: $18,330,662 for warfarin and $17,102,847 for the second drug. Fatalities for 5 years were 123 for warfarin and 101 for the non-warfarin drug. Varying assumptions for nursing home care and numbers of ischemic strokes and hemorrhages generated the widest variation in costs. We did not account for out-of-pocket expenses, 'pain and suffering' costs, or variation across practice settings. There was substantial variation in numbers and costs of adverse events across 20 scenarios, and for fatalities between the two drugs, but variation in costs between the two drugs was modest.


Transgene expression up to 7 years in nonhuman primates following hepatic transduction with helper-dependent adenoviral vectors.
Helper-dependent adenoviral vectors (HDAd) have been shown to mediate a considerably longer duration of transgene expression than first-generation adenoviral vectors. We have previously shown that transgene expression from HDAd-transduced hepatocytes can persist at high levels for up to 2.6 years in nonhuman primates following a single-vector administration. Because duration of transgene expression and long-term toxicity are critical for risk:benefit assessment, we have continued to monitor these animals. We report here that transgene expression has persisted for the entire observation period of up to 7 years for all animals without long-term adverse effects. However, in all cases, transgene expression level slowly declined over time to less than 10% of peak values by the end of the observation period but remained 2.3-111-fold above baseline values. These results will provide important information for a more informed risk:benefit assessment before clinical application of HDAd.


The Basis for Strain-Dependent Rat Aldehyde Dehydrogenase 1A7 (ALDH1A7) Gene Expression.
Aldehyde hydrogenases (ALDHs) belong to a large gene family involved in oxidation of both endogenous and exogenous compounds in mammalian tissues. Among ALDHs, the rat ALDH1A7 gene displays a curious strain dependence in phenobarbital (PB)-induced hepatic expression: the responsive RR strains exhibit induction of both ALDH1A7 and CYP2B mRNAs and activities, whereas the nonresponsive rr strains show induction of CYP2B only. Here, we investigated the responsiveness of ALDH1A1, ALDH1A7, CYP2B1, and CYP3A23 genes to prototypical P450 inducers, expression of nuclear receptors CAR and pregnane X receptor, and structure of the ALDH1A7 promoter in both rat strains. ALDH1A7 mRNA, associated protein and activity were strongly induced by PB and modestly induced by pregnenolone 16α-carbonitrile in the RR strain but negligibly in the rr strain, whereas induction of ALDH1A1 and P450 mRNAs was similar between the strains. Reporter gene and chromatin immunoprecipitation assays indicated that the loss of ALDH1A7 inducibility in the rr strain is profoundly linked with a 16-base pair deletion in the proximal promoter and inability of the upstream DNA sequences to recruit constitutive androstane receptor-retinoid X receptor heterodimers. SIGNIFICANCE STATEMENT: Genetic variation in rat ALDH1A7 promoter sequences underlie the large strain-dependent differences in expression and inducibility by phenobarbital of the aldehyde dehydrogenase activity. This finding has implications for the design and interpretation of pharmacological and toxicological studies on the effects and disposition of aldehydes.


Mechanical Stabilization of the Glandular Acinus by Linker of Nucleoskeleton and Cytoskeleton Complex.
The nucleoskeleton and cytoskeleton are important protein networks that govern cellular behavior and are connected together by the linker of nucleoskeleton and cytoskeleton (LINC) complex. Mutations in LINC complex components may be relevant to cancer, but how cell-level changes might translate into tissue-level malignancy is unclear. We used glandular epithelial cells in a three-dimensional culture model to investigate the effect of perturbations of the LINC complex on higher order cellular architecture. We show that inducible LINC complex disruption in human mammary epithelial MCF-10A cells and canine kidney epithelial MDCK II cells mechanically destabilizes the acinus. Lumenal collapse occurs because the acinus is unstable to increased mechanical tension that is caused by upregulation of Rho-kinase-dependent non-muscle myosin II motor activity. These findings provide a potential mechanistic explanation for how disruption of LINC complex may contribute to a loss of tissue structure in glandular epithelia.


Lack of association between intussusception and oral polio vaccine in Cuban children.
Two exploratory investigations found an increased risk of intussusception after oral polio vaccine (OPV). A large, national, population-based study was undertaken in Cuba to investigate a possible association. Three hundred and thirty-five cases of intussusception in children under 2 years of age occurring in 1995-2000 were identified and their OPV records retrieved. The relative incidence (RI) of intussusception in defined periods up to 42 days after OPV in children under 1 year was estimated using the self-controlled case series method, controlling for age and season. The RI was not significantly raised in any of the time intervals examined within the 0-42 day period after OPV. For the period 0-42 days as a whole the RI was 1.11, 95% CI 0.74-1.67. This study does not support the hypothesis that OPV causes intussusception.


[Reticulohistiocyte system activity of the liver in patients with nephrotuberculosis].
The reticuloendothelial system of the liver was studied in 27 patients with nephrotuberculosis by means of radionuclide hepatography with colloid solution of Au-198. It was found that all these patients showed a reduction of the activity of stellate reticuloendotheliocytes of the liver. These conclusions were made on the basis of distinctly changed values of the period of half-purification of the blood, constants of blood purification, coefficient of liver retention, etc. There was also a distinct reduction of fractional blood flow of the liver.


Manipulation of beclomethasone-hydrofluoroalkane interactions using biocompatible macromolecules.
The aim of this work was to physically stabilise beclomethasone dipropionate (BDP) microparticles within a hydrofluoroalkane (HFA) propellant using biocompatible polymers in order to allow the efficient delivery of the steroid to the airways from a pressurised metered dose inhaler (pMDI). BDP microparticles were coated with a number of different "amphiphilic" macromolecular excipients by spray-drying an aqueous BDP suspension in which the excipients were dissolved. The physical stability of the coated BDP microparticles was assessed both indirectly using a twin-stage impinger (TSI) and directly using "in-situ" laser diffraction particle size analysis in a range of nonpolar solvents. The solubility of the formulation excipients within a number of the nonpolar vehicles was determined using an internally manufactured filtration rig and the influence of zeta potential within the microparticle suspensions measured in a series surrogate nonpolar systems. The size of the pure BDP microparticles increased significantly (p < 0.05, ANOVA) from 3.13 +/- 0.15 microm to 9.86 +/- 0.50 microm upon suspension within a nonpolar HFA solvent. However, the addition of poly(vinyl alcohol) (PVA) and poly(vinyl pyrrolidone) (PVP) to the BDP microparticles dramatically reduce this aggregation leading to the production of physically stable suspensions with excellent aerosolisation properties (Stage 2 deposition >40% in the twin-stage impinger). It is postulated that the enhanced physical stability observed when PVA and PVP are coated onto BDP microparticles is partially as a result of steric stabilisation in HFA solvents. However, the large zeta potential associated with the nonpolar microparticle suspensions suggest that charge stabilisation may also influence the physical stability within these systems.


