Human pulmonary microvascular endothelial cells respond to DAMPs from injured renal tubular cells.

Acute kidney injury (AKI) causes distant organ dysfunction through yet unknown mechanisms, leading to multiorgan failure and death. The lungs are one of the most common extrarenal organs affected by AKI, and combined lung and kidney injury has a mortality as high as 60%-80%. One mechanism that has been implicated in lung injury after AKI involves molecules released from injured kidney cells (DAMPs, or damage-associated molecular patterns) that promote a noninfectious inflammatory response by binding to pattern recognition receptors (PRRs) constitutively expressed on the pulmonary endothelium. To date there are limited data investigating the role of PRRs and DAMPs in the pulmonary endothelial response to AKI. Understanding these mechanisms holds great promise for therapeutics aimed at ameliorating the devastating effects of AKI. In this study, we stimulate primary human microvascular endothelial cells with DAMPs derived from injured primary renal tubular epithelial cells (RTECs) as an ex-vivo model of lung injury following AKI. We show that DAMPs derived from injured RTECs cause activation of Toll-Like Receptor and NOD-Like Receptor signaling pathways as well as increase human primary pulmonary microvascular endothelial cell (HMVEC) cytokine production, cell signaling activation, and permeability. We further show that cytokine production in HMVECs in response to DAMPs derived from RTECs is reduced by the inhibition of NOD1 and NOD2, which may have implications for future therapeutics. This paper adds to our understanding of PRR expression and function in pulmonary HMVECs and provides a foundation for future work aimed at developing therapeutic strategies to prevent lung injury following AKI.

An interaction between OTULIN and SCRIB uncovers roles for linear ubiquitination in planar cell polarity.

Planar cell polarity (PCP) plays critical roles in developmental and homeostatic processes. Membrane presentation of PCP complexes containing Van Gogh-like (VANGL) transmembrane proteins is central to PCP and can be directed by the scaffold protein scribble (SCRIB). The role atypical linear ubiquitin (Met1-Ub) chains might play in PCP is unknown. Here, HEK293 cell-based interactomic analyses of the Met1-Ub deubiquitinase OTULIN revealed that OTULIN can interact with SCRIB. Moreover, Met1-Ub chains associated with VANGL2 and PRICKLE1, but not SCRIB, can direct VANGL2 surface presentation. Mouse embryos lacking Otulin showed variable neural tube malformations, including rare open neural tubes, a deficit associated with PCP disruption in mice. In Madin-Darby canine kidney cells, in which the enrichment of VANGL2-GFP proteins at cell-cell contacts represents activated PCP complexes, endogenous OTULIN was recruited to these sites. In the human MDA-MB-231 breast cancer cell model, OTULIN loss caused deficits in Wnt5a-induced filopodia extension and trafficking of transfected HA-VANGL2. Taken together, these findings support a role for linear (de)ubiquitination in PCP signaling. The association of Met1-Ub chains with PCP complex components offers new opportunities for integrating PCP signaling with OTULIN-dependent immune and inflammatory pathways.

The Application of GHRH Antagonist as a Treatment for Resistant APL.

GHRH is a hypothalamic peptide shown to stimulate the proliferation of malignant cells in humans. We have previously shown that the use of GHRH antagonist MIA-602 successfully suppressed the growth of many human cancer cell lines, spanning more than 20 types of cancers. In this study, we demonstrate the presence of GHRH-R in the NB4, NB4-RAA, and K-562 model cell lines. Furthermore, we demonstrate the inhibited proliferation of all three cell lines in vitro after incubation with MIA-602. The treatment of xenografts of human APL cell lines with MIA-602 led to a significant reduction in tumor growth. Additionally, combination therapy with both doxorubicin (DOX) and MIA-602 showed a marked synergistic effect in reducing the proliferation of the K-562 AML cell line. These findings suggest that MIA-602 could be utilized to address resistance to all-trans retinoic acid (ATRA) and arsenic trioxide (ATO) therapies, as well as in augmenting anthracycline-based regimens.

Low-Level Light Therapy and Minoxidil Combination Treatment in Androgenetic Alopecia: A Review of the Literature.

