Activism, affect, identification: trans documentary in France and Spain and its reception

This article explores the documentation of trans activism in France and Spain since the 2000s. The first part addresses questions surrounding the place of affect and narrative in documentary film, particularly in relation to trans issues. The second part o f the article analyses an audience case study from a screening at the International Gay and Lesbian Film Festival in Barcelona of Valérie Mitteaux's Girl or Boy, My Sex is not my Gender (2011), considering how different viewers respond to the representatio n of trans identities. The article builds on qualitative research whilst extending the exploration of sexuality and gender in previous audience studies to a consideration of documentary film, seeking to provide a more nuanced understanding of what audience claims for identification in politicised contexts mean

Mutations in ELAC2 associated with hypertrophic cardiomyopathy impair mitochondrial tRNA 3'-end processing

Mutations in either the mitochondrial or nuclear genomes are associated with a diverse group of human disorders characterized by impaired mitochondrial respiration. Within this group, an increasing number of mutations have been identified in nuclear genes involved in mitochondrial RNA metabolism, including ELAC2. The ELAC2 gene codes for the mitochondrial RNase Z, responsible for endonucleolytic cleavage of the 3' ends of mitochondrial pre-tRNAs. Here, we report the identification of sixteen novel ELAC2 variants in individuals presenting with mitochondrial respiratory chain deficiency, hypertrophic cardiomyopathy and lactic acidosis. We provide evidence for the pathogenicity of the novel missense variants by studying the RNase Z activity in an in vitro system. We also modelled the residues affected by missense mutation in solved RNase Z structures, providing insight into enzyme structure and function. Finally, we show that primary fibroblasts from the affected individuals have elevated levels of unprocessed mitochondrial RNA precursors. Our study thus broadly confirms the correlation of ELAC2 variants with severe infantile-onset forms of hypertrophic cardiomyopathy and mitochondrial respiratory chain dysfunction. One rare missense variant associated with the occurrence of prostate cancer (p.Arg781His) impairs the mitochondrial RNase Z activity of ELAC2, suggesting a functional link between tumorigenesis and mitochondrial RNA metabolism

Monografies del Montseny - 35

Des de la primera edició de 1985 comandada per Ramon Bofill Portabella i Antoni Pladevall Font han escrit a les Monografies del Montseny un total de 324 autors diferents, d’arreu del Montseny, molts de Barcelona, però també de la resta del país, inclòs forans. Podeu consultar-los tots a l’índex actualitzat que publiquem cada any. Entre aquest llarg llistat podem trobar escriptors, poetes, científics, polítics, cineastes, esportistes, masovers, propietaris forestals, arquitectes, professors, fotògrafs, llicenciats, advocats, diplomàtics, historiadors, catedràtics, pagesos, filòsofs, guardes forestals, geògrafs, químics, doctors, pintors... Molts dels autors han comandat importants institucions i associacions del nostre país, força són membres de la Reial Acadèmia de Bones Lletres, la Reial Acadèmia Catalana de Belles Arts, l’Institut d’Estudis Catalans... i també hi ha autors que són posseïdors de la Creu de Sant Jordi, la Medalla d’Or de la Generalitat, i els més variats reconeixements. Tot i això, des d’un bon principi els fundadors de Monografies del Montseny van voler que les col·laboracions no fossin tancades a ningú, mentre els treballs tinguessin un mínim de solvència i aportessin valor a la col·lecció, independentment del curriculum vitae particular. Tots els autors han contribuït de manera desinteressada amb un o més articles a la col·lecció Monografies del Montseny; de fet és normal trobar a l’índex històric autors amb una desena de col·laboracions, però aquí cal destacar que Antoni Pladevall Font ha participat a totes les Monografies amb més de cinquanta escrits. Com a trets curiosos podem trobar tres articles d’autors de finals de segle XIX i principis del XX publicats en versió facsímil. Monografies del Montseny acumula avui 591 articles, cosa que les converteixen en “la Bíblia del Montseny”, el lloc necessari on acudir per completar qualsevol recerca sobre un tema montsenyenc. Cada volum s’ha publicat amb una mitja variable d’entre 250-300 pàgines, però en algun cas extraordinari hem arribat fins les 400. Cal tenir en compte que des de l’any 1995 les Monografies incorporen un apèndix amb els versos guardonats als premis de poesia Montseny, i les obres guanyadores del premis de fotografia Montseny mentre va durar el certamen (avui extingit). Això suposa que el conjunt de pàgines publicades durant aquests 35 anys arriba a les 9.295.Postprint (published version

RNAi of cyclin D2 and cyclin D3 differently impact viral replication depending on MDM type.