Na+-K+-ATPase in rat vascular smooth muscle cell grown in vitro.
This study has focused on the characteristics of the Na+-K+-ATPase in in vitro preparations of vascular smooth muscle cells (VSMCs) derived from the rat carotid artery. The maximum velocity of enzyme reaction (Vmax) for the specific activity of the enzyme in the VSMCs' preparations was 2.36 +/- 0.04 (SE) mumol Pi X mg cell protein-1 X h-1 or 0.82 +/- 0.02 mumol Pi X 10(6) cells-1 X h-1. The activation of the enzyme by potassium, sodium and ATP has been investigated. The half-maximal values for potassium and sodium activation of the enzyme in the preparations were 1.18 and 10-20 meq/l, respectively. The respective Vmax values for potassium and sodium activation were reached at concentrations of 4-10 and 80-100 meq/l. The Michaelis constant for ATP was 0.83 mM. Calcium exerted a potent inhibition on the activity of the enzyme (I50 at 1 mM). It has been concluded that the Na+-K+-ATPase kinetic pattern in in vitro preparations of VSMCs is quite similar to that observed in homogenates or subcellular fractions of other tissues.


Effect Of Age, Gender And Type Of Varicosity On The Post-Operative Pain Score After Seven-Day Compression Dressing After Varicose Vein Surgery.
Great saphenous vein insufficiency is linked to 80% of all remarkable lower limb varicosities. A total of 30 patients were enrolled from OPD fulfilling the inclusion criteria after the approval of the hospital's ethical committee. Patients had compression dressing for seven days after surgery. The patients were divided into two groups-Group-A (Compression dressing for two days) and Group-B (Compression dressing for seven days). Stratification of pain score was done against age, gender, and grades of varicose veins, and after dividing into groups t- test was put into use. A p 20 value ≤0.05 was contemplated to be remarkable. Thirty patients with primary varicose vein were enrolled in this study. The mean age of patients was 35.4±9.9 years. Mean pain score in these patients was 2.9±0.8 years. Pain score after seven days of compression dressing after the surgery for varicose veins depend upon the gender, age, and grades of the varicosity of the veins. It is lesser in the females, younger age groups, and in those who had initially lesser severity of the varicose veins.


Late swing running mechanics influence hamstring injury susceptibility in elite rugby athletes: A prospective exploratory analysis.
Hamstring injuries are one of the most prevalent injuries in rugby union and many other running-based sports, such as track sprinting and soccer. The majority of these injuries occur during running; however, the relationship between running mechanics and hamstring injury is unclear. Obtaining large samples of prospective injury data to examine this relationship is difficult, and therefore exploratory analysis frameworks may assist in deriving valuable information from studies with small but novel samples. The aim of this study was to undertake a prospective exploratory analysis of the relationship between running mechanics and hamstring injury. Kinematic and kinetic data of the trunk, pelvis and lower limbs were collected during maximal overground running efforts for ten elite rugby union athletes. Subsequently, hamstring injury occurrence was recorded for the following Super Rugby season, during which three athletes sustained a running-based hamstring injury. Functional principal component analysis was used to visualise patterns of variability in running mechanics during the late swing phase between athletes. Results indicated that subsequently injured athletes demonstrated a tendency for greater thoracic lateral flexion, greater hip extension moments and greater knee power absorption, compared to uninjured athletes. All variables demonstrated an ability to descriptively differentiate between injured and uninjured athletes at approximately 60% of the late swing phase. Therefore, we hypothesize that greater thoracic lateral flexion, a greater hip extension moment and greater knee power absorption between peak hip flexion and peak knee extension during the late swing phase may put rugby athletes at greater risk of running-based hamstring injury.


Intra-arterial chemotherapy as second-line treatment for advanced retinoblastoma: A 2-year single-center study in China.
To evaluate the effectiveness and complications of intra-arterial chemotherapy (IAC) for treating advanced refractory retinoblastoma (RB) in a large single-center cohort. Eighty-four eyes of 62 consecutive patients with advanced refractory RB who received IAC were included in the study during January 2013 and April 2015. These patients failed to respond adequately to a standard systemic chemotherapy (i.e., carboplatin, vincristine, and etoposide) with or without local therapy. Clinical outcomes and complications of these patients were reviewed. All of these patients received IAC of melphalan combined with topotecan. The mean follow-up period was 14.2 months after final IAC (ranged from 3 to 28 months). The rate of eye preservation was 41.67% in Group D and 20.83% in Group E of this study. Short-term ocular adverse events included eyelid edema (n = 12, 14.29%), bulbar conjunctiva congestion (n = 25, 29.76%), and excessive tearing (n = 10, 11.90%). Long-term complications included vitreous hemorrhage (n = 7, 8%), subretinal hemorrhage (n = 9, 11%), retinal vasculopathy (n = 6, 7%), and ophthalmic artery spasm with reperfusion (n = 11, 13%). Fever was observed after IAC in 14 patients; transient vomiting was observed in 17 patients; there were 8 cases of transient myelosuppression. IAC can be an optional treatment to save eyes of Group D RB that failed in systemic chemotherapy and were destined for enucleation. However, it should be cautioned for Group E. Both the ocular and systemic toxicities of IAC were within tolerance.


Application of mendelian randomization to study the causal relationship between smoking and the risk of chronic obstructive pulmonary disease.
Smoking is a risk factor for chronic obstructive pulmonary disease (COPD). Few studies have assessed the causal relationship between smoking and COPD using Mendelian randomization. Exposure and outcome datasets were obtained from the IEU Open GWAS project (https://gwas.mrcieu.ac.uk/). The exposure data set includes smoking (ever smoke, smoking/smokers in household, exposure to tobacco smoke at home). The outcome data set includes COPD susceptibility and acute COPD admissions. The main methods of Mendelian randomization analysis are weighted median method and MR-Egger method. Heterogeneity and polymorphism analyses were performed to ensure the accuracy of the results. ever smoke increased the risk of COPD prevalence, and ever smoke and smoking/smokers in household increased the risk of acute COPD admission. Conclusion Therefore, we should enhance the management of nonpharmacological prescription of COPD to reduce the individual incidence.


Oral cobalamin supplementation in cats with hypocobalaminaemia: a retrospective study.
Objectives The objective of the study was to evaluate whether oral cobalamin supplementation can restore normocobal-aminaemia in cats with hypocobalaminaemia and clinical signs of gastrointestinal disease. Methods This was a retrospective study based on a computerised database search for client-owned cats treated at Evidensia Specialist Animal Hospital, Helsingborg, Sweden, during the period December 2013 to August 2016. Inclusion criteria were cats with clinical signs of chronic enteropathy, an initial serum cobalamin concentration ⩽250 pmol/l (reference interval 214-738 pmol/l) and oral treatment with cobalamin tablets. Results Twenty-five cats met the inclusion criteria. The cats were treated with 0.25 mg cyanocobalamin tablets once daily. Serum cobalamin concentration was rechecked 27-94 days after continuous oral cobalamin supplementation. All cats had serum cobalamin concentrations above the reference interval after oral cobalamin supplementation. Median (range) serum cobalamin concentration was 128 pmol/l (111-250 pmol/l) prior to treatment and 2701 pmol/l (738-16,359 pmol/l) after supplementation. This difference was statistically significant ( P <0.0001). Conclusions and relevance Our results suggest that oral cobalamin supplementation is effective in increasing serum cobalamin to supranormal concentrations in cats with hypocobalaminaemia. Thus, oral cobalamin supplementation is a promising alternative to parenteral administration. Prospective comparative studies in cats being treated with parenteral vs oral cobalamin supplementation in a larger number of patients are warranted before oral supplementation can be recommended for routine use.