Introduction: We analyzed randomized clinical trials (RCTs) evaluating the efficacy of combined therapy with low-level light therapy (LLLT) and topical minoxidil for treatment of androgenetic alopecia (AGA). Methods: A literature search within PubMed identified RCTs evaluating hair regrowth following LLLT and minoxidil. Selection criteria were 600-1,100 nm wavelengths, treatment time ≥16 weeks, and objective evaluation for hair regrowth. Results: Five RCTs compared LLLT with minoxidil (2% or 5%) to 5% minoxidil treatment or LLLT treatment. One study showed combination therapy of LLLT, and 5% minoxidil improved hair density more than monotherapy. Another found combination LLLT with 2% minoxidil induced hair regrowth equivalent to 5% minoxidil. Similarly, another study described LLLT with 5% minoxidil versus minoxidil monotherapy to increase the number of hairs with no statistical difference between groups. One trial found that combination group increased hair regrowth in the first 2 months. The last study found a statistically significant increase in hair density with combined therapy compared to monotherapy. Conclusion: The studies describe either superiority or equivalence of combination therapy to minoxidil monotherapy for AGA. Early outcomes appear to support the superiority of combination therapy, but this advantage wanes at the end of the study periods.

Investigation of cytokine polymorphisms on viral infections after renal transplantation exhibit association between IFN-γ +874 A > T and CMV manifestations.

We investigated the effects of TNF-α, IFN-γ, IL-10 polymorphisms on viral infections (CMV, BKPyV, HHV-6, EBV) after renal transplantation. IFN-γ+874 A > T (lower IFN production) was associated with CMV disease (p = .039) in patients under mycophenolate-based therapy and graft failure (p = .025). This study underscores the role of IFN-γ+874 SNP in CMV infection.

Helminth infection modulates number and function of adipose tissue Tregs in high fat diet-induced obesity.

BACKGROUND: Epidemiological and experimental studies have shown a protective effect of helminth infections in weight gain and against the development of metabolic dysfunctions in the host. However, the mechanisms Treg cells exert in the helminth-obesity interface has been poorly investigated. The present study aimed to verify the influence of Heligmosomoides polygyrus infection in early stages of high fat diet-induced obesity. PRINCIPAL FINDINGS: The presence of infection was able to prevent exacerbated weight gain in mice fed with high fat diet when compared to non-infected controls. In addition, infected animals displayed improved insulin sensitivity and decreased fat accumulation in the liver. Obesity-associated inflammation was reduced in the presence of infection, demonstrated by lower levels of leptin and resistin, lower infiltration of Th1 and Th17 cells in adipose tissue, higher expression of IL10 and adiponectin, increased infiltration of Th2 and eosinophils in adipose tissue of infected animals. Of note, the parasite infection was associated with increased Treg frequency in adipose tissue which showed higher expression of cell surface markers of function and activation, like LAP and CD134. The infection could also increase adipose Treg suppressor function in animals on high fat diet. CONCLUSION: These data suggest that H. polygyrus modulates adipose tissue Treg cells with implication for weight gain and metabolic syndrome.

A systematic review of the treatment of residual below the knee venous reflux after endovenous thermal ablation of the great saphenous vein.

