Blood and cerebellar abundance of ATXN3 splice variants in spinocerebellar ataxia type 3/Machado-Joseph disease.

Spinocerebellar ataxia type 3 (SCA3)/Machado-Joseph disease (MJD) is a heritable proteinopathy disorder, whose causative gene, ATXN3, undergoes alternative splicing. Ataxin-3 protein isoforms differ in their toxicity, suggesting that certain ATXN3 splice variants may be crucial in driving the selective toxicity in SCA3. Using RNA-seq datasets we identified and determined the abundance of annotated ATXN3 transcripts in blood (n = 60) and cerebellum (n = 12) of SCA3 subjects and controls. The reference transcript (ATXN3-251), translating into an ataxin-3 isoform harbouring three ubiquitin-interacting motifs (UIMs), showed the highest abundance in blood, while the most abundant transcript in the cerebellum (ATXN3-208) was of unclear function. Noteworthy, two of the four transcripts that encode full-length ataxin-3 isoforms but differ in the C-terminus were strongly related with tissue expression specificity: ATXN3-251 (3UIM) was expressed in blood 50-fold more than in the cerebellum, whereas ATXN3-214 (2UIM) was expressed in the cerebellum 20-fold more than in the blood. These findings shed light on ATXN3 alternative splicing, aiding in the comprehension of SCA3 pathogenesis and providing guidance in the design of future ATXN3 mRNA-lowering therapies.

Blood transcriptome sequencing identifies biomarkers able to track disease stages in spinocerebellar ataxia type 3.

Transcriptional dysregulation has been described in spinocerebellar ataxia type 3/Machado-Joseph disease (SCA3/MJD), an autosomal dominant ataxia caused by a polyglutamine expansion in the ataxin-3 protein. As ataxin-3 is ubiquitously expressed, transcriptional alterations in blood may reflect early changes that start before clinical onset and might serve as peripheral biomarkers in clinical and research settings. Our goal was to describe enriched pathways and report dysregulated genes, which can track disease onset, severity or progression in carriers of the ATXN3 mutation (pre-ataxic subjects and patients). Global dysregulation patterns were identified by RNA sequencing of blood samples from 40 carriers of ATXN3 mutation and 20 controls and further compared with transcriptomic data from post-mortem cerebellum samples of MJD patients and controls. Ten genes-ABCA1, CEP72, PTGDS, SAFB2, SFSWAP, CCDC88C, SH2B1, LTBP4, MEG3 and TSPOAP1-whose expression in blood was altered in the pre-ataxic stage and simultaneously, correlated with ataxia severity in the overt disease stage, were analysed by quantitative real-time PCR in blood samples from an independent set of 170 SCA3/MJD subjects and 57 controls. Pathway enrichment analysis indicated the Gαi signalling and the oestrogen receptor signalling to be similarly affected in blood and cerebellum. SAFB2, SFSWAP and LTBP4 were consistently dysregulated in pre-ataxic subjects compared to controls, displaying a combined discriminatory ability of 79%. In patients, ataxia severity was associated with higher levels of MEG3 and TSPOAP1. We propose expression levels of SAFB2, SFSWAP and LTBP4 as well as MEG3 and TSPOAP1 as stratification markers of SCA3/MJD progression, deserving further validation in longitudinal studies and in independent cohorts.

Multifractal fluctuations in zebrafish (Danio rerio) polarization time series.

In this work, we study the polarization time series obtained from experimental observation of a group of zebrafish (Danio rerio) confined in a circular tank. The complex dynamics of the individual trajectory evolution lead to the appearance of multiple characteristic scales. Employing the Multifractal Detrended Fluctuation Analysis (MF-DFA), we found distinct behaviors according to the parameters used. The polarization time series are multifractal at low fish densities and their average scales with ρ - 1 / 4 . On the other hand, they tend to be monofractal, and their average scales with ρ - 1 / 2 for high fish densities. These two regimes overlap at critical density ρ c , suggesting the existence of a phase transition separating them. We also observed that for low densities, the polarization velocity shows a non-Gaussian behavior with heavy tails associated with long-range correlation and becomes Gaussian for high densities, presenting an uncorrelated regime.