A cohort study of the ocular safety of anti-ulcer drugs.
1. Recently, some cases have been reported where intravenous use of omeprazole was followed by loss of vision. We followed up a cohort of close to 140,000 persons during periods of treatment and non-treatment with five anti-ulcer drugs. 2. The relative risk of vascular disorders of the eye during use of omeprazole compared with non-use was 1.8 (95% CI 0.5-6.0). Use of other anti-ulcer drugs was associated with a similar risk of vascular disorders. The relative risk associated with current use of any anti-ulcer drug was 1.9 (95% CI 1.1-3.4). We did not find a single case of optic inflammatory disorder during treatment with any of the five anti-ulcer drugs. 3. These results do not suggest a major increased risk for vascular or inflammatory disorders of the eye associated with use of omeprazole or other anti-ulcer drugs.


Baclofen suppresses motivation to consume alcohol in rats.
Recent studies demonstrated that treatment with the gamma-aminobutyric acid (GABA)(B) receptor agonist baclofen reduced alcohol intake in selectively bred Sardinian alcohol-preferring (sP) rats tested under the home-cage, two-bottle choice regimen. The present study investigated the effect of baclofen on the appetitive, rather than consummatory, aspects of alcohol ingestion in sP rats. Rats were trained to lever-press for oral alcohol (10%, v/v) or sucrose (3%, w/v) under a fixed-ratio schedule of 4. Once self-administration behavior was established, alcohol intake averaged approximately 0.7 g/kg over the 30-min session. Subsequently, the effect of the acute administration of baclofen (0, 1, 2 and 3 mg/kg, i.p.) on the extinction responding for alcohol and sucrose (defined as the maximal number of lever responses reached in the absence of reinforcement and used as index of motivation to consume alcohol and sucrose) was evaluated. RESULTS. All doses of baclofen produced a marked suppression of extinction responding for alcohol. Conversely, only the 3-mg/kg baclofen dose significantly affected extinction responding for sucrose. A separate open-field test indicated that baclofen (0, 1, 2 and 3 mg/kg, i.p.) did not affect spontaneous motor activity in sP rats. These results suggest that baclofen may specifically reduce the motivational properties of alcohol; further, these results are in agreement with the recently reported anti-craving potential of baclofen in alcoholics.


The universe: a cryogenic habitat for microbial life.
Panspermia, an ancient idea, posits that microbial life is ubiquitous in the Universe. After several decades of almost irrational rejection, panspermia is at last coming to be regarded as a serious contender for the beginnings of life on our planet. Astronomical data is shown to be consistent with the widespread distribution of complex organic molecules and dust particles that may have a biological provenance. A minuscule (10(-21)) survival rate of freeze-dried bacteria in space is all that is needed to ensure the continual re-cycling of cosmic microbial life in the galaxy. Evidence that terrestrial life may have come from elsewhere in the solar system has accumulated over the past decade. Mars is seen by some as a possible source of terrestrial life, but some hundreds of billions of comets that enveloped the entire solar system, are a far more likely primordial reservoir of life. Comets would then have seeded Earth, Mars, and indeed all other habitable planetary bodies in the inner regions of the solar system. The implications of this point of view, which was developed in conjunction with the late Sir Fred Hoyle since the 1970s, are now becoming amenable to direct empirical test by studies of pristine organic material in the stratosphere. The ancient theory of panspermia may be on the verge of vindication, in which case the entire universe would be a grand crucible of cryomicrobiology.


Experience of using the CRQ (Chronic Respiratory Questionnaire).
We have examined the role of the Chronic Respiratory Questionnaire in practice. The questionnaire covers aspects of disability in patients with chronic lung disease in terms of dyspnoea, fatigue, emotional function and mastery. It therefore provides a measure of physical disability and quality of life in patients with lung disease. The questionnaire was validated in older patients with chronic obstructive lung disease and is particularly responsive to changes within an individual. Its major disadvantages include the lack of sensitivity in patients with minor symptoms or possibly those in a younger age group and that it is also not possible to make comparisons between populations with the CRQ. It is, however, particularly good at demonstrating changes in disability in older patients with chronic airways disease.


A comparison of the kinematics, kinetics and muscle activity between pneumatic and free weight resistance.
Pneumatic devices provide a resistance comprising minimal mass, possibly affording greater movement velocities, compared to free weight, while reducing the influence of momentum. Thirty men completed three testing sessions [free weight (FW), ballistic (BALL) and pneumatic (P)] each consisting of a one repetition maximum (1RM) and six sets (15, 30, 45, 60, 75 and 90% 1RM) of four explosive repetitions of a bench press. Dependent variables were expressed as mean and as a percentage of the concentric displacement. Significant differences (P < 0.05) were evaluated using two way repeated measures ANOVAs with Holm-Sidak post hoc comparisons. On average, the mean and peak P velocity were 36.5 and 28.3% higher than FW, and 22.9 and 19.1% higher than the BALL movements. The FW and BALL peak force were both significantly higher than the P (26.3 and 22.7% for FW and BALL, respectively). BALL mean power output was significantly higher than the FW and P at loads of 15 and 30% 1RM; however, between loads of 60-90% 1RM the highest mean power was produced with a P resistance. A 15% 1RM load maximized the peak power for each condition and no significant differences were found between the P and BALL. For loads of 45-90% 1RM the force, power and muscle activity were higher during the last 10-20% of the concentric displacement when subjects employed the P resistance. In summary, pneumatic resistance may offer specific advantages over loads comprising only mass (FW and BALL), although not without its own limitations.


Examining racial disparities in colorectal cancer care.
African Americans are disproportionately burdened with colorectal cancer. Although incidence and mortality rates have declined in the past two decades, the disparity in health outcomes has progressively increased. This comprehensive review examines the existing literature regarding racial disparities in colorectal cancer screening, stage at diagnosis, and treatment to determine if differences exist in the quality of care delivered to African Americans. A comprehensive review of relevant literature was performed. Two databases (EBSCOHOST Academic Search Premier and Scopus) were searched from 2000 to 2007. Articles that assessed racial disparities in colorectal cancer screening, stage of disease at diagnosis, and treatment were selected. The majority of studies identified examined colorectal cancer screening outcomes. Although racial disparities in screening have diminished in recent years, African American men and women continue to have higher colorectal cancer incidence and mortality rates and are diagnosed at more advanced stages. Several studies regarding stage of disease at diagnosis identified socioeconomic status (SES) and health insurance status as major determinants of disparity. However, some studies found significant racial disparities even after controlling for these factors. Racial disparities in treatment were also found at various diagnostic stages. Many factors affecting disparities between African Americans and Whites in colorectal cancer incidence and mortality remain unexplained. Although the importance of tumor biology, genetics, and lifestyle risk factors have been established, prime sociodemographic factors need further examination to understand variances in the care of African Americans diagnosed with colorectal cancer.


