Smoking behavior and secondhand smoke exposure among university students in northern Portugal: Relations with knowledge on tobacco use and attitudes toward smoking.

OBJECTIVES: To assess the prevalence of active smoking and secondhand smoke (SHS) exposure among college students in the north of Portugal, and analyze the relationship between knowledge about tobacco use and attitudes toward smoking. MATERIALS AND METHODS: This a cross-sectional study with a representative sample of college students (n=840) in one university in Portugal. A validated self-reported questionnaire was administered to a proportional stratified random sample during the academic year of 2018/2019. We evaluated associations between smoking status, SHS exposure, smokers peers, knowledge and attitudes toward smoking and sociodemographic variables. RESULTS: The results showed that 20.1% of the students surveyed were current smokers (7.3% occasional smokers, 2.9% regular smokers and 9.9% daily smokers). Most current smokers started smoking before the age of 17 (61.4%) and reported never having tried to quit smoking (59.7%). Only 34.4% of students reported (almost) not having been in enclosed spaces with smokers in the past 7 days. Exposure to SHS and having smoker friends contributes to the prevalence of tobacco use. In general, students showed favorable attitudes toward smoking, especially those who are smokers, have smoking friends and are more exposed to SHS. The level of knowledge about tobacco was moderate, with a higher number of correct responses by former smokers. CONCLUSIONS: These results suggests an urgent need for socio-educational programs for counseling on smoking cessation. In addition, is also strongly recommended that, throughout academic training, students develop personal and social skills for dealing with the tobacco epidemic.

Taxonomy and phylogeny of cercosporoid ascomycetes on Diospyros spp. with special emphasis on Pseudocercospora spp.

A worldwide survey of cercosporoid ascomycete species on hosts of the genus Diospyros (persimmon) with key to the species based on characters in vivo is provided. Special emphasis is placed on species of the genus Pseudocercospora, which are in part also phylogenetically analysed, using a multilocus approach. Species of the latter genus proved to be very diverse, with a remarkable degree of cryptic speciation. Seven new species are described (Pseudocercospora diospyri-japonicae, P. diospyriphila, P. ershadii, P. kakiicola, P. kobayashiana, and P. tesselata), and two new names are introduced [P. kakiigena (≡ Cylindrosporium kaki, non Pseudocercospora kaki), and Zasmidium diospyri-hispidae (≡ Passalora diospyri, non Zasmidium diospyri)]. Six taxa are lectotypified (Cercospora atra, C. diospyri, C. diospyri var. ferruginea, C. flexuosa, C. fuliginosa, C. kaki), and Pseudocercospora kaki is epitypified.

Development of a non-viral gene delivery vector based on the dynein light chain Rp3 and the TAT peptide.

Gene therapy and DNA vaccination trials are limited by the lack of gene delivery vectors that combine efficiency and safety. Hence, the development of modular recombinant proteins able to mimic mechanisms used by viruses for intracellular trafficking and nuclear delivery is an important strategy. We designed a modular protein (named T-Rp3) composed of the recombinant human dynein light chain Rp3 fused to an N-terminal DNA-binding domain and a C-terminal membrane active peptide, TAT. The T-Rp3 protein was successfully expressed in Escherichia coli and interacted with the dynein intermediate chain in vitro. It was also proven to efficiently interact and condense plasmid DNA, forming a stable, small (∼100nm) and positively charged (+28.6mV) complex. Transfection of HeLa cells using T-Rp3 revealed that the vector is highly dependent on microtubule polarization, being 400 times more efficient than protamine, and only 13 times less efficient than Lipofectamine 2000™, but with a lower cytotoxicity. Confocal laser scanning microcopy studies revealed perinuclear accumulation of the vector, most likely as a result of transport via microtubules. This study contributes to the development of more efficient and less cytotoxic proteins for non-viral gene delivery.

Characterization of the human dynein light chain Rp3 and its use as a non-viral gene delivery vector.

Dynein light chains mediate the interaction between the cargo and the dynein motor complex during retrograde microtubule-mediated transport in eukaryotic cells. In this study, we expressed and characterized the recombinant human dynein light chain Rp3 and developed a modified variant harboring an N-terminal DNA-binding domain (Rp3-Db). Our approach aimed to explore the retrograde cell machinery based on dynein to enhance plasmid DNA (pDNA) traffic along the cytosol toward the nucleus. In the context of non-viral gene delivery, Rp3-Db is expected to simultaneously interact with DNA and dynein, thereby enabling a more rapid and efficient transport of the genetic material across the cytoplasm. We successfully purified recombinant Rp3 and obtained a low-resolution structural model using small-angle X-ray scattering. Additionally, we observed that Rp3 is a homodimer under reducing conditions and remains stable over a broad pH range. The ability of Rp3 to interact with the dynein intermediate chain in vitro was also observed, indicating that the recombinant Rp3 is correctly folded and functional. Finally, Rp3-Db was successfully expressed and purified and exhibited the ability to interact with pDNA and mediate the transfection of cultured HeLa cells. Rp3-Db was also capable of interacting in vitro with dynein intermediate chains, indicating that the addition of the N-terminal DNA-binding domain does not compromise its function. The transfection level observed for Rp3-Db is far superior than that reported for protamine and is comparable to that of the cationic lipid Lipofectamine™. This report presents an initial characterization of a non-viral delivery vector based on the dynein light chain Rp3 and demonstrates the potential use of modified human light chains as gene delivery vectors.

