Performance of collagen-based matrices from Nile tilapia skin: A pilot proteomic study in a murine model of wound healing.

Full-thickness cutaneous trauma, due to the lack of dermis, leads to difficulty in epithelialization by keratinocytes, developing a fibrotic scar, with less elasticity than the original skin, which may have disorders in predisposed individuals, resulting in hypertrophic scar and keloids. Biomedical materials have excellent characteristics, such as good biocompatibility and low immunogenicity, which can temporarily replace traditional materials used as primary dressings. In this work, we developed two dermal matrices based on Nile tilapia collagen, with (M_GAG) and without (M) glycosaminoglycans, using a sugarcane polymer membrane as a matrix support. To assess the molecular mechanisms driving wound healing, we performed qualitative proteomic analysis on the wound bed in an in vivo study involving immunocompetent murine models at 14 and 21 days post-full-thickness skin injury. Gene Ontology and Pathway analysis revealed that both skins were markedly represented by modulation of the immune system, emphasizing controlling the acute inflammation response at 14 and 21 days post-injury. Furthermore, both groups showed significant enrichment of pathways related to RNA and protein metabolism, suggesting an increase in protein synthesis required for tissue repair and proper wound closure. Other pathways, such as keratinization and vitamin D3 metabolism, were also enriched in the groups treated with M matrix. Finally, both matrices improved wound healing in a full post-thick skin lesion. However, our preliminary molecular data reveals that the collagen-mediated healing matrix lacking glycosaminoglycan (M) exhibited a phenotype more favorable to tissue repair, making it more suitable for use before skin grafts.

Role of nicergoline in corneal wound healing in diabetic rats.

BACKGROUND: To investigate the effect of nicergoline on the rate of complete corneal ulcer reepithelialization (CCUR) in diabetic rats with diabetic keratopathy. METHODS: Forty-eight streptozotocin-induced diabetic rats were randomly divided into two groups. The experimental group (n = 24) received nicergoline (10 mg.kg- 1.day- 1), while the control group (n = 24) received a placebo. A corneal epithelial defect was induced using a corneal diamond burr, and defect area was compared at time points of 0, 12, 24, 48 and 72 h after the injury using image analysis software. The probability of CCUR within 72 h was assessed using the Kaplan-Meier survival analysis log-rank test. RESULTS: When compared, 4 of the 24 rats (17%) in the placebo group and 12 of the 24 rats (50%) in the nicergoline group were found to have CCUR within 72 h (log-rank = 0.027). Cox regression analysis found no effect of the covariates blood glucose (P = 0.601) or weight (P = 0.322) on the corneal reepithelialization (survival) curve. CONCLUSIONS: Nicergoline increased wound healing rates relative to placebo and may therefore be investigated as a treatment option in diabetic keratopathy.

Influence of Butyrylcholinesterase in Progression of Mild Cognitive Impairment to Alzheimer's Disease.

BACKGROUND: Several demographic and genetic prognostic factors of conversion from mild cognitive impairment (MCI) to Alzheimer's disease (AD) have been recognized so far. The most frequent polymorphism of butyrylcholinesterase (BuChE), the K-variant, has been proposed as a risk factor for AD, but data regarding its influence on early disease progression is still limited. OBJECTIVE: To investigate the influence of the BuChE-K variant in MCI progression to AD. METHODS: 96 MCI patients were included in the study and were genotyped for BuChE-K variant and Apolipoprotein E (ApoE). Cerebrospinal fluid (CSF) BuChE activity, as well as the levels of AD biomarkers amyloid-ß 42 (Aß42), total and hyperphosphorylated tau (t-tau and p-tau) were also determined. RESULTS: No significant differences were found in either BuChE-K variant or BuChE activity between MCI patients that progressed to AD (MCI-AD) and patients that remained stable during clinical follow-up (MCI-St). As expected, baseline CSF levels of Aß42 were significantly lower and t-Tau, p-Tau, and ApoE ɛ4 allele frequency were significantly higher in MCI-AD patients. An association between the ApoE ɛ4 allele and the BuChE-K variant in MCI-AD, but not in MCI-St patients, was found with patients carrying both alleles presenting the highest incidence of progression and the lowest estimated time of progression to AD. CONCLUSION: Although BuChE-K alone does not seem to play a major role in progression to AD in MCI patients, a synergistic effect with the ApoE ɛ4 allele was found, highlighting the importance of assessing these combined genotypes for evaluating risk progression in MCI patients.