Thermal stress and comfort assessment in urban areas using Copernicus Climate Change Service Era 5 reanalysis and collected microclimatic data.

In this initial study of a research project, this paper seeks to understand the thermal conditions in the cities of Lisbon and Munich, specifically focusing on Urban Heat Island intensity and on thermal comfort using the Universal Thermal Climate Index modeling data at the Local Climate Zone scale. Based on these datasets, Munich has exhibited more unfavourable thermal conditions than Lisbon. In terms of UHII, both cities have shown that low, medium, and high rise compact urban areas and bare rock or paved areas have the highest values, while sparsely built areas have the lowest. These results differ from the UTCI, which indicates that in Lisbon and Munich, these sparsely built areas as well as areas with low plants and vegetation are the most uncomfortable. In Munich, the population was exposed to very strong heat stress, while Lisbon experienced strong heat stress conditions. Conversely, low, medium, and high rise compact urban areas and densely wooded areas in Munich, and scattered trees areas and large low-rise urban areas in Lisbon, have demonstrated the lowest monthly mean and average maximum values. These results will be further explored in future studies in the city of Lisbon and cross-checked with data obtained from roving missions. This will enable a more detailed temporal and local analysis.

Novel Machado-Joseph disease-modifying genes and pathways identified by whole-exome sequencing.

Machado-Joseph disease (MJD/SCA3) is a neurodegenerative polyglutamine disorder exhibiting a wide spectrum of phenotypes. The abnormal size of the (CAG)n at ATXN3 explains ~55% of the age at onset variance, suggesting the involvement of other factors, namely genetic modifiers, whose identification remains limited. Our aim was to find novel genetic modifiers, analyse their epistatic effects and identify disease-modifying pathways contributing to MJD variable expressivity. We performed whole-exome sequencing in a discovery sample of four age at onset concordant and four discordant first-degree relative pairs of Azorean patients, to identify candidate variants which genotypes differed for each discordant pair but were shared in each concordant pair. Variants identified by this approach were then tested in an independent multi-origin cohort of 282 MJD patients. Whole-exome sequencing identified 233 candidate variants, from which 82 variants in 53 genes were prioritized for downstream analysis. Eighteen disease-modifying pathways were identified; two of the most enriched pathways were relevant for the nervous system, namely the neuregulin signaling and the agrin interactions at neuromuscular junction. Variants at PARD3, NFKB1, CHD5, ACTG1, CFAP57, DLGAP2, ITGB1, DIDO1 and CERS4 modulate age at onset in MJD, with those identified in CFAP57, ACTG1 and DIDO1 showing consistent effects across cohorts of different geographical origins. Network analyses of the nine novel MJD modifiers highlighted several important molecular interactions, including genes/proteins previously related with MJD pathogenesis, namely between ACTG1/APOE and VCP/ITGB1. We describe novel pathways, modifiers, and their interaction partners, providing a broad molecular portrait of age at onset modulation to be further exploited as new disease-modifying targets for MJD and related diseases.

Prospective evaluation of immune haemolysis in liver transplantation.

BACKGROUND: Immune haemolysis in liver transplant (LT) can occur due to autoantibodies and alloantibodies. The aim of this study was to evaluate the prevalence and risk factors for immune haemolysis in LT. METHODS: A total of 175 consecutive patients were included. Multiorgan recipients were excluded. Samples, from before LT, seven consecutive days and weekly for 4 weeks, were evaluated for haemolysis and immunohaematological tests. SPSS 24 was used for statistical analysis. RESULTS: Nine patients (5·1%) presented positive antibody screen (AS) before LT, (2·3% clinically significant), more frequent in RhD-negative (P = 0·017). Positive DAT occurred in 53 (30·3%) and was related to high MELD score (P = 0·048), HCV (P = 0·005) and furosemide use (P = 0·001). Positive AS after LT occurred in 22 patients (12·5%), with nine (5·7%) clinically significant antibodies. Positive AS occurred more frequently in RhD negative (P = 0·021) and in those transfused (P = 0·022). Post-transplant positive DAT was associated with piperacillin-tazobactam use (P = 0·021) and minor ABO incompatibility (P = 0·0038). Five patients presented passenger lymphocyte syndrome (PLS), all received liver-graft O, four presented haemolysis, and three were transfused due to PLS. CONCLUSION: Auto- and alloantibodies against red blood cell antigens are frequent in LT, but the frequency of immune haemolysis was only 2·8%. The only risk factor for PLS was minor ABO mismatch.

