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Glioblastoma (GB) is one of the most lethal types of neoplasms. Its biologically aggressive nature and the presence of the blood-brain barrier (BBB) limit the efficacy of standard therapies. Several strategies are currently being developed to both overcome the BBB and deliver drugs site-specifically to tumor cells. This work hypothesizes a two‑pronged approach to tackle glioblastoma: drug repurposing with celecoxib and a nanoformulation using ultra-small nanostructured lipid carriers (usNLCs). Celecoxib antitumor druggable activity was inspected bioinformatically and screened in four glioma cell lines aiming at the comparison with temozolomide, as standard of care. Delving into formulation design, it was tailored aiming at (i) improving the drug solubility/loading properties, (ii) assigning a thermal-triggerable drug release based on a lipid matrix with a low melting point, and (iii) enhancing the cytotoxic effect by selecting a template targetable to tumor cells. For this purpose, an integrated analysis of the critical material attributes (CMAs), critical process parameters (CPPs) and critical quality attributes (CQAs) was conducted under the umbrella of a Quality by Design approach. CMAs that demonstrate a high-risk level for the final quality and performance of the usNLCs include the drug solubility in lipids (solid and liquid), the lipid composition (envisioning a thermoresponsive approach), the ratio between lipids (solid vs. liquid), and the surfactant type and concentration. Particle size was shown to be governed by the interaction lipid-surfactant followed by surfactant type. The drug encapsulation did not influence colloidal characteristics, making it a promising carrier for lipophilic drugs. In general,usNLCs exhibited a controlled drug release during the 72 hours at 37 ºC with a final release of ca. 25 %, while at 45 ºC this was doubled. The in vitro cellular performance depended on the surfactant type and lipid composition, with the formulations containing a sole solid lipid (Suppocire® NB) and Kolliphor® RH40 as surfactant being the most cytotoxic. usNLCs with an average diameter of ca. 70 nm and a narrow size distribution (PdI lower than 0.2) were yielded, exhibiting, high stability, drug protection, sustained and thermo‑sensitive release properties, and high cytotoxicity to glioma cells, meeting the suitable CQAs for parenteral administration. This formulation may pave the way to a multi-addressable purpose to improve address GB treatment.

ObjectiveTo investigate the perceptions of young people and adults, smokers and non-smokers about the current set of innovations introduced in 2018 into the Brazilian tobacco products’ health warnings.MethodsTwenty focus groups were conducted in five state capitals in Brazil. The participants (n=163) were segmented by smoking status, age (15–17 years, 18–55 years) and social grade (C, D–E classes) to examine cigarette packaging and explore the participants’ perceptions of health warnings.ResultsHealth warnings capture attention, eliciting apprehension, fear, disgust and concern about the negative consequences of cigarette consumption. The 2018 Brazil health warnings are spontaneously recalled by participants, even without the presence of cigarette packages. However, the analysis also reveals the challenges of overcoming communication barriers and distorted interpretations, especially among smokers. The inclusion of direct and provocative stimuli, such as the use of the word ‘you’, attracts attention and creates more proximity to the recipient of the message. The results also highlight the interest and fear elicited by warnings on toxic constituents and the importance of using contrasting colours in warnings, which differentiate them from the colours of cigarette packs.ConclusionIntroducing innovative components in health warnings can catch consumers’ attention but considering that the interviewees encountered difficulties interpreting textual warnings about toxic constituents in cigarettes, the study reinforces the importance of adopting direct language and pictures, instead of text, which can visually transmit the warning messages and the use of specific wording that generates proximity between the emitter and receiver.

