PreImplantation Factor bolsters neuroprotection via modulating Protein Kinase A and Protein Kinase C signaling

A synthetic peptide (sPIF) analogous to the mammalian embryo-derived PreImplantation Factor (PIF) enables neuroprotection in rodent models of experimental autoimmune encephalomyelitis and perinatal brain injury. The protective effects have been attributed, in part, to sPIF's ability to inhibit the biogenesis of microRNA let-7, which is released from injured cells during central nervous system (CNS) damage and induces neuronal death. Here, we uncover another novel mechanism of sPIF-mediated neuroprotection. Using a clinically relevant rat newborn brain injury model, we demonstrate that sPIF, when subcutaneously administrated, is able to reduce cell death, reverse neuronal loss and restore proper cortical architecture. We show, both in vivo and in vitro, that sPIF activates cyclic AMP dependent protein kinase (PKA) and calcium-dependent protein kinase (PKC) signaling, leading to increased phosphorylation of major neuroprotective substrates GAP-43, BAD and CREB. Phosphorylated CREB in turn facilitates expression of Gap43, Bdnf and Bcl2 known to have important roles in regulating neuronal growth, survival and remodeling. As is the case in sPIF-mediated let-7 repression, we provide evidence that sPIF-mediated PKA/PKC activation is dependent on TLR4 expression. Thus, we propose that sPIF imparts neuroprotection via multiple mechanisms at multiple levels downstream of TLR4. Given the recent FDA fast-track approval of sPIF for clinical trials, its potential clinical application for treating other CNS diseases can be envisioned.

H19 lncRNA alters stromal cell growth via IGF signaling in the endometrium of women with endometriosis

Endometriosis affects approximately 15% of reproductive aged women and is associated with chronic pelvic pain and infertility. However, the molecular mechanisms by which endometriosis impacts fertility are poorly understood. The developmentally regulated, imprinted H19 long noncoding RNA (lncRNA) functions to reduce the bioavailability of microRNA let-7 by acting as a molecular sponge. Here we report that H19 expression is significantly decreased in the eutopic endometrium of women with endometriosis as compared to normal controls. We show that decreased H19 increases let-7 activity, which in turn inhibits Igf1r expression at the post-transcriptional level, thereby contributing to reduced proliferation of endometrial stromal cells. We propose that perturbation of this newly identified H19/Let-7/IGF1R regulatory pathway may contribute to impaired endometrial preparation and receptivity for pregnancy in women with endometriosis. Our finding represents the first example of a lncRNA-based mechanism in endometriosis and its associated infertility, thus holding potential in the development of novel therapeutics for women with endometriosis and infertility.

Immune regulatory and neuroprotective properties of preimplantation factor: From newborn to adult.

Embryonic-maternal interaction from the earliest stages of gestation has a key, sustained role in neurologic development, persisting into adulthood. Early adverse events may be detrimental in adulthood. Protective factors present during gestation could significantly impact post-natal therapy. The role of PreImplantation Factor (PIF) within this context is herein examined. Secreted by viable early embryos, PIF establishes effective embryonic-maternal communication and exerts essential trophic and protective roles by reducing oxidative stress and protein misfolding and by blunting the nocive let-7 microRNA related pathway. PIF's effects on systemic immunity lead to comprehensive immune modulation, not immune suppression. We examine PIF's role in protecting embryos from adverse maternal environment, which can lead to neurological disorders that may only manifest post-nataly: Synthetic PIF successfully translates endogenous PIF features in both pregnant and non-pregnant clinically relevant models. Specifically PIF has neuroprotective effects in neonatal prematurity. In adult relapsing-remitting neuroinflammation, PIF reverses advanced paralysis while promoting neurogenesis. PIF reversed Mycobacterium smegmatis induced brain infection. In graft-vs.-host disease, PIF reduced skin ulceration, liver inflammation and colon ulceration while maintaining beneficial anti-cancer, graft-vs.-leukemia effect. Clinical-grade PIF has high-safety profile even at supraphysiological doses. The FDA awarded Fast-Track designation, and university-sponsored clinical trials for autoimmune disorder are ongoing. Altogether, PIF properties point to its determining regulatory role in immunity, inflammation and transplant acceptance. Specific plans for using PIF for the treatment of complex neurological disorders (ie. traumatic brain injury, progressive paralysis), including neuroprotection from newborn to adult, are presented.

