Stable silencing of SNAP-25 in PC12 cells by RNA interference

BACKGROUND: SNAP-25 is a synaptic protein known to be involved in exocytosis of synaptic vesicles in neurons and of large dense-core vesicles in neuroendocrine cells. Its role in exocytosis has been studied in SNAP-25 knockout mice, in lysed synaptosomes lacking functional SNAP-25 and in cells after treatment with botulinum toxins A or E that specifically cleave SNAP-25. These studies have shown that SNAP-25 appears to be required for most but not all evoked secretion. In order to further study the role of SNAP-25 in catecholamine secretion from PC12 cells we have used the recently developed technique of RNA interference to generate PC12 cell lines with virtually undetectable levels of SNAP-25. RNA interference is the sequence-specific silencing or knockdown of gene expression triggered by the introduction of double-stranded RNA into a cell. RNA interference can be elicited in mammalian cells in a number of ways, one of which is by the expression of small hairpin RNAs from a transfected plasmid. Selection of stably transfected cell lines expressing a small hairpin RNA allows one-time characterization of the degree and specificity of gene silencing and affords a continuing source of well-characterized knockdown cells for experimentation. RESULTS: A PC12 cell line stably transfected with a plasmid expressing an shRNA targeting SNAP-25 has been established. This SNAP-25 knockdown cell line has barely detectable levels of SNAP-25, but normal levels of other synaptic proteins. Catecholamine secretion elicited by depolarization of the SNAP-25 knockdown cells was reduced to 37% of control. CONCLUSION: Knockdown of SNAP-25 in PC12 cells reduces but does not eliminate evoked secretion of catecholamines. Transient expression of human SNAP-25 in the knockdown cells rescues the deficit in catecholamine secretion

An integrative approach to evaluating the implementation of social and emotional learning and gender-based violence prevention education

Evaluation studies often use stand-alone and summative assessment strategies to examine the impacts of Social and Emotional Learning (SEL) and Gender-based Violence (GBV) prevention education programs. However, implementation research is yet to offer an integrative framework that can be used to investigate the implementation drivers that lead to the uptake of programs that pursue SEL and GBV prevention agendas. We address this gap in research by presenting a framework developed to investigate factors affecting the implementation of the Resilience, Rights and Respectful Relationships program, an SEL and GBV prevention education program developed for primary and secondary schools in the state of Victoria, Australia. Drawing upon and advancing a conceptual framework for implementation fidelity proposed by Carroll and colleagues we discuss the iterative process designed to investigate the individual, school and system level factors within the wider political and ideological setting(s) of the program that impact on its implementation. Within this iterative process, we highlight the need to focus on ‘the ecology of relations’ that exists between various implementation elements, and their possible mediating impact on program delivery, uptake and outcomes.peer-reviewe

COVID-19 Risk Factors for Cancer Patients: A First Report with Comparator Data from COVID-19 Negative Cancer Patients