Return to work after an initial myocardial infarction and subsequent emotional distress.
We examined how return to work predicted subsequent change in emotional distress in 143 patients who had been employed at the time of initial myocardial infarction. Ninety patients (63%) returned to work by 4 months and remained employed at 12 months. There were no differences in mental health at baseline between those who returned to work and those who did not, but emotional distress decreased significantly between 4 and 12 months only in the group who returned to work. Emotional distress declined after resuming work even when employees returned to jobs with which they reported dissatisfaction at the time of the myocardial infarction. The relationship between return to work and decreasing emotional distress remained after controlling for initial physical and psychological adjustment as well as sociodemographic and social support characteristics. The improvements in mental health associated with return to work should reassure clinicians who emphasize the emotional as well as economic value of work after an initial myocardial infarction.


Biocatalysis with thermostable enzymes: structure and properties of a thermophilic 'ene'-reductase related to old yellow enzyme.
We report the crystal structure of a thermophilic "ene" reductase (TOYE) isolated from Thermoanaerobacter pseudethanolicus E39. The crystal structure reveals a tetrameric enzyme and an active site that is relatively large compared to most other structurally determined and related Old Yellow Enzymes. The enzyme adopts higher order oligomeric states (octamers and dodecamers) in solution, as revealed by sedimentation velocity and multiangle laser light scattering. Bead modelling indicates that the solution structure is consistent with the basic tetrameric structure observed in crystallographic studies and electron microscopy. TOYE is stable at high temperatures (T(m)>70 degrees C) and shows increased resistance to denaturation in water-miscible organic solvents compared to the mesophilic Old Yellow Enzyme family member, pentaerythritol tetranitrate reductase. TOYE has typical ene-reductase properties of the Old Yellow Enzyme family. There is currently major interest in using Old Yellow Enzyme family members in the preparative biocatalysis of a number of activated alkenes. The increased stability of TOYE in organic solvents is advantageous for biotransformations in which water-miscible organic solvents and biphasic reaction conditions are required to both deliver novel substrates and minimize product racemisation.


Distal myopathy due to digenic inheritance of TIA1 and SQSTM1 variants in two unrelated Spanish patients.
Welander distal myopathy typically manifests in late adulthood and is caused by the founder TIA1 c.1150G>A (p.Glu384Lys) variant in families of Swedish and Finnish descent. Recently, a similar phenotype has been attributed to the digenic inheritance of TIA1 c.1070A>G (p.Asn357Ser) and SQSTM1 c.1175C>T (p.Pro392Leu) variants. We describe two unrelated Spanish patients presenting with slowly progressive gait disturbance, distal-predominant weakness, and mildly elevated creatine kinase (CK) levels since their 6th decade. Electromyography revealed abnormal spontaneous activity and a myopathic pattern. Muscle magnetic resonance imaging (MRI) showed marked fatty replacement in distal leg muscles. A muscle biopsy, performed on one patient, revealed myopathic changes with rimmed vacuoles. Both patients carried the TIA1 p.Asn357Ser and SQSTM1 p.Pro392Leu variants. Digenic inheritance is supported by evidence from unrelated pedigrees and a plausible biological interaction between both proteins in protein quality control processes. Recent functional studies and additional case descriptions further support this. Clinical suspicion is necessary to seek both variants.


A crystallographic comparison between mutated glyceraldehyde-3-phosphate dehydrogenases from Bacillus stearothermophilus complexed with either NAD+ or NADP+.
Mutations have been introduced in the cytosolic glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Bacillus stearothermophilus in order to convert its cofactor selectivity from a specificity towards NAD into a preference for NADP. In the B-S mutant, five mutations (L33T, T34G, D35G, L187A, P188S) were selected on the basis of a sequence alignment with NADP-dependent chloroplastic GAPDHs. In the D32G-S mutant, two of the five mutations mentioned above (L187A, P188S) have been used in combination with another one designed from electrostatic considerations (D32G). Both mutants exhibit a dual-cofactor selectivity at the advantage of either NAD (B-S) or NADP (D32G-S). In order to analyse the cofactor-binding site plasticity at the molecular level, crystal structures of these mutants have been solved, when complexed with either NAD+ (D32G-Sn, resolution 2.5 A, R = 13.9%; B-Sn, 2.45 A, 19.3%) or NADP+ (D32G-Sp, 2.2 A, 19.2%; B-Sp, 2.5 A, 14.4%). The four refined models are very similar to that of the wild-type GAPDH and as expected resemble more closely the holo form than the apo form. In the B-S mutant, the wild-type low affinity for NADP+ seems to be essentially retained because of repulsive electrostatic contacts between the extra 2'-phosphate and the unchanged carboxylate group of residue D32. Such an antideterminant effect is not well compensated by putative attractive interactions which had been expected to arise from the newly-introduced side-chains. In this mutant, recognition of NAD+ is slightly affected with respect to that known on the wild-type, because mutations only weakly destabilize hydrogen bonds and van der Waals contacts originally present in the natural enzyme. Thus, the B-S mutant does not mimic efficiently the chloroplastic GAPDHs, and long-range and/or second-layer effects, not easily predictable from visual inspection of three-dimensional structures, need to be taken into account for designing a true "chloroplastic-like" mutant of cytosolic GAPDH. In the case of the D32G-S mutant, the dissociation constants for NAD+ and NADP+ are practically reversed with respect to those of the wild-type. The strong alteration of the affinity for NAD+ obviously proceeds from the suppression of the two wild-type hydrogen bonds between the adenosine 2'- and 3'-hydroxyl positions and the D32 carboxylate group. As expected, the efficient recognition of NADP+ is partly promoted by the removal of intra-subunit electrostatic repulsion (D32G) and inter-subunit steric hindrance (L187A, P188S). Another interesting feature of the reshaped NADP+-binding site is provided by the local stabilization of the extra 2'-phosphate which forms a hydrogen bond with the side-chain hydroxyl group of the newly-introduced S188. When compared to the presently known natural NADP-binding clefts, this result clearly demonstrates that an absolute need for a salt-bridge involving the 2'-phosphate is not required to switch the cofactor selectivity from NAD to NADP. In fact, as it is the case in this mutant, only a moderately polar hydrogen bond can be sufficient to make the extra 2'-phosphate of NADP+ well recognized by a protein environment.


[Relation between the expression of hypoxia inducible factor-1alpha and angiogenesis in ovarian cancer using tissue microarray].
To study the relation between the expression of hypoxia inducible factor-1alpha (HIF-1alpha)and angiogenesis in ovarian cancer. The expressions of HIF-1alpha mRNA, vascular endothelial growth factor (VEGF), and CD(34) in 295 patients with epithelial ovarian tumors were analyzed by tissue microarray technology, in situ hybridization and immunohistochemistry, and compared with those of 13 normal ovarian tissue samples. The expressions of HIF-1alpha mRNA were observed in 0, 13.2%, 42.1% and 81.9% of normal ovarian tissue, benign, borderline and malignant ovarian tumors respectively. Expression rates of HIF-1alpha mRNA in borderline and invasive tumors were significantly higher than those in normal ovarian tissues and benign tumors (P < 0.01). Statistical analysis revealed that the expression of HIF-1alpha mRNA was not related to surgical stages and histological subtypes. Close positive relation was observed between the expression of HIF-1alpha mRNA and tumor histological grade (r = 0.246, P < 0.01). There were significant correlations between the expression of HIF-1alpha mRNA and VEGF (r = 0.206, P = 0.01) and microvessel density (MVD) level (r = 0.451, P < 0.01). HIF-1alpha may play a role in angiogenesis of ovarian carcinoma, and may promote the development of the carcinoma.