BACKGROUND: Great saphenous vein (GSV) antireflux procedures have evolved during the past few decades to reduce elevated venous pressure. Untreated reflux in the below knee (BK) GSV (BK-GSV) can lead to persistent venous hypertension and deterioration of the venous circulation. The purpose of the present systematic review was to study the influence of BK-GSV intervention on venous disease progression. METHODS: A search was conducted, adhering to the PRISMA (preferred reporting items for systematic reviews and meta-analyses) guidelines. The PubMed and Embase databases were searched and cross-referenced. Studies were included if they had met the inclusion criterion of BK-GSV disease as a primary or secondary outcome. Two of the authors independently determined the eligibility and extracted the relevant data. RevMan, version 5.3 (Cochrane Training, London, UK), and SPSS (IBM Corp, Armonk, NY) were used for statistical computation. RESULTS: Fifteen studies that had assessed BK-GSV reflux recurrence after ablative intervention were included in our analysis. Of the 15 studies, 6 had assessed patients after above knee (AK) high ligation and stripping (HLS), 7 after AK endovenous laser ablation (AK-EVLA), and 2 after AK- and BK-EVLA (AK+BK EVLA). In total, 525 limbs had undergone HLS, 696 AK-EVLA, and 147 AK+BK EVLA. AK+BK EVLA was associated with significantly lower odds of BK-GSV reflux recurrence compared with AK-EVLA only (odds ratio [OR], 0.1857; 95% confidence interval [CI], 0.076-0.4734; P < .0001). Although the odds of recurrent BK-GSV reflux appeared to be greater for patients who had undergone AK-HLS compared with AK+BK HLS, the difference was not statistically significant (OR, 0.62; CI, 0.27-1.43; P = .69). Finally, no statistically significant difference was observed in BK-GSV reflux recurrence between patients receiving AK-EVLA and those receiving AK-HLS (OR, 0.85; 95% CI, 0.52-1.39; P = .31). CONCLUSIONS: Axial hydrostatic reflux from the groin to ankle is best controlled with AK+BK-GSV ablation. However, GSV ablation can result in saphenous nerve injury. For C4 to C6 disease, more aggressive treatment of the AK+BK-GSV is justified if the duplex ultrasound findings demonstrate groin to ankle reflux. Thermal ablation of the BK-GSV has a lower incidence of saphenous nerve injury than does BK saphenous stripping. More randomized controlled trials are needed to answer questions involving disease recurrence and the best techniques to mitigate these recurrences.

Screening for BKV-DNAEMIA after renal transplantation in a resource limited setting.

Costs may hinder the implementation of BK polyomavirus (BKV)-DNAemia screening in resource-limited kidney transplant (KT) centers. We analyzed data from two studies to assess the performance and potential cost saving of a dual-step screening strategy based on the use of a preliminary qualitative semi-nested PCR (snPCR) assay followed by BKV-DNAemia quantification after KT. In the preliminary study, in which 130 samples from 33 KT recipients were screened for BKV-DNAemia, the estimated positive and negative predictive values of snPCR, as compared to quantitative PCR (qPCR), were 88% and 99%, respectively. In the second study, which included 84 KT recipients, BKV-DNAemia was detected by snPCR in 28/472 (5.9%) samples and confirmed by qPCR in 26 samples of 21 (25%) subjects. No graft loss occurred among KT recipients who developed BKV-DNAemia. Cost analyses suggested that this strategy might be a cost saving alternative for BKV-DNAemia screening for some resource-limited settings.

Impact assessment and investigation of factors associated with herpesviruses viremia in the first year of renal transplantation.

The increased risk for opportunistic infections after a renal transplant requires monitoring of viral infections to avoid future complications. Our goal was to investigate the impact and factors associated with Epstein-Barr virus (EBV), human cytomegalovirus (HCMV) and human herpesvirus type 6 (HHV-6) viremia in renal transplant recipients. Whole blood samples were collected monthly from 82 patients during the first semester and then quarterly up to 1 year after transplantation. EBV, HCMV, and HHV-6 were detected and quantified by TaqMan real-time polymerase chain reaction. The results showed that EBV and HCMV viremia were detected in 32 patients (39% each), while HHV-6 viremia in only 3 patients (3.7%). EBV was significantly associated with age (P = .050), thymoglobuline induction (P = .019), mTOR inhibitor-based therapy (P = .003), and female gender (P = .044). HCMV was significantly associated with basiliximab induction (P = .015), mycophenolate mofetil (MMF)-based therapy (P = .003) and allograft acute rejection (P = .033). Moreover, HCMV-disease was correlated with MMF-based therapy (P = .021) and female gender (P = .003). In conclusion, EBV and HCMV viremia were associated with different immunosuppressive induction and maintenance strategies. Additionally, higher HCMV viremia (> 10 4 copies/mL) was related to acute allograft rejection.

A globally applicable PCR-based detection and discrimination of BK and JC polyomaviruses.