Excess winter mortality and morbidity before, during, and after the Great Recession: the Portuguese case.

Although winter mortality and morbidity are phenomena common to most European countries, their magnitude varies significantly from country to country. The geographical disparities among regions with similar climates are the result of several social, economic, demographic, and biological conditions that influence an individual's vulnerability to winter conditions. The impact of poor socioeconomic conditions may be of such magnitude that an economic recession may aggravate the seasonal mortality pattern. This paper aims to measure the seasonal winter mortality, morbidity, and their related costs during the Great Recession (2009-2012) in mainland Portugal and its Regional Health Administrations (RHAs) and to compare it with the periods preceding and following it. Monthly mortality and morbidity data were collected and clustered into three periods: Great Recession (2009-2012), Pre-Recession (2005-2008), and Post-Recession (2013-2016). The impact of seasonal winter mortality and morbidity during the Great Recession in Portugal and its Regional Health Administrations was measured through the assessment of age-standardized excess winter (EW) death and hospital admissions rate and index, expected life expectancy gains without EW deaths, EW rate of potential years of life lost, and EW rate of emergency hospital admission costs. Important increases of winter deaths and hospital admissions were identified, resulting in an important number of potential years of life lost (87 years of life lost per 100,000 inhabitants in 2009-2012), life expectancy loss (1 year in 2009-2012), and National Health Service costs with explicit temporal and spatial variations. These human and economic costs have decreased consistently during the analyzed periods, while no significant increase was found during the Great Recession. Despite its reduction, the winter excess morbidity and mortality highlight that Portugal still faces substantial challenges related to a highly vulnerable population, calling for investments in better social and health protection.

[Benign Presentation of a Potentially Fatal Disease]. / Ateroembolismo cutâneo: relato de três casos clínicos.

Atheroembolism is a rare multisystemic disorder that is characterized by release of cholesterol crystals and particles from atheromatous plaques, which can occlude distal vessels and induce an inflammatory response. Most affected individuals are males, older than 60 years of age, with advanced atherosclerotic disease. The abdominal aorta is the most common origin of cholesterol emboli, being the peripheral arteries a rarer source. Cholesterol embolization syndrome is often associated with invasive vascular procedures, although, more rarely, it may occur spontaneously. In this paper, the authors present three cases of spontaneous atheroembolism with cutaneous manifestations and their clinical management. Being an underdiagnosed pathology, knowledge about its clinical manifestations is essential in order to allow an early diagnosis and treatment, to ensure a better prognosis for the patient.

O ateroembolismo é uma doença multissistémica rara caraterizada pela libertação de cristais de colesterol e partículas de placas ateroscleróticas, que podem ocluir vasos sanguíneos periféricos e induzir uma resposta inflamatória. A maioria dos indivíduos afetados é do sexo masculino, com idade superior a 60 anos e doença aterosclerótica avançada. A origem mais frequente de embolização de colesterol é a aorta abdominal, sendo as artérias periféricas uma fonte mais rara. A síndrome de embolização por colesterol surge frequentemente associada a procedimentos vasculares invasivos, embora, mais raramente, possa ocorrer de forma espontânea. Neste artigo os autores apresentam três casos clínicos de ateroembolismo espontâneo com envolvimento cutâneo e respetiva abordagem clínica. Sendo uma patologia subdiagnosticada, torna-se fundamental o conhecimento acerca das suas manifestações clínicas, para permitir um diagnóstico e tratamento precoces de forma a garantir um melhor prognóstico para o doente.