<p><b>(A)</b> Effective and specific knockdown of <i>CCND2</i> and <i>CCND3</i> expression by siRNA. Relative mRNA expression of <i>CCND2</i> (left panel) and <i>CCND3</i> (right panel) in M-CSF (white bars) and GM-CSF (black bars) macrophages. mRNA of the corresponding cyclin was measured by quantitative PCR and normalized to GAPDH expression. Data represents mean ± SD of 3 different donors and is normalized to Mock-transfected M-CSF macrophages. <b>(B)</b> Protein expression and SAMHD1 activation in cyclin D2 and cyclin D3 knockdown macrophages. Western blot showing cyclin protein expression, SAMHD1 expression and activation in siRNA-treated M-CSF and GM-CSF macrophages. SAMHD1 inactivation is reduced in siCCND3 (D3) M-CSF macrophages and increased in siCCND2 (D2) GM-CSF macrophages compared to the corresponding non-targeting siRNA (NT). Hsp90 was used as loading control. A representative donor is shown. <b>(C)</b> Evaluation of cell proliferation in cyclin D2 and cyclin D3 knockdown macrophages. Ki67 staining of siRNA-treated M-CSF (upper panels) and GM-CSF (lower panels) derived macrophages. Histograms from a representative donor are shown. <b>(D)</b> HIV-1 replication in siRNA-treated M-CSF or GM-CSF macrophages. Transfected MDM were infected with a VSV-pseudotyped, GFP-expressing HIV-1 and infection measured 72h later by flow cytometry. Data represent percentage replication relative to mock-transfected cells in M-CSF (white bars) or GM-CSF (black bars) macrophages. Mean ± SD of 3 different donors performed in triplicate is shown. * p<0.05; ** p<0.005; *** p<0.0005.</p

Cyclin D2 expression controls subsequent activation of CDK1 and CDK2.

<p><b>(A)</b> CDK1 and CDK2 gene expression in <i>CCND2</i> and <i>CDKN1A</i> knockdown GM-CSF macrophages. <i>CDK1</i> and <i>CDK2</i> gene expression was measured by quantitative PCR and normalized to GAPDH expression. Data represents mean ± SD of 3 different donors and is normalized to Mock-transfected macrophages. <b>(B)</b> Western blot showing CDK1 and CDK2 expression and phosphorylation of CDK2 in siRNA-treated GM-CSF macrophages. CDK1 expression is upregulated after cyclin D2 knockdown as well as CDK2 activity measured as phosphorylation at Thr130, compared to Mock-transfected or macrophages treated with a non-targeting siRNA (siNT). <b>(C)</b> Proposed regulatory model of Cyclin D2 mediated control of HIV-1 restriction in non-proliferating cells. The Cyclin D2/CDK4/p21 complex is responsible for the lack of active CDK2 and CDK1 expression in GM-CSF macrophages. This situation is reversed in the absence of cyclin D2, leading to the activation of CDK2 and CDK1 with the subsequent phosphorylation of its substrates, including SAMHD1.</p

M-CSF and GM-CSF macrophages present differential expression of cell cycle-related proteins and SAMHD1 activation.

