TERPENE AND TERPENOID EMISSIONS AND SECONDARY ORGANIC AEROSOL PRODUCTION

Approximately 90% of fine aerosol in the Midwestern United States has a regional component with a sizable fraction attributed to secondary production of organic aerosol (SOA). The Ozark Forest is an important source of biogenic SOA precursors like isoprene (> 150 mg m-2 d-1), monoterpenes (10-40 mg m-2 d-1), and sesquiterpenes (10-40 mg m-2d-1). Anthropogenic sources include secondary sulfate and nitrate and biomass burning (51-60%), vehicle emissions (17-26%), and industrial emissions (16-18%). Vehicle emissions are an important source of volatile and vapor-phase, semivolatile aliphatic and aromatic hydrocarbons that are important anthropogenic sources of SOA precursors. The short lifetime of SOA precursors and the complex mixture of functionalized oxidation products make rapid sampling, quantitative processing methods, and comprehensive organic molecular analysis essential elements of a comprehensive strategy to advance understanding of SOA formation pathways. Uncertainties in forecasting SOA production on regional scales are large and related to uncertainties in biogenic emission inventories and measurement of SOA yields under ambient conditions. This work presents a bottom-up approach to develop a conifer emission inventory based on foliar and cortical oleoresin composition, development of a model to estimate terpene and terpenoid signatures of foliar and bole emissions from conifers, development of processing and analytic techniques for comprehensive organic molecular characterization of SOA precursors and oxidation products, implementation of the high-volume sampling technique to measure OA and vapor-phase organic matter, and results from a 5 day field experiment conducted to evaluate temporal and diurnal trends in SOA precursors and oxidation products. A total of 98, 115, and 87 terpene and terpenoid species were identified and quantified in commercially available essential oils of Pinus sylvestris, Picea mariana, and Thuja occidentalis, respectively, by comprehensive, two-dimensional gas chromatography with time-of-flight mass spectrometric detection (GC × GC-ToF-MS). Analysis of the literature showed that cortical oleoresin composition was similar to foliar composition of the oldest branches. Our proposed conceptual model for estimation of signatures of terpene and terpenoid emissions from foliar and cortical oleoresin showed that emission potentials of the foliar and bole release pathways are dissimilar and should be considered for conifer species that develop resin blisters or are infested with herbivores or pathogens. Average derivatization efficiencies for Methods 1 and 2 were 87.9 and 114%, respectively. Despite the lower average derivatization efficiency of Method 1, distinct advantages included a greater certainty of derivatization yield for the entire suite of multi- and poly-functional species and fewer processing steps for sequential derivatization. Detection limits for Method 1 using GC × GC- ToF-MS were 0.09-1.89 ng μL-1. A theoretical retention index diagram was developed for a hypothetical GC × 2GC analysis of the complex mixture of SOA precursors and derivatized oxidation products. In general, species eluted (relative to the alkyl diester reference compounds) from the primary column (DB-210) in bands according to n and from the secondary columns (BPX90, SolGel-WAX) according to functionality, essentially making the GC × 2GC retention diagram a Carbon number-functionality grid. The species clustered into 35 groups by functionality and species within each group exhibited good separation by n. Average recoveries of n-alkanes and polyaromatic hydrocarbons (PAHs) by Soxhlet extraction of XAD-2 resin with dichloromethane were 80.1 ± 16.1 and 76.1 ± 17.5%, respectively. Vehicle emissions were the common source for HSVOCs [i.e., resolved alkanes, the unresolved complex mixture (UCM), alkylbenzenes, and 2- and 3-ring PAHs]. An absence of monoterpenes at 0600-1000 and high concentrations of monoterpenoids during the same period was indicative of substantial losses of monoterpenes overnight and the early morning hours. Post-collection, comprehensive organic molecular characterization of SOA precursors and products by GC × GC-ToFMS in ambient air collected with ~2 hr resolution is a promising method for determining biogenic and anthropogenic SOA yields that can be used to evaluate SOA formation models.