The influence of the winter North Atlantic Oscillation index on hospital admissions through diseases of the circulatory system in Lisbon, Portugal.

The aim of this paper is to analyze the relationship between North Atlantic Oscillation (NAO), meteorological variables, air pollutants, and hospital admissions due to diseases of circulatory systems in Lisbon (Portugal) during winter months (2003-2012). This paper is one of the few studies analyzing the impact of NAO on health through its influence on thermal stress and air pollution and is the first to be conducted in Lisbon. This study uses meteorological data (synthetized into a thermal comfort index), air pollutant metrics, and the NAO index (all clustered in 10-day cycles to overcome daily variability of the NAO index). The relationship between morbidity, thermal comfort index, NAO index, and air pollutants was explored through several linear models adjusted to seasonality through a periodic function. The possible indirect effect between the NAO index and hospital admissions was tested, assuming that NAO (independent variable) is affecting hospital admissions (outcome variable) through thermal discomfort and/or pollution levels (tested as individual mediators). This test was conducted through causal mediation analysis and adjusted for seasonal variation. The results from this study suggest a possible indirect relationship between NAO index and hospital admissions. Although NAO is not significantly associated with hospital admissions, it is significantly associated with CO, PM2.5, NO, and SO2 levels, which in turn increase the probability of hospitalization. The discomfort index (built with temperature and relative humidity) is significantly associated with hospital admissions, but its variability is not explained by the NAO index. This study highlights the impacts of the atmospheric circulation patterns on health. Furthermore, understanding the influence of the atmospheric circulation patterns can support the improvement of the existing contingency plans.

Deep Vein Thrombosis as First Manifestation of Hibernoma - A Case Report.

INTRODUCTION: Hibernoma is a very rare benign tumor that arises from vestigial remnants of fetal brown adipose cells and usually manifests as a slowly growing, painless soft-tissue mass. It mainly occurs in adults, in the third and fourth decade of life, slightly more in women and is commonly seen in the subcutaneous regions of the back, neck, thighs and retroperitoneum. It was originally described in 1906 by Merkel, who named it "pseudolipoma". In 1914, Gery derived the name hibernoma from the tumor's histological similarity to brown fat in hibernating animals. A hibernoma may be confused with a lipoma clinically and cannot be completely distinguished from hypervascular lesions such as lipossarcoma. METHODS: A 36-year-old woman presented with pain and edema of the left leg. It was diagnosed with non-recent femoro- -popliteal venous thrombosis, was medicated with rivaroxaban and prescribed compression stocking. RESULTS: The edema subsided after 2 weeks but she still complained of pain in the thigh several weeks after first visit along with subtle localized soft enlargement in the upper thigh. It was requested a CT scan that showed a nodular image with 60X 47 mm medially to vastus intermedius and beneath the sartorius and rectus femoris muscles, which was suspected to be a lipossarcoma. In this context, a magnetic ressonance imaging was requested and showed contact with femoral vessels with no cleavage plan, suspected to be a mixoid lipossarcoma. The biopsy didn't show malignancy. She was operated with local excision of the mass and preservation of adjacent structures. Pathologic evaluation revealed a hibernoma with 11.5 cm, PS100 positive and MDM2 negative. The patient was evaluated at outpatient clinic 6 months after surgery and had no evidence of relapse. CONCLUSION: The first clinical manifestation of this patient was a deep vein thrombosis and the diagnosis of the lipomatous tumor was delayed. Clinical awareness of less frequent causes of DVT is a key point to timely detection of this lesions that are rare and curable. The increased vascularity of this lesion raised suspicion of malignancy. Malignancy potential is perhaps the most difficult aspect to ascertain in this patient, being only completely disclosed after surgical excision. Optimal treatment is complete surgical resection. Local recurrence does not occur with complete excision. No reports of metastases of malignant transformation have been identified in the reviewed literature.