Background and Aims: Colorectal cancer (CRC) is a heterogeneous disease with distinctive genetic pathways, such as chromosomal instability, microsatellite instability and methylator pathway. Our aim was to correlate clinical and genetic characteristics of CRC patients in order to understand clinical implications of tumour genotype. Methods: Single-institution retrospective cohort of patients who underwent curative surgery for CRC, from 2012 to 2014. RAS and BRAF mutations were evaluated with the real-time PCR technique Idylla®. Mismatch repair deficiency (dMMR) was characterized by absence of MLH1, MSH6, MSH2 and/or PMS2 expression, evaluated by tissue microarrays. Overall survival (OS) and disease-free survival (DFS) were assessed using survival analysis. Results: Overall, 242 patients were included (males 57.4%, age 69.3 ± 12.9 years; median follow-up 49 months). RAS-mutated tumours were associated with reduced DFS (p = 0.02) and OS (p = 0.045) in stage I–III CRC. BRAF-mutated tumours were more predominant in females and in the right colon, similarly to dMMR tumours. BRAF status did not influence OS (4 years)/DFS (3.5 years) in stage I–III disease. However, after relapse, length of survival was 3.5 months in BRAF-mutated tumours in contrast to 18.6 months in BRAF wild-type tumours (p = NS). No germline mutations in mismatch repair genes were so far identified in the patients with dMMR tumours. Molecular phenotype (RAS, BRAF and MMR) did not influence OS in metastatic patients. Our small sample size may be a limitation of the study. Conclusion: In our cohort, RAS-mutated tumours were associated with worse DFS and OS in early-stage CRC, whereas the remaining molecular variables had no prognostic influence.

Background: The rate of diabetic ketoacidosis in new-onset type 1 diabetes mellitus depends on multiple factors and is very heterogeneous between countries. An inverse relationship between the incidences of T1D and DKA seems to exist. Moreover, DKA is reported to be more common in children under 5 years old. Current data on Portuguese pediatric DKA incidence in new-onset T1D is limited. We aimed to determine the rate of DKA incidence in this population, as well as risk and protective factors.Methods: Review of data from children aged 18 years or younger with new onset T1D, referred to a pediatric endocrinology unit between January 1st, 2013 and December 31th, 2020 (8 years).Results: 276 patients were included with a median age of 9.6 years-old (0.5-17.9 years): 15,6% aged between 2 and 5 years-old and 3,6% under two years. A mean incidence of 34,5 new cases per year was observed, with an upward trend. One hundred and five (38%) presented with DKA, which was considered severe in 9,4%; 50 patients (18,1%) presented only with hyperglycemia. Non-DKA at presentation was associated with family history of T1D (p=0.016). DKA at presentation was more frequent in the age group under 2 years old (p=0.005).Conclusions: DKA’s frequency at new onset T1D is still high, although only a small proportion of cases was considered severe. A family history of T1D correlated with a non-DKA presentation, while an age under 2 years presented as a risk factor for DKA.

This chapter examines the engagement of social work academics in the policy process in Portugal. It begins by presenting an overview of social policy and the historical development of the Portuguese welfare state and by discussing the emergence of the social work profession in that country. The development of social work education in Portugal and its contemporary features are depicted. Following these, the methodology and the findings of a study of the policy engagement of Portuguese social work academics are then presented. The findings relate to the levels of engagement in policy and the forms that this takes. The study also offers insights into various factors that are associated with these, such as perceptions, capabilities, institutional support and the accessibility of the policy process. The chapter concludes with an analysis of the findings and their implications.

Introduction: Febrile neutropenia (FN) is a potentially life-threatening complication of systemic chemotherapy (CT) that often requires hospital admission. Delay in diagnosis and treatment are associated with higher morbidity and mortality. Objective: We aimed to determine the factors that influence FN episodes outcomes in the emergency room (ER). Methods: This was a retrospective study of all FN episodes (with a collected blood culture [BC]) that occurred between 2012 and 2016 at our institution. FN was defined as a temperature ≥38°C and an absolute neutrophil count (ANC) Results: Between 2012 and 2016, there were 173 FN episodes in 153/1,947 patients treated with intravenous CT. Most of these episodes (n = 121, 70%) were diagnosed in the ER, 29 in the outpatient clinic, and 23 as inpatients. In the ER, the median time was 36 min from hospital nurse triage to medical observation, and 52 min from medical observation to complete blood count specimen collection. There was a positive BC in 33 FN episodes, 72% with Gram-negative bacteria. A total of 160 FN episodes led to hospital admission and 13 were treated as outpatients. Mortality associated with the FN episode was 15% and an ANC Conclusion: This study confirms that FN is a serious and common complication of IV CT which must be diagnosed and treated promptly. Profound neutropenia was the only predictive factor of mortality.

