Alzheimer's disease-like pathology has transient effects on the brain and blood metabolome

The pathogenesis of Alzheimer's disease (AD) is complex involving multiple contributing factors. The extent to which AD pathology impacts upon the metabolome is still not understood, nor is it known how disturbances change as the disease progresses. For the first time we have profiled longitudinally (6, 8, 10, 12 and 18 months) both the brain and plasma metabolome of APP/PS1 double transgenic and wild type (WT) mice. A total of 187 metabolites were quantified using a targeted metabolomics methodology. Multivariate statistical analysis produced models that distinguished APP/PS1 from WT mice at 8, 10 and 12 months.Metabolic pathway analysis found perturbed polyamine metabolism in both brain and blood plasma. There were other disturbances in essential amino acids,branched chain amino acids and also in the neurotransmitter serotonin.Pronounced imbalances in phospholipid and acylcarnitine homeostasis was evident in two age groups. AD-like pathology therefore impacts greatly on both the brain and blood metabolomes, although there appears to be a clear temporal sequence whereby changes to brain metabolites precede those in blood.

Wavelet entropy of Doppler ultrasound blood velocity flow waveforms distinguishes nitric oxide-modulated states

Wavelet entropy assesses the degree of order or disorder in signals and presents this complex information in a simple metric. Relative wavelet entropy assesses the similarity between the spectral distributions of two signals, again in a simple metric. Wavelet entropy is therefore potentially a very attractive tool for waveform analysis. The ability of this method to track the effects of pharmacologic modulation of vascular function on Doppler blood velocity waveforms was assessed. Waveforms were captured from ophthalmic arteries of 10 healthy subjects at baseline, after the administration of glyceryl trinitrate (GTN) and after two doses of N(G)-nitro-L-arginine-methyl ester (L-NAME) to produce vasodilation and vasoconstriction, respectively. Wavelet entropy had a tendency to decrease from baseline in response to GTN, but significantly increased after the administration of L-NAME (mean: 1.60 ± 0.07 after 0.25 mg/kg and 1.72 ± 0.13 after 0.5 mg/kg vs. 1.50 ± 0.10 at baseline, p < 0.05). Relative wavelet entropy had a spectral distribution from increasing doses of L-NAME comparable to baseline, 0.07 ± 0.04 and 0.08 ± 0.03, respectively, whereas GTN had the most dissimilar spectral distribution compared with baseline (0.17 ± 0.08, p = 0.002). Wavelet entropy can detect subtle changes in Doppler blood velocity waveform structure in response to nitric-oxide-mediated changes in arteriolar smooth muscle tone.

Acid-labile protein-adducted heterocyclic aromatic amines in human blood are not viable biomarkers of dietary exposure: A systematic study.

Heterocyclic aromatic amines (HCA) are carcinogenic mutagens formed during cooking of protein-rich foods. HCA residues adducted to blood proteins have been postulated as biomarkers of HCA exposure. However, the viability of quantifying HCAs following hydrolytic release from adducts in vivo and correlation with dietary intake are unproven. To definitively assess the potential of labile HCA-protein adducts as biomarkers, a highly sensitive UPLC-MS/MS method was validated for four major HCAs: 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx) and 2-amino-3,7,8-trimethylimidazo[4,5-f]quinoxaline (7,8-DiMeIQx). Limits of detection were 1e5 pg/ml plasma and recoveries 91e115%. Efficacy of hydrolysis was demonstrated by HCA-protein adducts synthesised in vitro. Plasma and 7-day food diaries were collected from 122 fasting adults consuming their habitual diets. Estimated HCA intakes ranged from 0 to 2.5 mg/day. An extensive range of hydrolysis conditions was examined for release of adducted HCAs in plasma. HCA was detected in only one sample (PhIP, 9.7 pg/ml), demonstrating conclusively for the first time that acid-labile HCA adducts do not reflect dietary HCA intake and are present at such low concentrations that they are not feasible biomarkers of exposure. Identification of biomarkers remains important. The search should concentrate on stabilised HCA peptide markers and use of untargeted proteomic and metabolomic approaches.