CD8+ T cells and NK cells from blister fluid of an EEM/SJS overlap highly express Fas ligand and Granulysin

INTRODUCTION Erythema exsudativum multiforme majus (EEMM) and Stevens-Johnson Syndrome (SJS) are severe cutaneous reaction patterns caused by infections or drug hypersensitivity. The mechanism by which widespread keratinocyte death is mediated by the immune system in EEMM/SJS are still to be elucidated. Here, we characterized the blister cells isolated from a patient with EEMM/SJS overlap and investigated its cause. METHODS Clinical classification of the cutaneous eruption was done according to the consensus definition of severe blistering skin reactions and histological analysis. Common infectious causes of EEMM were investigated using standard clinical techniques. T cell reactivity for potentially causative drugs was assessed by lymphocyte transformation tests (LTT). Lymphocytes isolated from blister fluid were analyzed for their expression of activation markers and cytotoxic molecules using flow cytometry. RESULTS The healthy 58 year-old woman suffered from mild respiratory tract infection and therefore started treatment with the secretolytic drug Ambroxol. One week later, she presented with large palmar and plantar blisters, painful mucosal erosions, and flat atypical target lesions and maculae on the trunc, thus showing the clinical picture of an EEMM/SJS overlap (Fig. 1). This diagnosis was supported by histology, where also eosinophils were found to infiltrate the upper dermis, thus pointing towards a cutaneous adverse drug reaction (cADR). Analysis of blister cells showed that they mainly consisted of CD8+ and CD4+ T cells and a smaller population of NK cells. Both the CD8+ T cells and the NK cells were highly activated and expressed Fas ligand and the cytotoxic molecule granulysin (Fig. 2). In addition, in comparison to NK cells from PBMC, NK cells in blister fluids strongly upregulated the expression of the skin-homing chemokine receptor CCR4 (Fig 4). Surprisingly, the LTT performed on PBMCs in the acute phase was positive for Ambroxol (SI=2.9) whereas a LTT from a healthy but exposed individual did not show unspecific proliferation. Laboratory tests for common infectious causes of EEMM were negative (HSV-1/-2, M. pneumoniae, Parvovirus B19). However, 6 weeks later, specific proliferation to Ambroxol could no longer be observed in the LTT (Fig 4.).

ΔNp63 REGULATES A COMPLEX NETWORK OF TARGET GENES IN LIMB AND EPIDERMAL DEVELOPMENT

The skin is composed of two major compartments, the dermis and epidermis. The epidermis forms a barrier to protect the body. The stratified epithelium has self-renewing capacity throughout life, and continuous turnover is mediated by stem cells in the basal layer. p63 is structurally and functionally related to p53. In spite of their structural similarities, p63 is critical for the development and maintenance of stratified epithelial tissues, unlike p53. p63 is highly expressed in the epidermis and previously has been shown to play a critical role in the development and maintenance of the epidermis. The study of p63 has been complicated due to the existence of multiple isoforms: those with a transactivation domain (TAp63) and those lacking this domain (ΔNp63). Mice lacking p63 cannot form skin, have craniofacial and skeletal defects and die within hours after birth. These defects are due to the ability of p63 to regulate multiple processes in skin development including epithelial stem cell proliferation, differentiation, and adherence programs. To determine the roles of these isoforms in skin development and maintenance, isoform specific p63 conditional knock out mice were generated by our lab. TAp63-/- mice age prematurely, develop blisters, and display wound-healing defects that result from hyperproliferation of dermal stem cells. That results in premature depletion of these cells, which are necessary for wound repair, that indicates TAp63 plays a role in dermal/epidermal maintenance. To study the role of ΔNp63, I generated a ΔNp63-/- mouse and analyzed the skin by performing immunofluorescence for markers of epithelial differentiation. The ΔNp63-/- mice developed a thin, disorganized epithelium but differentiation markers were expressed. Interestingly, the epidermis from ΔNp63-/- mice co-expressed K14 and K10 in the same cell suggesting defects in epidermal differentiation and stratification. This phenotype is reminiscent of the DGCR8fl/fl;K14Cre and Dicerfl/fl;K14Cre mice skin. Importantly, DGCR8-/- embryonic stem cells (ESCs) display a hyperproliferation defect by failure to silence pluripotency genes. Furthermore, I have observed that epidermal cells lacking ΔNp63 display a phenotype reminiscent of embryonic stem cells instead of keratinocytes. Thus, I hypothesize that genes involved in maintaining pluripotency, like Oct4, may be upregulated in the absence of ΔNp63. To test this, q-RT PCR was performed for Oct4 mRNA with wild type and ΔNp63-/- 18.5dpc embryo skin. I found that the level of Oct4 was dramatically increased in the absence of ΔNp63-/-. Based on these results, I hypothesized that ΔNp63 induces differentiation by silencing pluripotency regulators, Oct4, Sox2 and Nanog directly through the regulation of DGCR8. I found that DGCR8 restoration resulted in repression of Oct4, Sox2 and Nanog in ΔNp63-/- epidermal cells and rescue differentiation defects. Loss of ΔNp63 resulted in pluripotency that caused defect in proper differentiation and stem cell like phenotype. This led me to culture the ΔNp63-/- epidermal cells in neuronal cell culture media in order to address whether restoration of DGCR8 can transform epidermal cells to neuronal cells. I found that DGCR8 restoration resulted in a change in cell fate. I also found that miR470 and miR145 play a role in the induction of pluripotency by repressing Oct4, Sox2 and Nanog. This indicates that ΔNp63 induces terminal differentiation through the regulation of DGCR8.