A 56-year-old man with good performance status was diagnosed with plurimetastatic colon adenocarcinoma 1 year before. He is currently under second-line chemotherapy with 5-FU+oxaliplatin (FOLFOX) and bevacizumab. Due to a catheter-related thrombosis, the subclavian vein central venous

In cystic fibrosis (CF), respiratory failure caused by progressive airway obstruction and tissue damage is primarily a result of the aberrant inflammatory responses to lung infections with Pseudomonas aeruginosa. Despite considerable improvement in patient survival, conventional therapies are mainly supportive. Recent progress toward gene therapy for CF has been encouraging; however, several factors such as immune response and transduced cell turnover remain as potential limitations to CF gene therapy. As alternative gene therapy vectors for CF, we examined the feasibility of using recombinant SV40-derived vectors (rSV40s), which may circumvent some of these obstacles. To accommodate the large cystic fibrosis transmembrane conductance regulator (CFTR) cDNA, we removed not only SV40 Tag genes, but also all capsid genes. We, therefore, tested whether 'gutless' rSV40s could be packaged and were able to express a functional human CFTR cDNA. The results from our in vitro analysis determined that rSV40-CFTR was able to successfully result in the expression of CFTR protein, which localized to the plasma membrane and restored channel function to CFTR-deficient cells. Similarly, in vivo experiments delivering rSV40-CFTR to the lungs of Cftr-/- mice resulted in a reduction of the pathology associated with intra-tracheal P. aeruginosa challenge. rSV40-CFTR-treated mice had less weight loss when compared with control-treated mice as well as demonstrably reduced lung inflammation as evidence by histology and reduced inflammatory cytokines in the broncho-alveolar lavage. The reduction in inflammatory cytokine levels led to an evident decrease in neutrophil influx to the airways. These results indicate that further study of the application of rSV40-CFTR to CF gene therapy is warranted.

Many applications of gene delivery require long-term transgene expression. In dividing cells, this result necessitates vector genome persistence, usually by integrating into cellular DNA. Since recombinant gene delivery vectors derived from tag-deleted, replication-incompetent simian virus-40 (SV40) provide for long-term transgene expression in resting and dividing cells, we tested whether such enduring transgene expression reflected integration into cellular genomes. Several lines of evidence suggested this likelihood. After transduction in vitro, continuously dividing cell lines and continuously stimulated primary cells uniformly showed transgene expression for many months. Mice whose livers were transduced in vivo, partially resected, and allowed to regenerate showed comparable levels of transgene expression in regenerated and preoperative livers. Thus, replicationincompetent SV40 vectors (rSV40) persist in vitro and in vivo despite extensive cell division. We tested the possibility that this persistence reflected integration directly. Southern blot analyses of genomic DNA from transduced 293 cells showed that vector genome incorporation into cell DNA happened within days of transduction. Episomal vector DNA was barely detectable 96 hours post-transduction. Inverted PCR, used to characterize vector integration points, showed vector DNA integrated randomly into the cell genome. The circular rSV40 genome opened at different points in each integrand. A significant proportion of the integrands did not contain the entire vector sequence, but rather only portions thereof. Quantitative Southern blot analysis showed approximately 3.05 transgene copies per cell. Therefore, recombinant SV40 gene delivery vectors integrate into the cellular DNA of both resting and dividing cells, and do so randomly and within days of transduction. This integration may explain long-term transgene expression.

Compared with single agents, combination antilentiviral pharmacotherapy targets multiple HIV-1 functions simultaneously, maximizing efficacy and decreasing chances of escape mutations. Combination genetic therapy could theoretically enhance efficacy similarly, but delivery of even single genes to high percentages of hematopoietic cells or their derivatives has proven problematic. Because of their high efficiency of gene delivery, we tested recombinant SV40-derived vectors (rSV40s) for this purpose. We made six rSV40s, each carrying a different transgene that targeted a different lentiviral function. We tested the ability of these constructs, individually and in double and triple combinations, to protect SupT1 human T lymphoma cells from HIV-1 challenge. Single chain antibodies (SFv) against CXCR4 and against HIV-1 reverse transcriptase (RT) and integrase (IN) were used, as were polymeric TAR decoys (PolyTAR) and a dominant-negative mutant of HIV-1 Rev (RevM10). Immunostaining showed that virtually all doubly treated cells expressed both transgenes. All transgenes individually protected from HIV-1 but, except for anti-CXCR4 SFv, their effectiveness diminished as challenge doses increased from 40 through 2500 tissue culture infectious dose(50) (TCID(50))/10(6) cells. However, all combinations of transgenes protected target cells better than individual transgenes, even from the highest challenge doses. Thus, combination gene therapies may inhibit HIV-1 better than single agents, and rSV40s may facilitate delivery of multigene therapeutics.