Association between butyrylcholinesterase and cerebrospinal fluid biomarkers in Alzheimer's disease patients.

The deficit of cholinergic activity is one of the main findings in Alzheimer's disease (AD), and is related to the synthesis of acetylcholine, and the hydrolysing enzymes, acetylcholinesterase and butyrylcholinesterase (BuChE). Together with the Apolipoprotein E-ε4 allele (ApoE-ε4), the BuChE-K variant has been proposed to increase AD risk in certain populations. In addition, this polymorphism has been associated with a lower capacity to attenuate ß-amyloid aggregation. In the present study we explored the interaction of the BuChE-K variant with its activity in CSF, conventional AD biomarkers and ApoE genotype. 217 AD patients and 200 age-matched controls were genotyped for the ApoE and the BuChE-K variant. BuChE activity in CSF, as well as the levels of the CSF-AD biomarkers amyloid-beta 42 (Aß42), total and hyperphosphorylated tau (t-tau and p-tau) were determined in 88 of these patients. The results showed no significant differences in the BuChE-K variant distribution between patients and controls. No influence of the BuChE-K variant was seen neither in CSF BuChE activity, nor in the levels of Aß42, t-tau and p-tau in AD patients. ApoE genotype also did not seem to influence CSF BuChE activity. Interestingly, in AD patients, an association between high CSF BuChE activity and increased levels of CSF Aß42 was shown, particularly in ApoE-ε4 allele carriers. In our population, the BuChE-K variant does not seem to confer risk for AD or to influence the activity of the enzyme in CSF. However, we demonstrated an association between BuChE activity, ApoE-ε4 genotype and CSF Aß42 levels, highlighting the importance of assessing BuChE activity as a possible modulator of Aß load in the brain.

Prognosis of Early-Onset vs. Late-Onset Mild Cognitive Impairment: Comparison of Conversion Rates and Its Predictors.

BACKGROUND: Despite having the same histopathological characteristics, early-onset and late-onset Alzheimer's disease (AD) patients show some distinct clinical and neuropsychological profiles. Early Onset Mild Cognitive Impairment (EOMCI) is a less characterized group. The aim of this study is to characterize MCI probably due to AD in terms of the clinical, genetic, Cerebrospinal fluid (CSF) biomarkers profile and conversion rate of EOMCI, compared to the late-onset form (LOMCI). METHODS: 159 MCI patients were divided in two groups: 52 EOMCI (onset < 65 years) and 107 LOMCI (onset ≥ 65 years). We investigated differences in neuropsychological scores, clinical variables, ApoE genotype, CSF biomarkers (Aß42, t-Tau and p-Tau) in both groups. Conversion was ascertained during follow-up. RESULTS: EOMCI showed a longer duration of symptoms prior to the first evaluation (EOMCI = 4.57 vs. LOMCI = 3.31, p = 0.008) and scored higher on the subjective memory complaints scale (9.91 vs. 7.85, p = 0.008), but performed better in brief cognitive tests (27.81 vs. 26.51, p < 0.001 in Mini-Mental State Examination; 19.84 vs. 18.67, p = 0.005 in Montreal Cognitive Assessment) than LOMCI. ApoE genotype distribution and CSF biomarker profile were similar in both groups, as was the conversion risk. Lower Aß42 (Hazard ratio (HR): 0.998, 95% Confidence Interval (CI) = [0.996⁻1.000], p = 0.042), higher t-Tau levels (HR: 1.003, 95%CI = [1.000⁻1.005], p = 0.039) and higher scores in the Alzheimer Disease Assessment Scale-Cognitive (HR: 1.186, 95%CI = [1.083⁻1.299], p = 0.002) increased the risk of conversion. DISCUSSION: Despite differences in memory performance and memory complaints, EOMCI and LOMCI seem to represent indistinct biological groups that do not have a higher risk of conversion to AD or differ in risk factors for conversion.