EGFR inhibitors erlotinib and lapatinib ameliorate epidermal blistering in pemphigus vulgaris in a non-linear, V-shaped relationship.

Novel insights into intra-cellular signalling involved in pemphigus vulgaris (PV), an autoimmune blistering disease of skin and mucous membranes, are now revealing new therapeutic approaches such as the chemical inhibition of PV-associated signals in conjunction with standard immunosuppressive therapy. However, extensive inhibition of signalling molecules that are required for normal tissue function and integrity may hamper this approach. Using a neonatal PV mouse model, we demonstrate that epidermal blistering can be prevented in a dose-dependent manner by clinically approved EGFR inhibitors erlotinib and lapatinib, but only up to approximately 50% of normal EGFR activity. At lower EGFR activity, blisters again aggravated and were highly exacerbated in mice with a conditional deletion of EGFR. Statistical analysis of the relation between EGFR activity and the extent of skin blistering revealed the best fit with a non-linear, V-shaped curve with a median break point at 52% EGFR activity (P = 0.0005). Moreover, lapatinib (a dual EGFR/ErbB2 inhibitor) but not erlotinib significantly reduced blistering in the oral cavity, suggesting that signalling mechanisms differ between PV predilection sites. Our results demonstrate that future clinical trials evaluating EGFR/ErbB2 inhibitors in PV patients must select treatment doses that retain a specific level of signal molecule activity. These findings may also be of relevance for cancer patients treated with EGFR inhibitors, for whom skin lesions due to extensive EGFR inhibition represent a major threat.

Epidermolysis bullosa in Danish Hereford calves is caused by a deletion in LAMC2 gene.

BACKGROUND Heritable forms of epidermolysis bullosa (EB) constitute a heterogeneous group of skin disorders of genetic aetiology that are characterised by skin and mucous membrane blistering and ulceration in response to even minor trauma. Here we report the occurrence of EB in three Danish Hereford cattle from one herd. RESULTS Two of the animals were necropsied and showed oral mucosal blistering, skin ulcerations and partly loss of horn on the claws. Lesions were histologically characterized by subepidermal blisters and ulcers. Analysis of the family tree indicated that inbreeding and the transmission of a single recessive mutation from a common ancestor could be causative. We performed whole genome sequencing of one affected calf and searched all coding DNA variants. Thereby, we detected a homozygous 2.4 kb deletion encompassing the first exon of the LAMC2 gene, encoding for laminin gamma 2 protein. This loss of function mutation completely removes the start codon of this gene and is therefore predicted to be completely disruptive. The deletion co-segregates with the EB phenotype in the family and absent in normal cattle of various breeds. Verifying the homozygous private variants present in candidate genes allowed us to quickly identify the causative mutation and contribute to the final diagnosis of junctional EB in Hereford cattle. CONCLUSIONS Our investigation confirms the known role of laminin gamma 2 in EB aetiology and shows the importance of whole genome sequencing in the analysis of rare diseases in livestock.