Impaired orthostatic blood pressure recovery and cognitive performance at two-year follow up in older adults: The Irish Longitudinal Study on Ageing

Background: Prospective investigations of the association between impaired orthostatic blood pressure (BP) regulation and cognitive decline in older adults are limited, and findings to-date have been mixed. The aim of this study was to determine whether impaired recovery of orthostatic BP was associated with change in cognitive function over a 2-year period, in a population based sample of community dwelling older adults. 

Methods: Data from the first two waves of the Irish Longitudinal Study on Ageing were analysed. Orthostatic BP was measured during a lying to standing orthostatic stress protocol at wave 1 using beat-to-beat digital plethysmography, and impaired recovery of BP at 40 s post stand was investigated. Cognitive function was assessed at wave 1 and wave 2 (2 years later) using the Mini-Mental State Exam (MMSE), verbal fluency and word recall tasks. 

Results: After adjustment for measured, potential confounders, and multiple imputation for missing data, the change in the number of errors between waves on the MMSE was 10 % higher [IRR (95 % CI) = 1.10 (0.96, 1.26)] in those with impaired recovery at 40 s. However, this was not statistically significant (p = 0.17). Impaired BP recovery was not associated with change in performance on any of the other cognitive measures. 

Conclusions: There was no clear evidence for an association between impaired recovery of orthostatic BP and change in cognition over a 2-year period in this nationally representative cohort of older adults. Longer follow-up and more detailed cognitive testing would be advantageous to further investigate the relationship between orthostatic BP and cognitive decline.

Urinary thrombomodulin levels were significantly higher following occupational exposure to chemicals, in the presence of dipstick protein, but not in the presence of dipstick blood

Currently, there are no biomarkers which can identify patients with an increased risk of developing urothelial cancer as a result of occupational chemical exposure. The aim of this study was to evaluate the relationships between final diagnosis and 22 biomarkers measured in urine, serum and plasma collected from 156 hematuric patients. Fourteen of the 80 patients (17.5%) with urothelial cancer and 13/76 (17.1%) of the controls were deemed to have a history of chemical exposure. We applied Fisher's exact tests to explore associations between chemical exposure and final diagnosis, and tumor stage and grade, where applicable; ANOVA and t-test to compare age across patients with and without chemical exposure; and Zelen's exact test to evaluate relationships across final diagnosis, chemical exposure and smoking. Following pre-selection of biomarkers using Lasso, we identified biomarkers with differential levels across patients with and without chemical exposure using Welch's t-test. Using a one-sided t-test and considering multiple testing using FDR, we observed that TM levels in urine were significantly higher in samples from patients with a history of chemical exposure regardless of their diagnosis as control or urothelial cancer (one-sided t-test, p_UC = 0.014 and p_CTL = 0.043); in the presence of dipstick protein and when urinary pH levels ≤ 6 (p = 0.003), but not in the presence of dipstick blood (p = 0.115). Urothelial cancer patients with a history of chemical exposure were significantly younger (64.1 years) than those without chemical exposure (70.2 years) (one-sided t-test p-value = 0.012); and their tumors were higher grade (Fisher's exact test; p = 0.008). There was a strong association between a history of chemical exposure and smoking in urothelial cancer patients (Zelen's exact test; p = 0.025). Elevated urinary thrombomodulin levels could have the potential to identify chemical exposure in hematuric patients at high risk of developing urothelial cancer.

A spectral estimator using parallel-processing for use in a doppler blood-flow instrument

The work described here is part of a research program aiming to increase the sensitivity to disease detection using Doppler ultrasound by reducing the effects to the measurement procedure on the estimation of blood velocity and detection of flow disturbance.

Tissue temperature estimation with pulse-echo in blood flow presence

Aiming at time-spatial characterization of tissue temperature when ultrasound is applied for thermal therapeutic proposes two experiments were developed considering gel-based phantoms, one of them including an artificial blood vessel. The blood vessel was mimicking blood flow in a common carotid artery. For each experiment phantoms were heated by a therapeutic ultrasound (TU) device emitting different intensities (0.5, 1, 1.5, 1.8 W/cm2). Temperature was monitored by thermocouples and estimated through imaging ultrasound transducer's signals within specific special points inside the phantom. The temperature estimation procedure was based on temporal echo-shifts (TES), computed based on echo-shifts collected through image ultrasound (IU) transducer. Results show that TES is a reliable non-invasive method of temperature estimation, regardless the TU intensities applied. Presence of a pulsatile blood flow vessel in the focal point of TU transducer reduces thermal variation in more than 50%, also affecting the temperature variation in the surrounding area. In other words, vascularized tissues require longer ultrasound thermal therapeutic sessions or higher TU intensities and inclusion of IU in the therapeutic procedure enables non-invasive monitoring of temperature. © 2013 IEEE.

