In situ delivery of nanoparticles formulated with micron-sized crystals protects from murine melanoma.

INTRODUCTION Intratumoral injections of novel therapeutics can activate tumor antigen-specific T cells for locoregional tumor control and may even induce durable systemic protection (against distant metastases) via recirculating T cells. Here we explored the possibility of a universal immunotherapy that promotes T-cell responses in situ and beyond, upon intratumoral injection of nanoparticles formulated with micron-sized crystals. METHODS Cucumber mosaic virus-like particles containing a tetanus toxin peptide (CuMVTT) were formulated with microcrystalline tyrosine (MCT) adjuvant and injected directly in B16F10 melanoma tumors. To further enhance immunogenicity, we loaded the nanoparticles with a TLR7/8 ligand and incorporated a universal tetanus toxin T-helper cell peptide. We assessed therapeutic efficacy and induction of local and systemic immune responses, including RNA sequencing, providing broad insight into the tumor microenvironment and correlates of protection. RESULTS MCT crystals were successfully decorated with CuMVTT nanoparticles. This 'immune-enhancer' formed immunogenic depots in injected tumors, enhanced polyfunctional CD8+ and CD4+ T cells, and inhibited B16F10 tumor growth locally and systemically. Local inflammation and immune responses were associated with upregulation of genes involved in complement activation and collagen formation. CONCLUSIONS Our new immune-enhancer turned immunologically cold tumors into hot ones and inhibited local and distant tumor growth. This type of immunotherapy does not require the identification of (patient-individual) relevant tumor antigens. It is well tolerated, non-infectious, and affordable, and can readily be upscaled for future clinical testing and broad application in melanoma and likely other solid tumors

A video guide of five access methods to the splenic flexure: the concept of the splenic flexure box

Aim: The aim of this study was to describe all the possible approaches for laparoscopic splenic flexure mobilization (SFM), each suitable for specific situations, and create an illustrated system to show SFM approaches in an easy and practical way to make it easy to learn and teach. Methods: Two different phases. First part: Cadaver-based study of the colonic splenic flexure anatomy. In order to demonstrate the different approaches, a balloon was placed through the colonic hepatic flexure in the lesser sac without sectioning any of the fixing ligaments of the splenic flexure. Second part: A real case series of laparoscopic SFM. Results: First part: 11 cadavers were dissected. Five potential approaches to SFM were found: anterior, trans-omentum, lateral, medial infra-mesocolic, and medial trans-mesocolic. The illustrative system developed was named: Splenic Flexure “Box”(SFBox). Second part: One of the types of SFM described in first part was used in five patients with colorectal cancer. Each laparoscopic approach to the splenic flexure was illustrated in a video accompanied by illustration aids delineating the access. Conclusion: With the cadaver dissection and subsequent demonstration in real-life laparoscopic surgery, we have shown five types of laparoscopic splenic flexure mobilization. The Splenic Flexure “Box” is a useful way to learn and teach this surgical maneuver