Definitions and outcome measures for mucous membrane pemphigoid: recommendations of an international panel of experts

Mucous membrane pemphigoid encompasses a group of autoimmune bullous diseases with a similar phenotype characterized by subepithelial blisters, erosions, and scarring of mucous membranes, skin, or both. Although knowledge about autoimmune bullous disease is increasing, there is often a lack of clear definitions of disease, outcome measures, and therapeutic end points. With clearer definitions and outcome measures, it is possible to directly compare the results and data from various studies using meta-analyses. This consensus statement provides accurate and reproducible definitions for disease extent, activity, outcome measures, end points, and therapeutic response for mucous membrane pemphigoid and proposes a disease extent score, the Mucous Membrane Pemphigoid Disease Area Index.

Pemphigus. S2 Guideline for diagnosis and treatment--guided by the European Dermatology Forum (EDF) in cooperation with the European Academy of Dermatology and Venereology (EADV)

BACKGROUND Pemphigus encompasses a group of life-threatening autoimmune bullous diseases characterized by blisters and erosions of the mucous membranes and skin. Before the era of immunosuppressive treatment, the prognosis of pemphigus was almost fatal. Due to its rarity, only few prospective controlled therapeutic trials are available. OBJECTIVES For this reason, a group of European dermatologists with a long-standing interest and expertise in basic and clinical pemphigus research has sought to define diagnostic and therapeutic guidelines for the management of patients with pemphigus. RESULTS This group identified the statements of major agreement or disagreement regarding the diagnostic and therapeutic management of pemphigus. The revised final version of the pemphigus guideline was finally passed on to the European Dermatology Forum (EDF) for a final consensus with the European Academy of Dermatology and Venereology (EADV) and the European Union of Medical Specialists (UEMS).

Improvement of the mechanical properties of 2124 Al/SiCp MMC plate by optimisation of the solution treatment

Static mechanical properties of 2124 Al/SiCp MMC have been measured as a function of solution temperature and time. An optimum solution treatment has been established which produces significant improvements in static mechanical properties and fatigue crack growth resistance over conventional solution treatments. Increasing the solution treatment parameters up to the optimum values improves the mechanical properties because of intermetallic dissolution, improved solute and GPB zone strengthening and increased matrix dislocation density. Increasing the solution treatment parameters beyond the optimum values results in a rapid reduction in mechanical properties due to the formation of gas porosity and surface blisters. The optimum solution treatment improves tensile properties in the transverse orientation to a greater extent than in the longitudinal orientation and this results in reduced anisotropy. © 1996 Elsevier Science Limited.

Air sensitivity of MoS2, MoSe2, MoTe2, HfS2 and HfSe2

A surface sensitivity study was performed on different transition-metal dichalcogenides (TMDs) under ambient conditions in order to understand which material is the most suitable for future device applications. Initially, Atomic Force Microscopy and Scanning Electron Microscopy studies were carried out over a period of 27 days on mechanically exfoliated flakes of 5 different TMDs, namely, MoS2, MoSe2, MoTe2, HfS2, and HfSe2. The most reactive were MoTe2 and HfSe2. HfSe2, in particular, showed surface protrusions after ambient exposure, reaching a height and width of approximately 60 nm after a single day. This study was later supplemented by Transmission Electron Microscopy (TEM) cross-sectional analysis, which showed hemispherical-shaped surface blisters that are amorphous in nature, approximately 180–240 nm tall and 420–540 nm wide, after 5 months of air exposure, as well as surface deformation in regions between these structures, related to surface oxidation. An X-ray photoelectron spectroscopy study of atmosphere exposed HfSe2 was conducted over various time scales, which indicated that the Hf undergoes a preferential reaction with oxygen as compared to the Se. Energy-Dispersive X-Ray Spectroscopy showed that the blisters are Se-rich; thus, it is theorised that HfO2 forms when the HfSe2 reacts in ambient, which in turn causes the Se atoms to be aggregated at the surface in the form of blisters. Overall, it is evident that air contact drastically affects the structural properties of TMD materials. This issue poses one of the biggest challenges for future TMD-based devices and technologies.