Interdigitated residues within a small region of VP16 interact with Oct-1, HCF, and DNA.
Upon infection, the herpes simplex virus (HSV) activator of immediate-early (IE) gene transcription VP16 forms a multiprotein-DNA complex with two cellular proteins, Oct-1 and HCF. First, VP16 associates with HCF independently of DNA, and this association stimulates subsequent association with Oct-1 on the DNA target of VP16 activation, the TAATGARAT motif found in HSV IE promoters. We have analyzed the involvement of VP16 residues lying near the carboxy-terminal transcriptional activation domain of VP16 in associating with HCF, Oct-1, and DNA. To assay VP16 association with HCF, we developed an electrophoretic mobility retardation assay in which HCF is used to retard the mobility of a hybrid VP16-GAL4 DNA-binding domain fusion protein bound to a GAL4 DNA-binding site. Analysis of an extensive set of individual and combined alanine substitutions over a 61-amino-acid region of VP16 shows that, even within a region as small as 13 amino acids, there are separate residues involved in association with either HCF, DNA, or Oct-1 bound to DNA; indeed, of two immediately adjacent amino acids in VP16, one is important for DNA binding and the other is important for HCF binding. These results suggest that a small region in VP16 is important for linking in close juxtaposition the four components of the VP16-induced complex and support the hypothesis that the structure of the Oct-1-VP16 interaction in this complex is similar to that formed by the yeast transcriptional regulatory proteins MATa1 and MAT alpha2. We propose that HCF stabilizes this Oct-1-VP16 interaction.


Degradation of Polysaccharides from Grateloupia filicina and Their Antiviral Activity to Avian Leucosis Virus Subgroup J.
In this study, polysaccharides from Grateloupia filicinia (GFP) were extracted and several low molecular weight (Mw) G. filicina polysaccharides (LGFPs) were prepared by the hydrogen peroxide (H₂O₂) oxidation method. Additionally, the effect of different experimental conditions on the degradation of GFP was determined. Results showed that the GFP degradation rate was positively related to H₂O₂ concentration and temperature, and negatively related to pH. Chemical analysis and Fourier transform infrared spectra (FT-IR) of GFP and LGFPs showed that the degradation caused a slight decrease of total sugar and sulfate content. However, there was no obvious change for monosaccharide contents. Then, the anti-ALV-J activity of GFP and LGFPs were determined in vitro. Results revealed that all of the samples could significantly inhibit ALV-J and lower Mw LGFPs exhibited a stronger suppression, and that the fraction LGFP-3 with Mw 8.7 kDa had the best effect. In addition, the reaction phase assays showed that the inhibition effect was mainly because of the blocking virus adsorption to host cells. Moreover, real-time PCR, western-blot, and IFA were further applied to evaluate the blocking effects of LGFP-3. Results showed that the gene relative expression and gp85 protein for LGFPS-3 groups were all reduced. Data from IFA showed that there was less virus infected cells for 1000 and 200 μg/mL LGFPS-3 groups when compared to virus control. Therefore, lower Mw polysaccharides from G. filicina might supply a good choice for ALV-J prevention and treatment.


Optimization and Predictive Modeling of Reinforced Concrete Circular Columns.
Metaheuristic optimization techniques are widely applied in the optimal design of structural members. This paper presents the application of the harmony search algorithm to the optimal dimensioning of reinforced concrete circular columns. For the objective of optimization, the total cost of steel and concrete associated with the construction process were selected. The selected variables of optimization include the diameter of the column, the total cross-sectional area of steel, the unit costs of steel and concrete used in the construction, the total length of the column, and applied axial force and the bending moment acting on the column. By using the minimum allowable dimensions as the constraints of optimization, 3125 different data samples were generated where each data sample is an optimal design configuration. Based on the generated dataset, the SHapley Additive exPlanations (SHAP) algorithm was applied in combination with ensemble learning predictive models to determine the impact of each design variable on the model predictions. The relationships between the design variables and the objective function were visualized using the design of experiments methodology. Applying state-of-the-art statistical accuracy measures such as the coefficient of determination, the predictive models were demonstrated to be highly accurate. The current study demonstrates a novel technique for generating large datasets for the development of data-driven machine learning models. This new methodology can enhance the availability of large datasets, thereby facilitating the application of high-performance machine learning predictive models for optimal structural design.


Development of Oxytolerant Salmonella typhimurium Using Radiation Mutation Technology (RMT) for Cancer Therapy.
A critical limitation of Salmonella typhimurium (S. typhimurium) as an anti-cancer agent is the loss of their invasive or replicative activities, which results in no or less delivery of anti-cancer agents inside cancer cells in cancer therapy. Here we developed an oxytolerant attenuated Salmonella strain (KST0650) from the parental KST0649 (ΔptsIΔcrr) strain using radiation mutation technology (RMT). The oxytolerant KST0650 strain possessed 20-times higher replication activity in CT26 cancer cells and was less virulent than KST0649. Furthermore, KST0650 migrated effectively into tumor tissues in mice. KST0650 was further equipped with a plasmid harboring a spliced form of the intracellular pro-apoptotic protein sATF6, and the expression of sATF6 was controlled by the radiation-inducible recN promoter. The new strain was named as KST0652, in which sATF6 protein expression was induced in response to radiation in a dose-dependent manner. This strain was effectively delivered inside cancer cells and tumor tissues via the Salmonella type III secretion system (T3SS). In addition, combination treatment with KST0652 and radiation showed a synergistic anti-tumor effect in murine tumor model with complete inhibition of tumor growth and protection against death. In conclusion, we showed that RMT can be used to effectively develop an anti-tumor Salmonella strain for delivering anti-cancer agents inside tumors.


Patient care services in rural Minnesota community pharmacies.
To compare the availability of medication therapy management (MTM) services, point-of-care (POC) testing, and disease management resources between rural and urban Minnesota community pharmacies. Cross-sectional study. Minnesota in 2006. Owners and operational managers of 564 Minnesota community pharmacies. Mail survey containing structured, quantitative questions. Resulting data were separated to evaluate urban and rural area community pharmacies. Staffing trends, MTM services, and patient care services of urban compared with rural community pharmacies in Minnesota. Urban and rural pharmacies reported allocating nearly the same percent of a typical day to filling and dispensing prescriptions (approximately 70%). A higher percent of rural community pharmacies offered patient care services in 5 of 15 categories, including drug information services (55.7% vs. 45.6%), provision of durable medical equipment (43.4% vs. 32.6%), dyslipidemia management (7.8% vs. 3.8%), hypertension management (14.6%% vs. 7.3%), and MTM (29.4% vs. 18.7%). Although the time allocated to dispensing medication was approximately 70% for both urban and rural pharmacies, a significantly higher proportion of rural pharmacies reported providing MTM and other direct patient care services. This may be a result of geographic isolation and greater use of pharmacists as providers of first-contact care in these areas.