Cerebrospinal fluid Aß40 is similarly reduced in patients with Frontotemporal Lobar Degeneration and Alzheimer's Disease.

Cerebrospinal fluid (CSF) biomarkers have been increasingly studied for dementia diagnosis, however the accuracy to distinguish between different forms of dementia is still unsatisfactory. In this study, the added value of another CSF Aß-peptide (Aß40), along with the core CSF markers t-Tau, p-Tau, and Aß42, in the discrimination between two large dementia groups of Frontotemporal Lobar Degeneration (FTLD; n=107), Alzheimer's Disease (AD; n=107) and non-demented subjects (n=33) was evaluated. In FTLD, t-Tau and p-Tau were significantly increased in relation to controls, but lower than in AD, while Aß42 was similar in FTLD and controls, but higher than in AD. Equally reduced Aß40 levels were seen in both dementia groups, and therefore the combination of Aß40 with core CSF biomarkers optimally discriminated FTLD and AD patients from controls. Aß42 and t-Tau were selected as the best biomarker subset to differentiate FTLD from AD, with no added value of Aß40 to the model. Diagnostic accuracy between FTLD and AD was still sub-optimal, with a significant percentage (23%) of FTLD patients, in particularly women, carrying an ApoE-ε4 allele, showing a CSF-AD biomarkers profile. Although CSF Aß40 does not appear to have an additional value in the distinction between FTLD and AD, it increases the discrimination between subjects with dementia from controls. A CSF-AD biomarker profile can be seen in patients with a clinical phenotype of FTLD, reinforcing the need for autopsy confirmation.

Progranulin peripheral levels as a screening tool for the identification of subjects with progranulin mutations in a Portuguese cohort.

BACKGROUND: Progranulin (PGRN) mutations are associated with different clinical phenotypes, including frontotemporal lobar degeneration (FTLD), corticobasal syndrome (CBS) and Alzheimer's disease (AD). As all pathogenic PGRN mutations identified so far cause disease through haploinsufficiency, determination of PGRN levels has been proposed as a reliable method to identify mutation carriers. OBJECTIVE: To evaluate the accuracy of peripheral PGRN levels in the identification of the PGRN mutation carriers detected thus far in our Portuguese cohort. METHODS: Serum PGRN levels were measured in 244 subjects (124 patients in the spectrum of FTLD, 2 asymptomatic descendants of a FTLD patient, 56 AD patients and 64 controls) by a novel commercial ELISA kit. RESULTS: Low PGRN levels were detected in 7 individuals (5 behavioral variant frontotemporal dementia, 1 CBS, and 1 still clinically unaffected) that constituted the group of the null PGRN mutation carriers previously identified in our molecular diagnostic laboratory. The pathogenic mutations found consisted of 4 insertion-deletions, causing frameshifts resulting in premature stop codons, 3 of which were novel. In addition, a normal PGRN level was found in a patient harboring a novel missense variant. For this novel ELISA kit, we established a PGRN cut-off level that identified with 100% accuracy the pathogenic mutation carriers. CONCLUSION: This study supports the use of a novel assay for the determination of PGRN levels as a screening procedure to identify patients harboring null PGRN mutations. This approach would significantly decrease the required PGRN mutation analysis workload and should be extended to other clinical phenotypes than behavioral variant frontotemporal dementia and to apparently sporadic cases.

Microparticles of Aloe vera/vitamin E/chitosan: microscopic, a nuclear imaging and an in vivo test analysis for burn treatment.

