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A giant congenital melanocytic nevus (GCMN) is a malformation of the pigment cells. It is a distress to the patients for two reasons: one is disfigurement, and the other is the possibility of malignant changes. However, the underlying mechanisms of the development of GCMN and melanotumorigenesis in GCMN are unknown. Hence, the aim of this study was to identify the proteomic alterations and associated functional pathways in GCMN.
Proteomic differences between GCMN (n=3) and normal skin samples (n=3) were analyzed by one-dimensional-liquid chromatography-tandem mass spectrometry Relative levels of the selected proteins were validated using western blot analysis. The biological processes associated with the abundance modified proteins were analyzed using bioinformatic tools. Among the 46 abundance modified proteins, expression of 4 proteins was significantly downregulated and expression of 42 proteins was significantly upregulated in GCMN compared to normal skin samples (p<0.05). More importantly, 31% of the upregulated proteins were implicated in various cancers, with five proteins being specifically related with melanoma. The abundance modified proteins in GCMN were involved in the biological processes of neurotrophin signaling, melanosome, and downregulated of MTA-3 in ER-negative breast tumors. In particular, an increase in the expression of the 14-3-3 protein family members appeared to be associated with key cellular biological functions in GCMN. Western blot analysis confirmed the upregulation of 14-3-3epsilon, 14-3-3 tau, and prohibitin in GCMN.
These findings suggest that GCMN exhibits potential proteomic alterations, which may play a role in melanotumorigenesis, and the significant alteration of 14-3-3 family proteins could be a key regulator of the biological pathway remodeling in GCMN.
Congenital melanocytic nevi (CMN) are pigment cell malformations that are visible at birth, or are nevi showing congenital features that become clinically obvious shortly after birth [1]. CMN is caused by abnormal melanocyte differentiation, migration, and deposition in the dermis during the early stages of embryogenesis [2,3]. It is a distress to patients for two reasons: one is disfigurement, and the other is the increased risk of developing malignant melanoma [3,4], especially in individuals with giant congenital melanocytic nevi (GCMN; over 20 cm in diameter) [5]. Several genomic and proteomic studies have been performed to elucidate the mechanism of melanotumorigenesis arising from CMN. Gene-based analyses have revealed that the oncogenic BRAF[6] and NRAS[7] mutations are frequently seen in CMN. Additionally, increased Bcl-2 expression in CMN has been suggested to suppress apoptosis, which otherwise plays an important role in the maintenance of nevocytes [8]. In spite of these findings, the major biological processes and pathways of melanotumorigenesis remain unclear. Therefore, the complete proteomic characterization of GCMN and related biological pathways through comparative protein profiling is essential to understand the underlying mechanism of the origin of GCMN and the subsequent process of melanotumorigenesis.
In this study, the proteomic alteration and systemic properties of GCMN were assessed, with the aim of gaining an insight into the functional association between GCMN and melanotumorigenesis. We used label-free liquid chromatography-mass spectrometry (LC-MS) and established bioinformatic tools to identify the proteins that may play a key role in the malignant transformation of GCMN. We found that the 46 proteins were changed in protein abundance levels between normal skin and GCMN samples, and these proteins belonged to tightly organized functional clusters. Moreover, we found that five of the identified proteins were implicated in melanoma. The results of this study will improve our understanding in the biological identification of GCMN and the possible mechanisms that give rise to GCMN-associated melanotumorigenesis.
The clinical characteristics of patients who provided GCMN and normal skin samples are listed in Table Table1.1. All GCMN were over 20 cm in diameter. A representative GCMN lesion in a patient is shown in Figure Figure1A.1A. To minimize environmental bias between individuals, we collected three paired normal and GCMN skin samples from three GCMN donors for one-dimensional-liquid chromatography-tandem mass spectrometry (1D-LC-MS/MS) analysis (Table (Table1).1). Seven unpaired samples from normal skin and GCMN were used for western blot analysis to validate the proteomic results. The average age of GCMN patients who donated samples for western blot analysis was lower (5.57±1.90) than that of the normal skin donors (22.71±11.01, p=0.0058, n=7).
The same amount (20 μg) of soluble proteins obtained from GCMN and normal skin samples were analyzed on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE; Figure Figure1B).1B). The Coomassie blue-stained gels were sliced into 15 uniform slices. The total ion current chromatogram of each slice was acquired (Additional file 1: Figure S1), which led to the identification of 438 non-redundant proteins (Additional file 2: Table S2). Among these, 343 proteins were commonly identified in both the groups, 62 proteins were detected only in GCMN samples, and 33 proteins were detected only in normal skin samples (Figure (Figure1C).1C). Forty-six abundance modified proteins were identified whose protein abundance level was significantly (Student's t-test with Bonferroni correction, p<0.05) different between GCMN and normal skin (Figure (Figure1D).1D). The large majority of these abundance modified proteins were upregulated (42 of 46, 91.3%), and very few (4 proteins, 8.7%) were downregulated in GCMN (Figure (Figure2,2, Table Table2).2). The most upregulated protein in GCMN was cathepsin D (CTSD) [GenBank Gene ID: 1509], which was expressed at a 6.8-fold higher level than in normal skin. In addition, protein disulfide-isomerase A3 precursor (PDIA3, 5.4-fold) [GenBank Gene ID: 2923], prohibitin (PHB, 5.3-fold) [GenBank Gene ID: 5245], heat-shock protein 70 (5.2-fold) [GenBank Gene ID: 3312], D-3-phosphoglycerate dehydrogenase (5-fold) [GenBank Gene ID: 26227], and adenosine-5'-triphosphate synthase subunit beta (5-fold) [GenBank Gene ID: 506] were highly (>4-fold) upregulated in GCMN. The 4 proteins downregulated in GCMN were cytokeratin-1 (−1.4-fold) [GenBank Gene ID: 3848], filaggrin 2 (−4.7-fold) [GenBank Gene ID: 388698], hornerin (−6.0-fold) [GenBank Gene ID: 388697], and alcohol dehydrogenase 1B (−6.2-fold) [GenBank Gene ID: 125]. Interestingly, we found that 33% (14 of total 42) of the significantly upregulated proteins in GCMN samples were implicated in various kinds of cancers (Table (Table2).2). Five of the 14 proteins, namely CTSD [GenBank Gene ID: 1509][9,10], PHB [GenBank Gene ID: 5245][11,12], chondroitin sulfate proteoglycan 4 [GenBank Gene ID: 1464] [13], phosphatidylethanolamine-binding protein 1 [GenBank Gene ID: 5037] [14], and ribosomal protein SA [GenBank Gene ID: 3921] [15] were related with melanoma.
Proteomic profiling of certain diseases has been increasingly used to acquire data on a large number of proteins to identify potential biomarkers and to gain insight into the underlying mechanisms of a variety of diseases. However, the massive amount of generated information often only increases the complexity of these analyses. To address this issue, we developed a systemic approach to analyze the proteomic data generated in the present study. To gain an insight into the systemic properties of the GCMN proteome, the 46 abundance modified proteins were categorized based on molecular function, biological process, and Clusters of Orthologous Groups of proteins (COG) analysis. The proteins were classified into the following classes according to their molecular functions: chaperones (16% of total), oxidoreductases (8%), and cytoskeletal elements (7%) (Figure (Figure3A).3A). Proteins associated with the biological processes of signal transduction (15%), cell structure and motility (10%), protein metabolism and modification (10%), and cell cycle (10%) were markedly altered in their expression. The COG analysis showed that the altered proteins in GCMN samples were involved in posttranslational modification, turnover, chaperones (30%), and the cytoskeleton (12%; Figure Figure3C),3C), which was consistent with the molecular function and biological process classification.
Further network analysis was carried out using the software tool, ClueGO, to identify enriched functional groups in the GCMN samples and obtain detailed information on each one. Gene Ontology (GO)_biological process, GO_cellular component, GO_molecular function, KEGG pathway, and Reactome_biocarta were selected as ontology sources. The specific functional groups identified as significantly enriched in GCMN were melanosome (GO_cellular component), neurotrophin signaling pathway (KEGG pathway), downregulated of MTA-3 in ER-negative breast tumors (Biocarta), cell cycle (KEGG pathway), phospholipase inhibitor activity (GO_molecular function), and glycolysis/gluconeogenesis (KEGG pathway; Table Table3,3, Additional file 3: Figure S2).
Because proteins rarely act alone, but rather interact as a group, thus comprising a functional cluster, a protein-protein interacting network was constructed using STRING 8.0. This network consisted of 45 of the 46 abundance modified proteins, with 52 interactions between them. In the GCMN network, the upregulated proteins were tightly linked with each other, forming a large cluster. The biological meaning of the cluster was determined by overlaying the COG of each protein on the network, which revealed shared COGs between close neighboring proteins (Figure (Figure4).4). Moreover, the GCMN network contained a highly specific protein cluster, identified as the 14-3-3 protein family, whose members were expressed at significantly higher levels than in normal skin as follows: 14-3-3 sigma (or SFN, 1.7-fold) [GenBank Gene ID: 2810], 14-3-3 protein beta/alpha (YWHAB, 2.2-fold) [GenBank Gene ID: 7529], 14-3-3 protein epsilon (YWHAE, 4.2-fold) [GenBank Gene ID: 7531], 14-3-3 protein gamma (YWHAG, 2.1-fold) [GenBank Gene ID: 7532], 14-3-3 protein theta (YWHAQ, 3.3-fold) [GenBank Gene ID: 10971], and 14-3-3 protein zeta/delta (YWHAZ, 2.6-fold) [GenBank Gene ID: 7534].
To investigate the inter-relationship between the proteomic characteristics of GCMN and melanoma, we compared our GCMN proteome network with a recently reported proteome of metastatic melanoma cell line that contained 110 non-redundant proteins [16]. We selected 63 of the total melanoma proteins that had been included in the two high-score biological networks and analyzed protein-protein interactions between the GCMN and melanoma proteomes (Figure (Figure5A).5A). Five proteins including PHB, CTSD, elongation factor 1-alpha 1 (EEF1A1), Annexin A2 (ANXA2), and heat-shock protein beta-1 (HSPB1) were similarly upregulated in both proteome sets. Subsequently, we found that 217 interactions existed between the GCMN and melanoma proteomes. To estimate the biological importance of the 14-3-3 family proteins in the process of melanotumorigenesis, we next analyzed the protein-protein interactions of 14-3-3 family proteins with the GCMN and melanoma proteomes. We identified a total of 23 proteins, out of which 6 proteins in the GCNM and 17 proteins in the melanoma proteome possibly interacted with the 14-3-3 family proteins (Figure (Figure5B).5B). Two proteins interacting with 14-3-3, PHB and EEF1A1, were co-detected in both GCMN and melanoma proteomes. Additionally, the average interaction number of the 14-3-3 family proteins (7.66) was 2-fold higher than the average interaction number of other proteins (3.80). These results suggest that there are similar proteomic alterations in both GCMN and metastatic melanoma and the 14-3-3 family proteins may play an important role in melanotumorigenesis and tumor progression.
To further validate the proteomic results, we evaluated the altered expression of 14-3-3 alpha/beta, sigma, epsilon, zeta, tau, and of prohibitin in GCMN and unpaired normal skin samples (n=7) using semi-quantitative western blots. The integrated values of each protein band are reported in supplementary Additional files 4: Table S2 and Additional files 5.: Table S3 Because it was difficult to obtain skin specimens from normal children younger than 10 years of age, we could not use age-matched normal skin samples. However, we used comparatively young skin samples obtained from subjects whose average age was validated as 22.7 years. As seen in the representative western blots (Figures (Figures6A6A and B), the protein expression of 14-3-3 epsilon, 14-3-3 tau, and prohibitin in GCMN samples was 1.55-fold, 2-fold, and 2.53-fold, respectively, which was significantly higher than that in normal skin samples. However, the expression of 14-3-3alpha/beta, sigma, and zeta were not significantly different in GCMN and normal skin samples. One possible explanation for the unmatched results between the results of the proteomic and western blot analysis might be the diversity of samples used in the western blot analysis, which unlike the paired samples utilized in the proteomic analysis, were collected from normal skin samples of seven non-GCMN patients.