Perceções dos enfermeiros sobre os erros de medicação no pré-hospitalar

Quase metade da totalidade de eventos adversos evitáveis é consequência de erros de medicação (EM), contudo, não sendo possível evitá-los completamente, estes podem ser minorados. Esta problemática em contexto pré-hospitalar (PH) tem sido pouco estudada a nível internacional e nunca foi abordada em Portugal. O objetivo deste estudo é relacionar as variáveis sociodemográficas, socioprofissionais, formação, conhecimentos e experiências com EM com a perceção dos enfermeiros que exercem no PH relativamente à frequência da ocorrência dos tipos e causas de EM, dos obstáculos ao relato de EM, dos fatores facilitadores do relato de EM e com o grau de concordância sobre divulgação de EM. Métodos: Trata-se de um estudo analítico, descritivo, transversal e correlacional. A amostra é composta por 107 enfermeiros do PH (método snowball), dos quais 56.1% são do sexo masculino. Foi aplicado um questionário eletrónico constituído por uma componente sociodemográfica, escala de conhecimentos, perceções e experiência com erros de medicação (Raimundo, 2011; Maurer, 2010; Bohomol & Ramos, 2006; Mayo & Duncan, 2004; Osborne, Blais & Hayes, 1999; Gladstone, 1995). Resultados: Dos inquiridos 60.7% apresentam fracos a razoáveis conhecimentos sobre EM; mais de 54% perceciona a sua formação académica/contínua sobre EM como sendo inexistente/insuficiente e 52.3% não recebe formação sobre farmacologia há pelo menos 6 anos; 45.8% diz ter experienciado no PH um ou mais EM sem dano para o doente e apenas 14.9% relatou um ou mais EM sem dano para o doente. Os tipos e as causas de EM identificadas ocorrem com uma frequência elevada para mais de 39% dos inquiridos. A maioria dos inquiridos (47.7%) considera que no PH existem grandes obstáculos ao relato de EM e os fatores facilitadores do relato de EM apresentados são considerados por 49.5% dos enfermeiros como altamente prováveis de facilitar o relato. 52,3% dos enfermeiros do PH discordam de uma forma global com a divulgação de EM. O sexo feminino apresenta uma perceção mais elevada da ocorrência das causas primárias de EM (MF=2.68, Dp= 0.60 vs MM=2.36, Dp=0.66) e uma perceção mais elevada dos fatores facilitadores ao relato dos EM (MF=4.40, Dp= 0.64 vs MM=4.12, Dp=0.74). Os enfermeiros que exercem exclusivamente no PH possuem uma melhor perceção da frequência de ocorrência das causas primárias de EM. Quanto maior o conhecimento dos enfermeiros sobre EM, maior é a perceção destes relativamente aos tipos de erros e maior o grau de concordância com a divulgação dos EM. Existe evidência estatisticamente significativa (p<0.05) de que os enfermeiros que experienciaram a ocorrência de pelo menos 1 erro com dano para o doente possuem melhor perceção dos tipos, causas primárias e obstáculos ao relato dos EM, assim como apresenta um maior grau de concordância com a divulgação de EM. Conclusão: A perceção dos enfermeiros sobre a frequência dos tipos e das causas de EM, assim como dos obstáculos e dos fatores facilitadores do relato de EM por parte dos enfermeiros no PH não tem, de uma forma geral, relação com as características sociodemográficas e socioprofissionais, o que demonstra a transversalidade desta problemática. Tão ou mais importante do que avaliar a dimensão e caracterizar a tipologia, causas, obstáculos e fatores facilitadores ao relato dos EM será, com base no conhecimento obtido, definir e implementar ações de gestão de risco que permitam a sua redução ou mesmo a sua supressão. PALAVRAS-CHAVE: Erros de Medicação, Perceção dos Enfermeiros, Pré- Hospitalar.ABSTRACT Almost half of all preventable adverse events is the result of medication errors (ME), however, it’s not possible to avoid them completely, but they can be mitigated. This problem in the prehospital setting (PH) has been little studied internationally and has never been approached in Portugal. The objective of this study is to relate the socio-demographic, professional, training, knowledge and experience variables with ME, with the PH nurse’s perception of the occurrence frequency of the types and causes of ME, obstacles to the reporting of ME, facilitating factors of ME and with the degree of agreement on the disclosure of ME. Methods: Analytical, descriptive, cross-sectional and correlational study. Sample with 107 PH nurses (snowball method), of which 56.1% are male gender. Application of an electronic survey composed by a socio-demographic component, a knowledge scale on ME and a scale on perceptions and experiences on ME (Raymond, 2011; Maurer, 2010; Bohomol & Ramos, 2006; Mayo & Duncan, 2004; Osborne, Blais & Hayes, 1999 ; Gladstone, 1995). Results: 60.7% of the nurses inquired have a poor or reasonable knowledge about ME; over 54% consider their academic/continuous training on ME as inexistent/insufficient and 52.3% have not had any continuous training on this subject for at least 6 years; 45.8% claim to have experienced one or more PH ME with no damage for the patient and only 14.9% reported one or more reported one or ME without any harm to the patient. The identified types and causes for the ME occur with a high frequency for over 39% of the enquired people. Most respondents (47.7%) consider that in the PH setting there are considerable obstacles to the reporting of ME and the facilitating factors of ME reports presented are considered by 49.5% of the nurses as highly likely to facilitate reporting. 52.3% of the PH nurses disagree with the disclosure of ME. Females have a higher perception of the occurrence of the primary causes of ME (MF = 2.68, SD = 0.60 vs MM = 2.36, SD = 0.66) and a higher perception of the facilitating factors for reporting ME (MF = 4.40, SD = 0.64 vs MM = 4.12 SD = 0.74). Nurses that work exclusively in the PH have a better perception of the frequency of occurrence of the primary causes of ME. The higher nurses' knowledge of ME matches to a higher perception of the types of ME and to a higher degree of agreement with the disclosure of ME. There’s statistically significant evidence (p <0.05) that nurses who experienced the occurrence of at least one error with damage to the patient has better perception of types, primary causes and obstacles to reporting of ME, as well as presents a higher degree of agreement with the disclosure of ME Conclusion: The PH nurses’ perception on the frequency of the types and causes of ME, as well on the obstacles and facilitating factors for ME reporting is generally not related to the socio-demographic and socio-professional characteristics, demonstrating the transversality of this issue. Equally or more important than assessing the extent and characterize the types, causes, obstacles and facilitating factors for reporting ME, will be define and implement risk management actions that allow its reduction or even its suppression, based on the knowledge gained. KEY WORDS: Medication Errors; Nurse’s Perception; Prehospital