MicroRNA Regulation of Prostate Cancer Stem/Progenitor Cells and Prostate Cancer Development

Most human tumors contain a population of cells with stem cell properties, called cancer stem cells (CSCs), which are believed to be responsible for tumor establishment, metastasis, and resistance to clinical therapy. It’s crucial to understand the regulatory mechanisms unique to CSCs, so that we may design CSC-specific therapeutics. Recent discoveries of microRNA (miRNA) have provided a new avenue in understanding the regulatory mechanisms of cancer. However, how miRNAs may regulate CSCs is still poorly understood. Here, we present miRNA expression profiling in six populations of prostate cancer (PCa) stem/progenitor cells that possess distinct tumorigenic properties. Six miRNAs were identified to be commonly and differentially expressed, namely, four miRNAs (miR-34a, let-7b, miR-106a and miR-141) were under-expressed, and two miRNAs (miR-301 and miR-452) were over-expressed in the tumorigenic subsets compared to the corresponding marker-negative subpopulations. Among them, the expression patterns of miR-34, let-7b, miR-141 and miR-301 were further confirmed in the CD44+ human primary prostate cancer (HPCa) samples. We then showed that miR-34a functioned as a critical negative regulator in prostate CSCs and PCa development and metastasis. Over-expression of miR-34a in either bulk or CD44+ PCa cells significantly suppressed clonal expansion, tumor development and metastasis. Systemic delivery of miR-34a in tumor-bearing mice demonstrated a potent therapeutic effect again tumor progression and metastasis, leading to extended animal survival. Of great interest, we identified CD44 itself as a direct and relevant downstream target of miR-34a in mediating its tumor-inhibitory effects. Like miR-34a, let-7 manifests similar tumor suppressive effects in PCa cells. In addition, we observed differential mechanisms between let-7 and miR-34a on cell cycle, with miR-34a mainly inducing G1 cell-cycle arrest followed by cell senescence and let-7 inducing G2/M arrest. MiR-301, on the other hand, exerted a cell type dependent effect in regulating prostate CSC properties and PCa development. In summary, our work reveals that the prostate CSC populations display unique miRNA expression signatures and different miRNAs distinctively and coordinately regulate various aspects of CSC properties. Altogether, our results lay a scientific foundation for developing miRNA-based anti-cancer therapy.

Post-transcriptional Regulation of Mammalian Gene Expression in Non-coding Region of Target RNA

Tumor Suppressor Candidate 2 (TUSC2) is a novel tumor suppressor gene located in the human chromosome 3p21.3 region. TUSC2 mRNA transcripts could be detected on Northern blots in both normal lung and some lung cancer cell lines, but no endogenous TUSC2 protein could be detected in a majority of lung cancer cell lines. Mechanisms regulating TUSC2 protein expression and its inactivation in primary lung cancer cells are largely unknown. We investigated the role of the 5’- and 3’-untranslated regions (UTRs) of the TUSC2 gene in the regulation of TUSC2 protein expression. We found that two small upstream open-reading frames (uORFs) in the 5’UTR of TUSC2 could markedly inhibit the translational initiation of TUSC2 protein by interfering with the “scanning” of the ribosome initiation complexes. Site-specific stem-loop array reverse transcription-polymerase chain reaction (SLA-RT-PCR) verified several micoRNAs (miRNAs) targeted at 3’UTR and directed TUSC2 cleavage and degradation. In addition, we used the established let-7-targeted high mobility group A2 (Hmga2) mRNA as a model system to study the mechanism of regulation of target mRNA by miRNAs in mammalian cells under physiological conditions. There have been no evidence of direct link between mRNA downregulation and mRNA cleavages mediated by miRNAs. Here we showed that the endonucleolytic cleavages on mRNAs were initiated by mammalian miRNA in seed pairing style. Let-7 directed cleavage activities among the eight predicted potential target sites have varied efficiency, which are influenced by the positional and the structural contexts in the UTR. The 5’ cleaved RNA fragments were mostly oligouridylated at their 3’-termini and accumulated for delayed 5’–3’ degradation. RNA fragment oligouridylation played important roles in marking RNA fragments for delayed bulk degradation and in converting RNA degradation mode from 3’–5’ to 5’–3’ with cooperative efforts from both endonucleolytic and non-catalytic miRNA-induced silencing complex (miRISC). Our findings point to a mammalian miRNA-mediated mechanism for the regulation of mRNA that miRNA can decrease target mRNA through target mRNA cleavage and uridine addition