Repeats of Unusual Size in Plant Mitochondrial Genomes: Identification, Incidence and Evolution.
Plant mitochondrial genomes have excessive size relative to coding capacity, a low mutation rate in genes and a high rearrangement rate. They also have abundant non-tandem repeats often including pairs of large repeats which cause isomerization of the genome by recombination, and numerous repeats of up to several hundred base pairs that recombine only when the genome is stressed by DNA damaging agents or mutations in DNA repair pathway genes. Early work on mitochondrial genomes led to the suggestion that repeats in the size range from several hundred to a few thousand base pair are underrepresented. The repeats themselves are not well-conserved between species, and are not always annotated in mitochondrial sequence assemblies. We systematically identified and compared these repeats, which are important clues to mechanisms of DNA maintenance in mitochondria. We developed a tool to find and curate non-tandem repeats larger than 50bp and analyzed the complete mitochondrial sequences from 157 plant species. We observed an interesting difference between taxa: the repeats are larger and more frequent in the vascular plants. Analysis of closely related species also shows that plant mitochondrial genomes evolve in dramatic bursts of breakage and rejoining, complete with DNA sequence gain and loss. We suggest an adaptive explanation for the existence of the repeats and their evolution.


[Aromatase expression in invasive and in situ ductal carcinoma present in the same breast].
to evaluate the expression of aromatase in simultaneously invasive ductal carcinoma (IDC) and ductal carcinoma in situ (DCIS). forty-five surgical samples were obtained from mastectomy and quadrantectomy with simultaneous IDC and DCIS of stage I and II patients. Aromatase was evaluated using antibodies anti-aromatase and the samples classified in accordance with the number and intensity of stained cells. Aromatase was expressed positively in 32(71%) and negatively in 13(29%) of the cases in the IDC. The same results were obtained in the DCIS showing a perfect positive correlation. In the normal epithelium,aromatase was positive in 19(42.2%) and negative in 26 (57.8%) and a positive correlation, statistically significant was obtained when compared with IDC and DCIS(p<0.01). Concerning the normal stroma, positivity was only 7 (15.5%) showing no correlation with aromatase expression. Aromatase was positive in 36(80%) of the tumor stroma and this result was statistically significant as in the IDC and DCIS. Comparing results of aromatase expression with nuclear grade, histological grade, tumor size and age no difference was found. our results demonstrated high correlation between aromatase expression in IDC, DCIS, normal epithelium and tumor stroma showing a possible autocrine and paracrine mechanism of this enzyme in breast cancer.


New metabolites produced by endophyte Clonostachys rosea B5 - 2.
The endophytic fungus, Clonostachys rosea B5-2 was isolated from mangrove plants and subjected to the one strain many compounds (OSMAC) methodology. By this approach, it was found that modification of the culture media enhanced the production of secondary metabolites by C. rosea B5-2. The apple juice supplemented solid rice media led to significant changes in the secondary metabolism of the fungus, C. rosea B5-2, and induced the production of four new compounds, (-)-dihydrovertinolide (2), and clonostach acids A (3), B (4), and C (5) together with the known compound, (-)-vertinolide (1). The new compound (-)-dihydrovertinolide (2) exhibited phytotoxicity against lettuce seedlings at a concentration of 50 mg L-1.


Health promotion research and practice require sound policy analysis models: the case of Quebec's Tobacco Act.
In this paper we illustrate how policy analysis models can deepen our understanding of the challenges facing health promoters advocating for policy change. Specifically we describe the factors underpinning the adoption of Québec's Tobacco Act (1998) and the role played by actors from governmental public health agencies (GPHAs). Data were collected through interviews (n=39), newspapers articles (n=569) and documents (n > 200) from GPHAs, NGOs, the Québec National Assembly, and opponents to the legislative measures. Data collection and analysis were based on Sabatier and Jenkins-Smith's Advocacy Coalition Framework (1999) and Lemieux's theorization of coalition structuring (1998). We explain the adoption of the Act by: (1) the broad recognition within the policy elite of the main parameters of tobacco use (i.e. lethality, addictive properties, and legitimacy of governmental intervention), (2) the impacts of a series of events (e.g. cigarette contraband crisis) that enabled tobacco control advocates to influence public debates, and the governmental agenda, (3) the critical contribution of a coalition of GPHAs pooling resources to address both the sanitary and economic aspects of the legislation while countering the opposition's strategy, and (4) the failure of the opponents to present an unified voice on the definition of the tobacco policy. This study illustrates the merits of applying a policy-change model to grasp the complexity of the process. Our findings call for the development of permanent policy analysis capabilities within public health agencies and for a broader scrutiny of the non-health-related dimensions of policy debates.


Electrical features, liquid composition and toxicant emissions from 'pod-mod'-like disposable electronic cigarettes.
Use of flavoured pod-mod-like disposable electronic cigarettes (e-cigarettes) has grown rapidly, particularly among cost-sensitive youth and young adults. To date, little is known about their design characteristics and toxicant emissions. In this study, we analysed the electrical and chemical characteristics and nicotine and pulmonary toxicant emission profiles of five commonly available flavoured disposable e-cigarettes and compared these data with those of a JUUL, a cartridge-based e-cigarette device that pod-mod-like disposables emulate in size and shape. Device construction, electrical power and liquid composition were determined. Machine-generated aerosol emissions including particulate matter, nicotine, carbonyl compounds and heavy metals were also measured. Liquid and aerosol composition were measured by high-performance liquid chromatography, gas chromatography-mass spectrometry/flame ionisation detection, and inductively coupled plasma mass spectrometry. We found that unlike JUUL, disposable devices did not incorporate a microcontroller to regulate electrical power to the heating coil. Quality of construction varied widely. Disposable e-cigarette power ranged between 5 and 9 W and liquid nicotine concentration ranged between 53 and 85 mg/mL (~95% in the protonated form). In 15 puffs, total nicotine yield for the disposables ranged between 1.6 and 6.7 mg, total carbonyls ranged between 28 and 138 µg, and total metals ranged between 1084 and 5804 ng. JUUL emissions were near the floors of all of these ranges. Disposable e-cigarettes are designed with high nicotine concentration liquids and are capable of emitting much higher nicotine and carbonyl species relative to rechargeable look-alike e-cigarettes. These differences are likely due to the lower quality in construction, unreliable labelling and lack of temperature control regulation that limits the power during operation. From a public health perspective, regulating these devices is important to limit user exposure to carbonyls and nicotine, particularly because these devices are popular with youth and young adults.