The next generation virus-like particle platform for the treatment of peanut allergy.

BACKGROUND Allergy to peanut is one of the leading causes of anaphylactic reactions among food allergic patients. Immunization against peanut allergy with a safe and protective vaccine holds a promise to induce durable protection against anaphylaxis caused by exposure to peanut. A novel vaccine candidate (VLP Peanut), based on virus-like particles (VLPs), is described here for the treatment of peanut allergy. METHODS AND RESULTS VLP Peanut consist of two proteins: a capsid subunit derived from Cucumber mosaic virus engineered with a universal T cell epitope (CuMVTT ) and a CuMVTT subunit fused with peanut allergen Ara h 2 (CuMVTT -Ara h 2), forming mosaic VLPs. Immunizations with VLP Peanut in both naïve and peanut-sensitised mice resulted in a significant anti-Ara h 2 IgG response. Local and systemic protection induced by VLP Peanut were established in mouse models for peanut allergy following prophylactic, therapeutic and passive immunizations. Inhibition of FcγRIIb function resulted in a loss of protection, confirming the crucial role of the receptor in conferring cross protection against peanut allergens other than Ara h 2. CONCLUSION VLP Peanut can be delivered to peanut-sensitized mice without triggering allergic reactions, whilst remaining highly immunogenic and offering protection against all peanut allergens. In addition, vaccination ablates allergic symptoms upon allergen challenge. Moreover, the prophylactic immunization setting conferred the protection against subsequent peanut-induced anaphylaxis, showing the potential for preventive vaccination. This highlights the effectiveness of VLP Peanut as a prospective break-through immunotherapy vaccine candidate towards peanut allergy. VLP Peanut has now entered clinical development with the study PROTECT

The next generation virus‐like particle platform for the treatment of peanut allergy

Background: Allergy to peanut is one of the leading causes of anaphylactic reactions among food allergic patients. Immunization against peanut allergy with a safe and protective vaccine holds a promise to induce durable protection against anaphylaxis caused by exposure to peanut. A novel vaccine candidate (VLP Peanut), based on virus‐like particles (VLPs), is described here for the treatment of peanut allergy. Methods and Results: VLP Peanut consists of two proteins: a capsid subunit derived from Cucumber mosaic virus engineered with a universal T‐cell epitope (CuMV$_{TT}$) and a CuMV$_{TT}$ subunit fused with peanut allergen Ara h 2 (CuMV$_{TT}$‐Ara h 2), forming mosaic VLPs. Immunizations with VLP Peanut in both naïve and peanut‐sensitized mice resulted in a significant anti‐Ara h 2 IgG response. Local and systemic protection induced by VLP Peanut were established in mouse models for peanut allergy following prophylactic, therapeutic, and passive immunizations. Inhibition of FcγRIIb function resulted in a loss of protection, confirming the crucial role of the receptor in conferring cross protection against peanut allergens other than Ara h 2. Conclusion: VLP Peanut can be delivered to peanut‐sensitized mice without triggering allergic reactions, while remaining highly immunogenic and offering protection against all peanut allergens. In addition, vaccination ablates allergic symptoms upon allergen challenge. Moreover, the prophylactic immunization setting conferred the protection against subsequent peanut‐induced anaphylaxis, showing the potential for preventive vaccination. This highlights the effectiveness of VLP Peanut as a prospective break‐through immunotherapy vaccine candidate toward peanut allergy. VLP Peanut has now entered clinical development with the study PROTECT