BKV and JCV belong to the Polyomaviridae family and are opportunistic agents associated with complications in immunocompromised individuals. Although a single screening assay for both viruses would be convenient, the diversity of BKV and JCV serotypes and genotypes is a methodological challenge. In this paper, we developed a PCR method able to detect and segregate BKV and JCV, despite these genetic discrepancies. A duplex semi-nested PCR (duplex snPCR) was designed to target a conserved region (639nt-1516nt) within the VP2 gene. In the first PCR, a primer set common to all BKV and JCV serotypes/ genotypes was used, followed by a semi-nested PCR with internal primers for BKV and JCV segregation. The limit of detection of the duplex snPCR was as low as 10 copies of BKV or JCV plasmids/µL. Specific products were observed when JCV and BKV plasmids were mixed in the same reaction. In field sample testing, the duplex snPCR detected and distinguished both viruses in different biological samples. Results were confirmed by Sanger's sequencing. The geographical complexity of BKV and JCV serotypes and genotypes imposes limits to a simple and universal method that could detect each virus. However, we describe here a sensitive and reliable PCR technique for BKV and JCV diagnosis that overcomes these limitations and could be universally applied.

Cigarette Smoking and its Association with In-Hospital Mortality in Hemorrhagic Stroke Patients / Tabaquismo y su asociación con mortalidad intrahospitalaria en pacientes con ictus hemorrágico

Abstract Backgroud: Smoking is a well-known reversible risk factor associated with acute ischemic stroke (AIS), however, some data showed that in-hospital mortality rates among smoker's patients with hemorrhagic stroke is lower when compared with non-smokers patients. The objective of this analysis was to assess in-hospital patient mortality rate between smokers and non-smokers patients with confirmed diagnosis of hemorrhagic stroke. Methods and Results: We analyzed all the data from patients diagnosed with hemorrhagic stroke that were registered in the Florida Stroke database from 2008-2012. Among the 21,013 patients diagnosed with hemorrhagic stroke, 10.9% of patients were smokers. A Multivariable model was used to estimate the adjusted odds ratio of in-hospital mortality rate in smokers versus non-smokers. Smoking was associated with lower in-hospital mortality in hemorrhagic patients. (UOR= 0.71 vs. AOR= 0.75). Other risk factors like hypertension, BMI>40 and hyperlipidemia, remained significantly associated after modeling, and some others like gender, race, health insurance coverage, and diabetes became insignificant. The difference between unadjusted and adjusted odds ratios for smoking (0.71 versus 0.75) indicates no presence of substantial confounding by age and other control variables. Conclusions: Among patients hospitalized for hemorrhagic stroke, smoking is a risk factor for early age of onset, even among those with few vascular risk factors. The persistent association with lower in-hospital mortality after adjusted analyses probably represents other unmeasured confounding, although a biological effect of smoking cannot be excluded. Further clinical and prospective population-based studies are needed to explore variables that contribute to outcomes in these patients.

Resumen Antecedentes: El fumar es un factor de riesgo tradicional claramente identificado con un mayor riesgo de desarrollar accidente cerebrovascular (ACV). A pesar de existir un riesgo más elevado de desarrollar ACV, algunos reportes han demostrado que pacientes ya hospitalizados con diagnóstico de ACV hemorrágico tienen una mortalidad intrahospitalaria menor a sus pares no fumadores, situación conocida como la paradoja del tabaquismo. El objetivo de este estudio es analizar la mortalidad Intrahospitalaria de Pacientes con Accidente Cerebrovascular Hemorrágico entre aquellos pacientes fumadores versus los no fumadores. Métodos y resultados: Este estudio epidemiológico observacional analítico de casos y controles incluyó a todos los pacientes con diagnóstico de accidente cerebrovascular hemorrágico incluidos en el "Registro de Accidentes Cerebrovasculares de Florida" entre el año 2008 y 2012. Entre los 21.013 pacientes diagnosticados con accidente cerebrovascular hemorrágico, el 10,9% de los pacientes fueron fumadores. Se utilizaron modelos multivariables para estimar la tasa de probabilidad ajustada de mortalidad intrahospitalaria en fumadores versus no fumadores. El tabaquismo se asoció con una menor mortalidad hospitalaria en pacientes hemorrágicos. (UOR = 0,71 frente a AOR = 0,75). Como se observa en las tablas, muchos de los factores de confusión, la hipertensión, IMC> 40, y la hiperlipidemia, tuvieron asociación significativa después del modelamiento estadístico mientras que otros factores como el sexo, la raza, el tipo de seguro de salud y la presencia de diabetes no tuvieron una asociación estadísticamente significativa. La diferencia entre las tasas de probabilidad no ajustadas y ajustadas para el tabaquismo (0,71 frente a 0,75 respectivamente) indica que no hay presencia de factores de confusión sustanciales por edad y otras variables de control. Conclusiones: Entre los pacientes hospitalizados por accidente cerebrovascular hemorrágico, el tabaquismo es un factor de riesgo para el desarrollo del mismo, especialmente en pacientes jóvenes, incluso entre aquellos con pocos factores de riesgo vascular, sin embargo, la asociación persistente con menor mortalidad intrahospitalaria después de los análisis ajustados probablemente represente otra confusión no medida, aunque no se puede excluir un efecto biológico del tabaquismo. Se necesitan más estudios clínicos basados en la población para explorar las variables que contribuyen a los resultados en estos pacientes.