Estudo da prevalência e da gravidade da dermatite de contacto em 'frango do campo'

O sector avícola enfrenta atualmente dois desafios muito estimulantes. O primeiro decorre do aumento, que se prevê continuar a crescer, nos níveis de procura de carne de aves no mercado interno e internacional; o segundo decorre do facto da criação avícola ter adotado métodos de produção mais intensivos (kg peso vivo/m2/ano) e em maior escala, i.e. com maior concentração animal na mesma exploração. Este carácter vincadamente “industrial” tem merecido uma natural atenção das sociedades e das autoridades pecuárias no sentido desta economia de escala passar a ter num conjunto de instrumentos legais e técnicos o devido contrapeso para a salvaguarda das aves enquanto ser vivo. O presente trabalho tem como ponto de partida a Directiva 2007/43/CE do Conselho de 28 de Junho, relativa ao estabelecimento de regras mínimas para a proteção de frangos de carne. Em virtude de não existir ainda informação suficiente sobre a forma como a qualidade do maneio animal pode ser monitorizada, ao nível do abate, por médicos veterinários e auxiliares oficiais, em frangos de criação especial segundo os modelos definidos no Regulamento (CE) n.º 543/2008, urge realizar estudos neste domínio. O principal objetivo da realização do presente trabalho de campo foi o estudo da ocorrência das dermatites de contacto plantar (pododermatites) e da bolsa sinovial préesternal em frangos produzidos em sistemas de produção considerados “protetores” do bem-estar animal, designadamente os seguintes: i) ar livre; e, ii) extensivo de interior. O estudo foi efetuado num centro de abate de frangos do campo, em Oliveira de Frades, entre Maio de 20012 e Março de 2013. Os animais abatidos foram criados em explorações com contratos de integração situadas no Distrito de Viseu. Os dados foram recolhidos em 39 bandos diferentes da espécie Gallus domesticus, dos quais 1021 carcaças foram avaliadas após evisceração, o que correspondeu ao exame de uma a cada quinze aves da linha de abate. Para a avaliação da pododermatite foi utilizado o método adaptado pela DGAV, enquanto para a avaliação da bursite esternal foi efetuada tendo em conta o modelo aplicado em perus por Berk em 2002. Apesar do modelo estatístico desenvolvido para a análise dos resultados obtidos no presente trabalho exigir um maior número de observações, foi possível identificar com grande precisão alguns fatores de risco que devem ser realçados pela sua relevância no contexto dos sistemas produtivos escrutinados ou no mecanismo fisiopatológico da dermatite de contacto, nomeadamente os seguintes: (i) a idade das aves que, apesar de não ter sido identificada uma relação directa com os scores de pododermatite e bursite, verificou-se que a idade elevada que os animais tipicamente atingem nos sistemas de produção extensivos está associada a uma taxa superior de rejeições pela inspecção sanitária; (ii) o peso pré-abate que, independentemente da inconsistência defendida por diversos autores em relação à influência do peso vivo do frango industrial sobre a dermatite de contacto, nos animais produzidos em regime extensivo, esta variável pode desempenhar um fator chave para a ocorrência desta lesão. De facto, há que realçar que o peso destes animais tem uma importância fulcral na modelação da biomecânica da ave, incluindo na pressão exercida sobre a superfície plantar; (iii) o tipo de sistema de abeberamento, tendo ficado demonstrado que a selecção do tipo de bebedouro tem uma importância peculiar sobre a ocorrência de pododermatite em “frango de campo”, algo que está provavelmente relacionado com a influência exercida sobre o teor de humidade da cama. Globalmente, as frequências de pododermatite e bursite apuradas neste trabalho devem ser consideradas inquietantes. Esta preocupação eleva-se quando se toma consciência que as aves provieram de regimes considerados “amigáveis” e “sustentáveis”, pelo que urge monitorizar adequadamente aqueles sistemas produtivos, melhorar as suas condições e reanalisar os benefícios ao nível do bem-estar animal.

Electrical discharge machining of 6061 aluminium alloy

The wire electrical discharge machining (EDM) of 6061 aluminium alloy in terms of material removal rate, kerf/slit width, surface finish and wear of electrode wire for different pulse on time and wire tension was studied. Eight experiments were carried out in a wire EDM machine by varying pulse on time and wire tension. It is found that the material removal rate increases with the increase of pulse on time though the wire tension does not affect the material removal rate. It seems that the higher wire tension facilitates steady machining process, which generates low wear in wire electrode and better surface finish. The surface roughness does not change notably with the variation of pulse on time. The appearance of the machined surfaces is very similar under all the machining conditions. The machined surface contains solidified molten material, splash of materials and blisters. The increase of the pulse on time increases the wear of wire electrode due to the increase of heat input. The wear of wire electrode generates tapered slot which has higher kerf width at top side than that at bottom side. The higher electrode wear introduces higher taper.