[Endovascular Revascularization for Diabetic Foot Ulcer - Experience from a Dedicated Clinic]. / Revascularização Endovascular na Úlcera de Pé Diabético ­ experiência de uma Consulta Multidisciplinar.

OBJECTIVES: We aimed to assess the outcome of endovascular revascularization in Diabetic patients with Fontaine stage IV chronic ischaemia, at our Diabetic Foot Clinic. Primary outcomes were ulcer healing, major amputation and limb salvage. METHODS: Retrospective single center analysis of patients treated between January 2009 and May 2015. Time-dependent event rates were estimated by the Kaplan-Meier method. The differences between groups were evaluated with the chi-square test. A P value below 0,05 was considered statistically significant. RESULTS: There were 106 limbs (93 patients) revascularized during the study. The average age was 71 years, and PEDIS 3 or 4 infection was present in 45.3% on admission. 56.6% were treated for femoropopliteal injurie only, 17% infrapopliteal, and 26.4% for both levels. Technical success was achieved in 77.4%, and haemodynamic success in 58.3%. Ulcer healing was attained in 53.8%, with an average healing time of 8.4 months. The major amputation rate was 7.5%, with a limb salvage rate of 90.4% at 6, 12 and 24 months. Chronic pulmo- nary disease decreased the likelihood of healing (p=0,012). Restenosis was more likely to occur in patients with nephropathy (p=0,008) or insulin-treated diabetes (p=0,033). CONCLUSIONS: Dedicated multidisciplinary teams are key to successful treatment in diabetic foot disease. The good results achieved in our series arise from best medical treatment combined with timely revascularization in those ischemic.

Objetivos: Avaliar os resultados da revascularização endovascular em diabéticos com isquemia crónica grau IV de Lériche- -Fontaine, seguidos em Consulta Multidisciplinar de Pé Diabético na nossa instituição, relativamente a taxa de cicatrização, amputação major e salvamento de membro. Material e Métodos: Análise retrospetiva dos doentes tratados no período de janeiro de 2009 até maio de 2015. As taxas de eventos dependentes do tempo foram estimadas com recurso a curvas de Kaplan-Meier e as diferenças entre grupos investigadas pelo teste de qui-quadrado. Um valor de p < 0,05 foi considerado estatisticamente significativo. Resultados: No período analisado foram revascularizados 106 membros (93 doentes). A idade média foi 71 anos, sendo que 45,3% apresentavam infeção PEDIS 3 ou 4 na apresentação. Das lesões intervencionadas, 56,6% eram suprageniculares, 17% infrageniculares, e 26,4% foram intervencionados nos dois setores. Foi obtido sucesso técnico em 77,4% e sucesso hemodinâmico em 58,3%. Foi conseguido o encerramento da úlcera em 53,8%, com tempo médio de cicatrização de 8,4 meses. A taxa de amputação major foi de 7,5%, verificando-se uma taxa de salvamento de membro de 90,4% aos 6, 12 e 24 meses. A doença pulmonar crónica influenciou negativamente a probabilidade de cicatrização (p=0,012). A reestenose foi mais frequente em doentes com nefropatia (p=0,008) ou DM insulino-tratada (p=0,033). Conclusões: A existência de equipas multidisciplinares dedicadas ao Pé Diabético são fundamentais para o sucesso do tratamento. Os bons resultados obtidos resultam da melhor otimização do tratamento médico combinada com a revasculari- zação no doente isquémico, associado a um seguimento apertado e à reintervenção precoce quando clinicamente justificado.

[Deep vein thrombosis in the pediatric population - a rare entity of uncertain prognosis]. / Trombose Venosa Profunda na População Pediátrica ­ Entidade Rara de Prognóstico Incerto.

Deep vein thrombosis, whose prevalence remains unknown in the pediatric population, is an increasingly common diag- nosis in this age group. This increased incidence makes the post-thrombotic syndrome a likely long-term complication, so there is a critical need to establish high quality evidence over its suitable approach in this population. We present two cases of deep vein thrombosis in children and their management in a hospital of the northern region of Portugal, reviewing the literature on this under researched subject. In conclusion, further investigation is needed to assess the incidence, the prevalence, the predictors and the safety and efficacy of therapies for the prevention and appropriate treatment of deep vein thrombosis in children and eventual post- -thrombotic syndrome.