The use of drug-loaded nanoparticles and microparticles has been increasing, especially for cosmetic and drug delivery purposes. In this work, a new microparticle formulation was developed for use in the healing process of skin burns in a composition of Aloe vera/vitamin E/chitosan. In order to observe the morphological properties, Raman and atomic force microscopy evaluation were performed. The biodistribution studies were analyzed by using a nuclear methodology, labeling the microparticles with Technetium-99m and in vivo test was procedure to analyzed the cicatrization process. The results of AFM analysis show the formation and the adherence property of the microparticles. Raman analyses show the distribution of each component in the microparticle. The nuclear method used shows that the biodistribution of the microparticles remained in the skin. The in vivo cicatrization test showed that the poloxamer gel containing the microparticles make a better cicatrization in relation to the other formulations tested.

Evaluating the effectiveness of gel formulation of irradiated seed lectin Cratylia mollis during bone repair in rats.

CONTEXT: Regeneration corresponds to the replacement of damaged cells with ones that have the same morphology and function. For experimental evaluation of materials that may favor the process of bone healing, defects are created with dimensions that prevent spontaneous regeneration. For the development and use of new drugs, it is necessary to study its effects in vitro, which depends on the formulation, concentration, and rate of irradiation in vivo and the route and frequency of administration; thus, it is possible to characterize the physiological and molecular mechanisms involved in the response and cellular effects. OBJECTIVE: The objective of this study was to assess the effect of Cramoll-1,4 on the process of bone repair. MATERIALS AND METHODS: A formulation of biopharmaceutical lectin Cramoll-1,4 at a concentration of 300 mg/100 mL was applied in a single application via gamma radiation and its effect on the process of bone repair in rats was assessed. RESULTS: Histologically, it was observed that the bone defect is coated by loose connective tissue rich in fibroblasts, providing a range similar to the thick bone original and competing with site of new bone formation. This prevented direct contact between the formulation and experimental bone tissue, as, despite its proven effectiveness in experiments on the repair of skin lesions, the formulation used did not promote bone stimulation that would have promoted the tissue repair process. CONCLUSION: Because of the direct interference of loose tissue repair that prevented direct contact of the implant with the bone interface, the formulation did not promote bone stimulation.

Importância da Ortodontia no tratamento multidisciplinar da avulsão dentária: relato de caso clínico / Importance of orthodontics in the multidisciplinary treatment of thooth avulsion: a case report

Entre os tipos de traumatismos dentários, o que geralmente possui o prognóstico mais desfavorável é a avulsão dentária, principalmente por depender dos cuidados e medidas tomadas no local do acidente. Além disso, como consequência da avulsão, há o rompimento das fibras do ligamento periodontal e, a partir daí, segue-se uma série de eventos biológicos que podem resultar em necrose pulpar, anquilose e reabsorção da raiz. Nesses casos, a movimentação dentária deve ser protelada até que se complete um ano após o evento traumático. A aplicação de matriz derivada do esmalte tem sido indicada para esses casos como fator de regeneração periodontal e como medida profilática ao desenvolvimento de reabsorção radicular e anquilose. O presente trabalho tem a finalidade de apresentar um caso multidisciplinar com histórico de avulsão traumática e reimplante, que foi submetido a tratamento ortodôntico e aplicação de Emdogain. A abordagem multidisciplinar para esses casos é de fundamental importância para o sucesso do tratamento, ou seja, para a manutenção do dente no alvéolo, proporcionando um sorriso mais harmonioso e preservando tecido ósseo e tecido mole até que seja possível o tratamento reabilitador com implante e prótese.

Development of animal model for studying deep second-degree thermal burns.

Thermal lesions were produced in 12 male Wistar rats, positioning a massive aluminum bar 10 mm in diameter (51 g), preheated to 99°C ± 2°C/10 min. on the back of each animal for 15 sec. After 7, 14, 21, and 28 days, animals were euthanized. The edema intensity was mild, with no bubble and formation of a thick and dry crust from the 3rd day. The percentage of tissue shrinkage at 28 days was 66.67 ± 1.66%. There was no sign of infection, bleeding, or secretion. Within 28 days reepithelialization was incomplete, with fibroblastic proliferation and moderate fibrosis and presence of modeled dense collagen fibers. It is concluded that the model established is applicable in obtaining deep second-degree thermal burns in order to evaluate the healing action of therapeutic agents of topical use.