Construct validity of the UCSD performance-based skills assessment-brief version (UPSA-B) in HIV disease.
Among individuals living with HIV disease, approximately 60% experience problems with everyday functioning. The present study investigated the utility of the UCSD Performance-based Skills Assessment-Brief Version (UPSA-B) as a measure of functional capacity in HIV. We utilized a cross-sectional three-group design comparing individuals with HIV- associated neurocognitive disorder (HAND) (HIV + HAND+; n = 27), HIV+ neurocognitively normal individuals (HIV + HAND-; n = 51), and an HIV- comparison group (HIV-; n = 28) with broadly comparable demographics and non-HIV comorbidities. Participants were administered the UPSA-B, the Medication Management Test-Revised (MMT-R), and were assessed for manifest everyday functioning and quality of life, as part of a standardized clinical neurocognitive research battery. Results indicated that the HIV + HAND+ group had significantly lower UPSA-B scores than the HIV + HAND-group, but did not differ from the HIV- group. Among HIV+ individuals, UPSA-B scores were significantly related to MMT-R scores, all neurocognitive domains assessed, and education, but the UPSA-B was not related to manifest everyday functioning (e.g., unemployment), health-related quality of life, or HIV disease variables. Findings provide mixed support for the construct validity of the UPSA-B in HIV. Individuals impaired on the UPSA-B may be at increased risk for HAND, but the extent to which it detects general manifest everyday functioning problems is uncertain.


Ceramide induces apoptosis in human lung adenocarcinoma A549 cells through mitogen-activated protein kinases.
To provide experimental data for further research on the signal transduction of apoptosis in lung adenocarcinoma cells, we examined the effects of exogenous C2-ceramide administration on several members of the mitogen-activated protein kinase (MAPK) superfamily and caspase-3 in A549 cells. Cell viability and apoptosis were analyzed by cell counting kit-8 assay and flow cytometry. Various MAPK and caspase-3 proteins were detected by Western blotting. C2-ceramide selectively altered the phosphorylation state of members of the MAPK superfamily, causing hyperphosphorylation of mitogen-activated protein kinase kinase (MEK)1/2 and the p38 MAPK, but not affecting the phosphorylation of extracellular signal-regulated kinase 1/2 and the c-Jun N-terminal kinase. SB-203580 (a p38 MAPK inhibitor) and p38 siRNA, but not U0126 (a MEK inhibitor), partially rescued cell death induced by C2-ceramide. C2-ceramide promoted the activation of caspase-3. Exogenous C2-ceramide induced apoptosis in human lung adenocarcinoma A549 cells. The activation of MAPK and caspase-3 were involved in the mechanisms of C2-ceramide-induced apoptosis in A549 cells.


Cannabinoids: Possible agents for treatment of psoriasis via suppression of angiogenesis and inflammation.
Psoriasis is a chronic skin disease also affecting other sites such as joints. This disease highly depends on inflammation and angiogenesis as well as other pathways. At each step of the psoriasis molecular pathway, different inflammatory cytokines and angiogenic growth factors are involved such as hypoxia inducible factor-1 α (HIF-1 α), vascular endothelial growth factor (VEGF), matrix metalo proteinases (MMPs), basic fibroblast growth factor (bFGF), Angiopoitin-2, interleukin-8 (IL-8), IL-17, and IL-2. Beside the mentioned growth factors and cytokines, cellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) which play roles in both angiogenesis and inflammation are also involved in the pathogenesis. Cannabinoids are active compounds of Cannabina Sativa inducing their effects through cannabinoid receptors (CBs). JWH-133 is a synthetic cannabinoid with strong anti-angiogenic and anti-inflammatory activities. This agent is able to inhibit HIF-1 α, VEGF, MMPs, bFGF, IL-8, IL-17, and other mentioned cytokines and adhesion molecules both in vivo and in vitro. Altogether, authors suggest using this cannabinoid for treatment of psoriasis due to its potential in suppressing the two main steps of psoriatic pathogenesis. Of course complementary animal studies and human trials are still required.


Human retinal pigment epithelial cells express scavenger receptors BI and BII.
The retinal pigment epithelial (RPE) cells that underlie the retina ingest and metabolize thousands of lipid-rich photoreceptor outer segments (POS) every day. The scavenger receptor (SR) CD36 and integrin alphavbeta5 have been shown to participate in POS binding and internalization by RPE cells. The objective of the current study was to determine whether RPE cells express SRs other than CD36. Primary cultures of human RPE cells express both mRNA and protein for SR-BI and SR- BII. SR-BI and SR-BII mRNAs were detected by reverse transcription-polymerase chain reaction. SR-BI protein was detected by immune precipitation of [(35)S]methionine-labeled crude cellular extracts. SR-BII was detected by Western blotting of immune precipitated crude cellular extracts. SR-BI and SR-BII proteins were also detected by immunofluorescence staining of RPE cells in culture. The results suggest that these SRs may play a role in POS lipid binding and uptake by RPE cells in the eye.


Distinct Compartmentalization of Microbial Community and Potential Metabolic Function in the Fruiting Body of Tricholoma matsutake.
The uniquely compartmentalized fruiting body structure of the ectomycorrhizal fungus (EMF) Tricholoma matsutake, is a hotspot of microbial habitation and interaction. However, microbial diversity within this microniche structure of the EMF is rarely investigated. Furthermore, there is limited information concerning microbiomes associated with sporomes belonging to the ubiquitous fungal phylum Basidiomycota, particularly with respect to fungus-EMF interactions. In this study, we conducted high throughput sequencing, using ITS (fungal) and 16S rRNA (bacterial) marker genes to characterize and compare fruiting body microbiomes in the outer (pileipellis and stipitipellis) and inner layers (pileum context, stipe context, and lamellae) of the fruiting body of T. matsutake. Our results show the number of unique bacterial operational taxonomic units (OTUs) among the different compartments ranged from 410 to 499 and was more than double that of the shared/common OTUs (235). Micrococcales, Bacillales, Caulobacter, and Sphingomonas were the primary significant bacterial taxa within the different compartments of the dissected T. matsutake fruiting body. Non-parametric multivariate analysis of variance showed significant compartmental differences for both the bacterial and the fungal community structure within the T. matsutake fruiting body. The metabolic profiling revealed putative metabolisms (of amino acids, carbohydrates, and nucleotides) and the biosynthesis of secondary metabolites to be highly enriched in outer layers; in the inner parts, the metabolisms of energy, cofactors, vitamins, and lipids were significantly higher. This study demonstrates for the first time the distinct compartmentalization of microbial communities and potential metabolic function profiles in the fruiting body of an economically important EMF T. matsutake.


The splice variant of the V2 vasopressin receptor adopts alternative topologies.
The V2 receptor gene encodes two receptor variants by alternative splicing, the canonical V2 receptor (V2a receptor) and V2b. The V2b variant has an amino acid sequence identical to that of the V2a receptor up to the sixth transmembrane domain, but the V2b sequences corresponding to the putative seventh transmembrane domain and the carboxyl terminus are different from those of the V2a receptor. Here we investigate the topology and subcellular distribution of the V2b variant. We found that, in contrast to the V2a receptor, the V2b adopted two topologies: one with six transmembrane segments with the C-terminus on the extracellular side of the membrane and another with seven transmembrane segments with the C-terminus on the intracellular side, similar to typical G-protein-coupled receptors. Furthermore, we observed that both topological isoforms oligomerized with the V2a canonical receptor. Unlike the V2a receptor, V2b did not move to the plasma membrane, but it is retained in the ER--Golgi compartments. These findings indicate that the C-terminal sequence beyond the sixth transmembrane of the V2a is required for the stabilization of the seven-transmembrane topology of the receptor and is also essential for the trafficking of the receptor to the plasma membrane.