<p><b>(A)</b> Gene expression of cell cycle-related genes and SAMHD1 restriction pathway. mRNA levels of <i>SAMHD1</i>, <i>CDK1</i>, <i>CDK2</i>, <i>CDK4</i> and <i>CDK6</i>, the corresponding cyclins (<i>A2</i>, <i>B1</i>, <i>B2</i>, <i>D1</i>, <i>D2</i>, <i>D3</i>, <i>E1</i> and <i>E2</i>) and CDK2 inhibitor p21 (<i>CDKN1A</i>) was quantified in M-CSF (white bars) and GM-CSF (black bars) differentiated macrophages. Data is normalized to M-CSF relative expression. Mean ± SD of 3 independent donors is shown. ** p<0.005; *** p<0.0005. <b>(B)</b> Western blot showing protein expression of different cell cycle proteins, SAMHD1 expression and activation and Hsp90 as loading control. Two representative donors are shown. M; M-CSF MDM, GM; GM-CSF MDM. <b>(C)</b> Induction of gene expression following LPS (100 ng/ml) treatment of M-CSF and GM-CSF macrophages. Expression of <i>IFNB1</i>, <i>CCL-2</i> and <i>IL-10</i> were evaluated. Data is normalized to untreated M-CSF condition. Mean ± SD of 3 independent donors is shown.</p

Susceptibility to HIV-1 infection in primary macrophages depends on differentiation stimuli.

<p><b>(A)</b> Evaluation of cell proliferation by Ki67 staining. Histograms of a representative donor showing Ki67 staining in M-CSF and GM-CSF derived macrophages. Percentage of positive cells is shown in each case. <b>(B)</b> Dot plots representing cell cycle profile showing DNA (7-AAD) and RNA (Pyronin Y) content of M-CSF and GM-CSF differentiated macrophages quantified by flow cytometry. Data from a representative donor is shown. <b>(C)</b> Dot plots of infected M-CSF macrophages (upper panels) or GM-CSF macrophages (lower panels) untreated (left) or after treatment (right) with VLP_Vpx. Data from a representative donor is shown. <b>(D)</b> Percentage replication in M-CSF MDM and GM-CSF MDM untreated (white bars) or transduced with VLP_Vpx (black bars). Mean ± SD of 4 different donors performed in triplicate is shown. <b>(E)</b> and <b>(F)</b> Drug sensitivity in M-CSF and GM-CSF differentiated macrophages. Antiviral activity of zidovudine (AZT, 1 μM), nevirapine (NVP, 5 μM) and palbociclib (PD, 4 μM) was evaluated in M-CSF (E) and GM-CSF (F) MDM, untreated (white bars) or transduced with VLP_Vpx (black bars). Antiviral potency was decreased with AZT and completely lost with PD after degradation of SAMHD1 with VLP_Vpx. Mean ± SD of at least 3 different donors performed in triplicate is shown. ND; no-drug, ns; not significant, * p<0.05; ** p<0.005.</p

Cyclin D2 expression restricts HIV-1 replication by controlling SAMHD1 activation in GM-CSF macrophages.

<p><b>(A)</b> Effective and specific knockdown of <i>CCND2</i>, <i>CCND3</i>, <i>CDKN1A</i> and <i>CDK4</i> expression by siRNA in GM-CSF macrophages. mRNA of the corresponding gene was measured by quantitative PCR and normalized to GAPDH expression. Data represents mean ± SD of at least 3 different donors and is normalized to Mock-transfected macrophages. <b>(B)</b> Protein expression and SAMHD1 activation in siRNA-treated GM-CSF macrophages. Western blot showing cyclin D2, cyclin D3, CDK4 and p21 protein expression and SAMHD1 expression and activation in siRNA-treated GM-CSF macrophages. SAMHD1 inactivation by phosphorylation is increased in cyclin D2 and p21 knockdown macrophages, compared to Mock-transfected or macrophages treated with a non-targeting siRNA (siNT). <b>(C)</b> Evaluation of cell proliferation and cell cycle analysis in siRNA-treated GM-CSF macrophages. Histograms showing Ki67 staining (upper panels) and dot plots representing cell cycle profile by DNA (7-AAD) and RNA (Pyronin Y) staining (lower panels) in siRNA-treated macrophages. Percentage of positive cells in a representative donor is shown in each case. <b>(D)</b> HIV-1 replication in siRNA-treated GM-CSF macrophages. Transfected MDM were infected with a VSV-pseudotyped, GFP-expressing HIV-1 and infection measured 72h later by flow cytometry. Data represent percentage replication relative to mock-transfected macrophages. Mean ± SD of at least 3 different donors performed in duplicate is shown. <b>(E)</b> Proviral DNA formation after 16h infection with HIV-1 BaL of GM-CSF macrophages transfected with the indicated siRNA or treated with AZT (3 μM) or raltegravir (RAL; 2 μM). Proviral DNA was normalized to mock-treated macrophages. Mean ± SD of at least 3 different donors is shown. * p<0.05; ** p<0.005; *** p<0.0005.</p