Prognostic role of the LCS6 KRAS variant in locally advanced rectal cancer: results of the EXPERT-C trial.

Background: Lethal-7 (let-7) is a tumour suppressor miRNA which acts by down-regulating several oncogenes including KRAS. A single-nucleotide polymorphism (rs61764370, T > G base substitution) in the let-7 complementary site 6 (LCS-6) of KRAS mRNA has been shown to predict prognosis in early-stage colorectal cancer (CRC) and benefit from anti-epidermal growth factor receptor monoclonal antibodies in metastatic CRC. Patients and methods: We analysed rs61764370 in EXPERT-C, a randomised phase II trial of neoadjuvant CAPOX followed by chemoradiotherapy, surgery and adjuvant CAPOX plus or minus cetuximab in locally advanced rectal cancer. DNA was isolated from formalin-fixed paraffin-embedded tumour tissue and genotyped using a PCR-based commercially available assay. Kaplan–Meier method and Cox regression analysis were used to calculate survival estimates and compare treatment arms. Results: A total of 155/164 (94.5%) patients were successfully analysed, of whom 123 (79.4%) and 32 (20.6%) had the LCS-6 TT and LCS-6 TG genotype, respectively. Carriers of the G allele were found to have a statistically significantly higher rate of complete response (CR) after neoadjuvant therapy (28.1% versus 10.6%; P = 0.020) and a trend for better 5-year progression-free survival (PFS) [77.4% versus 64.5%: hazard ratio (HR) 0.56; P = 0.152] and overall survival (OS) rates (80.3% versus 71.9%: HR 0.59; P = 0.234). Both CR and survival outcomes were independent of the use of cetuximab. The negative prognostic effect associated with KRAS mutation appeared to be stronger in patients with the LCS-6 TT genotype (HR PFS 1.70, P = 0.078; HR OS 1.79, P = 0.082) compared with those with the LCS-6 TG genotype (HR PFS 1.33, P = 0.713; HR OS 1.01, P = 0.995). Conclusion: This analysis suggests that rs61764370 may be a biomarker of response to neoadjuvant treatment and an indicator of favourable outcome in locally advanced rectal cancer possibly by mitigating the poor prognosis of KRAS mutation. In this setting, however, this polymorphism does not appear to predict cetuximab benefit.

Generation and Characterisation of a Canine EGFP-HMGA2 Prostate Cancer In Vitro Model