We further analyzed the expression levels of 14-3-3 epsilon, 14-3-3 tau, and prohibitin in normal skin fibroblast cell line (Detroit 551) and three kinds of melanoma cell lines (SK-MEL-2, SK-MEL-5, and SK-MEL-28) to validate whether our proteomic findings are truly relevant to clinical melanoma. Our results showed that the protein levels of 14-3-3 epsilon were significantly increased in all melanoma cell lines, and the levels of 14-3-3 tau were significantly increased in the SK-MEL-2 and SK-MEL-28 cell lines. Additionally, the protein levels of prohibitin were increased in the SK-MEL-2 and SK-MEL-5 cells, but were decreased in the SK-MEL-28 cell line (Figures (Figures7A7A and B).
In the present study, the proteomic composition of GCMN was compared with that of normal skin. A major aim of the study was the identification of proteins whose expression is altered in GCMN, which will help understand the altered biological processes in GCMN and help gain an insight into the mechanism of melanotumorigenesis in these malformations. LC-MS/MS analysis showed that 46 of the 438 identified proteins changed in their abundance levels between the normal skin and GCMN samples. In the GCMN samples, 92% of the abundance modified proteins were upregulated, but only 8% were downregulated (Figure (Figure22 and Table Table2).2). The use of different bioinformatic tools showed that GCMN clearly differed from normal skin in terms of protein expression patterns, which suggested that specific biological processes are altered in GCMN. As derived from the GO categories, KEGG pathways, and Reactome_biocarta, these processes were shown to encompass several major biological functions, namely the neurotrophin signaling pathway, downregulated of MTA-3 in ER-negative breast tumors, the cell cycle, phospholipase inhibitor activity, and glycolysis/gluconeogenesis. Strikingly, among these, neurotrophin signaling [17,18], MTA-3 downregulation (Table (Table3)3) [19], cell cycle deregulation [20], and glycolysis/gluconeogenesis [21] have been implicated in the development and progression of melanoma and other cancers.
Comparison of systemic properties of the GCMN and metastatic melanoma proteomes revealed that these two different disease proteomes shared at least five proteomic alterations in common and their abundance modified proteins closely interacted with each other (Figure (Figure5A).5A). Because closely related diseases are known to share common proteins or common interactions [22], our results suggested the close relationship between GCMN and melanoma.
Our proteomic analysis also revealed the significantly increased expression of 14 cancer-related proteins in GCMN compared to normal skin samples. Among them, PHB is a molecular maker of malignant cancers, and overexpression of PHB has been reported in melanoma [11,12] and various kinds of cancers, including gastric carcinoma [23], thyroid cancer [24], and hepatocellular carcinoma [25]. This significant upregulation of cancer-related proteins in GCMN, specifically which of melanoma-implicated proteins, strengthened the possible risk of melanotumorigenesis in GCMN.
The 14-3-3 proteins comprise a highly conserved family of proteins whose members are found in both plants and mammals. They mediate signal transduction by binding to phosphoserine-containing proteins and are involved in many biological cellular processes, such as metabolism, protein trafficking, signal transduction, apoptosis, and cell cycle regulation, through interaction with various phosphoserine-containing proteins, such as CDC25 phosphatases, RAF1, and IRS1 proteins. In the present study, 14-3-3 family proteins were estimated to interact with 23 proteins in GCMN and melanoma (Figure (Figure5B),5B), and their average number of interactions was about 2-fold higher than the average number of interactions of other abundance modified proteins. These results suggested that 14-3-3 family proteins could play an important role in the alteration of biological processes in GCMN and melanoma.
The 14-3-3 family proteins consist of seven isoforms: beta, gamma, epsilon, sigma, zeta, tau, and eta. The alpha and sigma isoforms are the phosphoforms of 14-3-3 beta and zeta, respectively. All 14-3-3 proteins are ubiquitously expressed, with the exception of 14-3-3 sigma, which is exclusively expressed in epithelial cells [26]. Among the 14-3-3 family members, the overexpression of the 14-3-3 sigma gene or its respective protein is frequently found in cancers such as ovarian carcinomas [27], pancreatic cancer [28], papillary thyroid carcinoma [29], hepatocellular carcinoma [29], and breast cancer [30]. In our proteomic analysis, the expression of 14-3-3 proteins was significantly higher in GCMN than in normal skin samples, which strongly supports the greater tendency toward melanotumorigenesis in GCMN. In particular, the enhanced expression of 14-3-3 epsilon and tau proteins was clearly shown in western blot analysis (Figure (Figure66).
Compared to the other isoforms, little is known about the molecular and biological role of 14-3-3 tau and epsilon proteins. Like other isoforms, 14-3-3 tau is also involved in cell death and survival processes. For example, 14-3-3 tau binds to ataxia telangiectasia-mutated (ATM)-phosphorylated E2F1 during DNA damage and promotes E2F1 stability, leading to the induction of apoptosis [31], and the deletion of 14-3-3 tau leads to embryonic lethality in a mouse model [32]. Interestingly, a recent study suggested that 14-3-3 tau exhibits an oncogenic role by downregulating p21 in breast cancer [33].
14-3-3 epsilon has been shown to play an essential role in cell development. Studies in Drosophila showed that 14-3-3 epsilon is required for the correct timing of mitosis in undisturbed post-blastoderm cell cycle [34]. More recently, defects in neuronal migration during the development of 14-3-3 epsilon-knockout mice were reported [35].
The phosphorylation-induced binding of 14-3-3 epsilon to the pro-apoptotic transcription factor forkhead transcription factor-like 1 (FKHRL1 or FOXO3a) leads to structural changes in 14-3-3 epsilon and inhibits its pro-apoptotic activity [36]. In inflammation and carcinogenesis, 14-3-3 epsilon interacts with key molecules of the mitogen-activated protein kinase signaling module to selectively modulate tumor necrosis factor-alpha-induced nuclear factor-kappa-beta activity [37]. The function and regulatory mechanism of 14-3-3 epsilon in carcinogenesis is controversial and appears to be tumor-specific. Expression of the protein is higher in renal cell carcinoma than that in normal kidney [38]. Moreover, on the basis of their involvement in the tumorigenesis of meningioma, 14-3-3 epsilon, zeta, and theta are thought to be efficient markers for predicting the degree of malignancy of these tumors [39]. In contrast, mRNA and protein expression of 14-3-3 epsilon in laryngeal squamous cell carcinoma tissues was shown to be significantly lower than that in normal tissues [40]. An early role of 14-3-3epsilon in tumorigenesis is suggested by the observation that 14-3-3 epsilon expression is increased in intrinsically aged and photoaged human skin [41]. Interestingly, we found even higher protein levels of 14-3-3 epsilon, 14-3-3 tau, and PHB in GCMN than those in aged skin samples. This result suggested that GCMN may have a higher risk of tumorigenesis than aged skin. Because of the limitation in sample availability, we could not directly determine the expression level of 14-3-3 proteins and PHB in malignant melanoma tissue; however, we demonstrated significantly increased protein expression of 14-3-3 epsilon and tau in two different melanoma cell lines, SK-MEL-2 and SK-MEL-28, compared to normal skin cell line (Detroit 551). This result might support the association of 14-3-3 epsilon and tau upregulation with clinical melanotumorigenesis (Figures (Figures7A7A and B).
Nevertheless, further studies are needed to validate the functional role of 14-3-3 proteins in melanotumorigenesis through the proteomic comparison of different malignant melanoma patients with giant congenital melanocytic nevi. Furthermore, it is also necessary to carefully validate the biological meaning of the upregulation of melanoma-implicated proteins in GCMN and their role in melanotumorigenesis.
Taken together, our data suggest that proteomic modifications with tumorigenic potential are present in GCMN, and these proteomic alterations possibly modify six important biological processes or pathways that include melanosome, neurotrophin signaling pathway, downregulated of MTA-3 in ER-negative breast tumors, cell cycle, phospholipase inhibitor activity, and glycolysis/gluconeogenesis These pathways may be significantly altered in GCMN skins. The intensive alteration of 14-3-3 family proteins and PHB possibly acts as a central regulator of GCMN biological pathway remodeling, which may have an important role in the development of GCMN and could be associated with melanotumorigenesis.
A total of 10 normal and GCMN skin samples, which were defatted, were obtained from patients who underwent excision procedures at the Department of Plastic Surgery, Inje University Ilsan Paik Hospital, Korea. The collection and use of the samples were approved by the Institutional Review Board of Inje University Ilsan Paik Hospital (IRB No. IB-0902-015). The present study was carried out in accordance with The Code of Ethics of the World Medical Association (Declaration of Helsinki) for experiments involving humans.
The human embryo skin cell line Detroit 551 and the human malignant melanoma cell lines SK-MEL-2, SK-MEL-5, and SK-MEL-28 were obtained from the American Type Culture Collection (ATCC; Rockville, MD). The culture medium used throughout these experiments was RPMI-1640 (Lonza, Verviers, Belgium) containing 10% fetal bovine serum (PAA, Pasching, Austria) and 100 μg/ml penicillin-streptomycin (Lonza). The cells were incubated at 37°C in a humidified atmosphere of 5% CO2.
Three paired normal and GCMN skin samples were selected for 1D-LC-MS/MS proteomic analysis to exclude environmental bias. Excised skin samples were ground to a powder in liquid nitrogen, dissolved in lysis buffer (9 M urea, 2 M thiourea, 4% CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate), 40 mM dithiothreitol (DTT), and 1% protease inhibitor cocktail), vortexed, and incubated on ice for 1 h. The mixture was then centrifuged (10,000×g, 30 min, 4°C), and the total proteins contained in the supernatant were used for the experiments. The total protein content of the solution was determined using the 2D Quant kit (GE Healthcare, Milwaukee, WI), with bovine serum albumin (0–50 mg/ml) as the standard.
Protein separation and LC-MS analysis were performed as previously described [42]. Briefly, dissolved skin proteins were separated on a 12% polyacrylamide gel by SDS-PAGE. The gels were washed three times with ddH2O for 5 min each and stained with Bio-Safe Coomassie stain solution (Coomassie G250 stain; Bio-Rad, Hercules, CA) for 1 h, with gentle shaking at room temperature. The Coomassie-stained gels were evenly sliced into 15 slices and then destained by incubation in 75 mM ammonium bicarbonate/40% ethanol (1:1). Disulfides were reduced by treatment with 5 mM DTT/25 mM ammonium bicarbonate at 60°C for 30 min, followed by alkylation with 55 mM iodoacetoamide at room temperature for 30 min. The gel pieces were then dehydrated in 100% acetonitrile (ACN), dried, and swollen overnight at 37°C in 10 μl 25 mM ammonium bicarbonate buffer containing 20 μg modified sequencing-grade trypsin (Roche Applied Science, Indianapolis, IN) per ml. The tryptic peptide mixture was eluted from the gel using 0.1% formic acid. LC-MS/MS analysis was performed using a ThermoFinnigan ProteomeX workstation LTQ linear ion trap MS (Thermo Electron, San Jose, CA) equipped with a nanospray ionization (NSI) source (Thermo Electron). Briefly, 12 μl peptide sample obtained from the in-gel digestion was injected and loaded onto a peptide trap cartridge (Agilent, Palo Alto, CA). Trapped peptides were eluted onto a 10-cm reversed-phase PicoFrit column packed in-house with 5-μm, 300-Å pore size C18 and separated by gradient elution. The mobile phases consisted of H2O and ACN, both containing 0.1% v/v formic acid. The flow rate was maintained at 200 nl/min. The gradient started at 2% ACN, then reached 60% ACN in 50 min, 80% ACN in the next 5 min, and 100% H2O in the final 15 min. Data-dependent acquisition (m/z 400–1800) was enabled, and each MS survey scan was followed by five MS/MS scans within 30 s, with the dynamic exclusion option enabled. The spray voltage was 1.9 kV, the temperature of the ion transfer tube was 195°C, and the normalized collision energy was 35% [42].