Topical application effect of the isolectin hydrogel (Cramoll 1,4) on second-degree burns: experimental model.

This study aimed at evaluating the use of hydrogel isolectin in the treatment of second-degree burns. Twenty male rats were randomly divided into two groups (G1 = treatment with hydrogel containing 100 µg/mL Cramoll 1,4 and G2 = Control, hydrogel). After 7, 14, 21, 28, and 35 days, animals were euthanized. On the 7th day, G1 showed intense exudates, necrosis and edema. On the 14th day, G1 showed tissue reepithelialization and moderate autolysis. On the 21st day, G1 showed intense fibroblastic proliferation, presence of dense collagen, and moderate fibrosis. On the 28th day, G1 showed complete tissue epithelialization. On the 35th day, G1 showed modeled dense collagen. The significant wound contraction was initiated from day, 14 in the G1. There were no significant differences in biochemical and hematological parameters analyzed. These results extend the potential of therapeutic applications for Cramoll 1,4 in the treatment of thermal burns.

Radiolabeling of cramoll 1,4: evaluation of the biodistribution.

The cramoll 1,4 is a well-studied lectin. However, few studies about its biodistribution have been done before. In this study, we radiolabeled the cramol 1,4 with Tc-99m and analyzed the biodistribution. The results showed that the cramol has an abnormal uptake by the bowel with reflections on its clearance mechanism.

Lasertherapy efficacy in temporomandibular disorders: control study.

UNLABELLED: Temporomandibular dysfunction is characterized by the presence of painful joint/muscular symptoms muscle in the face. The main justification for the use of lasers in laser therapy dysfunction is its analgesic effect, which was observed in most studies in the literature. AIM: We evaluated the effectiveness of laser therapy in the treatment of temporomandibular disorders. METHODS: 50 volunteers with temporomandibular disorders were divided into two groups (control and experimental) had amplitudes of movements of mouth opening, right and left laterality recorded before and after laser application. Was also recorded, the score the individual gave to pain by visual analog scale and, through physical examination, the pain points. We used the AsGaAl laser with a 40mW power, with 80J/cm(2) for 16 seconds at four selected points for just one session with reassessment after a week. STUDY DESIGN: Clinical. RESULTS: It was noted that laser therapy increased the mean amplitude of mandibular movements (p = 0.0317) and decreased significantly (43.6%) the pain intensity measured by the visual analog scale. CONCLUSIONS: The laser decreases the painful symptoms of the patient after application through its analgesic and/or a placebo effect.

Analysis of plasma citrulline and intestinal morphometry in mice with hepatosplenic schistosomiasis.

BACKGROUND: Portal hypertension in the mucosa of the intestine and the presence of granulomas in the wall of this organ can alter digestive function in patients with schistosomiasis. Citrulline is a potential marker of intestinal function in some diseases that affect the morphometry of the mucosa because of its close association with enterocytes. The aims of the present study were to determine serum citrulline concentrations in mice with hepatosplenic schistosomiasis, analyze the morphologic repercussions for the mucosa of the small intestine, correlate citrulline concentrations with morphometric changes in the intestinal mucosa, and evaluate the effect of splenectomy on citrulline concentration. METHODS: After approval from the local ethics committee, 46 adult female albino Swiss mice were divided into two groups: Control (23 healthy mice) and experimental (23 mice with hepatosplenic schistosomiasis). Blood samples were collected for the analysis of plasma citrulline before and after splenectomy. A segment of the jejunum was resected for morphometric analysis. RESULTS: The average body mass in the control group was greater than that in the experimental group (p = 0.00062). The average citrulline concentration in the control group was greater than that in the experimental group both before and after splenectomy (p < 0.001). In the experimental group, the villi had less height and area, and there was a smaller perimeter of the mucosal surface (p = 0.003, <0.001, and p = 0.001, respectively). There was a direct correlation between citrulline concentration and the height and area of the villi (p = 0.003 and 0.04, respectively). There was no correlation between citrulline concentration and the perimeter of the surface of the jejunal mucosa. After splenectomy, there was a reduction in the mean citrulline concentration in the experimental group (p = 0.009). CONCLUSIONS: Serum citrulline concentrations were reduced in mice with schistosomiasis, and a direct correlation was found between the citrulline concentration and the morphometry of the jejunal villi. Moreover, there was a reduction in the plasma concentration of citrulline after splenectomy.