The architectural transcription factor HMGA2 is abundantly expressed during embryonic development. In several malignant neoplasias including prostate cancer, high re-expression of HMGA2 is correlated with malignancy and poor prognosis. The let-7 miRNA family is described to regulate HMGA2 negatively. The balance of let-7 and HMGA2 is discussed to play a major role in tumour aetiology. To further analyse the role of HMGA2 in prostate cancer a stable and highly reproducible in vitro model system is precondition. Herein we established a canine CT1258-EGFP-HMGA2 prostate cancer cell line stably overexpressing HMGA2 linked to EGFP and in addition the reference cell line CT1258-EGFP expressing solely EGFP to exclude EGFP-induced effects. Both recombinant cell lines were characterised by fluorescence microscopy, flow cytometry and immunocytochemistry. The proliferative effect of ectopically overexpressed HMGA2 was determined via BrdU assays. Comparative karyotyping of the derived and the initial CT1258 cell lines was performed to analyse chromosome consistency. The impact of the ectopic HMGA2 expression on its regulator let-7a was analysed by quantitative real-time PCR. Fluorescence microscopy and immunocytochemistry detected successful expression of the EGFP-HMGA2 fusion protein exclusively accumulating in the nucleus. Gene expression analyses confirmed HMGA2 overexpression in CT1258-EGFP-HMGA2 in comparison to CT1258-EGFP and native cells. Significantly higher let-7a expression levels were found in CT1258-EGFP-HMGA2 and CT1258-EGFP. The BrdU assays detected an increased proliferation of CT1258-HMGA2-EGFP cells compared to CT1258-EGFP and native CT1258. The cytogenetic analyses of CT1258-EGFP and CT1258-EGFP-HMGA2 resulted in a comparable hyperdiploid karyotype as described for native CT1258 cells. To further investigate the impact of recombinant overexpressed HMGA2 on CT1258 cells, other selected targets described to underlie HMGA2 regulation were screened in addition. The new fluorescent CT1258-EGFP-HMGA2 cell line is a stable tool enabling in vitro and in vivo analyses of the HMGA2-mediated effects on cells and the development and pathogenesis of prostate cancer.

Estudo da participação de microRNAs na regulação da resposta imune inata de macrófagos murinos à infecção por Paracoccidioides brasiliensis

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Biológicas, Departamento de Biologia Celular, Pós-Graduação em Biologia Molecular, 2016.

Let the journey begin : cultivating student identity and a sense of belonging through a formal academic welcome

Queensland University of Technology [QUT] Caboolture campus is a satellite campus co-located with Brisbane North Institute of TAFE. Building a sense of belonging on-campus continues to be challenging, with anecdotal evidence suggesting that QUT Caboolture students feel like second-class students, isolated from the prestige and excitement of the city campuses. A student identity and fostering a sense of belonging are fundamental to on-campus engagement and have been linked to retention (Field & Morgan-Klein, 2010). A formal welcome ceremony, including an academic procession in full regalia, was a new inclusion for QUT Caboolture’s 2013 Orientation Day. The ritual was intended to be a temporal mark for students to recognise their personal transition and emerging identity as a university student. Cultural capital is one ingredient in a complex mix of interactions and relationships to build a sense of identity. (Ecclestone, Biesta & Hughes, 2010). Fostering a sense of belonging and a connection to the campus and its staff was also anticipated. Students responded positively to the event, reporting feelings of excitement, inspiration and being welcomed into the university culture. The ceremony marked the beginning of the students’ journey together as QUT Caboolture students.

Review: Let's talk kinship: Innovating Australian social work education, theory, research and practice through Aboriginal knowledge

Review(s) of: Let's talk kinship: Innovating Australian social work education, theory, research and practice through Aboriginal knowledge, by Christine Fejo-King, published by Christine Fejo-King Consulting, ISBN: 978-0-9922814-0-3.

不同LET碳离子对V79细胞辐射敏感性的影响

以中国仓鼠肺V79细胞为材料 ,利用兰州近代物理研究所重离子研究装置 (HIRFL)产生的碳离子 ,研究了不同线性能量传递 (LET)的重离子对体外培养细胞的存活效应 ,并与γ射线的结果作了比较。结果表明 ,不同LET碳离子引起细胞失活效应由大到小的顺序依次为 12 5、2 0 0、70 0keV/μm。碳离子表现为无肩区的存活曲线 ,属单靶单击模型 ,γ射线表现为有肩区的存活曲线 ,属多靶单击模型。LET值为 12 5、2 0 0、70 0keV/μm时得到的失活截面分别为 35、12、8μm2 。当细胞存活比率为 0 .1和 0 .37,在LET为 12 5keV/μm时得到相对生物学效应 (RBE)值为1.4 7和 2 .19。

Show me, help me, let me : supporting teachers' changing conceptions of reading assessment and reading instruction