Caracterización de las condiciones de salud y de trabajo y su relación con lesiones cutáneas en los orfebres artesanales de Mompox Bolivar-Colombia 2015

INTRODUCCIÓN Las alteraciones de la salud relacionadas con el trabajo (enfermedades y accidentes), pueden prevenirse desde las actividades bien enfocadas del Sistema de Gestión de Seguridad y Salud en el Trabajo (SGSST), realizando la identificación y control de los agentes causales en el ambiente de trabajo y la vigilancia de la salud de la población trabajadora. El proyecto desarrollado va dirigido a un grupo específico de artesanos orfebres en la ciudad de Mompox, Bolivar, en la que esta labor artesanal se centra en el arte de la filigrana, transmitido generacionalmente desde antaño En esta ciudad los artesanos orfebres, aunque corresponden a un sector informal de la economía, se encuentran agremiados principalmente en tres asociaciones ubicadas en la cabecera municipal. En el desempeño de sus labores, estos profesionales del arte de la filigrana manipulan agentes químicos como el ácido nítrico, el ácido sulfúrico, la plata y el mercurio, los cuales se utilizan en las diferentes etapas del proceso que incluye técnicas propias de esta labor. Teniendo en cuenta que la información disponible en la literatura científica describe principalmente los efectos de agentes químicos en otros oficios diferentes a la población orfebre y conociendo que la exposición a estas sustancias químicas puede generar variados efectos en el organismo, el interés de este proyecto se centra específicamente en las alteraciones cutáneas posiblemente relacionadas con las condiciones de trabajo de esta población del sector informal. MATERIALES Y METODOS La presente investigación es un estudio de corte trasversal, el cual realizó una selección por conveniencia de 114 trabajadores de orfebres Momposinos con el fin de identificar la relación de las condiciones de trabajo con la presencia de alteraciones cutáneas de los trabajadores que laboran en la orfebrería artesanal en la ciudad de Mompox, departamento de Bolívar, en el año 2015, de tal manera que dejando a consideración la descripción de los hallazgos encontrados, se posibiliten futuras y precisas investigaciones e intervenciones en este colectivo de trabajadores. Los instrumentos empleados para la recolección de la información y para el cumplimento de los objetivos fueron la encuesta Nacional de Condiciones de Trabajo del Instituto Nacional de Seguridad e Higiene en el trabajo de España (INSHT) que permite recolectar información sobre la caracterización de la población a nivel sociodemográfico y ocupacional, y para la determinación de patologías dermatológicas relacionadas con el trabajo se utilizó el Cuestionario NOSQ-2002 Nórdico- Enfermedades Cutáneas de origen Laboral, en su versión validada en español. Se describieron las variables categóricas con porcentajes y las continuas (cuantitativas) con medidas de tendencia central y dispersión La asociación entre los hallazgos de exposición ocupacional y los síntomas y signos en piel, fue estimada mediante riesgos relativos. RESULTADOS El 75,4% del total de la población correspondió al género masculino y el 67,5% reportó realizar sus labores como trabajadores independientes. Respecto a la identificación de condiciones de salud, la percepción por parte de los orfebres fue positiva, reportando muy buena salud en el 34% de los mismos. El 8% de la población manifestó alteraciones dermatológicas tipo eczema en los últimos seis meses y el 11% las presentó principalmente en manos. Respecto de la iniciación del eczema, el 97% de los trabajadores reportó que se iniciaba al contacto con sustancias químicas y el 98,7% manifestó que se encontraban realizando la labor de orfebrería cuando inició el eczema. La lesión prevalente fue mancha roja sin edema (8%), seguida de ronchas o manchas y ampollas pequeñas (3%) y de piel seca con escamas (2%). CONCLUSIONES Los resultados de la presente investigación mostraron la prevalencia de alteraciones cutáneas principalmente en las manos, tipo eczema (manchas rojas) o prurito (picor). Se recomienda la disminución de los tiempos de exposición, adecuación de jornadas y tiempos de descanso, sistemas de protección personal adecuados y la implementación de un programa de educación y participación para el control integral del riesgo.