Data-analyzed tandem mass spectra were extracted, and the charge state was deconvoluted and deisotoped using the Sorcerer 3.4 beta2 platform (Sorcerer software 3.1.4, Sorcerer Web interface 2.2.0 r334, and Trans-, Proteomic Pipeline 2.9.5). All MS/MS samples were analyzed using SEQUEST (version v.27, rev. 11; ThermoFinnigan, San Jose, CA), which was set to search the ipiHuman 3.29 database (IPI ver.3.29, 40131 entries), with semitrypsin as the digestion enzyme. The search used a fragment-ion mass tolerance of 1.00 Da and a parent-ion mass tolerance of 1.5 Da. Iodoacetamide-derivatized cysteine was specified as a fixed modification. Methionine oxidation, iodoacetamide derivatizion of cysteine, and phosphorylation of serine, threonine, and tyrosine were specified as variable modifications. The Scaffold software (version Scaffold-2.0; Proteome Software Inc., Portland, OR) was used to validate MS/MS-based peptide and protein identifications. Peptide identifications were accepted if their probability was >95.0%, as specified by the Peptide Prophet algorithm, and if they contained at least one identified peptide. Protein probabilities were assigned by the Protein Prophet algorithm. Proteins containing similar peptides such that they could not be differentiated based on MS/MS analysis alone were grouped to satisfy the principles of parsimony. After identifying the proteins, each dataset was used for a subtractive analysis by semi-quantitative normalized spectral counts, which were normalized by total spectral counts in the Scaffold program [43].
A systemic bioinformatics analysis of the GCMN proteome was conducted using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING 8.3) [44], the Protein Analysis Through Evolutionary Relationships classification system (PANTHER 7.0) [45], the National Center for Biotechnology Information (NCBI) COG database [46], Cytoscape, and ClueGO [47].
Protein expressions of 14-3-3 alpha+beta (30 kDa), 14-3-3 epsilon (29 kDa), 14-3-3 zeta (28 kDa), 14-3-3 sigma (25 kDa), 14-3-3 tau (31 kDa), and prohibitin (30 kDa) in normal (n=7) and GCMN (n=7) samples were analyzed by western blots to confirm the proteomic results. In addition, protein expressions of 14-3-3 epsilon, 14-3-3 tau, and prohibitin in a normal cell line (Detroit 551) and melanoma cell lines (SK-MEL-2, SK-MEL-5, and SK-MEL-28) were analyzed by western blots. Relative expression of each protein was normalized to an internal standard protein, β-tubulin (55 kDa) or glyceraldehyde-3-phosphate dehydrogenase (37 kDa) (Abcam, Cambridge, MA). The values were expressed as mean±standard error.
Two-tailed unpaired Student's t-test in the Scaffold (version 2.06.02) software was used to compare abundance of each protein in the normal and GCMN skin samples. Using the Origin software (version 7.0220, OriginLabs, MA, USA), Bonferroni correction was applied to control the rate of false positives in the comparison of means of each protein's abundance. A two-tailed unpaired Student's t-test was used to compare the results of western blot analysis between the normal and GCMN skin samples. p<0.05 was considered statistically significant.
GCMN: Giant congenital melanocytic nevus; CTSD: cathepsin D; PDIA3: Protein disulfide-isomerase A3 precursor; PHB: Prohibitin; HSPA8: Heat shock protein 70; PHGDH: D-3-phosphoglycerate dehydrogenase; ATP5B: ATP synthase subunit beta; KRT1: Cytokeratin-1; FLG2: Filaggrin; HRNR: Hornerin; ADH1B: Alcohol dehydrogenase 1B; CSPG4: Chondroitin sulfate proteoglycan 4; PEBP1: Phosphatidylethanolamine-binding protein; RPSA: Ribosomal protein SA; COG: Clusters of Orthologous Groups; STRING: Search Tool for the Retrieval of Interacting Genes/Proteins; PANTHER: Protein Analysis Through Evolutionary Relationships; KEGG: Kyoto Encyclopedia of Genes and Genomes; GO: Gene Ontology.
The authors declare that they have no competing interests.
HKK performed the proteomic analysis including its design, coordination, analysis of the data, and drafted the manuscript. YKK provided GCMN and normal skin samples and was largely involved in the design of the study and in writing the manuscript. ISS, SRL, SHJ, MHK, and DYS performed sample preparation and western blot analysis. NK, BDR, KSK, KCT, and CGP critically revised the clinical aspect of the manuscript. JYC carried out the MS/MS proteomic analysis. JH conceived the overall experimental design and manuscript preparation. All authors read and approved the final manuscript.
Figure S1. Peptide mass peak of 15 slices dissected from 1D gels of normal skin and GCMN.
Table S1. Identified proteins in skin samples of Normal and GCMN patients.
Figure S2. The functional network and enriched functional group of GCMN was analyzed using ClueGO, a biological term enrichment analyzer. The following proteins were significantly enriched in GCMN: melanosome (GO_cellular component), neurotrophin signaling pathway (KEGG pathway), downregulated of MTA-3 in ER-negative breast tumors (Biocarta), cell cycle (KEGG pathway), phospholipase inhibitor activity (GO_molecular function), and glycolysis/gluconeogenesis (KEGG pathway).
Table S2. Integrated densitometry value of Western blot band.
Table S3. Statistical analysis result of western blot.
This work was supported by Priority Research Centers Program and Basic Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (353-2009-1-E00002 and 2010–0020224).
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Real Life Evaluation of Rheumatoid Arthritis in Canadians Taking HUMIRA (REACH)
This study has been completed.
AbbVie (prior sponsor, Abbott)
ClinicalTrials.gov Identifier:
First Posted: May 5, 2010
Last Update Posted: February 24, 2016
University Health Network, Toronto
Information provided by (Responsible Party):
AbbVie ( AbbVie (prior sponsor, Abbott) )
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Detailed Description:
The purpose of this study is to evaluate the clinical effectiveness and safety of adalimumab as used in routine clinical practice in adult participants with moderate to severe rheumatoid arthritis (RA) in Canada.
|Moderate-to-severe Rheumatoid Arthritis
|Study Type:
|Study Design:
|Observational Model: Cohort
Time Perspective: Prospective
|Official Title:
|Real Life Evaluation of Rheumatoid Arthritis in Canadians Taking HUMIRA (REACH)
Resource links provided by NLM:
Genetics Home Reference related topics: rheumatoid arthritis
MedlinePlus related topics: Arthritis Rheumatoid ArthritisU.S. FDA Resources
Further study details as provided by AbbVie ( AbbVie (prior sponsor, Abbott) ):
Primary Outcome Measures:
- Percentage of Participants That Achieved a Disease Activity Score 28 (DAS28) < 2.6 [ Time Frame: Month 0, 6, 12, 18 and 24 ]The DAS28 is a validated index of rheumatoid arthritis disease activity. Twenty-eight tender joint counts, 28 swollen joint counts, erythrocyte sedimentation rate (ESR), and the Subject's Global Assessment of Disease Activity (subject rates disease activity using a likert scale from 0 [low activity] to 10 [high activity]) are included in the DAS28 score. Scores on the DAS28 range from 0 to 10. A DAS28 score >5.1 indicates high disease activity, a DAS28 score <3.2 indicates low disease activity, and a DAS28 score <2.6 indicates clinical remission.
Secondary Outcome Measures:
- Mean Change From Baseline (Month 0) in Health Assessment Questionnaire (HAQ) [ Time Frame: Month 0, 6, 12, 18 and 24 ]Physical function was evaluated using the Health Assessment Questionnaire - Disability Index (HAQ-DI), a participant-reported questionnaire specific for rheumatoid arthritis. It consists of 20 questions referring to eight domains: dressing/grooming, arising, eating, walking, hygiene, reach, grip, and daily activities. Participants assessed their ability to do each task over the past week using the following response categories: without any difficulty (0); with some difficulty (1); with much difficulty (2); and unable to do (3). Scores on each task were summed and averaged to provide an overall score ranging from 0 to 3, where zero represents no disability and three very severe, high-dependency disability. Negative mean changes from baseline in the disability index of the HAQ-DI indicated improvement.
- Mean Change From Baseline (Month 0) in Rheumatoid Arthritis Disease Activity Index (RADAI) [ Time Frame: Month 0, 6, 12, 18 and 24 ]The RADAI is a questionnaire for participants used for measuring disease activity. The index consists of 6 questions. The items ask the participants about (1) global disease activity in the last 6 months, (2) disease activity in terms of current swollen and tender joints, (3) arthritis pain, (4) the current status of health, (5) duration of morning stiffness and (6) tender joints to be rated in a joint list. The joint list asks about pain in the left and right shoulders, elbows, wrists, fingers, hips, knees, ankles and toes. The first 3 items are all rated on a numeric rating scale from 0 to 10, where higher scores indicate more disease activity. The RADAI total score is the sum of individual items divided by 5 (range 0-10), with a higher score signifying more disease activity.
|Study Start Date:
|July 2005
|Study Completion Date:
|December 2014
|Primary Completion Date:
|December 2014 (Final data collection date for primary outcome measure)
Moderate-to-severe rheumatoid arthritis
Participants with moderate-to-severe rheumatoid arthritis treated with adalimumab in routine clinical practice
Detailed Description:
REACH is an observational survey of participants with moderate to severe rheumatoid arthritis taking adalimumab. Participants who volunteer will be asked to provide information about their medical history and experiences with adalimumab. No drug will be provided as a result of participation in the registry. All treatment decisions are independent of participation in the registry.
Information from the National Library of Medicine
Choosing to participate in a study is an important personal decision. Talk with your doctor and family members or friends about deciding to join a study. To learn more about this study, you or your doctor may contact the study research staff using the contacts provided below. For general information, Learn About Clinical Studies.
|Ages Eligible for Study:
|18 Years and older (Adult, Senior)
|Sexes Eligible for Study:
|Accepts Healthy Volunteers:
|Sampling Method:
|Probability Sample
Study Population
Participants with moderate-to-severe rheumatoid arthritis
Inclusion Criteria:
- Participant is eligible to take part in the registry as per the product monograph.
- Participants that are naïve to adalimumab therapy / or participants that have been receiving adalimumab therapy for less than 4 months.
- Participant has moderately to severely active RA.
- Participant who has had an inadequate response to one or more Disease Modifying Anti-Rheumatic Drugs (DMARDs).
- Participant received provincial or private (insurance companies) approval for adalimumab.
- Participant is able to give written informed consent and to understand the survey requirements.
Exclusion Criteria:
- Participant to whom a traditional DMARD had never been tried.
- Participant with a known hypersensitivity to adalimumab, or any of its components.
- Participant is receiving free adalimumab as part of a compassionate program or an early access drug distribution program.
- Participant with clinically significant concurrent medical or psychiatric disorders that may influence survey outcomes.
- Participant with any condition that would prevent participation in the survey and completion of the survey procedures including language limitation or possibility that the participant will not be available for a period of time (> 12 months) while being enrolled in the survey.
Contacts and Locations
Information from the National Library of Medicine
To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor.
To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor.
Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT01117480
Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT01117480
Sponsors and Collaborators
AbbVie (prior sponsor, Abbott)
University Health Network, Toronto
|Study Director:
|Linda Assouline, PhD
|AbbVie Corporation
More Information
Additional Information:
Keywords provided by AbbVie ( AbbVie (prior sponsor, Abbott) ):
Additional relevant MeSH terms:
Additional Information:
|Responsible Party:
|AbbVie (prior sponsor, Abbott)
|ClinicalTrials.gov Identifier:
|NCT01117480 History of Changes
|Other Study ID Numbers:
|First Submitted:
|May 4, 2010
|First Posted:
|May 5, 2010
|Results First Submitted:
|December 11, 2015
|Results First Posted:
|February 24, 2016
|Last Update Posted:
|February 24, 2016
|Last Verified:
|January 2016
Keywords provided by AbbVie ( AbbVie (prior sponsor, Abbott) ):
Moderate Rheumatoid Arthritis
Rheumatoid Arthritis
Severe Rheumatoid Arthritis
Clinical Effectiveness
Additional relevant MeSH terms:
Arthritis, Rheumatoid
Joint Diseases
Musculoskeletal Diseases
Rheumatic Diseases
Connective Tissue Diseases
Autoimmune Diseases
Immune System Diseases