Characterization of the aroma profile of Madeira wine by sorptive extraction techniques.

The characterization of the aroma profile of 33 samples of Madeira wine from five monovarieties (Sercial, Verdelho, Boal, Malvasia and Tinta Negra Mole) having different type and categories is presented, using solid phase microextraction and stir bar sorptive extraction techniques (SPME and SBSE) followed by capillary gas chromatography and mass spectrometry detection (GC-MS). Headspace SPME/GC-MS provided effectiveness to identify the major constituents of the aroma profile of Madeira wine, where no remarkable differences occur among the samples studied. The volatile compounds are mainly constituted by ethyl octanoate (11.3-256.9µgL-1), ethyl decanoate (21.5-210.5µgL-1), ethyl decenoate (0.1-112.8µgL-1), diethyl succinate (0.9-65.6µgL-1), ethyl dodecanoate (1.2-6.5µgL-1), ethyl nonanoate (0.6-5.2µgL-1), ethyl hexanoate (0.2-3.7µgL-1) and isoamyl octanoate (0-2.2µgL-1). C13 norisoprenoids such as vitispirane (0.9-7.0µgL-1) and 1,1,6-trimethyl 1,2-dihydro naphthalene (0.7-12.5µgL-1), as well as phenyl ethanol (0-8.1µgL-1), were also found in Madeira wine samples. The powerful capabilities of SBSE followed thermal desorption and GC-MS analysis allowed higher ability for profiling traces and ultra traces of compounds in Madeira wine samples, including esters (80.7-89.7%), carboxylic acids (1.6-4.2%), alcohols (3.5-8.2%), aldehydes (0.9-3.7%), pyrans (0.2-1.7%), lactones (<3%), monoterpenes (0.1-1.4%), sesquiterpenes (0.1-0.8%) and C13 norisoprenoids (1.7-6.5%), which some of them play a remarkable impact on the aroma complexity. C13 norisoprenoids in particular, seem to play an important role on Madeira wine bouquet since presenting very low sensorial threshold limits. Excellent correlation between Madeira wine ageing and the abundance of cis-oak lactone was attained showing to be an important chemical descriptor to characterize reserves and Vintages as well as a contributor to wine flavour. The differentiation between reserves, dry/medium dry and sweet/medium sweet young wines could be well established by means of chemometric analysis, using particular aroma compounds such as diethyl succinate, cis-oak lactone and ethyl octanoate as discriminating variables.

Measurement of body composition changes during hemodialysis by bioimpedance analysis.

Monitoring of fluid exchanges during hemodialysis is essential for the safety and efficacy of the procedure. Bioimpedance analysis has been used for the measurement of body compartments in many settings, but experience during hemodialysis is relatively scare. In a prospective study of 28 cases of chronic renal failure treated by long-term hemodialysis on an outpatient basis, body composition studies were done immediately before and after the dialysis session. Methods included body weight, serum albumin, hematocrit, hemoglobin, sodium and osmolality. These variables were compared with impedance (BIA) findings, namely lean body mass, body fat, total water, extra and intra-cellular water, third space, and exchangeable sodium/potassium ratio. Patients lost weight and improved hematocrit, hemoglobin, osmolality and serum sodium during the therapy, and BIA measurements were consistent with these changes. Lean body mass and body fat markedly changed along the dialysis session (respectively -24.5% and + 7.5%), and these results were out of proportion regarding known gains and losses in that period. It is concluded that BIA was useful for monitoring water fluxes during hemodialysis, but provided unreliable information about body fat and lean body mass.

Perimetria computadorizada em pacientes glaucomatosos com visäo sub-normal / Computerized perimetry in sub-normal vision glaucomatous patients

O objetivo deste estudo é analisar a possibilidade da utilização da perimetria computadorizada em pacientes glaucomatosos com acuidade visual inferior à 200/400. Foram examinados 33 olhos de 33 pacientes (21 com AV = CD e 12 com AV = MM), utilizando-se o programa 24-2 do perímetro da Humphrey, inicialmente com estímulo III e depois com estímulo V. Doze pacientes não conseguiram realizar nenhum dos exames (todos com AV = MM). Dezenove pacientes conseguiram realizar os dois exames, enquanto dois pacientes que realizando o exame com estímulo V. A conclusão deste estudo é que é possível a realização da perimetria computadorizada em pacientes glaucomatosos com defeitos avançados de campo visual e AV inferior à 20/400.