BK polyomavirus nephropathy in two kidney transplant patients with distinct diagnostic strategies for BK virus and similar clinical outcomes: two case reports.

BACKGROUND: BK polyomavirus-associated nephropathy is an important cause of post-transplantation renal failure. We present two cases of BK polyomavirus-associated nephropathy who were submitted to contrasting strategies of clinical follow-up to BK polyomavirus reactivation, but progressed to a similar final outcome. CASE PRESENTATION: Case 1 is a 37-year-old white man whose graft had never presented a good glomerular filtration rate function, with episodes of tacrolimus nephrotoxicity, and no urinary monitoring for BK polyomavirus; stage B BK polyomavirus-associated nephropathy was diagnosed by biopsy at 14 months post-transplant. Despite clinical treatment (dosage decrease and immunosuppressive drug change), he progressed to stage C BK polyomavirus-associated nephropathy and loss of graft function 30 months post-transplant. Case 2 is a 49-year-old mulatto man in his second renal transplantation who was submitted to cytological urinary monitoring for BK polyomavirus; he presented early, persistent, and massive urinary decoy cell shedding and concomitant tacrolimus nephrotoxicity. Even with decreasing immunosuppression, he developed BK polyomavirus-associated nephropathy 1-year post-transplant. Loss of graft function occurred 15 months post-transplant. CONCLUSIONS: Cytological urinary monitoring was an efficient strategy for monitoring BK virus reactivation. Decoy cell shedding may be related to BK polyomavirus-associated nephropathy when extensive and persistent. The presence of associated tacrolimus nephrotoxicity may be a confounding factor for the clinical diagnosis of BK polyomavirus-associated nephropathy.

Ensino e Aprendizagem em Odontologia: Análise de Sujeitos e Práticas / Teaching and Learning in Dentistry: Analysis of Individuals and Practices

RESUMO O presente estudo pedagógico objetivou investigar o processo de formação em Odontologia com base na visão dos gestores, docentes e discentes de um curso de Odontologia em uma instituição de ensino superior (IES) – Faculdade de Odontologia do Instituto de Saúde de Nova Friburgo da Universidade Federal Fluminense (FO/ISNF/UFF). A amostra não probabilística foi composta por 98 sujeitos (8 gestores, 28 docentes e 62 alunos), que foram entrevistados ou responderam a questionários semiestruturados. A análise dos resultados foi realizada por meio da técnica do Discurso do Sujeito Coletivo (DSC) e representações sociais. Os resultados apontaram que 49,0% e 76,5% do corpo docente e discente da IES não têm conhecimento sobre as Diretrizes Curriculares Nacionais (DCN). As sugestões de melhoria incluíram a oferta de cursos de formação continuada em docência, inclusive a respeito das DCN, melhorias na infraestrutura do campus universitário e uma relação professor/aluno mais compatível. O estudo permitiu verificar algumas deficiências nas práticas e posturas, percebidas pelos sujeitos entrevistados. Entretanto, parece haver uma relevante preocupação de alguns sobre um alinhamento do Projeto Político Pedagógico (PPP) com as DCN.