A trombose venosa profunda, cuja prevalência permanece desconhecida na população pediátrica, é um diagnóstico cada vez mais frequente neste grupo etário. Este aumento de incidência faz da síndrome pós-trombótica uma complicação provável a longo prazo, existindo uma necessidade crítica em estabelecer evidência de alta qualidade sobre a sua adequada abordagem nesta população. São apresentados dois casos clínicos pediátricos de trombose venosa profunda, e respetiva abordagem, num hospital da região Norte de Portugal. Em seguida, procede-se à discussão e revisão da literatura sobre o assunto. Em conclusão, são necessários estudos que avaliem a incidência, a prevalência, os fatores preditivos e a segurança e a eficácia de terapias para a prevenção e o tratamento adequados da trombose venosa profunda e da possível futura síndrome pós-trombótica associada em crianças.

Novel candidate blood-based transcriptional biomarkers of Machado-Joseph disease.

BACKGROUND: Machado-Joseph disease (or spinocerebellar ataxia type 3) is a late-onset polyglutamine neurodegenerative disorder caused by a mutation in the ATXN3 gene, which encodes for the ubiquitously expressed protein ataxin-3. Previous studies on cell and animal models have suggested that mutated ataxin-3 is involved in transcriptional dysregulation. Starting with a whole-transcriptome profiling of peripheral blood samples from patients and controls, we aimed to confirm abnormal expression profiles in Machado-Joseph disease and to identify promising up-regulated genes as potential candidate biomarkers of disease status. METHODS: The Illumina Human V4-HT12 array was used to measure transcriptome-wide gene expression in peripheral blood samples from 12 patients and 12 controls. Technical validation and validation in an independent set of samples were performed by quantitative real-time polymerase chain reaction (PCR). RESULTS: Based on the results from the microarray, twenty six genes, found to be up-regulated in patients, were selected for technical validation by quantitative real-time PCR (validation rate of 81% for the up-regulation trend). Fourteen of these were further tested in an independent set of 42 patients and 35 controls; 10 genes maintained the up-regulation trend (FCGR3B, CSR2RA, CLC, TNFSF14, SLA, P2RY13, FPR2, SELPLG, YIPF6, and GPR96); FCGR3B, P2RY13, and SELPLG were significantly up-regulated in patients when compared with controls. CONCLUSIONS: Our findings support the hypothesis that mutated ataxin-3 is associated with transcription dysregulation, detectable in peripheral blood cells. Furthermore, this is the first report suggesting a pool of up-regulated genes in Machado-Joseph disease that may have the potential to be used for fine phenotyping of this disease. © 2015 International Parkinson and Movement Disorder Society.

[An uncommon cause of lower limb oedema - Case report]. / Causa Incomum De Edema Do Membro Inferior - Caso Clínico.

The authors report the clinical case of a 50-year-old man who was observed in the emergency department because of a one week long, painless left lower limb swelling. At presentation, a gross edema was evident and peripheral pulses were present and symmetrical. The ultrasound scan findings suggested short occlusion in the transition of the common femoral (CFV) to the external iliac veins. A venography confirmed a filling defect in the above mentioned location, with normal findings throughout proximal and distal veins. On CT scans a cyst measuring 20mm of maximal diameter was disclosed, medial to the common femoral artery, compressing the CFV and seemingly continuous to the hip joint. He underwent surgical correction through open inguinal approach that allowed to confirm a complete compression of the CFV by the cyst against the inguinal ligament. The CFV recovered its normal aspect after cyst, removal and the patient was discarded home at day 2 after surgery, with complete regression of the oedema wich is maintained one year after the event, as well as absence of local ultrasound abnormalities. The hip joint is a rare location for synovial cysts. Synovial or adventitial cysts in that area may compress de CFV. The histological tests are useless for distinguishing between the two diagnosis. The clinical presentation consists of an inguinal mass or lower limb oedema due to CFV compression. Given its location underneath the vein, these masses are often difficult to detect. Vein decompression is suggested to avoid deep vein thrombosis. When a synovial cyst is present it is possible to find a dissecting plane between the two anatomical structures. Though feasible, percutaneous cyst aspiration is linked to high recurrence rates.