none32siSimple SummaryThe COVID-19 pandemic has had a detrimental impact on cancer patients globally. Whilst there are several studies looking at the potential risk factors for COVID-19 disease and related death, most of these include non-cancerous patients as the COVID-19 negative comparator group, meaning it is difficult to draw hard conclusions as to the implications for cancer patients. In our study, we utilized data from over 2000 cancer patients from a large tertiary Cancer Centre in London. In summary, our study found that patients who are male, of Black or Asian ethnicity, or with a hematological malignancy are at an increased risk of COVID-19. The use of cancer patients as the COVID-19 negative comparator group is a major advantage to the study as it means we can better understand the true impact of COVID-19 on cancer patients and identify which factors pose the biggest risk to their likelihood of infection with SARS-CoV2.Very few studies investigating COVID-19 in cancer patients have included cancer patients as controls. We aimed to identify factors associated with the risk of testing positive for SARS CoV2 infection in a cohort of cancer patients. We analyzed data from all cancer patients swabbed for COVID-19 between 1(st) March and 31(st) July 2020 at Guy's Cancer Centre. We conducted logistic regression analyses to identify which factors were associated with a positive COVID-19 test. Results: Of the 2152 patients tested for COVID-19, 190 (9%) tested positive. Male sex, black ethnicity, and hematological cancer type were positively associated with risk of COVID-19 (OR = 1.85, 95%CI:1.37-2.51; OR = 1.93, 95%CI:1.31-2.84; OR = 2.29, 95%CI:1.45-3.62, respectively) as compared to females, white ethnicity, or solid cancer type, respectively. Male, Asian ethnicity, and hematological cancer type were associated with an increased risk of severe COVID-19 (OR = 3.12, 95%CI:1.58-6.14; OR = 2.97, 95%CI:1.00-8.93; OR = 2.43, 95%CI:1.00-5.90, respectively). This study is one of the first to compare the risk of COVID-19 incidence and severity in cancer patients when including cancer patients as controls. Results from this study have echoed those of previous reports, that patients who are male, of black or Asian ethnicity, or with a hematological malignancy are at an increased risk of COVID-19.openRussell, Beth; Moss, Charlotte L; Palmer, Kieran; Sylva, Rushan; D'Souza, Andrea; Wylie, Harriet; Haire, Anna; Cahill, Fidelma; Steel, Renee; Hoyes, Angela; Wilson, Isabelle; Macneil, Alyson; Shifa, Belul; Monroy-Iglesias, Maria J; Papa, Sophie; Irshad, Sheeba; Ross, Paul; Spicer, James; Kordasti, Shahram; Crawley, Danielle; Zaki, Kamarul; Sita-Lumsden, Ailsa; Josephs, Debra; Enting, Deborah; Swampillai, Angela; Sawyer, Elinor; Fields, Paul; Wrench, David; Rigg, Anne; Sullivan, Richard; Van Hemelrijck, Mieke; Dolly, SaoirseRussell, Beth; Moss, Charlotte L; Palmer, Kieran; Sylva, Rushan; D'Souza, Andrea; Wylie, Harriet; Haire, Anna; Cahill, Fidelma; Steel, Renee; Hoyes, Angela; Wilson, Isabelle; Macneil, Alyson; Shifa, Belul; Monroy-Iglesias, Maria J; Papa, Sophie; Irshad, Sheeba; Ross, Paul; Spicer, James; Kordasti, Shahram; Crawley, Danielle; Zaki, Kamarul; Sita-Lumsden, Ailsa; Josephs, Debra; Enting, Deborah; Swampillai, Angela; Sawyer, Elinor; Fields, Paul; Wrench, David; Rigg, Anne; Sullivan, Richard; Van Hemelrijck, Mieke; Dolly, Saoirs

Igbo-English intrasentential codeswitching and the Matrix Language Frame model

This paper uses data from Igbo-English intrasentential codeswitching involving mixed nominal expressions to test the Matrix Language Frame (MLF) model. The MLF model is one of the most highly influential frameworks used in the study of grammatical aspects of codeswitching. Three principles associated with it, the Matrix Language Principle, the Asymmetry Principle and the Uniform Structure Principle, were tested on data collected from informal conversations by educated adult Igbo-English bilinguals resident in Port Harcourt. The results of the analyses suggest general support for the three principles and for identifying Igbo-English as a “classic” case of codeswitching

Diversity in African languages: Selected papers from the 46th Annual Conference on African Linguistics

Diversity in African Languages contains a selection of revised papers from the 46th Annual Conference on African Linguistics, held at the University of Oregon. Most chapters focus on single languages, addressing diverse aspects of their phonology, morphology, semantics, syntax, information structure, or historical development. These chapters represent nine different genera: Mande, Gur, Kwa, Edoid, Bantu, Nilotic, Gumuzic, Cushitic, and Omotic. Other chapters investigate a mix of languages and families, moving from typological issues to sociolinguistic and inter-ethnic factors that affect language and accent switching. Some chapters are primarily descriptive, while others push forward the theoretical understanding of tone, semantic problems, discourse related structures, and other linguistic systems. The papers on Bantu languages reflect something of the internal richness and continued fascination of the family for linguists, as well as maturation of research on the family. The distribution of other papers highlights the need for intensified research into all the language families of Africa, including basic documentation, in order to comprehend linguistic diversities and convergences across the continent. In this regard, the chapter on Daats’íin (Gumuzic) stands out as the first-ever published article on this hitherto unknown and endangered language found in the Ethiopian-Sudanese border lands

Diversity in African languages: Selected papers from the 46th Annual Conference on African Linguistics

Diversity in African Languages contains a selection of revised papers from the 46th Annual Conference on African Linguistics, held at the University of Oregon. Most chapters focus on single languages, addressing diverse aspects of their phonology, morphology, semantics, syntax, information structure, or historical development. These chapters represent nine different genera: Mande, Gur, Kwa, Edoid, Bantu, Nilotic, Gumuzic, Cushitic, and Omotic. Other chapters investigate a mix of languages and families, moving from typological issues to sociolinguistic and inter-ethnic factors that affect language and accent switching. Some chapters are primarily descriptive, while others push forward the theoretical understanding of tone, semantic problems, discourse related structures, and other linguistic systems. The papers on Bantu languages reflect something of the internal richness and continued fascination of the family for linguists, as well as maturation of research on the family. The distribution of other papers highlights the need for intensified research into all the language families of Africa, including basic documentation, in order to comprehend linguistic diversities and convergences across the continent. In this regard, the chapter on Daats’íin (Gumuzic) stands out as the first-ever published article on this hitherto unknown and endangered language found in the Ethiopian-Sudanese border lands