ABSTRACT This pedagogical study aimed to investigate the training process in dentistry, from the perspective of managers, teachers and students of a dentistry course at a Higher Education Institution (HEI) – the Fluminense Federal University (CNF-UFF) Dental School – Nova Friburgo Campus. A non-probabilistic sample was comprised of 98 subjects (8 managers, 28 teachers and 62 students) who completed questionnaires. The analysis was performed using the Collective Subject Discourse technique (DCS) and social representations. The results showed that 49% and 76.5% of the professors and students of HEI, respectively, do not know about the NCGs (National Curricular Guidelines). The suggestions for improvement included the provision of teacher training courses, including subjects for discussions such as NCGs, improvements in campus infrastructure and a more compatible student-teacher ratio. Therefore, the study showed some shortcomings in the practices and attitudes perceived by managers, teachers and students. However, some people seem to be duly concerned about the alignment of the Pedagogical Political Project (PPP) with the NCGs.

ASSOCIATION OF EPSTEIN-BARR VIRUS (EBV) BUT NOT HUMAN PAPILLOMAVIRUS (HPV) WITH GINGIVITIS AND/OR PERIODONTITIS IN TRANSPLANTED INDIVIDUALS.

The aim of this study was to investigate the association of EBV and HPV with gingivitis and/or periodontitis according to the immunologic status. To this end, 74 oral biopsies from transplanted and non-transplanted individuals with the abovementioned oral manifestations were submitted to a screening by PCR for both viruses. According to the results, EBV was strongly associated with gingivitis and/or periodontitis in transplanted individuals (p = 0.011) but not HPV (p = 0.766). EBV-HPV co-detections did not enhance the presence of tissue injury as well. Although a causal relationship was not investigated in this study, the higher frequency of these two oncoviruses in lesion tissues must be investigated in follow-up studies, especially among immunocompromised individuals.

Epidermoid Cyst Arising in the Buccal Mucosa: Case Report and Literature Review.

Epidermoid cysts are benign subcutaneous lesions, and the large majority of these cysts affect the floor of the mouth; however, the buccal mucosa is not a usual site of occurrence. To date, only 5 articles have been published with 6 cases of epidermoid cysts arising in the buccal mucosa. Therefore, the aim of this study was to describe the clinical, histopathological and immunohistochemical features of a case of epidermoid cyst located in the buccal mucosa. To our knowledge, this is the first report of an oral epidermoid cyst describing an intense foreign body gigantocellular inflammatory reaction against epithelial keratin component. Although the usual diagnosis for epidermoid cysts is based on histopathological findings, this case report addresses novel information regarding to the immunohistochemical pattern that may be found in these lesions.

Epidermoid Cyst Arising in the Buccal Mucosa: Case Report and Literature Review.

Epidermoid cysts are benign subcutaneous lesions, and the large majority of these cysts affect the floor of the mouth; however, the buccal mucosa is not the usual site of occurrence. To date, only 5 articles have been published with 6 cases of epidermoid cysts arising in the buccal mucosa. Therefore, the aim of this study was to describe the clinical, histopathological and immunohistochemical features of a case of an epidermoid cyst located in the buccal mucosa. To our knowledge, this is the first report of an oral epidermoid cyst describing an intense foreign body gigantocellular inflammatory reaction against epithelial keratin component. Although the usual diagnosis for epidermoid cysts is based on histopathological findings, this case report addresses novel information regarding the immunohistochemical pattern which may be found in these lesions.

Tratamento cirúrgico de fratura orbitária blow-out pura com tela de titânio: relato de caso clínico / Surgical treatment for blow-out pure orbital fracture using titanium mesh: clinical case report

As fraturas orbitárias do tipo blow-out são classificadas em dois tipos: pura e impura. O seu diagnóstico é realizado através do exame clínico auxiliado por exames de imagens. Dentre todas as imagens, a mais indicada é a tomografia computadorizada que oferece uma melhor visualização e avaliação do traço de fratura e possibilita um melhor planejamento cirúrgico.O tipo do tratamento, cirúrgico ou não-cirúrgico, assim como o momento mais oportuno para esse tratamento das fraturas orbitárias blow-out, imediato ou tardio, é controverso na literatura. O objetivo deste trabalho é apresentar um caso clínico de fratura orbitária blow-out pura, tratado cirurgicamente e de forma imediata com sucesso clínico.