subset: commoncrawl


text:
EOiNA: Gate Crasher
Wolfie finally gets to put that new form to use.
|‹‹ First
|‹ Prev
|Next ›
|Last ››
Wolfie finally gets to put that new form to use.
You think Loki and Hermes would LOVE gate crasher and ENCOURAGE THEM! And the guards would HAPPILY let him through!
And make the gate crashing less enjoyable?
this Hero really enjoys the powers of her new form.
… but we are all one of a kind. Therefore all of us are the most powerful?
someone answered this riddle before it was asked, all you have to is check the past.
Well t's pretty obvious, King Lycos is also known as King Lycaon. According to the legend he pissed of Zeus by serving him (aka offering as a sacrifice) the flesh of a child (depending on the legend, sometimes one of his own sons)
According to the legend, Zeus got mighty pissed off and transfomed him and all of his sons (he had a lot) into wolves. Thus founding lycantropy.
The first of her kind is noone else but King Lycos, aka herself, and she must have one big huge awesome morph version!
I hope Wolfie has tested out that alternate form as Lycos, because this could be embarrassing.


text:
Sliding window sweep lock
A quick learner from California says:
June 3, 2016
I need the opposite of the lock pictured. The sliding window in question opens left to right. 2.1/4" screw holes. Aluminum window. These locks are different depending on which way the window slides.
Tom from SWISCO responded:
June 3, 2016


text:
It's a holiday tradition to hunt for fun things at the SL Christmas Expo.
This year we introduce – Holiday Stocking Stuffers
Each stocking has something very special from the merchant. Bring your friends and family for a fun day at the Expo and hunt for the Holiday Stocking Stuffers
Below are some of the fabulous items available from participating Christmas Expo merchants.


text:
Results 1 to 2 of 2
Thread: Change instance owner id ?
01-14-10, 04:10 #1Registered User
- Join Date
- Nov 2007
- Posts
- 62
Unanswered: Change instance owner id ?
Running db2 v9.5 on AIX6.1 box.
I need to change db2 owner id.
So I asume I must change id and permissions on all db2 files.
Do I need some aditional actions ?
Or maybe there are some utilities made for this purpose ?
01-14-10, 04:39 #2Registered User
Provided Answers: 1
- Join Date
- Jan 2003
- Posts
- 1,636
I have changed IDs for DB2 users on Linux several months ago and got no problem at all. I just did
the following:
# log in with db2inst1 user and execute the following commands db2stop force exit # if you have DB2 admin server running, then log in with dasusr1 and execute the following commands db2admin stop exit # log in with root user and execute the following commands gawk -F: '{print $3, $1}' /etc/passwd | sort -n # Check IDs, you have to set new ID to some value that is not already assigned. usermod -u 999 db2inst1 usermod -u 998 dasusr1 usermod -u 997 db2fenc1
You don't need to change any files permissions because there are not ID dependent, but user_name dependent. Since you haven't change user_name no need of changing file permissions.
I haven't tested this on Unix, just assume it is the same as on Linux. I suggest you test this on test system before doing some change on production environment.
Last edited by grofaty; 01-14-10 at 04:45.


text:
Bounty Beach Cocobana Resort
Malapascua Island, Philippines
Bounty Beach Cocobana Resort Malapascua Island Reviews
18 reviews
Bounty Beach Cocobana Resort Sep 11, 2011
I stayed here for two nights in their superior room (5000p/night). The room is large and comes with a terrace. It has AC, a fridge, table and chairs, a satellite TV and a spacious bathroom. I enjoyed my stay here as the beach is just a few steps away. The hotel arranges land and boat transportation from Cebu City to the island for a fee. If you don't mind the hassle, you can take a bus from the city going to Maya wharf in Daan Bantayan, then take a boat to the island. Cocobana is very close to the dock point and its staff meet us and carry our bags. As this place is a little costly, I definitely recommend it to those who are willing to splurge a little and want to have a good view of the beach.
PS. I brought my dog to the island and the resort's staff didn't mind having him there.
1 / 1 TravBuddies found this review helpful/trustworthy
Join TravBuddy to leave comments, meet new friends and share travel tips!


text:
She lick and rub my pussy while hubby fuck her doggy
And I should be there fucking you as she licks your pussy juice from my shaft!
Yes please!!!!!!
@drkrod69 stick it into her mouth then in my cunt
Nice quality videos babe I need to come to Italy for a few months!
@superdick33 yes, we have better camera quality
@ziofester no doubt babe at least you have videos up to just like me and a few others here not camera shy lol I just need to get more women and produce more but America has a lot of flakes lol
Mmmm hott. Looks you're missing some chocolate!!
@Blackbone69 Yes, chocolate is so good for my skin...mmm
I agree. Good for more than just skin.
Works wonders in pussies!!
Hot as fuck that two tight white sluts need dark bigger meat though
A woman licks a pussy better than a man. She knows where, how and when put the tongue inside another pussy. I learned eat pussy with my wife.
Separate names with a comma.


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Changing your hair colour might seem like life-altering decision…because it is! Kind of!
After having the same hair colour and/or style for years on end, sitting in the salon chair is a weeeee bit nerve-wracking. I know. Last New Years Eve, I chopped all of my locks off! But, a little change goes a long way. Here are some styles you can try without giving yourself a mid-teen-life crisis.
Header image: www.bohemianbrighton.com
Grey Ombrè
Source: www.imagesoyl.com
If you have brunette hair, ombrè grey should definitely be an option. It's not too much, but it's just enough to give yourself a whole new look because of the contrast!
Pink Ombrè
Source: www.pointyshoecrew.com
If you're a blondie, try the pink ombrè for some dramatic but easy change.
Green Frost
Source: Jinx In The Sky
If you're feeling really daring, why not pair your lip liner with some green hair? The greatest thing about this colour is that it works for both summer and winter. #Score
Reverse Ombrè
Source: www.stylesbysteph.com
So *progressive*.
Colours of the Rainbow
Source: www.weheartit.com
If you're really, really daring, try the Katy Perry dark colours of the rainbow style! It never hurts to try new things!
Tagged Colour, fashion, hair, hairspiration, Kylie Jenner, style