Diversity in African languages: Selected papers from the 46th Annual Conference on African Linguistics

Diversity in African Languages contains a selection of revised papers from the 46th Annual Conference on African Linguistics, held at the University of Oregon. Most chapters focus on single languages, addressing diverse aspects of their phonology, morphology, semantics, syntax, information structure, or historical development. These chapters represent nine different genera: Mande, Gur, Kwa, Edoid, Bantu, Nilotic, Gumuzic, Cushitic, and Omotic. Other chapters investigate a mix of languages and families, moving from typological issues to sociolinguistic and inter-ethnic factors that affect language and accent switching. Some chapters are primarily descriptive, while others push forward the theoretical understanding of tone, semantic problems, discourse related structures, and other linguistic systems. The papers on Bantu languages reflect something of the internal richness and continued fascination of the family for linguists, as well as maturation of research on the family. The distribution of other papers highlights the need for intensified research into all the language families of Africa, including basic documentation, in order to comprehend linguistic diversities and convergences across the continent. In this regard, the chapter on Daats’íin (Gumuzic) stands out as the first-ever published article on this hitherto unknown and endangered language found in the Ethiopian-Sudanese border lands

Diversity in African languages: Selected papers from the 46th Annual Conference on African Linguistics

Diversity in African Languages contains a selection of revised papers from the 46th Annual Conference on African Linguistics, held at the University of Oregon. Most chapters focus on single languages, addressing diverse aspects of their phonology, morphology, semantics, syntax, information structure, or historical development. These chapters represent nine different genera: Mande, Gur, Kwa, Edoid, Bantu, Nilotic, Gumuzic, Cushitic, and Omotic. Other chapters investigate a mix of languages and families, moving from typological issues to sociolinguistic and inter-ethnic factors that affect language and accent switching. Some chapters are primarily descriptive, while others push forward the theoretical understanding of tone, semantic problems, discourse related structures, and other linguistic systems. The papers on Bantu languages reflect something of the internal richness and continued fascination of the family for linguists, as well as maturation of research on the family. The distribution of other papers highlights the need for intensified research into all the language families of Africa, including basic documentation, in order to comprehend linguistic diversities and convergences across the continent. In this regard, the chapter on Daats’íin (Gumuzic) stands out as the first-ever published article on this hitherto unknown and endangered language found in the Ethiopian-Sudanese border lands

Diversity in African languages: Selected papers from the 46th Annual Conference on African Linguistics

Diversity in African Languages contains a selection of revised papers from the 46th Annual Conference on African Linguistics, held at the University of Oregon. Most chapters focus on single languages, addressing diverse aspects of their phonology, morphology, semantics, syntax, information structure, or historical development. These chapters represent nine different genera: Mande, Gur, Kwa, Edoid, Bantu, Nilotic, Gumuzic, Cushitic, and Omotic. Other chapters investigate a mix of languages and families, moving from typological issues to sociolinguistic and inter-ethnic factors that affect language and accent switching. Some chapters are primarily descriptive, while others push forward the theoretical understanding of tone, semantic problems, discourse related structures, and other linguistic systems. The papers on Bantu languages reflect something of the internal richness and continued fascination of the family for linguists, as well as maturation of research on the family. The distribution of other papers highlights the need for intensified research into all the language families of Africa, including basic documentation, in order to comprehend linguistic diversities and convergences across the continent. In this regard, the chapter on Daats’íin (Gumuzic) stands out as the first-ever published article on this hitherto unknown and endangered language found in the Ethiopian-Sudanese border lands

Diversity in African languages: Selected papers from the 46th Annual Conference on African Linguistics

Diversity in African Languages contains a selection of revised papers from the 46th Annual Conference on African Linguistics, held at the University of Oregon. Most chapters focus on single languages, addressing diverse aspects of their phonology, morphology, semantics, syntax, information structure, or historical development. These chapters represent nine different genera: Mande, Gur, Kwa, Edoid, Bantu, Nilotic, Gumuzic, Cushitic, and Omotic. Other chapters investigate a mix of languages and families, moving from typological issues to sociolinguistic and inter-ethnic factors that affect language and accent switching. Some chapters are primarily descriptive, while others push forward the theoretical understanding of tone, semantic problems, discourse related structures, and other linguistic systems. The papers on Bantu languages reflect something of the internal richness and continued fascination of the family for linguists, as well as maturation of research on the family. The distribution of other papers highlights the need for intensified research into all the language families of Africa, including basic documentation, in order to comprehend linguistic diversities and convergences across the continent. In this regard, the chapter on Daats’íin (Gumuzic) stands out as the first-ever published article on this hitherto unknown and endangered language found in the Ethiopian-Sudanese border lands