Inactivation of aPKCλ Reveals a Context Dependent Allocation of Cell Lineages in Preimplantation Mouse Embryos

BACKGROUND:During mammalian preimplantation development, lineage divergence seems to be controlled by the interplay between asymmetric cell division (once cells are polarized) and positional information. In the mouse embryo, two distinct cell populations are first observed at the 16-cell stage and can be distinguished by both their position (outside or inside) and their phenotype (polarized or non-polarized). Many efforts have been made during the last decade to characterize the molecular mechanisms driving lineage divergence. METHODOLOGY/PRINCIPAL FINDINGS:In order to evaluate the importance of cell polarity in the determination of cell fate we have disturbed the activity of the apical complex aPKC/PAR6 using siRNA to down-regulate aPKClambda expression. Here we show that depletion of aPKClambda results in an absence of tight junctions and in severe polarity defects at the 16-cell stage. Importantly, we found that, in absence of aPKClambda, cell fate depends on the cellular context: depletion of aPKClambda in all cells results in a strong reduction of inner cells at the 16-cell stage, while inhibition of aPKClambda in only half of the embryo biases the progeny of aPKClambda defective blastomeres towards the inner cell mass. Finally, our study points to a role of cell shape in controlling cell position and thus lineage allocation. CONCLUSION:Our data show that aPKClambda is dispensable for the establishment of polarity at the 8-cell stage but is essential for the stabilization of cell polarity at the 16-cell stage and for cell positioning. Moreover, this study reveals that in addition to positional information and asymmetric cell divisions, cell shape plays an important role for the control of lineage divergence during mouse preimplantation development. Cell shape is able to influence both the type of division (symmetric or asymmetric) and the position of the blastomeres within the embryo

UV measurements at Marambio and Ushuaia during 2000–2010

Solar ultraviolet (UV) irradiances were measured with NILU-UV multichannel radiometers at Ushuaia (54°&thinsp;S) and Marambio (64°&thinsp;S) between 2000 and 2013. The measurements were part of the Antarctic NILU-UV network, which was started in cooperation between Spain, Argentina and Finland. The erythemally weighted UV irradiance time series of both stations were analysed for the first time. The quality assurance procedures included a travelling reference instrument to transfer the irradiance scale to the stations. The time series were homogenized and high quality measurements were available for the period 2000–2010. During this period UV indices of 11 or more were measured on 5 and 35 days at Marambio and Ushuaia, respectively. At Marambio, the peak daily maximum UV index of 12 and daily doses of around 7&thinsp;kJ&thinsp;m−2 were measured in November 2007. The highest UV daily doses at both stations were typically around 6&thinsp;kJ&thinsp;m−2 and occurred when the stations were inside the polar vortex, resulting in very low total ozone amount. At both stations, daily doses in late November could even exceed those in the summer. At Marambio, in some years, also daily doses in October can be as high as those during the summer. At Ushuaia, the peak daily maximum UV index of 13 was measured twice: in November 2003 and 2009. Also during those days, the station of Ushuaia was inside the polar vortex.</p

Clinical features and outcomes of patients with tubercular uveitis treated with antitubercular therapy in the collaborative ocular tuberculosis study (COTS)-1