text:
Traveling Luck for Elkhorn Ranger Station, Washington, United States
Where is Elkhorn Ranger Station?What's around Elkhorn Ranger Station?
Wikipedia near Elkhorn Ranger Station
Where to stay near Elkhorn Ranger Station
The timezone in Elkhorn Ranger Station is America/Whitehorse
Sunrise at 07:29 and Sunset at 16:29. It's light
Latitude. 47.8722°, Longitude. -123.4672° , Elevation. 457m
Weather near Elkhorn Ranger Station; Report from Port Angeles, William R. Fairchild International Airport, WA 32.2km away
Weather : mist
Temperature: 9°C / 48°F
Wind: 3.5km/h Southeast
Cloud: Broken at 200ft Broken at 3500ft Solid Overcast at 7500ft
Weather : mist
Temperature: 9°C / 48°F
Wind: 3.5km/h Southeast
Cloud: Broken at 200ft Broken at 3500ft Solid Overcast at 7500ft
Satellite map around Elkhorn Ranger Station
Loading map of Elkhorn Ranger Station and it's surroudings
Geographic features & Photographs around Elkhorn Ranger Station, in Washington, United States
- stream;
- a body of running water moving to a lower level in a channel on land.
- Stony Creek (0.2km)
- McCartney Creek (0.8km)
- Lost River (1.3km)
- Evergreen Creek (1.8km)
- Wildrose Creek (2.4km)
- Goldie River (4.1km)
- Prescott Creek (4.3km)
- Dorothy Creek (5.1km)
- Windfall Creek (5.5km)
- Bowman Creek (6.7km)
- Hayes River (8.3km)
- Lillian River (9km)
- Fairchild Creek (10km)
- Idaho Creek (11.2km)
- Antelope Creek (11.4km)
- mountain;
- an elevation standing high above the surrounding area with small summit area, steep slopes and local relief of 300m or more.
- Dodger Point (3.5km)
- Windfall Peak (4km)
- Ludden Peak (6.5km)
- Scott, Mount (6.6km)
- McCartney Peak (7.3km)
- Ferry, Mount (9.3km)
- Obstruction Peak (9.7km)
- Eagle Point (9.9km)
- Elk Mountain (10.3km)
- Dana, Mount (10.4km)
- Lost Cabin Mountain (10.7km)
- Stephen Peak (10.8km)
- South Dana (12km)
- Head Cameron (12.1km)
- Claywood, Mount (12.4km)
- Local Feature;
- A Nearby feature worthy of being marked on a map..
- Elkhorn Ranger Station (0km)
- Elkhorn Guard Station (0.1km)
- Stony Point Camp (0.2km)
- Canyon Camp (2km)
- Marys Falls Camp (4.2km)
- Tipperary Camp (6.6km)
- Chateau Camp (7.4km)
- Hayes River Guard Station (8km)
- Lillian Shelter (9.4km)
- Steeple Rock (11.4km)
- lake;
- a large inland body of standing water.
- Lillian, Lake (6.7km)
- Gladys Lake (9.5km)
- Moose Lake (10.2km)
- PJ Lake (10.5km)
- Grand Lake (10.8km)
- Frog Lake (11.2km)
- Cream Lake (11.3km)
- valley;
- an elongated depression usually traversed by a stream.
- Grand Canyon (10.8km)
- Grand Valley (11.1km)
- Badger Valley (11.4km)
Accommodation around Elkhorn Ranger Station
- trail;
- a path, track, or route used by pedestrians, animals, or off-road vehicles.
- Long Ridge Trail (4.7km)
- Elwha River Trail (4.8km)
- Lillian River Trail (9.6km)
- ridge(s);
- a long narrow elevation with steep sides, and a more or less continuous crest.
- Long Ridge (6km)
- Hurricane Ridge (9.2km)
- gap;
- a low place in a ridge, not used for transportation.
- Grand Pass (9.7km)
- Cameron Pass (11.1km)
- flat;
- a small level or nearly level area.
- Press Valley (6.2km)
- range;
- a series of associated ridges or seamounts.
- Bailey Range (10km)
- glacier(s);
- a mass of ice, usually at high latitudes or high elevations, with sufficient thickness to flow away from the source area in lobes, tongues, or masses.
- Lillian Glacier (8.2km)
- bench;
- a long, narrow bedrock platform bounded by steeper slopes above and below, usually overlooking a waterbody.
- Semple Plateau (3.7km)
Wikipedia entries close to
Elkhorn Ranger Station
- Mount Anderson (Washington) mountain mountain k
- Olympic Mountains mountain mountain g
- Mount Olympus (Washington) mountain mountain g
- Elwha, Washington city city o
- Elwha Dam hydroelectri
- Agnew, Washington city city o
- Port Angeles East, Washington city city o
- Port Angeles (Washington) city city i
- Ediz Hook Light tower
- William R. Fairchild International Airport airport airport
- Mount Constance mountain mountain k
- Carlsborg, Washington city city o
- River Road, Washington city city o
- Lake Crescent city city o
- Bell Hill, Washington city city o
- Sequim, Washington city city o
Airports close to Elkhorn Ranger Station
Port angeles cgas(NOW), Port angeles, Usa (34.4km)
Whidbey island nas(NUW), Whidbey island, Usa (91.8km)
Victoria international(YYJ), Victoria, Canada (98.1km)
Snohomish co(PAE), Everett, Usa (101.3km)
Boeing fld king co international(BFI), Seattle, Usa (109km)
Airfields or small airports close to Elkhorn Ranger Station
Photos provided by Panoramio are under the copyright of their owners.


text:
3 BHK Flats in Omaxe...
₹ 6,500,000.00
₹ 2,700,000.00
Panchsheel Greens is grand situated at Noida Extension, Having 80% green area approximately it. Located on 130mt wide road, it is surrounded by natural environment and fresh air. It is very close to many residential and commercial hubs. The floorplan of Panchsheel is great, having different size of floor plan which starts from 775 sq ft to 2350 sq. ft.
Panchsheel Greens Location Benefits -
· 5 km from crossing republik nh24
· 9 km from Noida Sector 71
· 5 km from noida sector 121
· 12 km from Noida City Center Metro station,
· 9.5 km from Sai mandir Noida
· 7.5 km from Indirapuram Ghaziabad
Panchsheel Greens Features –
Ground for Cricket, Football, 2 tennis courts, 2 basketball courts, Skating rink, Badminton court, Cycling, athletic track, Club with swimming pool, spa and gym, 2 exclusive swimming pools, Exclusive shopping plaza, Banquet hall and party lawns, Boating lake, 2 solariums, Separate Kids Play area, Multipurpose open play field Indoor restaurants, Food court etc.
Contact Us:
Contact No- +91 - 9560538123
Visit: - http://www.panchsheelgreens1.com/
Published date:
01/04/2016 2:59 pm
Expiration date:
Uttar Pradesh
City area:
Greater Noida West
Greater Noida West
|For sale
|New construction
|No. of bedrooms
|No. of bathrooms
|Square Meters
|semi furnished
Safety Tips
Contact publisher
You must log in or register a new account in order to contact the publisher
3 BHK Flats in Omaxe...
₹ 6,500,000.00
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₹ 4,000,000.00
Omaxe Residency – 2B...
₹ 12,000.00


text:
Holder as Kingmaker? Lawmakers in the Spotlight in Fight vs. Attorney General
The months of hearings that culminated in last week's contempt vote against Eric Holder have made the attorney general a villain in some eyes and a hero in others.
But the highly publicized oversight of the nation's top cop in the wake of Fast and Furious, voter ID, SB 1070 and other issues has also cast Holder in an unlikely role: kingmaker.
This isn't to say in any shape or form that the proceedings against Holder are rooted in simply generating publicity for lawmakers. It's just a natural effect that the bigger the case, the bigger the names, the greater the allegations, the more spotlight that's cast on the players involved.
Some members of Congress were simply destined to become bigger household names with the progression of Holder-related scandals, or even without the attorney general. House Oversight and Government Reform Committee Chairman Darrell Issa (R-Calif.) was raring to go after waste and abuse from the minute he knew he would take the gavel after the Republicans' 2010 midterm rout. A stick figure cop in front of the Capitol dome became his Twitter avatar, and the first hearing his committee held in January 2011 was on the TARP bailouts. Before Fast and Furious was on the radar, Issa was going after federal spending binges, regulatory impediments to job creation, and the failing U.S. Postal Service.
Issa, who did a triple-play on the Sunday shows this weekend, is no-holds-barred and you know that when he sets his mind to push forward with something no amount of arm-twisting will deter him. The Democrats brand him as a combative witch-hunter, which alone elevates him to most-favored-lawmaker status among many on the right, particularly in the grass-roots. His star won't necessarily be at the same height with Republican House leadership as it is with the conservative base, so it will be especially fascinating to see what lies in Issa's future, particularly as the end of the 112th Congress plays out.
Quotable: "No, Mr. Attorney General, you're not a good witness. A good witness answers the question asked. So let's go back again. Have you and your attorneys produced internally the materials responsive? In other words, have you taken the time to look up our subpoena and find out what material you have responsive to it, or have you simply invented a privilege that doesn't exist?" -- House Judiciary Committee oversight hearing with Holder testifying, June 7, 2012
Here are some others who have stood out as up-and-comers in the Holder proceedings.


text:
Pilot Study of External Hyperthermia and Intravesical Mitomycin-C To Treat Recurrent Bladder Cancer After Resection and Standard Adjuvant Therapy
It is well established that tumor cells are sensitive to heat and when combined with a
chemotherapeutic agent, drug uptake and intracellular distribution of drug within malignant
cells is improved due to increased cellular permeability. Further, hyperthermia inhibits
deoxyribonucleic acid (DNA) repair as a result of increased reaction between DNA and
chemotherapy. By heating bladder tissue and accelerating the necessary series of reactions
to link agents such as mitomycin C to cell DNA, this effect may be optimized. Depending on
the extent of resection (and location, size, and depth of invasion of remaining tumor) after
transurethral resection of the bladder tumor(TURBT), recommended adjuvant therapy consists
of intravesical chemotherapy. MMC has been studied at doses as high as 80 mg without
producing significant or irreversible side effects. The most commonly used dose of mitomycin
is 40 mg.2
This pilot study proposes to use Mitomycin C at a dose of 40 mg in conjunction with deep
hyperthermia to enhance the effect intravesical chemotherapy as second-line treatment of
recurrent TCC (Stage Ta, T1, or Tis) of the bladder after surgical resection and standard
adjuvant therapy.
Endpoint Classification: Safety/Efficacy Study, Intervention Model: Single Group Assignment, Masking: Open Label, Primary Purpose: Treatment
Number of patient treatments stopped due to safety concerns including side effect such as heat sensations or burning in the pelvis, intolerance to heat, weight of the water bolus on the abdomen, or supine position.
During Treatment Phase average 6 weeks
Zeljko Vujaskovic, MD, PhD
Principal Investigator
Duke University
United States: Food and Drug Administration
April 2008
March 2011
|Duke University Medical Center
|Durham, North Carolina 27710


text:
Sexual dimorphism in feeding structures is a common phenomenon in nature and more specifically in mammals. The purpose of this lab was to determine if canine size and bite force is sexually dimorphic in Arctic foxes (Vulpes lagopus). The canine width of female and male fox skulls were measured, as well photographs of fox skulls were used to measure bite force. The class data used in the study showed evidence that male Arctic foxes were found to have larger canine width as well as a larger bite force than females. The class data collected was analyzed using a t-test. The t-test was used to compare average canine width and average bite force between male and female Arctic foxes to determine if they are sexually dimorphic. For both average canine width and average bite force, the p value was less than 0.05, which showed evidence that the difference in male and female bite force is significant. As well, the difference in male and female canine width was significant. The data suggested that the differences in the feeding structures of the Arctic fox have resulted from sexual selection, food competition between sexes and dimorphic niche hypothesis. These three hypotheses help explain the results showing that bite force and canine width are sexually dimorphic in the Arctic fox.
Sexual dimorphism has been a widespread phenomenon and a major focus of evolutionary studies since Charles Darwin and his studies on natural selection in the animal kingdom. Males and females are known to have different reproductive organs; however other sexual differences in traits (odour, size, colour, etc.) of males and females do exist. The phenomenon of marked differences between the secondary sex characteristics of males and females is known as sexual dimorphism (Reece et al., 2012).
A peacock (Pavo cristatus) and a peahen are one of the extreme examples of sexual dimorphism. The peacock is known for his long upper tail covert feathers where he displays...