Fractures orbital blow-out are ranked in two types: pure and impure. Diagnosis is made through clinical examination aided by images. Among these images, the most recommended is CT, which provides better visualization and evaluation of the fracture and also allows better surgical planning. The treatment type, surgical or not, as well as timing, immediate or late on, of treatment for orbital blow-out fractures is controversial in the literature. The objective of this paper is to present a pure blow out fracture case surgically treated and immediately after.

The linear ubiquitin-specific deubiquitinase gumby regulates angiogenesis.

A complex interaction of signalling events, including the Wnt pathway, regulates sprouting of blood vessels from pre-existing vasculature during angiogenesis. Here we show that two distinct mutations in the (uro)chordate-specific gumby (also called Fam105b) gene cause an embryonic angiogenic phenotype in gumby mice. Gumby interacts with disheveled 2 (DVL2), is expressed in canonical Wnt-responsive endothelial cells and encodes an ovarian tumour domain class of deubiquitinase that specifically cleaves linear ubiquitin linkages. A crystal structure of gumby in complex with linear diubiquitin reveals how the identified mutations adversely affect substrate binding and catalytic function in line with the severity of their angiogenic phenotypes. Gumby interacts with HOIP (also called RNF31), a key component of the linear ubiquitin assembly complex, and decreases linear ubiquitination and activation of NF-κB-dependent transcription. This work provides support for the biological importance of linear (de)ubiquitination in angiogenesis, craniofacial and neural development and in modulating Wnt signalling.

The TRAF-interacting protein (TRIP) is a regulator of keratinocyte proliferation.

The TRAF-interacting protein (TRIP/TRAIP) is a RING-type E3 ubiquitin ligase inhibiting tumor necrosis factor-α (TNF-α)-mediated NF-κB activation. TRIP ablation results in early embryonic lethality in mice. To investigate TRIP function in epidermis, we examined its expression and the effect of TRIP knockdown (KD) in keratinocytes. TRIP mRNA expression was strongly downregulated in primary human keratinocytes undergoing differentiation triggered by high cell density or high calcium. Short-term phorbol-12-myristate-13-acetate (TPA) treatment or inhibition of phosphatidylinositol-3 kinase signaling in proliferative keratinocytes suppressed TRIP transcription. Inhibition by TPA was protein kinase C dependent. Keratinocytes undergoing KD of TRIP expression by lentiviral short-hairpin RNA (shRNA; T4 and T5) had strongly reduced proliferation rates compared with control shRNA. Cell cycle analysis demonstrated that TRIP-KD caused growth arrest in the G1/S phase. Keratinocytes with TRIP-KD resembled differentiated cells consistent with the augmented expression of differentiation markers keratin 1 and filaggrin. Luciferase-based reporter assays showed no increase in NF-κB activity in TRIP-KD keratinocytes, indicating that NF-κB activity in keratinocytes is not regulated by TRIP. TRIP expression was increased by ∼2-fold in basal cell carcinomas compared with normal skin. These results underline the important role of TRIP in the regulation of cell cycle progression and the tight linkage of its expression to keratinocyte proliferation.

Coping with youth suicide and overdose: one community's efforts to investigate, intervene, and prevent suicide contagion.

From 2000-2005, Somerville, MA, experienced a number of youth overdoses and suicides. The community response followed CDC recommendations for contagion containment. A community coalition, Somerville Cares About Prevention, became a pivotal convener of community partners and a local research organization, the Institute for Community Health, provided needed expertise in surveillance and analysis. Mayoral leadership provided the impetus for action while community activists connected those at risk with mental health resources. Using a variety of data sources (including death certificates, youth risk surveys, 911 call data, and hospital discharges) overdose and suicide activity were monitored. Rates of suicide and overdose for 10-24-year-olds were higher than in previous years. Using case investigation methods, the majority of suicide victims were found to be linked through common peer groups and substance abuse. Subsequent community action steps included: a community-based trauma response team, improved media relationships, focus groups for suicide survivors, and prevention trainings to community stakeholders. Youth suicide and overdose activity subsided in May of 2005. The community partnerships were critical elements for developing a response to this public health crisis. This collaborative approach to suicide contagion used existing resources and provides important lessons learned for other communities facing similar circumstances.