Eficácia da laserterapia nas disfunções têmporo-mandibulares: estudo controle / Lasertherapy efficacy in temporomandibular disorders: control study

Temporomandibular dysfunction is characterized by the presence of painful joint/muscular symptoms muscle in the face. The main justification for the use of lasers in laser therapy dysfunction is its analgesic effect, which was observed in most studies in the literature. AIM: We evaluated the effectiveness of laser therapy in the treatment of temporomandibular disorders. METHODS: 50 volunteers with temporomandibular disorders were divided into two groups (control and experimental) had amplitudes of movements of mouth opening, right and left laterality recorded before and after laser application. Was also recorded, the score the individual gave to pain by visual analog scale and, through physical examination, the pain points. We used the AsGaAl laser with a 40mW power, with 80J/cm² for 16 seconds at four selected points for just one session with reassessment after a week. Study design: Clinical. RESULTS: It was noted that laser therapy increased the mean amplitude of mandibular movements (p = 0.0317) and decreased significantly (43.6 percent) the pain intensity measured by the visual analog scale. CONCLUSIONS: The laser decreases the painful symptoms of the patient after application through its analgesic and/or a placebo effect.

A disfunção têmporo-mandibular é caracterizada pela presença de sintomatologia dolorosa articular/muscular na região da face. A principal justificativa do uso do laser da laserterapia na disfunção é seu efeito analgésico, fato observado na maioria dos estudos encontrados na literatura. OBJETIVO: Foi avaliar a eficácia da laserterapia no tratamento das disfunções têmporo-mandibulares. MATERIAL E MÉTODO: 50 voluntários com disfunção têmporo-mandibular foram divididos em dois grupos (controle e experimental) tiveram as amplitudes dos movimentos de abertura bucal, lateralidade direita e esquerda registrados, antes e após aplicação do laser. Foi registrada, também, a nota de dor do indivíduo através da escala analógica visual de dor e, através do exame físico, os pontos álgicos. Utilizou-se o laser de AsGaAl com potência de 40mW, com 80J/cm², por 16 segundos, em quatro pontos selecionados por apenas uma sessão com reavaliação após uma semana. Desenho Científico Utilizado: Clínico. RESULTADOS: Notou-se que a laserterapia promoveu aumento da média de amplitude dos movimentos mandibulares (p=0,0317) e houve redução significativa (43,6 por cento) da intensidade de dor dos pacientes medida através da escala analógica visual de dor...

Genetic screening of Alzheimer's disease genes in Iberian and African samples yields novel mutations in presenilins and APP.

Mutations in three genes (PSEN1, PSEN2, and APP) have been identified in patients with early-onset (<65 years) Alzheimer's disease (AD). We performed a screening for mutations in the coding regions of presenilins, as well as exons 16 and 17 of the APP gene in a total of 231 patients from the Iberian peninsular with a clinical diagnosis of early-onset AD (mean age at onset of 52.9 years; range 31-64). We found three novel mutations in PSEN1, one novel mutation in PSEN2, and a novel mutation in the APP gene. Four previously described mutations in PSEN1 were also found. The same analysis was carried in 121 elderly healthy controls from the Iberian peninsular, and a set of 130 individuals from seven African populations belonging to the Centre d'Etude du Polymorphisme Humain-Human Genome Diversity Panel (CEPH-HGDP), in order to determine the extent of normal variability in these genes. Interestingly, in the latter series, we found five new non-synonymous changes in all three genes and a presenilin 2 variant (R62H) that has been previously related to AD. In some of these mutations, the pathologic consequence is uncertain and needs further investigation. To address this question we propose and use a systematic algorithm to classify the putative pathology of AD mutations.