text:
This season we have been following Big Slugger and his outdoor medical cannabis garden of Blue Dream clones. Many of you have been patiently waiting for the updates. For those of you who have not been following along, the links in the following two paragraphs will take you to the earlier videos and get you up to speed.
Back in June we posted the first article and video Grow Like A Pro. We introduced you to the plants and showed as they moved from the grow room to the greenhouse. At that point they were only about 3 feet tall and looked pretty tiny in Sluggers huge greenhouse.
In August we posted the first update of Big Sluggers garden. The footage had been taken on July 19th and showed the Blue Dream clones doing nicely. The plants had received their seconds cages and the branches were being trained to maximize yield and support.
In this video we show you the plants as they begin to reach the end of their vegetative period and begin to flower. The plants are monsters averaging about 12 feet tall and 12 feet in diameter. The 25 Blue Dreams completely fill the 4000 square foot greenhouse. The branches are touching the exterior wall, rafters and are close to growing into each other. As they move further into flowering, their vegetative growth will slow as they divert energy to flower production. Whoever said clones don't grow big outdoors is going to have some explaining to do.
We hope you enjoy this video. Be sure to subscribe to our Youtube channel so you don't miss any of the updates. If you have questions or comments please post them at the end of this article in the comments section. We don't keep track of the comments on youtube as well as we should.
Monster Plants: Big Sluggers 2012 Outdoor Update,


text:
Do you need an advertising organization which will help you beat your rivals? Do you occasionally question what it really seems like to have your company rated at number 1? We're undoubtedly what you require.
At Bodcity SEO Consulting, we understand that researching your competitors is important if you wish to remain at the top of the game. In order to do this, we've our very own team of professionals in online marketing, Search engine optimization procedures, and also web design prepared to help your company if needed.
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We will undoubtedly have the ability to help with your internet business. Contact us t... more
Finally, a company that listens to what you need. Professional advice and great direction in helping make my website come alive. Highly recommended.


text:
1. Be more present
We live in a world of constant distraction. iPhones, laptops, radios, Twitter, Facebook, Instagram, the internet. The world is at our fingertips and it's never been full of so much crap. This year I resolve to be more present. This means if I am at work, I work. I actually had our IT person block Facebook from my computer. Facebook is nothing but a distraction. Rarely do I find it that interesting anymore. If I'm driving, I'm driving. I don't text while I drive but I do pick up my phone at long lights and check out the weather, etc. Enough. I need to just enjoy my time while driving (and listen to NPR of course). Most importantly, when I am with my daughter or Seth, I need to be fully present. Parents, you know how easy it is. You are playing on the floor with your kid, your phone beeps that you have a new email, Facebook notification, text message, etc. Rarely are these valuable things that should take us away from our sweet children. Let it go. Texts can wait. Groupon emails can wait. Instagram notifications don't matter. I will check my phone only when I'm alone. I'm tired of giving more attention to my iPhone than to my family and friends.
2. Stay active
I'm a better wife, mother, friend and coworker when I am exercising. I love when I am set in a routine. I don't want this year to be about losing a certain amount of weight. I just want it to be about staying active. Yoga, running, walking, etc., it doesn't matter. The important thing I that I keep moving. And I plan to. Hopefully I'll be pregnant at some point this year and I really want to stay active LONGER than I did last time around. Here's hoping.
3. Simplify
Our society makes us believe that we need to have a lot of "stuff'". My aunt and uncle recently lost nearly all of their possessions in a house fire. While my heart breaks for them, they have had the most amazing attitude about it. "It's just stuff." They are right. As horrible as it sounds to lose everything you have, how nice would it be? I am amazed at how much "stuff" we have. Closets full of things we never touch. Boxes in our garage. Old memorabilia that we honestly don't need or care much about but feel too guilty to give away? Stupid little gadgets that help accomplish one task (a mango cutter?). Enough. Time to simplify. I plan on making my way through my house this year. Yard sales, trips to the DI, posting items on KSL or Craigslist are all part of my future. None of this stuff really buys me happiness. I can't wait to feel less cluttered.
4. Budget
Going along with #3 is to budget. It's so easy to just buy crap that we don't need. Honestly I'm amazed every time I go to Costco, Target, Carters, even the grocery store at how much stuff I buy that I don't really need. I've never really budgeted and I feel that if I set a strict budget that I will spend less on stuff I don't need. I'm terrible at groceries. I buy a lot of food that ends up going bad. Another part of budgeting will hopefully make me use more of what I have in my cupboards. Seth and I want to start saving. We should be saving way more than we do. Now that I am down to working 30 hours a week, money will be tighter. I think budgeting will actually feel really great. Now someone teach me?
5. Stop Being So Critical
This goes not only being critical of others, but of myself. Seth and I realized the other day that we have been in a cycle of just nagging each other. For the most ridiculous things. We are stressed and tired and we are completely taking it out on each other. Being critical doesn't help our marriage. It doesn't help my friendships. It doesn't help my family. I am going to work on this.
6. Blog
Blogging is one of my only creative outlets. Last year I blogged very little. I felt like it was a burden instead of an escape. I was going months without blogging, instead of weeks and that added to the burden. I felt like I lost my creative energy. So as lame as this sounds, I am going to blog twice a month. Even if the topics are ridiculous, I will just start writing. Some I hope are more thoughtful. Others will hopefully make people laugh. Some may just be an overview of what's happening in my life. The important thing for me is to write. It helps me stay centered.
7. Try some new hobbies
Some ideas at the top of this list include: knitting, baking bread, gardening, crafting with Millie. I'm going to rely heavily on Pinterest and talented friends to help me with this one.
8. Don't chop off my hair
Damn it all I will have long hair! I must stay strong and not cut any length off this year. Two trims. That's it. This is really stupid, I know. But I am determined to make it work.
9. Plan ahead
I'm a planner. I am good at planning but man Christmas was stressful this year. Coworker gifts, daycare teacher gifts, homemade goodies, presents for family, Millie, Seth all took a toll on me. I tried to do too much and as a result got grumpy and overwhelmed. So I'm starting early. September suckers! I want to plan ahead for all major life events. To help keep my sanity.
10. Enjoy the ride
Juggling work, family, home, Millie, etc. It's hard and tiring but it's damn good. This year my goal is to just go with it. My life is never going to be perfect. Let go of unrealistic expectations and enjoy the ride.
Oh Becky. 10 resolutions? Really!? Hey better than not trying anything. Here goes nothing. Bring it on 2014!


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Create a small, scalable, low input/high output market garden, based on the support of local volunteers for the aid of food shelters and or profitable enterprise.
Making use of gardening techniques such as vertical growing, no till, companion planting while prolonging the growing season with cold frames, poly tunnells and greenhouses.
The purpose of this crowd funding page is to generate interest for volunteers and investors who are intrigued by such a project and would like to contribute their ideas time and resources towards food security in local communities.
I have been a gardener for 4 years and have wanted to become involved in a charitable concept regarding plants and people since I was at college. I have noticed a rise in food prices, a reduction in volume and quality, the manifesting of food shelters in the south of England, abnormal global weather and political uncertainty becoming a determining factor in how we may discuss food in the future. I am interested to see whether I can prompt a response and build a shared dream of bringing back the emphasis of good food, the value we place on the health benefits that results in balanced diets and shared surplus in the community.
I have been lucky enough to receive a sizeable amount of land in the back of a residence in Oakdale, Poole Dorset. The garden is roughly 35 foot long 25 feet wide. It has been used as a dumping ground for organic matter, the soil is fertile and no pesticides or chemicals have been used for the production or maintenance of the garden. The garden has an office/garage on the land. There is a possibility the neighbouring garden could be used, expansion may also be possible once we have a proven scalable concept.
I would like to turn a small amount of land into a productive scalable market garden, and spread the idea to neighbouring communities with the help of neighbouring communities.
Ideally we need the people with the following skills.
Maintenance Volunteers - People interested and willing to learn and manage crops
Hardlandscapers or building specialists able to make polytunnells or greenhouses , concrete bases and irrigation
Garden Planner/s - Able to design and adjust sowing and harvesting dates
Delivery drivers - To deliver goods to local food shelters
Marketing - Market the concept and spread the interest
The garden is currently being landscaped by myself, and is in its bare bones form. A perfect time to start a new project. I have bought seeds and have been building planter, I would use a certain amount of funding for a bee hive and to build a poly tunnell .
Ideally help from skilled and qualified, volunteers and people willing to spread the word.


text: Dont know if anyone else has had this problem but here goes. When I send an SMS it doesnt always log into the conversation, so I dont know if it has gone or not. I ultimately end up resending and then keep getting told that friends have received multiple SMS's. I'm on the latest version of Chomp on a UK Orange Hero ver 2.1 Anyone else? Woody


text:
This Hampton Bay 7-1/2 in. outdoor post lantern features a solid white finish and eight sides of clear, beveled glass. This simple, elegant design compliments any exterior decor. These fixtures are built with cast aluminum to last and they are suitable for use in wet locations.
- Protective white finish
- Sturdy cast aluminum construction; built to last
- Suitable for wet locations
- Post not included
- For outdoor use only
- No bulbs included
- Click here for more information on Electronic Recycling Programs


text:
Status: Closed
Know more about this business than we do? Cool! Please submit any corrections or missing details you may have.Help us make it right
Be the first OregonLive.com user to add photos or videos of Consulate Of Italy
Dear I would t make an appointment for have the chance for flight up to the Italy Roma cause we are Asylum
Consulate of Italy is located at 121 SW Salmon St, Portland, OR. This location is in the Downtown Portland neighborhood. This business specializes in Government and has 1 review(s) with a star rating of 5.0.