[Aneurysmal degeneration of a saphenous vein graft - Case report]. / Degenerescência Aneurismática De Conduto De Veia Safena - Caso Clínico.

Non-anastomotic degeneration of venous conduits is a rare condition, and its etiology is not fully understood. We report the clinical a case of an aneurysmal degeneration of a femoropopliteal bypass vein graft, undertook 19 years earlier for traumatic injury of the popliteal artery of the left lower limb. Following the resection of the aneurysm, a new femoro-tibioperoneal trunk vein graft was implanted. The choice of a new autologous graft was made based on the young age of the patient and the best outcomes in the long term, related to this conduits.

Nystagmus as an early ocular alteration in Machado-Joseph disease (MJD/SCA3).

BACKGROUND: Machado-Joseph disease (MJD), also named spinocerebellar ataxia type 3 (SCA3) is the most common autosomal dominant ataxia worldwide. Although nystagmus is one of the most frequently reported ocular alterations in MJD patients its behaviour during the course of the disease, namely in its early stages, has only recently started to be investigated. The main goal of this work was to characterize the frequency of nystagmus in symptomatic and presymptomatic carriers of the MJD mutation, and investigate its usefulness as an early indicator of the disease. METHODS: We conducted an observational study of Azorean MJD family members, comprising a total of 158 subjects which underwent neurological evaluation. Sixty eight were clinically and molecularly diagnosed with MJD, 48 were confirmed asymptomatic carriers and 42 were confirmed non-carriers of the MJD mutation. The frequency of nystagmus was calculated for the 3 groups. RESULTS: Nystagmus was present in 88% of the MJD patients. Seventeen percent of the at-risk subjects with a carrier result in the molecular test and none of the 42 individuals who received a non-carrier test result displayed nystagmus (p < 0.006). Although not reaching statistical significance, symptomatic subjects showing nystagmus had a tendency for a higher length of the CAG tract in the expanded allele, when compared to individuals who did not have nystagmus. CONCLUSIONS: The frequency of nystagmus in asymptomatic carriers and its absence in non-carriers of the mutation, suggests that nystagmus may appear before gait disturbance and can thus be considered an early sign of MJD.

Wound, Ischemia, Foot Infection (Wifi) Classification System And Its Predictive Ability Concerning Amputation-Free Survival, Mortality And Major Limb Amputation In A Portuguese Population: A Single Center Experience.

INTRODUCTION: Commonly used chronic limb-threatening ischemia (CLTI) classifications lack granularity and detail to precisely stratify patients according to risk of limb loss, expected revascularization benefit and mortality. The aim of this study is to evaluate in a Portuguese population the prognostic value of an updated CLTI classification based on Wound, Ischemia, and foot Infection (WIfI) proposed by the Society for Vascular Surgery. MATERIALS AND METHODS: Single-center retrospective evaluation of prospectively collected data of consecutive patients with CLTI submitted to lower limb revascularization from January to December of 2017. All consecutive patients with chronic peripheral artery disease with ischemic rest pain or tissue loss were included. The exclusion criteria were patients with intermittent claudication, vascular trauma, acute ischemia, non-atherosclerotic arterial disease and isolated iliac intervention. The primary end-point was major limb amputation, mortality and amputation-free survival (AFS) at 30 days, 1 year and 2 year follow-up. Secondary end-points were minor amputation, wound healing time (WHT) and rate (WHR). RESULTS: A total of 111 patients with CLTI were submitted to infra-inguinal revascularization: 91 endovascular and 20 open surgery. After categorizing them according to the WIfI: 20 had stage 1 (18.52%), 29 stage 2 (26.85%), 38 stage 3 (35.19%) and 21 stage 4 (19.44%). Overall mortality rate was 1.8%, 17% and 22.3% at 30 days, 1 year and 2 years follow-up. Major amputation rate was 0.9%, 2.7% and 2.7% at 30 days, 1 year and 2 years follow-up. AFS rate was 97.3%, 82.1%, and 76.8% at 30 days, 1 year, 2 years follow-up. In multi-variable analysis, higher WIfI score was the only predictive factor for mortality and AFS. WIfI 3 and 4 were also associated with increased risk of non-healing ulcer. CONCLUSION: This study proved the prognostic value of the WIfI classification in a Portuguese population by showing an association between higher scores and increased mortality, lower AFS and non-healing ulcer.