This qualitative inquiry used case study methodology to explore the change processes of 3 primary-grade teachers throughout their participation in 7 -month professional learning initiative focused on reading assessment and instruction. Participants took part in semimonthly inquiry-based professional learning community sessions, as well as concurrent individualized classroom-based literacy coaching. Each participant's experiences were first analyzed as a single case study, followed by cross-case analyses. While their patterns of professional growth differed, findings documented how all participants altered their understandings of the roles and relevancy of individual components of reading instruction (e.g., comprehension, decoding) and instructional approaches to scaffold students' growth (e.g., levelled text, strategy instruction), and experienced some form of conceptual change. Factors identified as affecting their change processes included; motivation, professional knowledge, professional beliefs (self-efficacy and theoretical orientation), resources (e.g., time, support), differentiated professional learning with associated goal-setting, and uncontrollable influences, with the affect of each factor compounded by interaction with the others. Comparison of participants' experiences to the Cognitive-Affective Model of Conceptual Change (CAMCC) and the Interconnected Model of Teacher Professional Growth (IMTPG) demonstrated the applicability of using both conceptual models, with the IMTPG providing macrolevel insights over time and the CAMCC microlevel insights at each change intervaL Recommendations include the provision of differentiated teacher professional learning opportunities, as well as research documenting the effects of teacher mentorship programs and the professional growth of teacher educators. ii

« Good night, sleep tight, don’t let the bed bugs bite» : exploring the mental health fallout of urban bed bug infestation in Montréal, Québec

Problématique : La littérature scientifique ne contient actuellement aucune étude épidémiologique portant sur la relation entre l’exposition à une infestation de punaises de lit (Cimex lectularis) et la santé mentale. L’objectif de cette étude est d’explorer la relation entre le statut d’exposition aux punaises de lit et des mesures de dépression, anxiété et perturbation du sommeil. Méthodes : Cette recherche est une étude transversale basée sur une analyse secondaire des données provenant de N=91 adultes locataires de logements insalubres qui ont répondu à un questionnaire de santé au moment d’interventions médico-environnementales menées par la direction de santé publique de Montréal entre janvier et juin 2010. Le questionnaire de santé inclut le « Questionnaire de santé du patient (QSP-9) », qui est un outil mesurant les symptômes associés à la dépression, l’outil de dépistage d’anxiété généralisée (GAD-7) et les items 1-18 de l’indice de la qualité du sommeil de Pittsburgh (PSQI). L’association entre une exposition autorapportée à une infestation de punaises de lit et le niveau de perturbation du sommeil selon la sous-échelle correspondante du PSQI, les symptômes dépressifs selon le QSP-9 et les symptômes anxieux selon le GAD-7, a été évaluée en utilisant une analyse de régression linéaire multivariée. Des données descriptives relatives aux troubles de sommeil autodéclarés et à des comportements reliés à un isolement social, dus à une exposition aux punaises de lit, sont aussi présentées. Résultats : L’échantillon comprenait 38 hommes et 53 femmes. Parmi les 91 participants, 49 adultes ont signalé une exposition aux punaises de lit et de ce nombre, 53,06% (26/49) ont déclaré des troubles de sommeil et 46,94% (23/49) des comportements d’isolement social, en raison de l’exposition. Les adultes exposés à des punaises de lit ont obtenu des résultats significativement plus élevés sur le QSP-9 (p=0,025), le GAD-7 (p=0,026) et sur la sous-échelle mesurant la perturbation du sommeil (p=0,003) comparativement à ceux qui n’étaient pas exposés. Conclusions : Cette première étude exploratoire met en évidence une association significative entre l’état d’exposition aux punaises de lit et des troubles de sommeil, ainsi que des symptômes anxieux et dépressifs, indiquant que ceux qui sont exposés aux punaises de lit représentent un groupe possiblement à risque de développer des problèmes de santé mentale. Ces résultats viennent en appui aux initiatives des décideurs pour organiser des efforts coordonnés d’éradication au niveau des villes et des provinces.