Diagnostic value of CSF protein profile in a Portuguese population of sCJD patients.

The clinical diagnosis of sporadic Creutzfeldt-Jakob disease (sCJD) is difficult, and reliable markers are highly desired. In this work we assess the value of several cerebrospinal fluid (CSF) markers for sCJD diagnosis. Within the framework of the Portuguese Epidemiological Surveillance Program for Human Prion Diseases, CSF samples from 71 patients with clinically suspected sCJD, 30 definite sCJD and 41 non-CJD patients, were analysed for the presence of 14-3-3 protein. CSF levels of tau (t-tau), and phosphorylated tau (p-tau181), S-100b and beta amyloid (Abeta42) proteins were determined. The influence of clinical and genetic characteristics on CSF markers sensitivity was also evaluated. Protein 14-3-3 was detected in 29/30 sCJD patients and 9/41 non-CJD patients. Extremely elevated t-tau and S-100b protein levels were found in sCJD patients, while p-tau181 levels were only slightly elevated and Abeta42 showed no differences compared to controls. 14-3-3 was the most sensitive parameter (97%), but its specificity was low (78%); sensitivity/specificity for other proteins were: S-100b-93/93%, t-tau-93/95%, with maximum accuracy being obtained by a combination of tests (14-3-3 combined with either t-tau or S-100b, or combining S-100b with t-tau/Abeta42 or p-tau/t-tau ratios). The sensitivity of 14-3-3, as well as of p-tau181/t-tau ratio, was decreased in younger patients with long disease duration, with the PrP-2 isotype and MV genotype. Both 14-3-3, t-tau and S-100b are sensitive markers for sCJD, but 14-3-3 specificity seems to be lower in this special clinical setting of rapidly progressing dementias. We propose that in cases with a 14-3-3 weak positive result, or in young patients with long disease duration, a second CSF marker would be valuable for the diagnosis of sCJD.

Complete screening for glucocerebrosidase mutations in Parkinson disease patients from Portugal.

Mutations in the gene encoding beta-glucocerebrosidase, a lysosomal degrading enzyme, have recently been associated with the development of Parkinson disease. Here we report the results found in a cohort of Portuguese Parkinson disease patients and healthy age-matched controls for mutations in the aforementioned gene. This screening was accomplished by sequencing the complete open-reading frame, as well as intron/exon boundaries, of the glucocerebrosidase gene, in a total of 230 patients and 430 controls. We have found an increased number of Parkinson disease patients presenting mutations in GBA when compared to controls. These results, together with recent literature, clearly suggest a role of glucocerebrosidase in the development of Parkinson disease.

Novel progranulin mutation: screening for PGRN mutations in a Portuguese series of FTD/CBS cases.

Mutations in the progranulin (PGRN) gene were recently described as the cause of ubiquitin positive frontotemporal dementia (FTD) in many families. Different frequencies of these genetic changes have been reported in diverse populations leading us to determine if these mutations were a major cause of FTD in the Portuguese population. The entire coding sequence plus exon 0 of PGRN were sequenced in a consecutive series of 46 FTD/CBS Portuguese patients. Two mutations were found: a novel pathogenic insertion (p.Gln300GlnfsX61) and a previously described point variant (p.T182M) of unclear pathogenicity. Pathogenic mutations in the PGRN gene were found in one of the 36 probands studied (3% of the probands in our series) who had a corticobasal syndrome presentation, indicating that in the Portuguese population, mutations in this gene are not a major cause of FTD.