Comparison of Non-Hilar Clamping Simple Enucleation and Enucleo-Resection of Exophytic Renal Tumors.
To retrospectively evaluate our institutional experience with non-hilar-clamping simple enucleation (SE) and enucleoresection (ER) for the treatment of exophytic renal tumors regarding their oncological outcomes. We retrospectively evaluated patients treated between 2006 and 2013 for clinical exophytic T1-T2a renal tumors using open nephron-sparing surgery. A total of 33 patients underwent SE and 39 underwent ER. The mean tumor size was 38.7 mm. None of the patients had positive surgical margins. No local recurrences were observed during the postoperative follow-up period (mean 40.7 ± 23.4 months); however, ipsilateral adrenal and contralateral kidney metastasis was detected in one of the patients. There was no statistically significant difference in the R.E.N.A.L Nephrometry Score, operative time, or intraoperative blood loss in the non-hilar-clamping SE and ER groups (P > .05). During the third postoperative month, the estimated glomerular filtration rate (eGFR) levels in the SE group were significantly reduced compared with the preoperative eGFR levels (P = .046). SE and ER with non-hilar clamping are safe, acceptable approaches for treating exophytic renal tumors.


Incorporating societal concerns for fairness in numerical valuations of health programmes.
The paper addresses some limitations of the QALY approach and outlines a valuation procedure that may overcome these limitations. In particular, we focus on the following issues: the distinction between assessing individual utility and assessing societal value of health care; the need to incorporate concerns for severity of illness as an independent factor in a numerical model of societal valuations of health outcomes; similarly, the need to incorporate reluctance to discriminate against patients that happen to have lesser potentials for health than others; and finally, the need to combine measurements of health-related quality of life obtained from actual patients (or former patients) with measurements of distributive preferences in the general population when estimating societal value. We show how equity weights may serve to incorporate concerns for severity and potentials for health in QALY calculations. We also suggest that for chronically ill or disabled people a life year gained should count as one and no less than one as long as the year is considered preferable to being dead by the person concerned. We call our approach 'cost-value analysis'.


In plants, decapping prevents RDR6-dependent production of small interfering RNAs from endogenous mRNAs.
Cytoplasmic degradation of endogenous RNAs is an integral part of RNA quality control (RQC) and often relies on the removal of the 5' cap structure and their subsequent 5' to 3' degradation in cytoplasmic processing (P-)bodies. In parallel, many eukaryotes degrade exogenous and selected endogenous RNAs through post-transcriptional gene silencing (PTGS). In plants, PTGS depends on small interfering (si)RNAs produced after the conversion of single-stranded RNAs to double-stranded RNAs by the cellular RNA-dependent RNA polymerase 6 (RDR6) in cytoplasmic siRNA-bodies. PTGS and RQC compete for transgene-derived RNAs, but it is unknown whether this competition also occurs for endogenous transcripts. We show that the lethality of decapping mutants is suppressed by impairing RDR6 activity. We establish that upon decapping impairment hundreds of endogenous mRNAs give rise to a new class of rqc-siRNAs, that over-accumulate when RQC processes are impaired, a subset of which depending on RDR6 for their production. We observe that P- and siRNA-bodies often are dynamically juxtaposed, potentially allowing for cross-talk of the two machineries. Our results suggest that the decapping of endogenous RNA limits their entry into the PTGS pathway. We anticipate that the rqc-siRNAs identified in decapping mutants represent a subset of a larger ensemble of endogenous siRNAs.


Social representations of health councilors regarding the right to health and citizenship.
To know the structure of the social representations of right to health and citizenship of health municipal councilors. This is a qualitative study, based on the central nucleus theory of social representations, carried out in eight municipalities of the Integrated Region for the Development of the Surroundings of the Federal District, Brazil. The intentional sample consisted of municipal health councilors. Between June and December 2012, free recall questionnaires were used, of which 68 were answered with the inducing term health, and 64 with the inducing term citizenship. Data were analyzed using EVOC software and Bardin's content analysis. The representational field of the right to health is associated with the idea of universal law guaranteed by the Constitution and the Unified Health System (SUS), and of citizenship linked to rights and duties. The conceptions of right to health are understood as a condition for reaching citizenship, and citizenship as social protection.


Silanized polymeric nanoparticles for DNA isolation.
The aim of this study is to prepare silanized polymeric nanoparticles for DNA isolation. Polymeric p(HEMA)-IMEO-PBA nanoparticles around 85.7 nm diameter, was obtained by surfactant free emulsion polymerization for DNA isolation. Synthesized nanoparticles for characterization studies were realized scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and Zeta-size. Surface area, average particle size and size distribution were also performed. The surface area of synthesized silanized polymeric nanoparticles was 2460 m(2)/g. Synthesized polymeric nanoparticles were silanized with 3-(2-imidazoline-1-yl)propyl (triethoxysilane) (IMEO). After that, phenylboronic acid (PBA) which is DNA specific ligand were covalently binded to silanized polymeric nanoparticles. The amount of DNA adsorbed onto the p(HEMA)-IMEO-PBA nanoparticles first increased and then reached a saturation value at around 14.0 mg/mL of DNA concentration. The maximum adsorption was 672.41 mg/g silanized polymeric nanoparticles in the optimum adsorption medium. The maximum DNA adsorption was achieved at 4°C. The overall recovery of DNA was calculated as 95%. In repetitive adsorption-desorption circles, it is observed not being important decrease in DNA adsorption capacities. The results were shown that silanized polymeric nanoparticles can be a good alternative for DNA isolation.


Upregulation of Calbindin-D-28k immunoreactivity by excitatory amino acids.
Excessive or prolonged exposure to excitatory amino acids (EAA) are thought to be neurotoxic by altering calcium homeostasis. A protective role of Calbindin-D-28 k (Calbindin) has been postulated due to its capacity to buffer calcium. Calbindin is highly expressed in the Purkinje cells (PCs), of the cerebellar cortex. Changes of the Calbindin immunoreactivity (IR) by the EAA has been here investigated in cerebellar slices maintained in vitro. It was found that at low temperature, PCs are very slightly immunoreactive and therefore the experiments were done at 22 degrees C. The results show that Calbindin-IR increases in PCs exposed to the neurotoxic agonists, Kainic acid (KA) and AMPA as well as to glutamate (Glu), the endogenous EAA. The increase is very rapid and slowly reversible; is induced by excitatory and excitotoxic concentrations of the agonists; is independent of the calcium influx. While KA- and AMPA-induced Calbindin-IR is blocked by CNQX, the KA/AMPA receptor antagonist, Glu-induced Calbindin-IR is only slightly decreased by CNQX and AP5, the NMDA receptor antagonist. It is concluded that Calbindin-containing neurons can increase their calcium buffering capacity in response to EAA binding to specific receptors, the response being independent of, but concomitant to calcium influx.