TCH codes are a class of binary nonlinear cyclic codes that have been used in several systems (e.g. mobile satellite, Ultra-Wideband) for tasks like coding and synchronization. The original generating method of this class of codes restricts the length (or period) of the codewords to the set {2, 4, 16, 256, 65536}. In this paper we use an alternative generation method based on the algebraic properties of Zech logarithms that allow us to generalize the construction in a systematic manner and improving the codeword length choices, code size and minimum distance. The performance of these generalized codewords are then evaluated using both AWGN and Rayleigh communication channels and qualitatively compared with the originally extended TCH codewords.

This paper presents an extension of the ASM (Accelerated Simulation Method) to be able to evaluate the performance of concatenated coding schemes, so as to get similar results to the MCSM (Monte Carlo Simulation Method) more efficiently, i.e. using less time and computational resources. The efficiency of the ASM is particularly useful to estimate the performance of complex or unknown coding schemes, so this optimised method is applied to estimate the performance of various TCH (Tomlinson, Cercas, Hughes) codes concatenated with RS (Reed-Solomon) codes, namely, TCH(16,6,2) with RS(63,45,9) and TCH(32,8,6) with RS(255,223,16). Both SD (Soft-Decision) and HD (Hard-Decision) decoding are considered and the overall performance takes into account modulation and channel conditions.

Experiments were undertaken to analyze proviral DNA in primary (wing web) and visceral sarcomas arising in FP chickens infected with BH-RSV(RAV-2). Using the degree of heterogeneity of BH-RSV proviral integration sites as a measure of the degree of polyclonality of sarcoma tissue, we observed that a high proportion of the visceral sarcomas examined comprised dominant clones, independently of whether these sarcomas were isolated from immune-suppressed or nonsuppressed infected chickens; by contrast, a marked heterogeneity of BH-RSV proviral integration sites was noted with primary sarcoma tissue. Several visceral sarcomas containing dominant clones were characterized by the integration of a deleted form of the BH-RSV provirus. In addition, all of the primary and visceral sarcomas exhibited sequences specific for the RAV-2 provirus, and both types of sarcoma tissue were competent for infectious sarcoma virus production.

Sarcomas arising in FP chickens infected with subgroup B avian sarcoma virus at increasing times posthatch were compared for their capacity to mediate recruitment of new sarcoma cells by infectious virus production. This comparison was premised on the observation that primary sarcoma growth is much slower when older chickens are infected with avian sarcoma virus. As measured in a transplantation assay, no age-related diminution in the recruitment capacity of sarcoma tissue from virus-infected transplant donors was observed, despite the marked age-related differences in their patterns of primary sarcoma growth. These observations establish that a diminished recruitment capacity is not required for slow primary sarcoma growth. The capacity to induce the formation of sarcomas at sites distal to the primary sarcoma was correlated with a strong recruitment capacity.

1. A case of essential osteolysis, progressing for thirteen years and involving the right femur and pelvis of a boy who was thirteen years old at the time of onset, is presented. 2. The histological appearance of fragments of bone and soft tissue removed from the affected thigh is described. 3. No evidence that might throw light on the etiology of the disease was obtained from biochemical investigations or from the histological studies. 4. None of the many lines of treatment that were tried had any effect upon the progress of the disease.

Tese de mestrado em Ecologia e Gestão Ambiental, Universidade de Lisboa, Faculdade de Ciências, 2021 Submitted by Cristina Manessiez (camanessiez@fc.ul.pt) on 2021-06-15T16:46:24Z No. of bitstreams: 1 TM_Francisco_Basilio.pdf: 1555227 bytes, checksum: e857c5bc48b386cde5b0dfa1bb22f099 (MD5) Made available in DSpace on 2021-06-15T16:46:33Z (GMT). No. of bitstreams: 1 TM_Francisco_Basilio.pdf: 1555227 bytes, checksum: e857c5bc48b386cde5b0dfa1bb22f099 (MD5) Previous issue date: 2021