IMPORTANCE Eradication of systemic tuberculosis (TB) has been limited by neglected populations and the HIV pandemic. Whereas ocular TB often presents as uveitis without any prior evidence of systemic TB, the existing uncertainty in the diagnosis of TB uveitis may perpetuate missed opportunities to address systemic TB. OBJECTIVE To examine the clinical features of TB uveitis and the associations with response to antitubercular therapy (ATT). DESIGN, SETTING, AND PARTICIPANTS This retrospective multinational cohort study included patients from 25 ophthalmology referral centers diagnosed with TB uveitis and treated with ATT from January 1, 2004, through December 31, 2014, with a minimum follow-up of 1 year. MAIN OUTCOMES AND MEASURES Treatment failure, defined as a persistence or recurrence of inflammation within 6 months of completing ATT, inability to taper oral corticosteroids to less than 10mg/d or topical corticosteroid drops to less than 2 drops daily, and/or recalcitrant inflammation necessitating corticosteroid-sparing immunosuppressive therapy. RESULTS A total of 801 patients (1272 eyes) were studied (mean [SD] age, 40.5 [14.8] years; 413 [51.6%] male and 388 [48.4%] female; 577 [73.6%] Asian). Most patients had no known history (498 of 661 [75.3%]) of systemic TB. Most patients had bilateral involvement (471 of 801 [58.8%]). Common clinical signs reported include vitreous haze (523 of 1153 [45.4%]), retinal vasculitis (374 of 874 [42.8%]), and choroidal involvement (419 of 651 [64.4%]). Treatment failure developed in 102 of the 801 patients (12.7%). On univariate regression analysis, the hazard ratios (HRs) associated with intermediate uveitis (HR, 2.21; 95%CI, 1.07-4.55; P = .03), anterior uveitis (HR, 2.68; 95%CI, 1.32-2.35; P = .006), and panuveitis (HR, 3.28; 95%CI, 1.89-5.67; P &lt; .001) were significantly higher compared with posterior distribution. The presence of vitreous haze had a statistically significant association (HR, 1.95; 95%CI, 1.26-3.02; P = .003) compared with absence of vitreous haze. Bilaterality had an associated HR of 1.50 (95%CI, 0.96-2.35) compared with unilaterality (HR, 1 [reference]), although this finding was not statistically significant (P = .07). On multivariate Cox proportional hazards regression analysis, the presence of vitreous haze had an adjusted HR of 2.98 (95%CI, 1.50-5.94; P = .002), presence of snow banking had an adjusted HR of 3.71 (95%CI, 1.18-11.62; P = .02), and presence of choroidal involvement had an adjusted HR of 2.88 (95%CI, 1.22-6.78; P = .02). CONCLUSIONS AND RELEVANCE A low treatment failure rate occurred in patients with TB uveitis treated with ATT. Phenotypes and test results are studied whereby patients with panuveitis having vitreous and choroidal involvement had a higher risk of treatment failure. These findings are limited by retrospectivemethods. A prospectively derived composite clinical risk score might address this diagnostic uncertainty through holistic and standardized assessment of the combinations of clinical features and investigation results that may warrant diagnosis of TB uveitis and treatment with ATT

A survey of the European Reference Network EpiCARE on clinical practice for selected rare epilepsies

Objective: Clinical care of rare and complex epilepsies is challenging, because evidence-based treatment guidelines are scarce, the experience of many physicians is limited, and interdisciplinary treatment of comorbidities is required. The pathomechanisms of rare epilepsies are, however, increasingly understood, which potentially fosters novel targeted therapies. The objectives of our survey were to obtain an overview of the clinical practice in European tertiary epilepsy centers treating patients with 5 arbitrarily selected rare epilepsies and to get an estimate of potentially available patients for future studies. Methods: Members of the European Reference Network for rare and complex epilepsies (EpiCARE) were invited to participate in a web-based survey on clinical practice of patients with Dravet syndrome, tuberous sclerosis complex (TSC), autoimmune encephalitis, and progressive myoclonic epilepsies including Unverricht Lundborg and Unverricht-like diseases. A consensus-based questionnaire was generated for each disease. Results: Twenty-six of 30 invited epilepsy centers participated. Cohorts were present in most responding centers for TSC (87%), Dravet syndrome (85%), and autoimmune encephalitis (71%). Patients with TSC and Dravet syndrome represented the largest cohorts in these centers. The antiseizure drug treatments were rather consistent across the centers especially with regard to Dravet syndrome, infantile spasms in TSC, and Unverricht Lundborg / Unverricht-like disease. Available, widely used targeted therapies included everolimus in TSC and immunosuppressive therapies in autoimmune encephalitis. Screening for comorbidities was routinely done, but specific treatment protocols were lacking in most centers. Significance: The survey summarizes the current clinical practice for selected rare epilepsies in tertiary European epilepsy centers and demonstrates consistency as well as heterogeneity in the treatment, underscoring the need for controlled trials and recommendations. The survey also provides estimates for potential participants of clinical trials recruited via EpiCARE, emphasizing the great potential of Reference Networks for future studies to evaluate new targeted therapies and to identify novel biomarkers