Clinical impact of unsuccessful subcutaneous administration of octreotide LAR instead of intramuscular administration in patients with metastatic gastroenteropancreatic neuroendocrine tumors.

Octreotide LAR is a long-acting somatostatin analogue (SSA) used in the management of metastatic gastroenteropancreatic neuroendocrine tumors (GEP NETs). It requires intramuscular (IM) injection. Missed IM injections cause subcutaneous nodules (SCNs) on radiologic images. We reviewed the rates of SCNs in a real-world cohort of GEP NETs receiving octreotide LAR and explored treatment outcomes. Patients commencing octreotide LAR between August 5, 2010 and March 8, 2018 at a single cancer center in Canada were identified from pharmacy records. Patients were included if they had a computed tomography (CT) scan performed at the time of progression and a preceding CT with pelvis included to enable assessment for the presence of nodules. Fisher's exact test was used to examine predictors of SCNs, and Kaplan-Meier curves summarized differences in progression free (PFS) and overall survival (OS) that were compared with log-rank tests. Of 243 patients receiving octreotide LAR, 45 had all required CT images available for central review. SCNs were found in 20/45 (44%) of patients on the last scan showing stable disease before progression and were numerically but not statistically more likely in females (OR: 2.36, 95% CI: 0.66-8.29, p = .23). There was an increased risk of SCNs in patients with a skin-to-muscle distance >38 mm (the length of an octreotide LAR needle) on CT (OR: 5.09, 95% CI: 1.39-16.6, p = .018) and a trend toward increased risk in obese patients (OR: 5.71, 95% CI: 1.26-23.4, p = .061). PFS (HR: 1.01, 95% CI: 0.56-1.78, p = .98) and OS (HR: 0.86, 95% CI: 0.41-1.8, p = .70) was similar between those with/without SCNs. In conclusion, almost half of patients receiving octreotide LAR had SCNs; however, missed administration of SSA did not appear to result in worse survival in this small study. Factors such as sex, younger age skin-to-muscle distance, and obesity may affect SCN development and should be considered when choosing an SSA.

Behavioral Effects of the Mixture and the Single Compounds Carbendazim, Fipronil, and Sulfentrazone on Zebrafish (Danio rerio) Larvae.

Pesticides are often detected in freshwater, but their impact on the aquatic environment is commonly studied based on single compounds, underestimating the potential additive effects of these mixtures. Even at low concentrations, pesticides can negatively affect organisms, altering important behaviors that can have repercussions at the population level. This study used a multi-behavioral approach to evaluate the effects of zebrafish larvae exposure to carbendazim (C), fipronil (F), and sulfentrazone (S), individually and mixed. Five behavioral tests, thigmotaxis, touch sensitivity, optomotor response, bouncing ball test, and larval exploratory behavior, were performed to assess potential effects on anxiety, fear, and spatial and social interaction. Significant changes were observed in the performance of larvae exposed to all compounds and their mixtures. Among the single pesticides, exposure to S produced the most behavioral alterations, followed by F and C, respectively. A synergistic effect between the compounds was observed in the C + F group, which showed more behavioral effects than the groups exposed to pesticides individually. The use of behavioral tests to evaluate pesticide mixtures is important to standardize methods and associate behavioral changes with ecologically relevant events, thus creating a more realistic scenario for investigating the potential environmental impacts of these compounds.