Mutations in ELAC2 associated with hypertrophic cardiomyopathy impair mitochondrial tRNA 3'-end processing

Mutations in either the mitochondrial or nuclear genomes are associated with a diverse group of human disorders characterized by impaired mitochondrial respiration. Within this group, an increasing number of mutations have been identified in nuclear genes involved in mitochondrial RNA metabolism, including ELAC2. The ELAC2 gene codes for the mitochondrial RNase Z, responsible for endonucleolytic cleavage of the 3' ends of mitochondrial pre-tRNAs. Here, we report the identification of sixteen novel ELAC2 variants in individuals presenting with mitochondrial respiratory chain deficiency, hypertrophic cardiomyopathy and lactic acidosis. We provide evidence for the pathogenicity of the novel missense variants by studying the RNase Z activity in an in vitro system. We also modelled the residues affected by missense mutation in solved RNase Z structures, providing insight into enzyme structure and function. Finally, we show that primary fibroblasts from the affected individuals have elevated levels of unprocessed mitochondrial RNA precursors. Our study thus broadly confirms the correlation of ELAC2 variants with severe infantile-onset forms of hypertrophic cardiomyopathy and mitochondrial respiratory chain dysfunction. One rare missense variant associated with the occurrence of prostate cancer (p.Arg781His) impairs the mitochondrial RNase Z activity of ELAC2, suggesting a functional link between tumorigenesis and mitochondrial RNA metabolism This article is protected by copyright. All rights reserved

Monografies del Montseny - 35

Des de la primera edició de 1985 comandada per Ramon Bofill Portabella i Antoni Pladevall Font han escrit a les Monografies del Montseny un total de 324 autors diferents, d’arreu del Montseny, molts de Barcelona, però també de la resta del país, inclòs forans. Podeu consultar-los tots a l’índex actualitzat que publiquem cada any. Entre aquest llarg llistat podem trobar escriptors, poetes, científics, polítics, cineastes, esportistes, masovers, propietaris forestals, arquitectes, professors, fotògrafs, llicenciats, advocats, diplomàtics, historiadors, catedràtics, pagesos, filòsofs, guardes forestals, geògrafs, químics, doctors, pintors... Molts dels autors han comandat importants institucions i associacions del nostre país, força són membres de la Reial Acadèmia de Bones Lletres, la Reial Acadèmia Catalana de Belles Arts, l’Institut d’Estudis Catalans... i també hi ha autors que són posseïdors de la Creu de Sant Jordi, la Medalla d’Or de la Generalitat, i els més variats reconeixements. Tot i això, des d’un bon principi els fundadors de Monografies del Montseny van voler que les col·laboracions no fossin tancades a ningú, mentre els treballs tinguessin un mínim de solvència i aportessin valor a la col·lecció, independentment del curriculum vitae particular. Tots els autors han contribuït de manera desinteressada amb un o més articles a la col·lecció Monografies del Montseny; de fet és normal trobar a l’índex històric autors amb una desena de col·laboracions, però aquí cal destacar que Antoni Pladevall Font ha participat a totes les Monografies amb més de cinquanta escrits. Com a trets curiosos podem trobar tres articles d’autors de finals de segle XIX i principis del XX publicats en versió facsímil. Monografies del Montseny acumula avui 591 articles, cosa que les converteixen en “la Bíblia del Montseny”, el lloc necessari on acudir per completar qualsevol recerca sobre un tema montsenyenc. Cada volum s’ha publicat amb una mitja variable d’entre 250-300 pàgines, però en algun cas extraordinari hem arribat fins les 400. Cal tenir en compte que des de l’any 1995 les Monografies incorporen un apèndix amb els versos guardonats als premis de poesia Montseny, i les obres guanyadores del premis de fotografia Montseny mentre va durar el certamen (avui extingit). Això suposa que el conjunt de pàgines publicades durant aquests 35 anys arriba a les 9.295