In vitro blood-brain barrier models to predict the permeation of gene therapy vectors into the brain

A terapia génica tem-se revelado uma alternativa relevante no tratamento de doenças neurodegenerativas (DN). Contudo, a entrega de vetores para transferência génica no cérebro representa ainda um enorme desafio devido à presença da barreira hemato-encefálica (BHE). A BHE é uma interface dinâmica e seletiva entre o sangue e o cérebro, constituída pelas células endoteliais cerebrais, astrócitos e pericitos, desempenhando um importante papel na regulação da homeostasia cerebral. A BHE representa um dos maiores obstáculos no tratamento de DN, uma vez que esta barreira impede o transporte para o cérebro da maioria das moléculas terapêuticas, incluindo os vetores para terapia génica. Embora tenham sido desenvolvidos diferentes modelos in vitro da BHE de forma a avaliar o transporte de fármacos através da BHE, muito poucos foram criados com o intuito de testar a permeabilidade desta barreira a vetores de terapia génica. O presente trabalho teve como objetivo principal o desenvolvimento e a avaliação de modelos in vitro de BHE que permitam a investigação da capacidade dos vetores de terapia génica de penetrarem no cérebro. No nosso estudo, foram testados diferentes modelos in vitro de BHE em monocultura, constituídos por células endoteliais de rato ou murganho (RBE4 e bEnd3, respetivamente), e modelos de co-cultura, que combinam células endoteliais com células neuronais (Neuro2a) ou astrócitos primários, cultivados num sistema transwell. Para caraterizar estes modelos foram realizados testes de permeabilidade e de resistência elétrica transendotelial, bem como estudos baseados na técnica de PCR quantitativo e na imunocitoquímica das proteínas das junções intercelulares. Verificámos que os modelos baseados na cultura de células bEnd3 e células neuronais ou astrócitos apresentavam as melhores propriedades de barreira. Posteriormente foi avaliada nos modelos selecionados a penetração de um vetor não-viral que reconhecidamente tem a capacidade de atravessar in vivo a BHE: o peptídeo da glicoproteína do vírus da raiva (RGV-9r). Os siRNAs marcados com um fluoróforo e acoplados ao peptídeo RVG-9r foram capazes de penetrar eficientemente as células bEnd3, localizadas no lado luminal do insert, via endocitose mediada por recetores, e ainda de penetrar os astrócitos ou células neuronais, previamente cultivadas no lado abluminal. Estes resultados correlacionam-se, de forma clara, com os resultados previamente descritos em estudos in vivo. Em conclusão, os modelos in vitro de BHE baseados na co-cultura de células bEnd3 com células Neuro2a ou astrócitos, têm grande potencial na seleção de candidatos a vetores de terapia génica para o cérebro, uma vez que apresentam importantes características da BHE e se baseiam num método fácil e reprodutível. Tal facto representa uma promessa significativa para a identificação de novas estratégias de terapia génica não invasiva para o tratamento de doenças neurológicas.

Study of the association of Gla Rich Protein (GRP) to human blood cells

Dissertação de mestrado, Ciências Biomédicas, Departamento de Ciências Biomédicas e Biomedicina, Universidade do Algarve, 2013

Hypothalamic and medullar mechanisms for long-term autonomic regulation of arterial blood pressure

Tese de doutoramento, Ciências Biomédicas (Fisiologia), Universidade de Lisboa, Faculdade de Medicina, 2014

Purity of Blood and Caste. Identity Narratives among Early Modern Goan Elites

This essay analyses the roles played by purity of blood and caste in seventeenth-and eighteenth-century identity narratives of Goan clites. Goa and its population are usually excluded from the mainstream literature of Indian social history, and seldom related to the early-modern Atlantic world, making this case study all the more valuable as a place to think the topic of blood and caste. The early establishment and the longevity of the Portuguese imperial presence (1510-1961) in Goa, its location at the crossroads of multiple cultural geographies (Iberian and Indian, and later, also Dutch, British and French), as well as the systematic process of religious conversion of its inhabitants and the questions of legal equality that conversion entailed, all intensified the types, textures, layers and meanings of experiences of social differentiation in this colonial context. This mapping of the experiences of purity of blood and caste in early-modem Goa therefore illuminates from a new angle the role of European imperial powers in the mUltiple expressions of racial classification.