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Assuring success through Learning & Development
What Do We Offer?
NTI offers an opportunity for those who have a desire to share knowledge and experience. The specialised Train-The-Trainer programs allows individuals to be competent trainers and will add value to the existing technical and academic qualifications.
NTI offers training courses in the following fields:
What Makes Us Different?
NTI offers multiple Certification Options apart from offering customised training sessions with industry expert trainers. The courses meet international standards and are designed for the local market.
Who is it Designed For?
Influenced by the local market requirements and combined with international standards, the Learning & Development training programmes aims at enhancing the teaching skills across all levels. Hence our training programmes are designed to meet the learners's specific needs, which in turn allow them to attain personal as well as organisational goals.


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1820s – British missionary Robert Moffat befriends Ndebele King Mzilikazi Khumalo in present-day South Africa.
1830s – The Ndebele flee South Africa and arrive in southwestern Zimbabwe; overthrow many Shona chieftaincies; establish the Matabeleland state.
1859 – Robert Moffat and the London Missionary Society (non-denominational Protestants) establish the first permanent white settlement in Zimbabwe, the Inyati Mission.
1860 – King Mzilikazi's son, Lobengula, becomes king of Matabeleland.
1879 – First Jesuit mission in Zimbabwe is established.
1888 – Lobengula signs Moffat Treaty, which prevents him from dealing with foreign powers other than England, and the Rudd Concession, which gives Cecil John Rhodes of the British South Africa Company (BSAC), "complete and exclusive charge of all metal and mineral rights," as well as commercial and legal powers in exchange for British protection, a monthly payment, and weapons.
1889 – Queen Victoria gives royal charter to Cecil John Rhodes and his British South Africa Company, effectively granting them complete imperial and colonial power.
September 1890 – Union Jack raised in Salisbury (present-day Harare, the setting of The Convert). Members of the British South African Company's paramilitary Pioneer Column occupy Mashonaland. In the "scramble for Africa," Zimbabwe is now in the British sphere.
1893 -First Anglo-Ndebele War breaks out when execution of treaties does not meet Lobengula's expectations. Ndebele lose and Lobengula flees north
1894 – Hut tax introduced. Most Africans in the region had previously held their wealth largely in cattle. The introduction of this tax forces many to work for colonists for currency so that they can pay taxes.
May 1895 – British South African Company officially adopts the name Southern Rhodesia for the region.
1896 – Rinderpest epidemic devastates cattle. African religious leaders and mediums include this as a sign of anger of the ancestors over the presence of the settlers. White forces kill off cattle to limit spread of disease, further enraging native people.
March 1896 – Ndebele uprising against the settlers begins. Beginning of siege of Bulawayo. Mlimo, the Ndebele spiritual leader, is one of the instigators of the rebellion.
June 1896 – Related Shona uprising begins. Shona traditional spiritual leaders, including spirit mediums of Kaguvi, Nehanda and Mwari, inspire rebels. Bernard Mzeki, African Episcopal catechist, killed by rebels.
September 1897 – Shona Chief Makoni executed after surrendering.
1922 – British South African Company rule ends. Rhodesia is formally annexed by British Government.
1930s – 1960s – Opposition to colonial rule increases; nationalist groups emerge.
1965 – The white-minority government, led by Ian Smith, unilaterally declares Rhodesia independent from British rule.
1972 – 1979 – Guerrilla war against white rule ensues.
1979 – British-brokered peace talks lead to peace agreement and new constitution.
1980 – Pro-independence leader Robert Mugabe wins independent elections; Zimbabwe is internationally recognized.
1998 – Economic crisis, strikes and riots.
2000 – Seizure of white-owned farms and accompanying violence.
2002 – Mugabe re-elected in elections criticized by foreign observers.
2003 – Opposition leader Morgan Tsvangirai arrested and charged with treason. (He is later acquitted.)
2006 – Inflation exceeds 1000%.
2008 – After charges of intimidation lead the opposition to pull out of the election, Mugabe and opposition leader Morgan Tsvangirai sign power-sharing agreement. Implementation is slow and problematic.
Courtesy of Carrie Hughes and the McCarter Theatre Center's Education Department


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DEBORAH AMOS, host:
It's MORNING EDITION from NPR News. I'm Deborah Amos.
STEVE INSKEEP, host:
And I'm Steve Inskeep. Good morning.
This week, we're giving some time to the opposition. From Africa to Russia and beyond, critics of governments have been making news or struggling to. Just yesterday, 100 students confronted the president of Iran, calling him a dictator. This week we will examine who the opposition is in several countries.
And we begin in Myanmar, the country that many still insist on calling Burma. Bridget Welsh of Johns Hopkins School of Advanced International Studies is a regular visitor there. She's been watching the career of the best-known opposition leader, Nobel Prize winner Aung San Suu Kyi.
Professor BRIDGET WELSH (Johns Hopkins School of Advanced International Studies): She's the daughter of the most important general in Burmese history. His name is Aung San. He was seen as the key independence fighter and he was assassinated.
INSKEEP: This is independence from?
Prof. WELSH: From the British.
INSKEEP: After World War II now?
Prof. WELSH: After World War II Burma was an important colony. It is an extension of India. And Aung San Suu Kyi is his daughter.
INSKEEP: The sons and daughters of major political figures do not automatically become major political figures themselves. What did she do to rise to prominence?
Prof. WELSH: Well, she began as a key leader, raising concerns about the government, overseas, where she studied in England. And she decided to leave England, where she was married to an Englishman, come back into Burma, and to lead the opposition. And in 1988, there was a key student movement and Aung San Suu Kyi came back. And because of her status as a daughter and an effective spokesman, she began to be the symbolic leader for the opposition movement. And they formed at that time the National League for Democracy.
INSKEEP: What is known about what she actually stands for them at this time? Is she a small-D democrat?
Prof. WELSH: Absolutely. She is seen as a person who wants to have dialogue and open change in the regime. Aung San Suu Kyi is never categorically come out and say that there can be no military government. What she has said is that we want a change of regime that is better for the people, and that it needs to be one that is democratically elected, and that the military may play a role in some sort of government, will not be its leader.
INSKEEP: Now, what is the connection between the monks who have demonstrated so prominently in recent weeks and the opposition group that we've just described?
Prof. WELSH: Well, definitely step back for a bit. There are three major opposition groups that one could to talk about in Burma. The first we've discussed, the National League for Democracy, and that particular party has really been massively repressed. The second group of opposition has traditionally been the students. They were the leaders of the 88 protests.
The third group is the one you just asked me about, which are the monks. And in this case we have a situation where the government has tried to co-opt the monasteries through the existing hierarchy in the monks - the monks at the very top by paying off towards monasteries or building pagodas. The monks, they are divided. There are some that people who are still collected and part of the government side. But the monks, they have formed a national organization and they are continuing in the most recent statements to come out and say that they will continue forward protesting the regime.
INSKEEP: So are the Buddhist monks and nuns as conflicted and as complicated a situation as you could imagine, the Catholic Church in any number of countries that have been under dictatorships over the year, some might end up being part of the power structure, some might end up being compromised, others stand up bravely against what they've seen as wrong.
Prof. WELSH: Absolutely. Although what has happened in the last month or so is a real tide has turned where more and more have become organized against the government.
INSKEEP: So given that this opposition is widespread - you mentioned three major groups - what is it that they're lacking that has made them unable so far, anyway, to dislodge the military government?
Prof. WELSH: It's not about what they're lacking; it's more about what they're facing. There is a very high level of fear in society. The military has used fear systemically to keep themselves in power. And the costs are not just about your own - to your own person, but to your family.
The second thing that's happened in Burma is the government has basically favored some groups over others. And in a situation of scarcity, they have actually allowed some people to have more access to economic gain through allegiance to the military. And so this kind of divide and rule strategy limits the way that people can organize.
INSKEEP: Have outside groups or countries such as China or the United States had any influence on the opposition movements inside what we know as Burma?
Prof. WELSH: I would say that the U.S. has had influence. When there's been particular incidents on human rights violations in the country, you've seen U.S. support to raise those things on the international agenda. So the U.S. has played a key role in trying to allow the opposition to get more space and to flower.
INSKEEP: There are some places where U.S. support of the opposition would actually discredit the local opposition.
Prof. WELSH: Yes, that's true. But I would argue that in the case of Burma, that's not one of them, that in fact despite all of the change towards the United States internationally, whenever I would go to Burma, there would still be a tremendous welcome because I was an American.
INSKEEP: Bridget Welsh works for the School of Advanced International Studies at Johns Hopkins University and she's a regular visitor. Thanks very much.
Prof. WELSH: You're welcome. Thank you.
Copyright © 2007 NPR. All rights reserved. Visit our website terms of use and permissions pages at www.npr.org for further information.
NPR transcripts are created on a rush deadline by Verb8tm, Inc., an NPR contractor, and produced using a proprietary transcription process developed with NPR. This text may not be in its final form and may be updated or revised in the future. Accuracy and availability may vary. The authoritative record of NPR's programming is the audio record.