A novel method to count the red blood cells in thin blood films

This paper describes a novel idea to identify the total number of red blood cells (RBCs) as well as their location in a Giemsa stained thin blood film image. This work is being undertaken as a part of developing an automated malaria parasite detection system by scanning a photograph of thin blood film in order to evaluate the parasitemia of the blood. Not only will this method eliminates the segmentation procedures that are normally used to segment the cells in the microscopic image, but also avoids any image pre-processing to deal with non uniform illumination prior to cell detection. The method utilizes basic knowledge on cell structure and brightness of the components due to Giemsa staining of the sample and detects and locates the RBCs in the image.

Habitual coffee and tea drinkers experienced increases in blood pressure after consuming low to moderate doses of caffeine; these increases were larger upright than in the supine posture

Caffeine users have been encouraged to consume caffeine regularly to maintain their caffeine tolerance and so avoid caffeine’s acute pressor effects. In controlled conditions complete caffeine tolerance to intervention doses of 250 mg develops rapidly following several days of caffeine ingestion, nevertheless, complete tolerance is not evident for lower intervention doses. Similarly complete caffeine tolerance to 250 mg intervention doses has been demonstrated in habitual coffee and tea drinkers’ but for lower intervention doses complete tolerance is not evident. This study investigated a group of habitual caffeine users following their self-determined consumption pattern involving two to six servings daily. Cardiovascular responses following the ingestion of low to moderate amounts caffeine (67, 133 and 200 mg) were compared with placebo in a double-blind, randomised design without caffeine abstinence. Pre-intervention and post-intervention (30 and 60 min) 90 s continuous cardiovascular recordings were obtained with the Finometer in both the supine and upright postures. Participants were 12 healthy habitual coffee and tea drinkers (10 female, mean age 36). Doses of 67 and 133 mg increased systolic pressure in both postures while in the upright posture diastolic pressure and aortic impedance increased while arterial compliance decreased. These vascular changes were larger upright than supine for 133 mg caffeine. Additionally 67 mg caffeine increased dp/dt and indexed peripheral resistance in the upright posture. For 200 mg caffeine there was complete caffeine tolerance. Cardiovascular responses to caffeine appear to be associated with the size of the intervention dose. Habitual tea and coffee drinking does not generate complete tolerance to caffeine as has been previously suggested. Both the type and the extent of caffeine induced cardiovascular changes were influenced by posture.

The Finometer can function as a standalone instrument in blood pressure variability studies and does not require support equipment to determine breathing frequency

Objective: The Finometer (FMS, Finapres Measurement Systems, Amsterdam) records the beat-to-beat finger pulse contour and has been recommended for research studies assessing shortterm changes of blood pressure and its variability. Variability measured in the frequency domain using spectral analysis requires that the impact of breathing be restricted to high frequency spectra (> 0.15 Hz) so data from participants needs to be excluded when the breathing impact occurs in the low frequency spectra (0.04 - 0.15 Hz). This study tested whether breathing frequency can be estimated from standard Finometer recordings using either stroke volume oscillation frequency or spectral stroke volume variability maximum scores. Methods: 22 healthy volunteers were tested for 270s in the supine and upright positions. Finometer recorded the finger pulse contour and a respiratory transducer recorded breathing. Stoke volume oscillation frequency was calculated manually while the stroke volume spectral maximums were obtained using the software Cardiovascular Parameter Analysis (Nevrokard Kiauta, Izola, Slovenia). These estimates were compared to the breathing frequency using the Bland-Altman procedures. Results: Stroke volume oscillation frequency estimated breathing frequency to <±10% 95% levels of agreement in both supine (-7.7 to 7.0%) and upright (-6.7 to 5.4%) postures. Stroke volume variability maximum scores did not accurately estimate breathing frequency. Conclusions: Breathing frequency can be accurately derived from standard Finometer recordings using stroke volume oscillations for healthy individuals in both supine and upright postures. The Finometer can function as a standalone instrument in blood pressure variability studies and does not require support equipment to determine breathing frequency.