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One Fordham Plaza, 4th Floor
Bronx, NY 10458
(718) 741-8402
Public comment will be received until: Monday, December 21st. (Preferable to submit by Friday, December 18th.)
Public comment will be received until: Monday, December 21st. (Preferable to submit by Friday, December 18th.)
By. Moonhawk River Stone
"For The Times They are a-Changin'"
-Bob Dylan (October 1964)…Indeed
One of favorite quotes comes from The Farthest Shore by Ursula K. LeGuin (this book is part of the Earthsea Trilogy). The young hero, Arren, is being taught by the Wizard of Earthsea, Ged, and it goes like this:
Do you see, Arren, how an act is not, as young
men think, like a rock that one picks up and throws,
and it hits or misses, and that's the end of it. When
that rock is lifted, the earth is lighter; the hand that
bears it heavier. When it is thrown, the circuits of the
stars respond, and where it strikes or falls the
universe is changed. On every act the balance of the
whole depends….We must learn to keep the balance.
This is only a small part of a much longer dialogue where the Wizard urges young Arren to do "only that which you must do and which you
cannot do in any other way."
I have carried these quotes around with me for over 35 years. They have become part of the foundation for me of how I do the work that is my life–professionally, in activism and in my personal life. Turning to Bob Dylan's song, now 50 years old and still ever so relevant today, we can see that those actions taken long ago, those actions that were what those early transgender pioneers felt was the only thing to do, have created, perhaps, what will be the most enduring and revolutionary changes about sex and gender we human beings will experience until we're growing our sex/genders in 3-D printers. No kidding. The emergence of the all things Trans* (new word being used to be ultra inclusive of the umbrella) has done more to revolutionize how we think of sex and gender than any other single event.
In 1975, a group of people, mostly male, mostly crossdressers and , in those days we called such folks transvestites, met in Provincetown, MA, post "the season" for the 1st Fantasia Fair, and yes, there were a few transsexual folks included, too. Back then, Provincetown shuttered itself up for the winter right after Labor Day as the summer people fled back to the cities for the winter. The summer season had very definitive boundaries and P'town was not a year round destination then. Provincetown in 1975 was perceived as a open and welcoming place for diverse folks, and this drove the Fantasia Fair folks' decision to locate the Fair in P'town. where a much safer environment for differently gendered people to be "out and about" existed and so they were out and about. I took my first "adult" vacation that October 1975 and Unbeknownst to me, I stumbled into the 1st Fantasia Fair while at a tres chic expensive French restaurant. The waiter was all nervous and upset, I remember scurrying around to make apologies for his unusual quests. I was not at all the flummoxed or disdainful guest he thought me to be. I just had this sense of curiosity and comfort. Not consciously then, but at some level I understood that whoever those folks were, I was, too and I was OK with them–and with me, all dressed up in my Sunday best male clothes, I don't suppose they thought twice about me. I remember being vaguely thrilled, and my extravagant dinner all the more delicious because of them. Re-balancing the whole of sex/gender diversity, changing that part of the Universe became real that first year of Fantasia Fair.
But the rock not thrown that evening was that I did not get up and go over to them and join them introducing myself, though I know in hindsight it would have been OK to do so. What was set in motion that night, was Fantasia Fair took off, and Hawk stayed still a while longer, each action the right one, the necessary one. Now, 40 years later, I go back Provincetown in October 2014 to celebrate Fantasia Fair's 40th anniversary and have the honor of giving one of the week's keynote addresses, having come full circle, the balance somehow having been struck, the Equilibrium reestablished. My speech is going to focus on just this column's theme how differently gendered people have changed the conversation about sex and gender not only in Trans* land, but among all human beings. How that moment where the rock thrown, lands in the ocean and the ripples have reached the farthest shore to only redouble and ripple back yet again and again.
In far off time when history takes measure of this era in America it will note that something changed. The divide between male and female crumbled a bit, and morphed into a continuum that became a garden of delight in which us humans could play. We will note it spurred the larger world to its own imaginings of what lay beyond the strict binary of male and female, though in many places that struggle would be decades or centuries longer than here in America. Dr. Milton Diamond, Ph.D., one of the leading researcher allies to the transgender and intersex communities is oft quoted as saying, "Nature loves diversity. Society does not." Suppose the rock that was thrown that October evening in 1975 obliterated the second sentence and we are just left with "Nature loves diversity." I want to be at Fantasia Fair 75 (in 2049) just past 101 to see what ripples then and how the journey unfolded. What new balance will be struck? And will all of us Trans* and cisgender be free to be ourselves and love one another? For one thing we know then as we do now, "For The Times They Are a-Changin'. Until next time…T
Today we're pleased to share a special complimentary online edition of our CommUNITY magazine feature, Trans View, by columnist Moonhawk River Stone. In keeping with CommUNITY's May theme of Vintage Pride, this month's column focuses on the topic of aging in the transgender community. We hope enjoy the read.
Trans View #122
by. Moonhawk River Stone© 2014
First off my heartfelt thanks to Loree Cook-Daniels for writing the Trans View guest column for the May 2014 CommUNITY. As an ally to transgender people Loree is without peer and has over the decades contributed mightily both to our knowledge about trans elders and services for them. I am so grateful for her, and for all of her steadfast hard work on behalf of LGBT people, especially transgender people. In my column, an online only companion column, I'm going to touch base about some issues beyond policy making for older transgender people. Times are changing rapidly for LGBT people with the increasing recognition of same-sex marriage, the Social Security Administration's (SSA) policy clarification for transgender marriages among them. I mean, who would have thought 10 years ago, or even 5 years ago, that SSA would be talking proactively about transgender marriages and clarifying policy so spouses receive the full benefits they are entitled to receive. I mean, really? For that we have the progress in same-sex marriage to thank as well as our own transgender advocates. Older transgender people come from basically four different cohorts: 1) people who transitioned medically and legally decades ago and have faded into the woodwork ; 2) people who did various kinds of transition related actions and never changed their documents, or had surgery (Billy Tipton, Virginia Prince come to mind here) and who lived very stealth (secret) lives; 3) people who transitioned medically and legally decades ago and who remained in the public eye; and 4) the ever increasing numbers of older transgender folks transitioning in their 60's, 70's and yes, 80's who are seeking access to care in their elder years. Finding the transgender individuals in cohorts #1 and #2 is proving to be quite a difficult challenge. They are vulnerable populations, not connected to the larger trans communities, the result is that they may suddenly be outed without their permission, they may be denied health care, denied access to partner benefits from government agencies, denied inheritance, and suddenly find themselves homeless and without financial resources or face a premature death because they have avoided being seen in a health care setting because of their transgender status. They are a high risk population. Transgender individuals in cohort #4 are the emerging population gaining much notice in the media. People who have yearned their whole lives to transition, people who have had failed or truncated transitions are increasingly transitioning. People who have waited until their partner of many decades has passed are transitioning. One of the challenges here has to do with surprising adult children and grandchildren with news that has been kept secret for a long time. Most elderly transgender people do not have access to genital reconstruction surgeries, either the out of pocket cost is a choice between maintaining a retirement fund–and a standard of living or spending it on surgery. Many elderly transgender people have their social security money as their sole income. Medicare does not yet cover the cost of genital reconstruction surgeries (of any kind), but will cover the costs of mental health counseling and hormone therapy. The Veterans Administration has similar coverage. There is also prejudice within medicine about giving hormone treatment or surgical treatment to elderly people. There is really not much known about the effects of hormones on elderly bodies, and treatment approaches are largely anecdotal. Elderly transgender individuals often have other health challenges which make surgery a more risky undertaking. Transition for older people brings a mixed blessing–getting to finally be who you are, but, maybe, not being able to surgically transition. Transgender elders also need to have access to proper legal advice in order to fully protect their assets and property and their identity. Many states have rigid rules about gender marker documentation and will not be flexible concerning that when a transgender person dies. Several high profile legal cases have involved the issue of late life transgender marriage, and inheritance with the transgender person being the one who suffers. Economic access to quality trans-legal services is also an issue in many localities.
For the larger community and society, we are quickly closing a window of opportunity to collect the wisdom, the stories and the videos of transgender elders. Transgender elders can give us a very unique glimpse of transgender history from an individual perspective that is so valuable. It is important that we capture and remember the time before it all changed and transgender became (not quite yet, but almost) a household word. It's something like the stories my grandfather told me about walking to school barefoot, and riding his horse when he got older to do errands for the family. Trans elders do really come from the last century one that saw tremendous change for transgender people. They have so much to offer us in stories of courage, survival, living one's truth, and just how risky and closeted things were back then. They are pioneers to whom we owe a great debt of gratitude. Until next time…T


text:
Recently I brought the B in for an inspection; all went well, except for my headlights are not adjusted to proper height or angle (passed anyway). I know that the adjustment screws are behind the chrome ring around the headlight, but what are the proper specs for height and for their left to right angling direction. If you could include a measurement also as to how far I need to be away from the wall when I do the adjustment, that would be much appreciated.
As always thanks for any help.
Adam West
1979 MGB Vermillion Red
MGB & GT Forum
Headlight adjustment and specs
Posted by awest
Jun 1, 2010 06:55 PM
Joined 11 years ago
931 Posts
Jun 1, 2010 07:00 PM
Joined 9 years ago
386 Posts
This is from a MGExperience member
sweep Chris W
Gosford, NSW, Australia
1966 MG MGB "Basil"
2013 Volkswagen Tiguan
2015 Audi A3
Jun 1, 2010 07:02 PM
Top Contributor
Joined 11 years ago
6,621 Posts
I asked this question a while ago and didn't get a definitive answer from the group - a few links to some generic auto electrical sites which suited me fine.
See here:
See here:
Jun 1, 2010 07:11 PM
Joined 11 years ago
931 Posts
Thank you Chris and Kent.
Fairfield, CA, USA
1973 MG MGB MkIII
Jun 1, 2010 07:18 PM
Top Contributor
Joined 12 years ago
23,180 Posts
Daniel Stern has a similar chart on his site as well.
1973 Pale Primrose Roadster. A nice 10-footer!
SUs, Datsun 5-speed
1973 Pale Primrose Roadster. A nice 10-footer!
SUs, Datsun 5-speed
Jun 2, 2010 01:11 AM
Top Contributor
Joined 14 years ago
13,919 Posts
As mentioned by Mac - DanielSternLighting.com has the necessary aiming procedure. Note that it differs depending on whether you have DOT style lenses or E-Code beams. If you don't have E-code, do yourself a favour and get some. I like Cibie, but there are other options as well.
Terry Ingoldsby
Terry Ingoldsby
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text:
I ate a bunch of Warheads and now my mouth is destroyed
By FunExplosions 1 Comments
No seriously. My tongue has lost all taste and is incredibly sore. The roof of my mouth feels like I took a nap with a scolding hot piece of cheese resting on it. The insides of my cheeks are horribly tender and for awhile chunks of blue-colored flesh were peeling off them.
How did we eat these things as a kid? Did we just not care? I can't imagine it was this bad, with how many I used to eat all the time. Now, a day and a half later, it's incredibly painful to even eat some soft bread. It's like someone beat up the inside of my mouth. I need some aftercare, Warheads.
For reals, bro, you best be playin' safe if you fi'in' ta suck some 'Heads.


text:
Eaton Hotel (Xianggang Yi Dong Jiudian) is located in the heart of Kowloon's famed shopping and commercial district on Nathan Road, just a few minutes' walk from Jordan MTR Station. Guests here are only steps away from many of Hong Kong's top attractions including Temple Street and the Hong Kong Jade Market.
This Hong Kong hotel's luxurious, smart and comfortable rooms all feature high quality bedding, LCD TV screens and broadband Internet access. The larger Club rooms offer more space with a lounge area and coffee table as well as access to the E-Club where continental breakfast, all-day tea, coffee and evening drinks are provided with guilty snacks.
In the Yat Tung Heen restaurant, guests can enjoy authentic Cantonese cuisine and dim sum. Other trendy dining venues offer Western and Japanese dishes while cocktails, select beers and light refreshments are on offer in the T Bar.
A fully equipped business center and meeting rooms are on-site to hold conferences and events.
Travelers looking to keep in shape will have no problem at this Hong Kong hotel with its modern 24-hour gym, Wii-Fit consoles and an attractive rooftop swimming pool. Basketball, tennis courts and a climbing wall can also be found in nearby King's Park.
Check in: from 15:00 Check out: before 12:00
The hotel does not accept guests below 18 years of age staying alone.
Buffet breakfast HKD 185
No pets allowed.
Payment can be made by credit card at the hotel. You can use the following credit cards:
All rates displayed are based on current exchange rate between CNY and the selected currency.
Children and Extra Beds
The hotel does not accept guests below 18 years of age staying alone.
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